1116 Manual 2017 PDF

LAB MANUAL 2017
EVOLUTION,
ECOLOGY
and
BIODIVERSITY
DEPARTMENT OF BIOLOGY
University of Winnipeg
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BIOL-1116 EVOLUTION, ECOLOGY, & BIODIVERSITY
LABORATORY ORGANIZATION
The organization of the labs in this course follows the same pattern as the Cells &
Cellular Processes labs (BIOL-1115). Laboratories are subdivided into practical and
demo/tutorial sections. These labs will take place in their respective practical and
tutorial rooms. The practical room is 0RC046, and the tutorial room is 0RC044. In
order to complete both theoretical and practical labs, students will be subdivided into
groups A and B, as shown in the table below. The specific laboratory schedule for each of
these two groups is shown on page 4.
GROUP A GROUP B
SECTION # DAY AND TIME SECTION # DAY AND TIME
071 Monday 2:30 070 Monday 2:30
073 Tuesday 8:30 072 Tuesday 8:30
078 Wednesday 8:30
081 Wednesday 2:30 080 Wednesday 2:30
082 Thursday 8:30
085 Thursday 11:30 084 Thursday 11:30
087 Thursday 2:30 086 Thursday 2:30
089 Friday 2:30 088 Friday 2:30
The content of this laboratory manual is also subdivided into practical laboratories, starting
on page 19 and tutorial laboratories, starting on page 145.
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EVOLUTION, ECOLOGY AND BIODIVERSITY
(BIOL-1116)
LABORATORY SCHEDULE WINTER 2017
Week # Practical room (0RC046) Demo/Tutorial room (0RC044)

Week 1 Practical Lab 1 (A)
Jan. 9-13 Bacteria, Protists, & Fungi
Week 2 Practical Lab 1 (B) Demo/Tutorial Lab 1 (A)
Jan. 16-20 Bacteria, Protists, & Fungi Bacteria, Protists, & Fungi
Week 3 Practical Lab 2 (A) Demo/Tutorial Lab 1 (B)
Jan. 23-27 Kingdom Plantae Bacteria, Protists, & Fungi
Jan. 30-Feb. 3 Kingdom Plantae Kingdom Plantae
Week 5 Practical Lab 3 (A) Demo/Tutorial Lab 2 (B)
Feb. 6-10 Animals I Kingdom Plantae
Week 6 EXAM I
Feb. 13-17 (Modules 1 & 2)
Week 7
READING WEEK (NO LABS)
Feb. 20-24
Feb. 27-Mar. 3 Animals I Animals I
Week 9 Practical Lab 4 (A) Demo/ Tutorial Lab 3 (B)
Mar. 6-10 Animals II Animals I
Week 10 Practical Lab 4 (B) Demo/ Tutorial Lab 4 (A)
Mar. 13-17 Animals II Animals II
Week 11 Demo/ Tutorial Lab 4 (B)
Mar. 20-24 Animals II
Week 12 EXAM II
Mar 27-31 (Modules 3 & 4)
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MARK BREAKDOWN:
THE LAB IS WORTH 40% OF YOUR MARK IN THIS COURSE:
Midterm Lab Exam (Exam I) = 12%; week of February 13, 2017
Final Lab Exam (Exam II) = 20%; week of March 27, 2017
Lab Assignments = 8%
TOTAL = 40
The general assignments will vary in format throughout the term. Some may be short
mini and spot quizzes done either at the beginning or at the end of the laboratory. Others
might be written assignments that will have to be handed in at the end of the lab period,
or the following week. Most assignments are marked out of 10, and at the end of the
term a mark out of 8% is calculated, based on the total value of all assignment grades.
For example, if there are 8 assignments, each of them is worth 1%.
INTRODUCTORY REMARKS:
The laboratory course in Biology is designed to serve several purposes:
1. Illustration of some of the lecture material.
2. Acquisition of information first hand.
3. Providing training in careful observation and deduction, the accurate

making of records and in the use of the tools of Biology: dissecting
instruments, microscopes, recording materials and other equipment.
Laboratory work is extremely important in Biology. Textbook, lecture

descriptions and diagrams cannot take the place of your own observations on the actual
material. Accurate observation is the basis of all scientific work. The laboratory
exercises, which follow are designed both to complement and supplement the material
covered in the lectures. In this course the laboratories will introduce you to the structure
and function of organisms that make up the living world. The only way to really
understand these organisms is to examine them first hand in the lab.
Although laboratories and lectures are integrated to some extent, do not expect
your lecturer to cover, in detail, what you do in the laboratory. The lecturers will often
discuss and illustrate the theoretical aspects of a topic in a general way. And, in
subsequent laboratories, certain aspects will be selected for more detailed investigation
by the student. With the theoretical background provided by your lecturer and the
textbook, and following the directions provided by the laboratory manual, you should be
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able to gain first hand knowledge of some of the structural and functional relationships in
organisms, and of the techniques by which they are studied.
The role of the laboratory Instructor and student demonstrators is to provide

general directions, and to assist you in following the directions given in the laboratory
manual, and in interpreting your results. Do not hesitate to ask for their help when you
need it, but do not expect them to do your work for you. They will be familiar with the
material and will be glad to discuss with you any phase of your work in the Biology
course.
MATERIALS SUPPLIED BY THE STUDENT:
1. Textbook: Campbell Biology in Focus, 2nd Ed. by Campbell et al.,

published by Pearson Education.
2. Laboratory Manual for BIOL-1116.
3. a) Dissecting Kit including:

surgical scissors (coarse and fine)
2 wood handle needles
forceps (coarse and fine)
1 probe
b) Pencil, blank white paper, eraser and ruler -- to be brought to every lab
period.
4. A lab coat is mandatory. Please be sure to bring a lab coat to every

Practical AND Tutorial lab. It will often save on laundry and cleaning
bills, especially in exercises involving dissection or the use of chemicals
and dyes.
NOTE: THE ABOVE ARE THE MINIMUM REQUIREMENTS.
Students who do not purchase the approved dissecting kit from the U of W
Bookstore should have an Instructor check their instruments and may be required to
purchase additional instruments separately if their sets do not meet minimum standards.
MATERIALS SUPPLIED BY THE UNIVERSITY:
All other materials and equipment required for the laboratory course are supplied
by the University. Most of the equipment provided is expensive and much is fragile and
easily damaged or broken. The student is responsible for any equipment in his/her care
and may be charged for damages.
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LABORATORY PROCEDURES:
1. Attendance Policy
Lab attendance in BOTH practical and theoretical labs is compulsory not

optional! If you know in advance that you will miss a laboratory period for some
unavoidable reason, you should make arrangements prior to this time to make up work
covered in your absence. There are no regularly scheduled make up laboratory periods.
Similarly, if a laboratory is missed due to illness, arrangements should be made to make
up the material. To do this, you should contact the coordinator of the BIOL-1116
Laboratory Program Carlton DuGuay; Office: 2RC027; Phone: 204-786-9727;
email: c.duguay@uwinnipeg.ca to obtain written permission to attend a different lab
section. Students who fail to contact the coordinator regarding missed laboratory
time will not be granted credit for the work that they have missed.
Missed Quizzes and Exams:

- contact your Instructor or the coordinator immediately.
- a doctors certificate (or other documentation) will be required.
- if you do not contact your Instructor or the coordinator promptly,
and do not have a medical certificate (or other documentation), you
will receive a grade of zero for the quiz and/or exam.
NO SPECIAL CONSIDERATIONS WILL BE MADE IF THE INSTRUCTOR (OR

COORDINATOR) IS NOTIFIED AFTER THE FACT.
Missed Assignments:
- if you have a legitimate excuse for a missed lab with a lab assignment and do
not make up this particular lab, you might be excused from handing in that
assignment. Appropriate documentation may be required.
Late assignments:
- 10% per day will be deducted for late assignments. Please note that if an
assignment is due at the beginning of the laboratory period but, is handed in either
during or at the end of laboratory, it will be considered late.
2. Laboratory Preparation
It is assumed that all students will have read the laboratory outline before coming
to the lab. In addition, students are expected to read the appropriate sections in their
textbook for further clarification. Students should always have their lab manuals with
them and should bring materials when needed -- i.e. text, dissecting kit.
For the purpose of laboratory exams, students are responsible for knowing all
bold faced and/or underlined structures.
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3. Laboratory Organization
In this course, laboratories are subdivided into practical and demo/tutorial

sections. The practical room is 0RC046, and the demo/tutorial room is 0RC044.
Both practical and theoretical labs are equally important and the material studied in both
of these types of labs will be included on lab exams I and II.
Depending on the nature of the exercises, students will work individually or in

groups in these two types of labs. Although co-operation and discussions between
students is encouraged in the belief that students learn a great deal from each other,
"freeloaders" who attempt to rely on other students to do their work for them will find
that laboratory tests and examinations are designed to test their knowledge of the material
first hand.
Most laboratory exercises include a number of demonstrations, specimens, or

experiments set up somewhere in the room for the entire class to observe. This material
is an integral part of the laboratory course, and for examination purposes, is considered
equally important to parts of the exercises or dissections performed individually or in
groups. In many of the laboratories during this term, you will dissect and examine
preserved specimens. For that purpose, you will need to follow the dissection techniques
described below.
Basic Dissection Techniques
1. Practice safe hygiene when dissecting. Wear appropriate protective clothing,

gloves and eyewear, and DO NOT place your hands near your mouth or eyes
while handling preserved specimens. While many of the preservatives currently
used are non-toxic to the skin, they may cause minor skin or eye irritations in
some individuals and should never be ingested. In general, the preservatives used
on these specimens are desiccants and may dry out your skin after prolonged
exposure. If fumes from your specimen irritate your eyes, ask your instructor
about the availability of goggles.
2. Read all instructions CAREFULLY before making any incisions. Make sure
you understand the direction and depth of the cuts to be made many important
structures may be damaged by careless or imprecise cutting. For instance, while
investigating the digestive system you do not want to damage vessels in the
circulatory system that you will need to identify later.
3. Use scissors, a teasing needle and a blunt dissecting probe whenever possible.
Despite their popularity, scalpels usually do more harm than good and should not
be relied upon as your primary dissection tool. Remember the purpose of blunt
dissection is to separate muscles, organs, and glands from one another without
cutting them.
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4. When instructed to expose or view an organ, you should attempt to remove
all of the membranous tissues that typically cover organs (fat, fascia, etc.).
This takes time, when done thoroughly. Your goal should be to expose the organ
or structure as completely as possible.
5. A good strategy to use if working in groups is to read aloud the directions

from the book while your partner(s) performs the dissection. These roles
should be traded from section to section to give both of you a chance to have
hands-on experience. In addition to simply identifying the organs and structures
from the photos and diagrams, make sure you read the descriptions of them in the
text. You should be able to recognize each organ or structure and describe the
function it performs in the body.
4. Slide Box Policy
In this course, you will be required to examine many slides. There is a slide box,
with a number of slides in it, located in every drawer. Each slide in these boxes has two
numbers on it:
a "B" number which states the box number to which the slide belongs.
an "F" number which indicates the position in which the slide is to be found
within the box.
Each student is held responsible for broken or lost slides in the box assigned to
him/her. Since there are numerous lab sections, each box is used by a number of students
during the term. Thus, every week before you start your observations, check if all of the
slides that you will require are present in your box, so you are not held responsible for
previously lost or broken slides.
Please return each slide to the correct position in the slide box when you are
finished using it, so that other students can find it quickly and easily when they need
it. If you realize that a slide is missing, report it immediately.
5. Safety
- All students are required to have a valid WHMIS certificate in order to participate.
- Do not eat or drink in the laboratory.
- No sandals allowed in the lab. Only closed-toe shoes are permitted.
- Report all spills and accidents to your instructor.
- Upon completion of the laboratory exercises, place all materials in their proper place
and clean up your lab desk (wipe it down, if needed).
- Place broken glass and sharp objects in the Broken Glass Garbage container; do not
place them in a regular garbage container where they might injure someone.
- Leave the lab clean and organized.
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The MSDS sheets for
0RC044 & 046 are
located in the SE corner
of 0RC046 (next to the
emergency shower).
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NOTE: The emergency shower, fire extinguisher, fire blanket, first aid kit, and main gas line shut-
off valve are all located in the SE corner of both 0RC044 & 046. There is an eye wash station in
the NE corner of 0RC044 as well as in the NE & SW corners of 0RC046.
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Anatomical Terminology
Anatomy is a highly descriptive science that has a language of its own. Many of the terms
used are derived from Latin or Greek words that describe the size, colour, shape, texture
or other physical characteristics of body structures. Other terms are used to indicate the
direction or position of the body part with respect to their parts. Read the definitions of
the terms below. Examine the diagrams and familiarize yourself with these terms so that
the dissection instructions will be easier to understand. This material could also be tested
on exams.
I Directional Terms
The terms used to indicate direction will mainly apply to animals whose vertebrae lie in
the horizontal plane e.g. most four legged (quadruped) animals. Most of the terms are the
same for humans but a few apply only to us because of our upright stance. Directional
terms refer to an animal in its normal body position. For humans, anatomical position is a
person standing erect with their arms at their sides, palms facing forward.
i) Cranial, caudal and rostral

In a quadruped, structures that are closer to the head end are cranial while those that are
closer to the tail are caudal. For structures just within the head region, the term rostral is
often used instead of cranial. Rostral means toward the nose. Anterior and posterior are
the same as cranial and caudal for quadrupeds but because they mean something different
in humans (see below), it is probably better to just use cranial and caudal to avoid
confusion. For humans the terms comparable to caudal and cranial are superior and
inferior.
ii) Dorsal and ventral

Dorsal refers to structures that are closer to the vertebral column or backbone and
ventral means closer to the belly in four-legged animals. In humans the back is the
posterior surface, while the stomach side of the body is anterior.
iii) Lateral and medial

Structures that are positioned more to the side of the body are termed lateral, while those
more toward the midline are termed medial.
iv) Proximal and distal

These terms refer to structures that are closer to and farther from the main part of the
body or the point of attachment of the limb, respectively. For instance, the elbow lies
proximal to the wrist because it is closer to the point where the limb attaches to the body
(the shoulder) than is the wrist. The wrist is distal to the elbow because it lies farther
from the shoulder joint than does the elbow. Proximal and distal are the best directional
terms to use for the appendages. The terms can also be used to indicate a position closer
to or farther form a particular reference point. E.g., a particular blood vessel that lies
distal to (farther from) the heart.
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v) Superficial and deep
Structures that are superficial lie closer to the skin surface, while those that are deep lie
closer to the interior of the body.
vi) Right and left

These terms are obvious, but it is important to remember that they always refer to the
specimens left or right, not yours.
Note: All of the above directional terms are used to describe a position of one body part
with respect to another (i.e. they are relative positions). Your neck is superior (or cranial)
to your chest, but is inferior (or caudal) to your head.
II Planes of Section
There are three commonly used planes of section with which you need to be familiar.
These can be used to indicate positions of body parts or you may be asked to cut
specimens along these planes.
1) Sagittal plane
A sagittal section passes through the body from dorsal to ventral. A midsagittal section
divides the body into equal right and left halves. Any sagittal section not in the midline
but off to one side is a parasagittal section. Parasagittal sections are parallel and lateral
to the midsagittal section.
2) Frontal plane
A section made in the frontal plane divides the body into dorsal and ventral parts,
longitudinally. For humans, this plane divides us into front and back halves. For
quadrupeds, the frontal plane separates the back from the belly.
3) Transverse plane
The transverse (or cross-sectional) plane divides the body into cranial and caudal parts
and occurs at right angles to the longitudinal axis.
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Figure 1: Directional terms and planes of section.
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PRACTICAL LABORATORIES
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PRACTICAL LABORATORY 1 BACTERIA, PROTISTS, & FUNGI:
INTRODUCTION TO THE LIVING WORLD

DOMAIN BACTERIA
DOMAIN EUKARYA: THE PROTISTS
DOMAIN EUKARYA: THE FUNGI
OBJECTIVES:
1. To briefly introduce the organization of the living world.
2. To describe the basic characteristics of the Domain

Bacteria.
3. To describe the basic characteristics of some

representative members of the Bacteria, Protists, and Fungi.
4. To examine asexual reproduction in the three main groups

of Fungi.
I THREE DOMAINS OF LIFE
Until recently, most biologists adopted a taxonomic scheme that divided the
diverse organisms into five Kingdoms, such as: Monera, Protista, Fungi, Plantae, and
Animalia. However, new methods such as, comparisons of DNA sequences from
different species have led to an ongoing reevaluation of the number and boundaries of
Kingdoms. Of the five Kingdoms previously recognized by taxonomists, most biologists
continue to recognize only Plantae, Fungi, and Animalia. But, as debate continues at the
Kingdom stage, there is a consensus that living things can now be grouped into higher
levels of classification called Domains. Presently, biologists recognize three Domains:
Archaea, Bacteria, and Eukarya. As discussed earlier, each of these three Domains is
further subdivided into Kingdoms, Phyla, Classes, Orders, etc. (Campbell, Reece, et. al.,
CDN ed., 2013). A more detailed explanation of this classification system can be found
in the Lab 1 Tutorial section of this manual.
The organisms making up Domain Archaea and Domain Bacteria are all
prokaryotic. Although both Bacteria and Archaea are prokaryotic, they differ in many
structural, biochemical, and physiological characteristics. They also tend to live in
different environments. Representatives of the Domain Archaea, for example, live in
Earths extreme locations, such as salty lakes and boiling hot springs. This Domain
consists of multiple Kingdoms. The Domain Bacteria includes the most diverse and
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widespread prokaryotes. This Domain is also divided into several Kingdoms. All
eukaryotic organisms are now grouped into Domain Eukarya. This domain includes
many groups of single-celled organisms as well as multicellular plants, fungi, and
animals which will be discussed in subsequent laboratories.
In this laboratory, we will study Domain Bacteria and start our investigations of
Domain Eukarya. Domain Archaea will not be studied in detail in the laboratory portion
of this course.
II DOMAIN BACTERIA
The diversity of the Bacteria Domain should not be surprising. Representatives of

this Domain are related to the oldest organisms on Earth and had more time to evolve and
differentiate than all others. They thrive nearly everywhere, from the depths of the ocean
to the upper atmosphere; they are on surfaces of everything you touch and in every type
of water. Some are dependent on oxygen, others indifferent to its presence, and still
others find oxygen toxic. They are also numerous. For example, if you are healthy, the
number of individual bacteria in your intestines exceeds the number of humans that have
ever lived (Campbell and Reece, 2011).
Members of this Domain are called "prokaryotes", from the Greek for
"prenucleus". Their one distinguishing characteristic is that they do not contain
membrane-bounded "organelles". Their genetic material consists of a single circular loop
of DNA located in a nucleoid region of the cytoplasm (Figure 1). In addition to this
single chromosome, a typical bacterial cell may also have much smaller rings of
separately replicating DNA called plasmids as well as ribosomes (Figure 1). The cell
walls of prokaryotes also differ from those of eukaryotes that you studied in the previous
semester. Most bacterial cell walls contain peptidoglycan, which is a network of sugar
polymers cross linked by short polypeptides. Furthermore, cell walls of bacteria may be
covered by a capsule, a sticky layer of polysaccharide or protein (Figure 1).
Bacteria reproduce by the process of "binary fission", where the cell duplicates
its components and divides into two cells (Figure 2). In other words, the cell pinches into
two without the complex movement of chromosomes seen in mitosis. Newly produced
cells usually become independent, but they may remain attached in linear chains or
grapelike clusters. In favourable environments, individual bacterial cells rapidly
proliferate, forming colonies consisting of millions of cells.
Binary fission is a form of asexual reproduction. However, simple sexual

reproduction also occurs. For example, in the process called conjugation, two cells form
a cytoplasmic bridge through which they pass at least a portion of a chromosome.
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Figure 1: Typical prokaryotic cell (non-photosynthetic bacterial cell).
Figure 2: Binary fission in bacteria.
Within their environments, some bacteria are virtually immobile, while others
come equipped with one or more flagella that whip back and forth, pushing them rapidly
along. Still others glide, using mechanisms that are poorly understood.
Most bacteria live as heterotrophs, which means that they derive their energy
from organic molecules made by other organisms. Many heterotrophic bacteria are
important in the ecosystem as decomposers because they feed on dead organic matter
and release nutrients locked in dead tissues. Specifically, they secrete enzymes that cause
the breakdown of organic matter in dead organisms and their wastes. This activity is
ecologically crucial and is undoubtedly the most important of all bacterial activities.
Decomposition releases such key ions as nitrates, phosphates, and sulphates, which then
become available to other organisms. Other heterotrophs are parasites, often referred to as
pathogens. They cause many of the diseases of plants and animals, including those of
humans. Other heterotrophic bacteria live as mutualistic symbionts. They form
partnerships with other organisms in which both benefit. Human intestines, for example,
are home to many species of bacteria. Many of these species digest food that our own
digestive system can not break down.
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Microscopic examination of bacterial cells reveals that most bacteria can be
classified according to three basic shapes: bacilli (rods), cocci (spheres), and spirilla
(spirals, or corkscrews).
Examine slide #89, which is a composite slide that shows each of these three major
bacterial types. You need to scan the whole slide in order to find all three shapes. It is
best to use the lowest magnification of the microscope to initially find these cells, and
then switch to the highest power to observe them in more detail. Compare your
microscopic observations to Figure 3, which shows each of these different bacterial types.
Figure 3: Bacterial types.
Using a technique called the Gram stain (developed by H. Gram), scientists can
classify bacteria into two groups based on differences in cell wall composition. Gram-
positive bacteria have walls with a relatively large amount of peptidoglycan. Gram-
negative bacteria, on the other hand, have less peptidoglycan in their cell walls. In Gram
staining, samples are first stained with crystal violet dye and iodine, then rinsed in
alcohol, and finally stained with a red dye. The structure of the cell wall determines the
staining response. Gram positive bacteria (with thicker peptidoglycan) trap crystal violet
in the cytoplasm. The subsequent rinse with alcohol does not remove the crystal violet
from the cytoplasm, hence, cells appear violet under the microscope. In Gram negative
bacteria (with thinner peptidoglycan) the crystal violet is easily rinsed from the cytoplasm
and the cells appear either pink or red under the microscope.
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Observe the demonstration slide of Gram stained bacteria and, based on the color, be able
to differentiate Gram-positive from Gram-negative bacteria.
Fig. 27-3c
Gram- Gram-
positive negative
bacteria bacteria
20 m
Figure 4: Example of Gram-positive and Gram-negative bacteria.
Cyanobacteria
Cyanobacteria are commonly known as blue-green algae. They are

autotrophic, which means that they derive their energy from photosynthesis or the
oxidation of inorganic molecules. In addition to chlorophyll a, they contain phycocyanin
(blue) and phycoerythrin (red). Because of various proportions of these pigments, only
about half of cyanobacteria are actually blue-green in colour. Many range in colour from
brown to olive green. They live in aquatic environments including oceans, ponds, lakes,
tidal flats, and moist soil. Cyanobacteria exist mostly as colonies and filaments, but
sometimes as single cells.
The cells of the cyanobacteria are prokaryotic but reveal a considerable level of
complexity (Figure 5). Their chlorophyll is integrated into thylakoids, extensions of the
cell membrane. Actually, the entire photosynthetic cell is comparable to a eukaryotic
chloroplast. Photosynthesis in the cyanobacteria is nearly identical, biochemically, to
that of the algae and the green plants. Like the algae and plants, their photosynthetic
pigments include chlorophyll a and the accessory pigment beta-carotene, although they
lack chlorophyll b. The glucose produced by the cyanobacteria in the process of
photosynthesis is stored in their own form of starch, which is similar to animal glycogen.
These characteristics make cyanobacteria predecessors of the eukaryotic chloroplasts.
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Figure 5: Typical cyanobacteria cell.
A number of cyanobacteria produce specialized, nitrogen-fixing cells called

heterocysts. Their role is to incorporate atmospheric nitrogen into a form useful for
producing amino acids and other nitrogen containing molecules. Some cyanobacteria
also produce spores (akinetes) that are resistant to drying. These spores allow
cyanobacteria to survive unfavorable environmental conditions.
Examine the prepared slide of Anbaena sp. (slide #76) and note the vegetative
(photosynthetic) cells and the heterocysts (Figure 6). Anabaena sp. is a very common
filamentous blue-green alga found in stagnant ponds late in the summer.
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Figure 6: Anabaena sp.
Examine the prepared slide of Gloeocapsa sp. (slide #78), which is another very common
colonial blue-green alga. It produces a thick gelatinous sheath (Figure 7).
Figure 7: Gloeocapsa sp.
III DOMAIN EUKARYA
The Domain Eukarya includes three very diverse Kingdoms of eukaryotic

organisms: Plantae, Fungi, and Animalia. In addition to these three Kingdoms, the
Domain Eukarya includes several Kingdoms of protists. In the era of the five-kingdom
scheme, most single-celled eukaryotic organisms were placed in the Kingdom Protista.
Many biologists, however, extended the boundaries of Kingdom Protista to include
some multicellular forms. Thus, the recent taxonomic trend has been to split the protists
into several Kingdoms (Campbell and Reece, 2008).
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A. THE PROTISTS
Organisms belonging to this group are eukaryotic and therefore their cells are
composed of a membrane-bound nucleus and other membrane-bound organelles. This is
a broad group that shows incredible variety. Some protists resemble animals, others
resemble plants, and still others resemble fungi. The major feature they all possess is a
eukaryotic cellular structure. In all other respects, such as cellular anatomy, life cycles,
and ecological roles, protists are difficult to describe. However, some general
characteristics can be cited. Most protists are single-celled (unicellular) organisms,
although some exist as colonies of cells and others are multicellular. The majority of
species are aquatic, living in freshwater or marine environments. Terrestrial protists
inhabit damp soil or leaf litter. The group is represented by both autotrophic and
heterotrophic species. Reproduction in the protists is also quite varied. All protists can
reproduce asexually, however, many species can also reproduce sexually.
Because protists are so diverse, we can study their characteristics effectively only
by dealing with some example organisms. For our purposes in the lab, the protists will be
divided into three large groups:
i) the algae (plant-like protists)

ii) the protozoa (animal-like protists)
iii) the fungal-like protists
In this lab, we will examine some characteristics of a few representatives of each

of these categories. In the Tutorial lab we will discuss the evolutionary history of these
organisms in more detail.
i) Algae (Plant-like protists)
The algae (singular, alga) are generally considered simple plant-like organisms,
both morphologically and anatomically. This is a reflection of the stable, aquatic
environment which they inhabit. Algae are found in waters of varying salinity, including
both fresh water and marine environments. Although quite simple in form, the algae are
an extremely diverse group. They vary in size from the minute (1-5 m) to the very large
species such as kelp, known to grow to 50 m or more in length. Taxonomists suggest
there are probably over 100,000 different algal species. This diversity has occurred over
2 to 3 billion years. These differences are mainly morphological, rather than anatomical,
and are a result of changes in cell differentiation and variations in planes of cell division.
Expressed in a different way, differences in algae are due to differences in shapes of cells
in single celled algae; number of cells in multicellular forms, and differences in the
planes of cell division. The results of the differences in planes of cell division are
illustrated in Figure 13.
The algae are an immensely diverse group of organisms. As a result, we can only
look at a few groups. These include: a) the Dinoflagellates; b) the Euglenids; c) the
Diatoms; and d) the Chlorophytes (Green Algae).
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Diversity in the Algae
a) Dinoflagellates
Most members of this group of algal protists are unicellular. They are abundant
in both fresh and marine waters. The cell wall of some dinoflagellates is composed of
plate-like cellulose segments called theca. Motile dinoflagellates possess two flagella
located in perpendicular grooves. The arrangement of the flagella causes the algae to
move through the water in a spinning fashion, much like a spinning top. The
dinoflagellates utilize chlorophyll a and b as well as carotenoids for photosynthesis.
Energy reserves are stored in the form of starch or oils. Reproduction is primarily
asexual. Dinoflagellates are an important component of both freshwater and marine
plankton. Rapid increases in the population of certain species can result in an "algal
bloom" which colours the water red or brown. These dense algal growths are referred to
as red tides and can cause the shellfish that eat these algae to accumulate a deadly toxin,
therefore making the shellfish themselves toxic to eat.
Examine slide #68 of the freshwater dinoflagellate Ceratium sp. Also, refer to Figure 8.
Figure 8: The freshwater dinoflagellate - Ceratium sp.
29
b) Euglenids
Almost all euglenids are unicellular flagellates which contain a spiral or

crystalline rod inside their flagella. They inhabit freshwater pools and possess either one
or two flagella. Some euglenids have a flexible layer of protein bands (located beneath
the plasma membrane) called a pellicle rather than a cell wall. The pellicle allows their
body shape to change as they swim. Many species of euglenids are mixotrophs, which
means that in sunlight they are autotrophic, but when sunlight is not available they can
become heterotrophic and absorb organic nutrients from their environment. Many other
euglenids can engulf prey by phagocytosis. Reproduction is asexual, by longitudinal cell
division. Sexual reproduction has not yet been observed in this group.
Examine slide #57 of Euglena sp.
Make a wet mount of Euglena sp., which is commonly found in pond water. Observe its
characteristic movement and its ability to change shape (due to the pellicle). Also, refer to
Figure 9.
Figure 9: The unicellular flagellate - Euglena sp.
c) Diatoms
Diatoms are unicellular algae that have a unique glass-like wall perforated by a
delicate lacework of holes and grooves (Figure 10). This wall is made of silica (silicon
dioxide) embedded in an organic matrix. The cells of diatoms are unique since they
30
consist of two parts that overlap like a shoe box and its lid. Such an arrangement
provides effective protection from the crushing jaws of predators.
Diatoms are a major component of plankton in oceans and lakes. Their

populations have a tendency to increase rapidly (bloom) when ample nutrients are
available. Massive accumulations of fossilized diatom walls are major components of the
sediment known as diatomaceous earth, which is mined for its quality as a filtering
medium, for example. Diatoms reproduce mainly asexually by mitosis.
Examine slide #72 of diatoms and note many different shapes.

Fig. 28-13
3 m
Figure 10: A freshwater diatom.
d) Chlorophytes (Green Algae)
This algal group, commonly known as the green algae, exhibits a great deal of
morphological and reproductive diversity. The green algae and the true plants share
many common features: both are photosynthetic, containing chlorophylls a and b as well
as carotenoids within their chloroplasts; their primary energy sources are stored as starch;
and both have cell walls composed of cellulose. Thus, scientists have little doubt that
green algae and green plants are closely related.
Within the green algae, three basic morphological types will be discussed. These
are the unicellular, the colonial, and the filamentous forms. Note the importance of the
basic morphological adaptations of these algae to their environment. Also, note that
some of the adaptations discussed are also utilized by the Dinoflagellate and Euglenid
groups.
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Unicellular Chlorophytes (Green Algae)
These are the simplest algae. Except for a few that live in damp terrestrial
habitats, these organisms are mainly confined to aquatic environments which provide an
adequate supply of nutrients, water, a means of dispersal, and a degree of support. Thus,
these simple organisms can survive and even thrive in this environment. Although they
consist of a single cell, botanists are able to place them in literally thousands of different
genera and species. The range of cellular morphology is, at least in part, an adaptation
for floatation or for predation avoidance. These organisms must have some ability to
float or move, at least through a portion of their life cycle, because they must remain near
enough to the surface to obtain sufficient light for photosynthesis. Larger and/or
irregularly shaped cells have the advantage of being avoided by herbivores in the water.
Most of the animals which eat algae are very tiny and these bigger and/or irregularly
shaped cells would be difficult for them to ingest.
Make a wet mount of Chlamydomonas sp. and Chlorella sp..
Observe the similarities and differences between these two organisms.
In addition, examine slide #77 of Chlamydomonas sp. Also refer to Figure 11.
Figure 11: Unicellular Algae. Chlamydomonas sp. (top) and Chlorella sp. (bottom).
32
Colonial Chlorophytes (Green Algae)
These organisms are also found primarily in the aquatic environment.

Morphological variation is due to differences in number and plane of cell division only.
These algae have only one basic vegetative cell type. Division in definite and consistent
planes results in formation of a regular colony while division in random planes results in
the formation of irregular colonies. Some colonial algae possess flagella for motility.
Again, the major environmental pressure is the need to remain in the photosynthetic zone.
Because of the potential increase in sinking rates due to the increase in mass, these algae
tend to inhabit shallow waters where the effective light penetration is to the bottom. In
addition, many colonial forms have small cells, which secrete mucilage. The mucilage is
less dense than water and aids in buoyancy.
Make a wet mount of Eudorina sp. Note that Eudorina sp. colonies are usually made up
of 16, 32, or 48 cells that resemble Chlamydomonas sp. Each spherical cell has two
flagella (visible as faint lines surrounding the colony in Figure 12). The combined
beating of all the flagella in the colony creates colonial movement.
Also, examine slide #79 of Scenedesmus sp. Also refer to Figure 12.
Figure 12: Colonial Algae. Eudorina sp. (top) and Scenedesmus sp. (bottom).
33
Filamentous Chlorophyta (Green Algae)
While the colonial body form appears to be a dead end from an evolutionary
viewpoint, the filamentous algae apparently had the morphological flexibility to develop
into more complex aquatic and terrestrial plants. The body of the simplest filamentous
algae consists of a single chain of cells. This is the result of cell division in one plane
only. Branching filaments occur when there is periodic division in a second plane
(Figure 13). Once this stage of development was reached, the next step was to evolve
more complex branching patterns. Some cells in the filaments divide several times in
different planes resulting in thick and thin portions. Finally, some filamentous algae
began to show some cellular differentiation. For example, thin portions penetrate the
substrate, thicker portions lie on the surface, and erect portions project about the mass
and bear reproductive structures. This could easily lead to a division of labour among the
three areas. The algae that evolved these features were ones that attached to the substrate
or lived on damp soils.
Examine the slide of Ulothrix sp. (#28).
Examine the slide of Cladophora sp. (#27).
Examine the demonstration slide of Fritschiella sp.
Also, refer to Figure 14 illustrating typical filamentous algae, from the simplest (A) to the
most complex (C).
Figure 13: Planes of cell division.
34
Figure 14A: Ulothrix sp.
Figure 14B: Cladophora sp.
Figure 14C: Fritschiella sp.
35
ii) Protozoa (Animal-like protists)
The protozoa are those protists that are animal-like in that they are heterotrophic
and motile. The majority are found as free-living organisms in freshwater or marine
environments, but there are many parasitic, commensal, and mutualistic species.
Protozoa obtain nutrients primarily by ingesting food. Digestion occurs intracellularly
within a food vacuole and the products of digestion are absorbed into the surrounding
cytoplasm. Protozoa have no special organelles for respiration; the cell membrane serves
as the site for gas exchange. Many species, particularly those living in freshwater,
possess one or more contractile vacuoles which function in osmoregulation. Contractile
vacuoles act as pumps to remove excess water from the cytoplasm of protozoa living in
the hypotonic freshwater environment. Small amounts of nitrogenous wastes may also be
expelled by the contractile vacuoles, although most protozoa usually depend on simple
diffusion through the cell membrane for excretion of these waste products. Contractile
vacuoles may be absent in marine protozoa, since their internal salt concentrations are
very similar to those of their environment. Reproduction in most protozoa is primarily
asexual.
As we have seen with the algae, the animal-like Protists or "protozoa" are so
varied that it would take several weeks to examine each group in detail. Thus, we will
study this group of protists based on the structures they use for locomotion.
Diversity in the Protozoa
a) Flagellated protozoans
Examine the demonstration slide of Trypanosoma sp., a flagellated genus that causes
several severe diseases in humans and domestic animals (sleeping sickness, for example
in humans). Most Trypanosoma sp. live in the blood or tissue fluids of their host. They
are common in the tropics and are spread by infection from biting insects such as
mosquitoes, sand flies, and tsetse flies.
Observe that this parasitic organism has an elongate body and a whip-like flagellum,
used for locomotion, which is located at the anterior end of the body (Figure 15). In
addition, it has a single, large mitochondrion that contains an organized mass of DNA
called a kinetoplast.
Figure 15: Diagram of Trypanosoma sp. (left) and a microscopic view of Trypanosoma
sp. in a blood smear. (right)
36
b) Pseudopodia-using protozoans
The amoeboid protozoa possess flowing extensions of the body called

pseudopodia (singular, pseudopodium). Pseudopodia are temporary projections of the
cytoplasm that are used to engulf food and serve as locomotory structures (Figure 16).
Examine the slide of Amoeba sp. (Slide #53). Observe that this protozoan has an
asymmetrical body that constantly changes shape as new pseudopodia may emerge from
virtually anywhere on the cell surface.
Figure 16: An amoeboid protozoan engulfing food (top) and a microscopic view of
Amoeba sp. (bottom).
c) Ciliated protozoans
Ciliated protozoans have large numbers of cilia that are used in movement and/or
feeding. The cilia may completely cover the body or may be clustered in rows or tufts.
They are the most highly specialized and complicated of the animal-like protists.
Paramecium sp., a representative of this group, is widely studied and easily examined
(Figure 17). It is a free-living, unicellular, freshwater organism. Like many protozoans,
Paramecium sp. reproduces asexually by binary or transverse fission, which is the
division of an organism into two (Figure 17). Fission results in two individuals, which
are genetically similar.
37
Examine the prepared slide (#86), which shows Paramecium sp. undergoing fission.
Make a wet mount from a culture of living Paramecium sp. and note the cilia.
Figure 17: Diagram of Paramecium sp. (left) and Paramecium sp. undergoing fission
(right).
Make a wet mount of hay infusion and try to find the specimens you were studying in
todays lab.
38
iii) Fungal-like Protists
The slime molds, water molds (and their relatives) have traditionally been
considered with the Kingdom Fungi, but in the modern system they are grouped with the
protists. In recent years it has become clear that there is very little evidence for a direct
relationship between the slime molds and water molds and the fungi. They are so distinct
from one another and from the fungi, that each of them is best regarded as a separate
evolutionary line among the eukaryotes.
Diversity in the Fungal-like Protists
a) Plasmodial slime molds
Plasmodial slime molds stream along the damp forest floor in a mass of brightly
colored cytoplasm called a plasmodium (plural, plasmodia). The plasmodium contains
many diploid nuclei, but is not divided by cell walls. It is therefore referred to as
coenocytic, or multinucleate (it is NOT multicellular). As it grows, the nuclei divide
repeatedly and synchronously (that is the nuclei of the slime mold divide at the same
time). During their non-reproductive stages the plasmodial slime molds are thin
streaming masses of protoplasm that creep along in an amoeboid fashion. They resemble
a moving mass of slime. As these plasmodia travel, they engulf (by phagocytosis) and
digest bacteria, yeast cells, fungal spores, and small particles of decayed plant and animal
matter.
Examine the live culture of the plasmodial slime mold Physarum polycephalum on
demonstration and see if you can detect the cytoplasmic streaming (Figure 18).
Remember to switch off the light and to cover the culture with foil after you are done
with your observations.
Figure 18: Picture of the plasmodial slime mold Physarum polycephalum.
39
b) Oomycetes (Water Molds and Downy Mildews)
Oomycetes are a very distinctive group of organisms. They include water molds
and downy mildews. The cell walls of this group of organisms are composed largely of
cellulose or cellulose-like polymers, thus differing markedly from the cell walls of the
fungi, which are made of another polysaccharide, chitin. They range from unicellular
forms to highly branched, coenocytic filamentous ones.
Most species can reproduce both sexually and asexually. Sexual reproduction is
via heterogamy and is said to be oogamous" (in which one of the gametes, the egg, is
large and non-motile, and the other gamete, the sperm, is smaller and biflagellate). Their
union results in the production of a thick-walled zygote, called the "oospore", which
serves as a resting spore. Asexual reproduction is by means of zoospores that have two
flagella.
The Water Molds
The water molds are principally aquatic, inhabiting fresh waters. They are
decomposers that grow as a cottony mass on dead algae and animals. Some are weak
parasites. They have profusely branched coenocytic hyphae (hyphae = slender filaments
of cytoplasm and nuclei) which form colonies around decaying organic material in water.
Asexual reproduction is by means of sporangia, which produce biflagellate zoospores.
Sexual reproduction is by means of gametangial contact (gametangia = gamete
producing structures).
You will examine Saprolegnia sp. as a representative of the water molds.

Members of this genus decompose dead fish, insects, and other organic matter in fresh
water. They can reproduce either asexually or sexually. We will concentrate on sexual
reproduction. For the purpose of sexual reproduction, compatible oogonia and
antheridia develop on the same diploid mycelium. Meiosis apparently occurs within
these gametangia. The oogonia are enlarged cells, in which a number of spherical eggs
(oospheres) are produced. The antheridia develop from the tips of other filaments of the
same individual and produce numerous male nuclei (Figure 20). In mating, antheridia
grow toward the oogonia and develop tubular processes called "fertilization tubes",
which penetrate the oogonia. Male nuclei travel down the fertilization tubes to the
female nuclei and fuse with them. Following each nuclear fusion, a thick-walled zygote,
called the oospore, is produced. On germination, the oospore develops into hyphae,
which then produces a sporangium, beginning the cycle anew (Figure 20).
Examine the prepared slide (slide #100) of Saprolegnia sp. and note: oogonia, oospheres
(light in colour), oospores (fertilized eggs which are dark in colour), antheridia (might be
difficult to find), and fertilization tubes (might be difficult to find) (Figures 19 & 20).
Also examine the demonstration slide of Achyla sp. (another water mold) and find
oogonia, oospheres, and oospores.
40
Examine the demonstration of the live culture of Saprolegnia sp. growing on decaying
matter in water (usually grains in this laboratory) and note its fuzzy appearance.
Figure 19: Microscopic view of the reproductive structures of Saprolegnia sp.
Figure 20: Life Cycle of Saprolegnia sp.
41
The Downy Mildews (Peronosporales)
Downy mildews generally live on land and are parasites of plants. Many have
significant impact on economic plants and cause tremendous losses of crops each year.
One species, Peronospora parasitica, causes infections in many plants. The diseased
plants are detected by their swollen and distorted stems bearing a white layer of powdery
(flower-like) sporangiophores. The sporangiophores (sporangia bearers) cover the
surface of the leaves and stems of the infected plants, giving them a downy appearance,
from which this group gets its common name. The sporangia are released and fall to the
ground to be picked up by emerging seedlings, or are carried by the wind to land on the
leaves of more mature plants. If free water is present, the sporangia germinate and
release flagellated zoospores. These zoospores swim and enter the plant tissue through
open stomata or any break in the cuticle. The zoospores then develop into hyphae, which
invade the host tissue. The mycelia feed on living cells by the use of haustoria (literally
"one who drinks"). The haustoria penetrate the living hosts cells, absorb nutrients, and
pass them to the growing hyphae (Figures 21 & 22).
Examine the demonstration slide of the cross section through a stem invaded by downy
mildew. Observe haustoria and sporangiophores.
Examine the preserved material and note the "downy" appearance of the plants caused by
the downy mildew.
(a) (b)
Figure 21: (a) View of haustoria feeding on living cells. (b) Sporangiophores.
Figure 22: Diagram of downy mildew (Peronospora parasitica) on a leaf.
42
B. KINGDOM FUNGI
For the rest of this lab we will examine the Kingdom Fungi. For many of you this
will be your only exposure to this fascinating group of organisms. The fungi are as
distinct from the mosses and higher vascular plants as they are from the animals and
therefore, are a distinct kingdom. Today you will examine some of the morphological
and structural features that make each of the major groups of fungi unique. In the
Tutorial lab you will examine, in detail, the life cycles of the major groups of fungi.
There are about 100,000 species of fungi that have been described, and it is
estimated that as many as 200,000 more may await discovery. There may actually be as
many species of fungi as there are species of plants, although far fewer have been
described thus far. The fungi have no direct evolutionary connection with the plants and
apparently were derived independently from a different group of single-celled
eukaryotes. The oldest fossils that resemble fungi occur in the strata about 900 million
years ago, but the oldest that have been identified with certainty as fungi are from the
Ordovician period, 450 to 500 million years ago. Representatives of Division
Zygomycota were associated with the underground portions of the earliest vascular plants
in the Silurian period, some 400 million years ago. Fungi may be among the oldest
eukaryotes; the four major groups were in existence by the close of the Carboniferous
period, some 300 million years ago.
The fungi, together with the heterotrophic bacteria and a few other groups of
organisms, are the decomposers of the biosphere. Their activities are as essential to the
continued functioning of the biosphere, as are those of the primary producers. They are
responsible for recycling essential chemical elements back to the environment. Fungi
help plant roots absorb water and minerals from the soil. The decomposition releases
carbon dioxide into the atmosphere and returns nitrogenous compounds and other
minerals to the soil where they can be used again by green plants and eventually animals.
It is estimated that, on average, the top 20 centimeters of fertile soil may contain nearly 5
metric tons of fungi and bacteria per hectare (2.47 acres).
As decomposers, fungi often come into direct conflict with human interests. A
fungus that feeds on wood makes no distinction between a fallen tree in the forest or a
railway tie. Equipped with a powerful arsenal of enzymes that break down organic
products, fungi are often nuisances and are sometimes highly destructive. This is
especially true in the tropics, where the warmth and dampness promote fungal growth.
Fungi attack cloth, paint, leather, jet fuel, insulation on cables and wires, photographic
film, and even the coating of the lenses of optical equipment - in fact almost any
conceivable substance. Even in temperate regions, they are the scourge of food
producers and sellers alike. They can grow on bread, fresh fruits, vegetables, meats, and
other products. Fungi reduce the nutritional value, as well as the palatability, of such
foodstuffs. A number of fungi also produce extremely poisonous toxins, some of which -
the aflotoxins - are extremely carcinogenic and show their effects at concentrations as
low as a few parts per billion. Many fungi are pathogenic, and attack living organisms,
rather than dead ones. They are the most important single cause of plant diseases. Well
43
over 5000 species of fungi attack economically valuable crop and garden plants, as well
as many wild plants. Other fungi are the cause of serious diseases in humans and other
animals.
However, many fungi are commercially valuable. For example, the yeasts are
useful because of their abilities to produce substances such as ethanol and carbon dioxide
(which plays a central roll in baking). Others are of interest as sources of antibiotics,
including penicillin, which was the first antibiotic to be widely used. Fungi are also good
potential sources of proteins.
Associations between fungi and other organisms are extremely diverse. For
example, about four-fifths of all land plants form associations between their roots and
fungi called mycorrhizae. Mycorrhizae increase the absorptive surfaces of the plant
roots and aid in mineral exchange between the soil and the plant. These associations play
a critical role in plant nutrition and distribution.
Basic Structure and Nutrition
Some fungi exist as single cells and are known as yeasts. However, most species
are multicellular. The basic structure of a fungus is the hypha (plural, hyphae) - a
slender filament of cytoplasm and nuclei enclosed by a cell wall (Figure 23). A mass of
these hyphae make up an individual organism, and is collectively called a mycelium. A
mycelium can permeate soil, water, or living tissue; fungi certainly seem to grow
everywhere. In all cases, the hyphae of a fungus secrete enzymes for extracellular
digestion of the organic substrate. Then, the mycelium and its hyphae absorb the
digested nutrients. For this reason, fungi are called absorptive heterotrophs.
Heterotrophs obtain their energy from organic molecules made by other organisms.
Fungi feed on many types of substrates. Most fungi obtain food from dead
organic matter and are called saprophytes. Other fungi feed on living organisms and are
parasites. Many of the parasitic fungi have modified hyphae called haustoria (Figure
23), which are thin extensions of the hyphae that penetrate living cells and absorb
nutrients.
Hyphae of some species of fungi have cross walls called septa that separate
cytoplasm and nuclei into cells. Hyphae of other species have incomplete or no septa
(i.e., are aseptate) and therefore are coenocytic (multinucleate). The cell walls of fungi
are made of chitin, the same polysaccharide that comprises the exoskeleton of insects and
crustaceans.
44
Figure 23: Basic structure of fungus.
Reproduction in Kingdom Fungi
Fungi reproduce by either sexual or asexual means. However, as you will see in
the Tutorial lab, as one moves up the evolutionary ladder of the fungi, sexual
reproduction becomes more important and asexual reproduction becomes less important.
All phyla, however, share similar patterns of reproduction and morphology. In this lab
we will examine the basic concepts and structures involved in asexual fungal
reproduction. In the Tutorial lab we will examine, in more detail, the specific sexual
reproductive life cycles of the three main groups of higher fungi. The following is a brief
introduction to both forms of reproduction.
a) Asexual Reproduction
Fungi commonly reproduce asexually by mitotic production of haploid vegetative

cells called spores in sporangia, and conidia on conidiophores. Spores are microscopic
and surrounded by a covering well suited for the rigors of distribution into the
environment.
Budding and fragmentation are two other methods of asexual reproduction.

Budding is mitosis with an uneven distribution of cytoplasm and is common in yeasts.
After budding, the cell with the lesser amount of cytoplasm eventually detaches and
matures into a new organism. Fragmentation is the breaking of an organism into one or
more pieces, each of which can develop into a new individual.
45
b) Sexual Reproduction
The basic sexual life cycle of fungi includes the familiar events of: vegetative growth,
meiosis (genetic recombination), and fertilization. However, the timing of these events is
unique in fungi. Fungi reproduce sexually when hyphae of two genetically different
individuals of the same species encounter each other. The following is a list of the four
important features of the sexual cycle of fungi.
Nuclei of a (monokaryotic) fungal mycelium are haploid during much of the life
cycle.
Gametes are produced by mitosis and differentiation of haploid cells.
Fertilization (syngamy) is a common feature of all sexual reproductive life cycles.
However in fungi there is a unique delay between the two processes involved in
fertilization. During fertilization the initial union of the cytoplasm of two
parent mycelia (gametes) is known as plasmogamy. The union of two haploid
nuclei contributed by two gametes is known as karyogamy. In the fungi there
may be a considerable delay between plasmogamy and karyogamy. During this
period of time between plasmogamy and karyogamy the resulting cell(s) contain
the two haploid nuclei contributed by the gametes. So there are actually two sets
of chromosomes, just not in the same nucleus. So the cell is not really diploid
but it does have two sets of homologous chromosomes. As you will see,
eventually the two haploid nuclei will fuse (karyogamy) and the cell (nucleus)
will be diploid. But in the interim we call the cell with two separate haploid
nuclei dikaryotic (or heterokaryotic). The dikaryotic stage in fungi can be a
very prominent part of their sexual life cycle.
Once a cell becomes diploid meiosis quickly follows, producing haploid spores
which act to disseminate the fungi to new locations. The haploid meiospores
germinate into haploid hyphae.
The three main groups of higher fungi are actually named after the structures (cells)
in which meiosis occurs. Consider the following diagram, which illustrates the
generalized life cycle of fungi.
46
Figure 24: Generalized life cycle of Fungi.
Classification of Fungi
Campbell and Reece (2011) report that the taxonomy of fungi is currently the
subject of much research. Although there is considerable discrepancy in the
classification of fungi, we will treat the kingdom as having 3 phyla: the Zygomycota,
Ascomycota, and Basidiomycota. There are actually 3 other phyla of fungi but we
wont be looking at them in this course. Where the Ascomycota and Basidiomycota are
considered the "higher fungi", the other phylum is known as the "lower fungi" because
they have retained many primitive characteristics. The separation of these 3 phyla in our
system is based on reproductive structures in which meiosis occurs. The Zygomycota
(or zygote fungi) produce diploid "zygospores" in which meiosis will eventually occur.
The Ascomycota (or sac fungi) produce a diploid cell called an ascus. Meiosis in the
ascus produces haploid "ascospores". The Basidiomycota (or club fungi) produce a
diploid structure called a basidium in which meiosis occurs to produce haploid
"basidiospores". This week, as you examine members of these major fungal groups,
carefully note variations in the fundamental structure of vegetative mycelia and
specialized structures associated with sexual and asexual reproduction.
a) Phylum Zygomycota
Most of the Zygomycota live on decaying plant or animal matter in the soil but
some are parasites on plants, insects, or small soil animals. There are approximately 750
described species of Zygomycota. The term "Zygomycota" refers to the chief
characteristic of the division; the production of sexual resting spores called
zygospores. Most zygomycetes are saprophytic and their vegetative haploid hyphae
47
lack septa (i.e., they are aseptate). In order to rapidly colonize new substrates, these
haploid zygomycota hyphae can produce very large numbers of asexual sporangia. These
sporangia produce large numbers of asexual spores by mitosis. These asexual spores are
very small and are easily carried long distances on the wind. This allows the fungi to
reproduce very quickly when growing conditions are good and the food supply is
plentiful.
In this laboratory, you will study one representative of this phylum, Rhizopus
stolonifera, a common black bread mold.
Rhizopus stolonifera (black bread mold)
This species is one of the most common members of this phylum. This organism
causes the black bread mold that forms cottony masses on the surface of moist bread
exposed to the air.
Asexual reproduction in Rhizopus occurs by spores (Figures 25 & 26). The
mycelium of R. stolonifera is composed of three different types of haploid hyphae
(another word for a filament). The bulk of the mycelium consists of rapidly growing
submerged hyphae that are coenocytic (multinucleate) and aseptate (not divided by cross
walls into cells or compartments). From the submerged hyphae, aerial hyphae called
stolons are formed. The stolons form rhizoids wherever their tips come in contact with
the substrate. Sporangia form on the tips of sporangiophores, which are erect branches
formed directly above the rhizoids (Figures 25 & 26). The sporangiophores support the
asexual reproductive structures: the sporangia. Within a sporangium, haploid nuclei
divide by mitosis and produce haploid spores. The cell wall that forms around each
spore is black, giving the mold its characteristic color. The spores are released to the
environment when the sporangium matures and breaks open. Each spore can germinate
to produce a new mycelium.
Figure 25: Diagram of the vegetative & asexual reproductive structures of Rhizopus sp.
48
Examine the prepared slide #83 of Rhizopus stolonifera, noting the mycelia differentiation
in stolons, rhizoids, sporangiophores, and the sporangia with spores. Also see Figure 26.
The life cycle diagram of Zygomycota can be seen in the Lab 1 Tutorial section of this
manual (Figure 10).
Figure 26: Picture illustrating a Rhizopus stolonifera sporangium.
b) Phylum Ascomycota
The Ascomycota comprise about 60,000 described species, including a number of

familiar and economically important fungi. Most of the blue-green, red, and brown
molds that cause food spoilage are Ascomycota. This includes the salmon-coloured
bread mold Neurospora sp., which has played an important role in the development of
modern genetics. Many Ascomycota are the cause of serious plant diseases, including
powdery mildews that attack fruits, chestnut blight, and Dutch Elm disease (caused by
Ceratocytis ulmi, a fungus native to certain European countries). Yeasts, as well as the
edible morels and truffles are also Ascomycota. This group of fungi, as a whole, is
relatively poorly known, and thousands of additional species await scientific description.
Ascomycota, with the exception of the unicellular yeasts, are hyphal. The hyphae
are septate, or divided by cross walls. The hyphal cells of the vegetative mycelium may
be either uninucleate or multinucleate. Some species of Ascomycota can self-fertilize
and produce sexual structures from a single genetic strain; others require a combination
of + and - strains.
Asexual reproduction in the majority of the Ascomycota occurs by the formation

of specialized spores, known as "conidia" (a Greek word for "fine dust"), which are cut
off from tips of modified hyphae called "conidiophores"("conidia bearers"). The
conidiophores partition conidia in longitudinal chains. Each conidium contains one or
more nuclei. Conidia form on the surface of conidiophores in contrast to spores that form
within sporangia in Rhizopus. When mature, conidia are released in large numbers and
49
germinate to produce new organisms. Pencillium sp., which you will examine in this
laboratory, is a common example of a fungus that forms conidia. Many "crops" of
conidia are produced during the growing season, and it is the conidia that are responsible
for propagation of the fungus.
Examples of Asexual Reproduction in the Ascomycota
i) Yeast
Yeast are somewhat atypical of most Ascomycota. They are predominantly

unicellular and reproduce asexually by fission or by budding (pinching-off of small
buds), rather than by spore or conidia formation.
Examine slide #82 of Saccharomyces sp. for the tiny asexual buds.
Figure 27: Yeast cells budding.
ii) Penicillium sp. (blue - green molds)
Penicillium sp. has become celebrated in connection with antibiotics. Penicillin, a

by-product of Penicillium notatum when liberated into the culture medium, inhibits the
growth of gram-positive bacteria. Penicillin was discovered by Sir Alexander Fleming in
1929, but was not exploited until World War II. The great importance of this substance is
that it represses bacterial growth without being toxic to animal tissues. Interest in
penicillin led to an intensive search for other antibiotics, and molds suddenly became of
considerable economic interest. However, Penicillium sp. is of economic importance in
other respects. For example, certain species give some types of cheese the flavour,
odour, and character so highly prized by gourmets. One such mold, P. roquefortii, was
first found in caves near the French village of Roquefort. Legend has it that a peasant
boy left his lunch, a fresh piece of mild cheese, in one of these caves and on returning
50
several weeks later found it marbled, tart, and fragrant. Only cheeses from the area
around these particular caves are permitted to bear the name of Roquefort. Another
species of this genus, P. camembertii, gives Camembert cheese its special qualities.
As mentioned in the introduction to this phylum of Fungi, Penicillium sp.

reproduces asexually by forming spores called conidia.
Examine the prepared slide #74 of the conidia of Penicillium sp. Note the conidiophores
arising from the host tissue, and the conidia (Figure 28).
Also, examine the demonstration of the culture of Penicillium growing on agar. The
conidia and conidiophores that you have observed under the microscope are part of this
culture.
If available, observe other specimens from this Genus on demonstration.
Note that Penicillium sp. can also reproduce sexually, by ascospores. However,
the sexual phase will not be examined in this course.
Figure 28: Penicillium sp. conidiophores and conidia.
c) Phylum Basidiomycota
The most familiar of all fungi are members of this large phylum. It includes some
25,000 described species including not only the mushrooms, toadstools, stinkhorns,
puffballs, and shelf fungi, but also two important plant pathogens: the rusts and smuts.
The vegetative portion of the fungus exists as a mycelial network, which grows
saprobically beneath the substrate, often as mycorrhizae with trees. The Basidiomycota
have no asexual reproductive structures. Recall that the Zygomycota produced large
numbers of asexual spores and the Ascomycota produced large numbers of asexual
conidia, but the Basidiomycota produce no asexual spores at all. The only spores they
51
produce are the sexual basidiospores produced by meiosis. The Basidiomycota are
distinguished from all other fungi by the production of these basidiospores. A great
example of this phylum is a typical mushroom that you would by at a grocery store.
(Figure 29) The mushroom itself is actually the reproductive structure of the fungus.
The feeding hyphae (mycelium) of the Basidiomycota is found underground, is always
septate, and can cover a very large area. The sexual life cycle, as well as some examples
of Basidiomycota, will be examined next week in the Lab 1 Tutorial section of this
manual.
Figure 29: A typical representative of the Phylum Basidiomycota.
52
PRACTICAL LABORATORY 2 KINGDOM PLANTAE:
INTRODUCTION TO THE KINGDOM PLANTAE

LAND PLANT EVOLUTION
LAND PLANT REPRODUCTION
OBJECTIVES:
1. To briefly introduce the Kingdom Plantae.
2. To introduce land plant evolution with regard to the

reproductive structures.
3. To examine reproduction in a few select representatives of

the Kingdom Plantae.
I INTRODUCTION
In the next two weeks we will introduce you to the plant kingdom. In a first year
course we cant possibly examine all groups of terrestrial plants so this will only be a
brief introduction. However, there will be a few underling themes to our examination of
the plants: (i) their origin from the aquatic green algae; (ii) their invasion of the
terrestrial environment; and (iii) their evolutionary history on land.
The land plants have become better and better adapted to the terrestrial
environment throughout their evolutionary history by improvements in their vegetative
morphologies and through evolutionary improvements in their abilities to reproduce. In
this weeks lab, we will examine the evolution of the reproductive structures and
processes in the land plants. Next week you will examine the evolution of the vegetative
structures.
The land plants are a very important group of organisms on earth. They constitute
the bulk of the primary producers for terrestrial ecosystems, and they have a tremendous
impact on the hydrological cycle as well as global climate. As you begin to think of life
from an evolutionary perspective, you should begin to see that natural selection acts on
morphological structures to produce well adapted morphologies. However, natural
selection also works on those structures and processes involved in reproduction. One
may have very well adapted morphologies, and be able to survive the current
environmental conditions, but if you cant reproduce, then the species is doomed to
extinction in that environment.
The major plant groups you will examine are mosses, ferns, and flowering plants.
There are many other groups of land plants, but the important evolutionary and ecological
concepts of terrestrial plants can be nicely explained using these three groups.
53
II PLANT EVOLUTION
It has become clear that the terrestrial plants have descended from a group of
green algae called the Charophytes. This explains why the basic cell structure,
photosynthetic apparatus and biochemistry, and basic patterns of cell division are
virtually the same in the Charophytes and the terrestrial plants. Some would go so far as
to say that terrestrial plants are nothing more than a special group of land adapted green
algae. But what adaptations! The terrestrial environment has imposed some spectacular
selective pressures on the terrestrial plants. Much of the evolutionary history of land
plants can only be understood by examining the problems these plant groups must have
overcome in order to survive in the terrestrial environment.
Aquatic green algae and land plants have three important things they need to do in
order to survive and reproduce:
1) They must be able to absorb light for photosynthesis.
2) They will need to absorb water from their surroundings and make sure
it is available to all the cells of the organism.
3) They will need a supply of carbon dioxide and nitrogen compounds
that are needed to build the cells organic molecules.
Once those issues have been dealt with, additional issues involving reproduction
have to be considered. These include the protection of the delicate gamete forming
structures, problems of gamete transfer, and survival of delicate young during
reproduction. The wet, stable aquatic environment meant that algae didnt need to
protect their reproductive structures. However, elaborate protection systems for the
delicate reproductive structures of the land plants had to evolve before successful
reproduction could be ensured in the terrestrial environment.
III REPRODUCTIVE DIVERSITY IN THE PLANT KINGDOM
A. THE CONCEPTS OF LIFE CYCLES & ALTERNATION OF GENERATIONS
In Practical Lab 1 we introduced you to the basic ideas of the sexual life cycle.
Fortunately, plants have a simpler life cycle than do the fungi. However, all the plants
we will examine show a distinct Alternation of Generations. That is, they spend part of
their life cycle as multicellular haploid gametophytes and part of their life cycle as
multicellular diploid sporophytes.
The basic form of the alternations of generations life cycle is illustrated in Figure
1. Variability in this cycle will occur from one group of plants to another. We will
examine some of this variation in morphology of the different developmental phases, the
time spent in each phase, and the actual mechanism of forming the reproductive cells for
the three major groups of plants: mosses, ferns, and flowering plants.
54
As illustrated in Figure 1, there are several features which are common
components of every plant life cycle. These are as follows:
1. Sexual reproduction always includes the processes of meiosis and

fertilization.
2. Asexual reproduction never includes either of the above processes.
3. Sexual reproduction always has a portion of the cycle in which all the cells
of the organism are haploid (gametophyte generation), alternating with a
portion of the cycle in which all cells of the organism are diploid
(sporophyte generation). This constitutes the concept of
ALTERNATION OF GENERATIONS (Figure 1).
4. The sporophyte portion of the cycle always ends with the production of
spores by meiosis (meiospores). The spores are produced in a structure
known as sporangium (plural, sporangia). The gametophyte portion of the
cycle terminates with the production of gametes by mitosis. The gametes
are produced in a structure known as gametangium (plural, gametangia).
There are two types of gametangia. The egg producing gametangia are
known as archegonia (singular, archegonium), while the sperm producing
ones are known as antheridia (singular, antheridium).
5. Asexual reproduction always results in the formation of new individuals

which are genetically identical to the parent organisms.
Figure 1: Generalized plant life cycle showing alternation of generations.
55
Both asexual and sexual reproduction provides a means of increasing the
population size. However, sexual reproduction also allows for variations in an
organism's genotype and phenotype. If an organism is well suited to its environment, and
if that environment is stable, then asexual reproduction may play an important role in
maintaining the species. In fact, genetic variation may be a short term detriment. On the
other hand, sexual reproduction becomes more important in less stable environments. As
you complete the exercises in this lab, you should observe how changes in reproductive
patterns parallel the evolution of plants towards a greater ability to withstand unstable
terrestrial conditions. The basic plant groups seem to illustrate this evolution. Also, you
should consider the environmental pressures placed on reproductive adaptations evolved
in plants to combat these pressures, the success of these adaptations, and the general
trends in the evolution of plant reproductive systems.
Two important concepts must be considered when studying reproduction in the

mosses, ferns, and seed plants. The first is the concept of life cycles or life histories. The
second is the concept of alternation of generations. Too often, life cycles are examined
as a series of somewhat unrelated static events. Ideally, they should be considered as a
series of phases in the development of the organism. Each phase is simply a transition
from the previous to the subsequent phase. Also, difficulty arises when one begins to
examine, in detail, all the morphological changes that occur from one phase to another.
In this case, the cycle itself may be ignored.
B. EVOLUTION OF REPRODUCTIVE STRATEGIES IN PLANTS
Natural selection has worked on not only the morphological characteristics of the
land plants, but also on the structures and strategies involved in sexual reproduction.
This makes sense given the ultimate definition of the fitness of a phenotype (and
ultimately a genotype). Fitness refers to the ability to produce the greatest number of
offspring that live to reproduce themselves. Therefore, the more fit a species is, the
more successful they will be. So what elements of sexual reproduction in land plants are
crucial to maximizing reproductive success?
Survival of the gamete forming organs On land, exposed to the dry air, the gametangia
(gamete forming organs) on the gametophyte need an outer layer of protective cells to
prevent them from drying out. This outer protective layer was not needed in the algae
that lived underwater.
Getting the gametes together Transfer of the male gametes from the gametangia
(antheridium) where they are made, to the female gametes (egg) inside the archegonia
can be a problem on land. The sperm in the mosses and ferns are flagellated, and require
free water for the sperm to swim through. The availability of water (rain) can severely
limit the transfer of gametes and ultimately the success of reproduction in mosses and
ferns. In the flowering plants natural selection has favored the loss of motility of the
sperm and a reduction in the size of the gametophytes so that water is no longer needed in
gamete transfer. This has allowed the flowering plants to reproduce successfully in much
dryer habitats than either the mosses or the ferns.
56
Protection of the delicate embryo Once fertilization has occurred, the dividing zygote is
very delicate. Algal zygotes, underwater, dont have any problem with drying out.
However the mosses, ferns, and flowering plants all need to protect these delicate
embryos during their early development. This is done by having the embryo develop
inside the female archegonia. The wall of the archegonia even grows along with the
embryo making sure it stays covered, until the embryo is sufficiently well developed to
protect itself from the dry air.
To be diploid, or not to be? That is the question When you examine the life cycles of
the mosses, ferns, and flowering plants you will notice an interesting trend. In the
mosses, the dominant generation in the life cycle is the haploid gametophyte. By
dominant we mean most obvious, longest lived, easiest to find generation. (not dominant
like in genetics. i.e., dominant vs. recessive). Recall that in the aquatic algae, the only
cell that was diploid was the zygote, and this single cell was where meiosis occurred to
produce many long lived haploid cells. In the ferns, the dominant generation becomes
the diploid sporophyte. The fern gametophyte is very small, delicate and lives only long
enough to produce the gametes and get the diploid embryo started. In the flowering
plants the male and female gametophytes have been reduced down to one cell and two
nuclei in the male gametophyte; and a 7 celled-8 nucleate female gametophyte. So why
has there been this evolutionary push towards a dominant diploid sporophyte generation
in land plants? What is so good about being diploid on land? Or what is so bad about
being haploid? Next week you will examine this evolutionary trend in Tutorial Lab 2.
i) Reproduction in Mosses
The mosses generally occupy habitats which are damp and shaded. These
habitats can be considered as transitional areas between aquatic and terrestrial
environments. Only a few genera exist in dry areas and these require free water for
reproduction. The Bryophytes are unique among land plants because of the dominant
gametophyte generation. The life cycles of the mosses are quite similar to each other.
Figure 2 represents a "typical" moss life cycle. As seen in Figure 2, the mosses have two
phases in their gametophyte generation. The first to develop is an algal-like filamentous
phase termed a protonema. The second phase is the mature gametophyte, which is green
and leafy. Also, note the presence of an embryo in the sporophyte phase. This is an
undifferentiated mass of cells which is nourished and protected within the archegonia by
the gametophyte.
57
Figure 2: Life Cycle of Polytrichum sp., a moss.
Asexual Reproduction
Mosses have not utilized asexual reproduction to the same extent as the algae.
This may be due to the instability of the transitional environment. This instability
demands that the organisms within the environment have a high degree of genetic
variability to allow adaptation to changing conditions.
Asexual reproduction occurs through the process of fragmentation. During

fragmentation, any part of the gametophyte plant is capable of regenerating the
protonemal phase followed by a mature gametophyte. Leaves, stems, and rhizoids will
all produce new protonemata (plural) if the environment is favourable.
58
Sexual Reproduction
The variability in sexual reproduction found in the algae is not found in the
mosses. There is only one pattern to the plant's development through the various phases
of its life. The only variation occurs in the actual vegetative morphology. Several parts
of the cycle are available for your examination. You should observe each phase in
sequence and then remember to look for features which show advancement over the algae
with regards to a terrestrial existence. Also, note any features of the cycle which would
be detrimental to a land plant.
1. Mature Gametophyte Observe this plant on demonstration. You will

examine this plant in more detail in Tutorial Lab 2. For now, be certain that
you are able to recognize male and female mature gametophytes. In male
plants, the leaves at the top are arranged to form a cup-shaped structure,
whereas in female plants, the leaves at the top are all standing straight up.
(Figure 2)
2. Gametangia (singular, gametangium) These are the gamete-producing

structures on the gametophyte plant. They can be separated according to sex
with the male gametangium being an antheridium and the female
gametangium being an archegonium. The gametangia are found at the top of
the gametophyte amongst the leaves.
a) Examine the prepared slide of a male Polytrichum sp. (Slide #9). Locate
the antheridia containing the sperm cells at the top of the gametophyte.
The antheridium has an outer protective cell layer which does not produce
gametes. This layer provides some protection from desiccation.
b) Examine the prepared slide of a female Minium sp. (Slide #6). Locate the
archegonia and the egg cell (female gamete) within it. Again, note that the
gamete is surrounded by a protective layer of cells. Refer to Figure 2 to
help you visualize where these two gametangia are located in the
gametophyte plant.
c) Observe the preserved plants of Polytrichum sp., which should be on your

desks.
d) Observe the demonstration of mosses and make sure that you are able to
differentiate between the sporophyte and gametophyte generations
described below.
3. Sporophyte Examine the gametophyte plant with a sporophyte growing

out from the top. The point where the sporophyte is attached is called the
foot. Note the long stalk or seta of the sporophyte (Figure 3). This serves to
aid in wind dispersal of the spores by elevating the spore-bearing structure
above the clump of mosses.
59
Figure 3: Diagram of a sporophyte plant (left); and picture (right).
4. Capsule Examine the prepared slide of Polytrichum sp. capsule (Slide #5).
Compare this slide with Figure 4 and note the operculum, spores, and seta.
The operculum provides a protective cover to the spore-producing region until
the spores are mature. At this time the operculum will fall off and the spores
will escape from the top of the capsule.
Figure 4: Longitudinal section of moss capsule (sporangium).
60
5. Protonema Examine the culture of living protonema. These filamentous
structures grew from spores which were planted on the medium. Note the
structural resemblance of protonema to algae.
a) Examine the prepared slides of moss protonema (Slides #3 & #4) and
Figure 5. Note the young developing leafy gametophyte phase growing
from the filamentous mass.
Figure 5: Moss protonema (left); and moss protonema with leafy gametophyte (right).
ii) Reproduction in the Ferns
Ferns are located in a range of terrestrial habitats. Most are in areas of damp soil,
high humidity, and low light. Several vegetative features of the sporophyte generation,
such as the presence of true vascular tissue, permit this existence. Ferns exhibit an
alternation of two free living generations. The sporophyte generation is the dominant and
obvious generation. The gametophyte generation is a small vulnerable transitory phase.
Figure 6 illustrates the life cycle of most ferns. Compare this cycle with the cycle of
mosses. These cycles are used to point out the similarities in reproductive patterns
throughout the plant kingdom. You should now realize the inadequacies of such cycles
when morphological comparisons are to be made. They also give no indication as to the
survival capacities of each phase of the plant's development. Only an actual examination
of each phase will reveal these answers.
Asexual reproduction by the ferns is even more restricted than in the mosses. They
produce no specific asexual reproductive structures. Asexual reproduction occurs
through extensively spreading and branching rhizomes (underground stems).
Multiplication is accomplished after death of older parts of the rhizomes at points of
branching. Separate plants are then established.
Examine the fern plants on demonstration and find rhizomes.
61
Sexual Reproduction
The pattern of sexual reproduction is very similar to that of the mosses. The
major differences you should be aware of include:
- the increase in number of meiospores.
- the independence of the mature stages of both generations.
- the dominance of the sporophyte generation.
The major similarities are:

- the presence of flagellated sperm cells.
- the dependence on free water for fertilization.
- the formation of an embryo which is protected and nourished
by the gametophyte plant.
- the need to establish the gametophyte generation from a single
unprotected spore.
Examine the following phases of fern development and note how they relate to Figure 6.
Also compare the structures with similar phases in the moss life cycle. Attempt to find
portions of the fern life cycle which are detrimental to a terrestrial environment.
62
Figure 6: Life Cycle of a Fern.
1. Mature Gametophyte Plant (Prothallus) Examine mature gametophyte

plants using the dissecting microscope. Note their heart shape. (Figure 7)
63
Figure 7: Prothallus Mature Gametophyte.
2. Gametangia Examine the prepared slide of the fern prothallus (Slide #24).
Locate the antheridia and archegonia. Refer to Figure 8. Also examine the
prepared slides of separated antheridia and archegonia (Slides #52 & #25).
(a)
(b)
Figure 8: (a) Fern Antheridia. (b) Fern Archegonia.
64
3. Young Sporophyte Examine the culture of gametophyte prothalli
(singular, prothallus) with new sporophytes using the dissecting microscope.
Identify the initial frond. The embryo is located in the region of the rhizome.
Note that there is vascular tissue in the sporophyte, but not in the
gametophyte.
Examine the prepared slide #55 illustrating the gametophyte and attached
sporophyte. Also refer to Figure 9.
Figure 9: Fern Gametophyte and Attached Young Sporophyte.
4. Mature Sporophyte Compare the complexity of this generation in the

ferns with the comparable generation in the mosses.
5. Sporangium Examine the underside of a mature fern frond and note the
brown capsules, known as sori (singular, sorus). Each sorus is a cluster of
sporangia.
Prepare a wet mount of one sorus and examine it under the light compound
microscope. Because the sorus will be broken during the slide preparation, you
should be able to identify individual sporangia. On each sporangium, try to
identify the stalk, annulus, and meiospores. (Figure 10)
Examine the prepared slide of the fern frond x.s. (Slide #56) and note the sorus,
sporangia, annulus, and spores (Figure 10).
65
Figure 10: Fern Sporangia in a Sorus.
iii) Reproduction in the Flowering Plants

Flowering plants are by far the most successful land plants. They can be found in
nearly every terrestrial habitat. There are several evolutionary adaptations that have
occurred in the history of the flowering plants that have led to their dominance on land.
Of interest to us in this lab are the reproductive adaptations. The first adaptation is the
continuation of a trend that started with the ferns. Recall that an important distinction
between mosses and ferns is a gametophyte-dominated life cycle for mosses, and a
sporophyte-dominant life cycle for ferns. Continuing the trend of sporophyte dominance,
the gametophytes of flowering plants are even more reduced than those of ferns. In
flowering plants, the delicate female gametophyte and young embryos are protected from
many environmental stresses because they are retained within the moist sporangia of the
parental sporophyte. For the gametophyte to exist within the sporophyte has required
extreme miniaturization of the gametophyte of flowering plants.
The second reproductive adaptation involves the advent of the seed as the
resistant stage of the life cycle. That is, in mosses and ferns, spores from the sporophyte
are the resistant stage in the life cycle. They can survive environments too extreme for
the moss (or fern) plants themselves to survive. The seed represents a different solution
to resisting harsh environments and dispersing offspring. In contrast to a single-celled
spore, a multi-cellular seed is a more complex, resistant structure. A seed consists of a
sporophyte embryo packaged along with a food supply within a protective coat.
The last adaptation revolves around the development of a system to disperse the
male gametes through the harsh terrestrial environment in a safe way. In the mosses and
ferns, flagellated sperm cells were used for dispersal of the male gametes. These relied
on the availability of free water. The flowering plants solved the problem of requiring
water by developing the pollen grain. Pollen grains are extremely tough structures that
can be dispersed by the wind or animals. Because water was no longer needed for
dispersal, flowering plants could move away from the waters edge.
66
Asexual reproduction in flowering plants is common. Many different seed plants

utilize one of a number of different methods of this form of reproduction. There are
several reasons why seed plants may find this form of reproduction advantageous. If the
environment has been stable for many generations, variability may not be as essential to
the survival of the species. Asexual reproduction, which is not as complex and requires
far less energy, would be preferable. When colonizing a new area, finding a mate for
sexual reproduction may be difficult or impossible. If the environment is particularly
harsh, the more delicate or susceptible organs or stages of sexual reproduction may not be
able to survive. Many plants which inhabit such areas (e.g., deserts or arctic tundra) only
reproduce asexually. In this laboratory, you will examine various types of asexual
reproduction, which are described in the following paragraphs.
Types of Asexual Reproduction
1) Rhizomes Plants such as the grasses, cattails, and sedges produce underground
stems or rhizomes. As these stems grow through the soil, they periodically produce
adventitious roots and a new above ground shoot. If the rhizome subsequently dies, a
new separate plant will have been formed (Figure 11).
Figure 11: Plant reproduction by the use of rhizomes.
2) Tubers Tubers are actually modified rhizomes. They are formed in plants such as
Irish potatoes. They develop when specialized stem branches grow down into the ground
and swell up with starch-containing cells. Buds on the tubers will grow into new plants.
Examine the potato tuber and note the buds which are commonly termed "eyes" (Figure
12).
67
Figure 12: Plant reproduction by the use of tubers.
3) Runners (Stolons) These are horizontally growing stems that produce few, if any,
leaves. At the spot where a leaf would normally develop (a node) these plants will
produce adventitious roots down into the soil and new above ground shoots instead.
Examine the strawberry plant or spider plant. Note the runner and the new shoots (Figure
13).
Figure 13: Plant reproduction by the use of runners/stolons.
4) Plantlets A few seed plants such as the duckweed and Kalanchoe sp. produce
miniature plants on the margin of their leaves. These drop off and develop into mature
plants. The duckweed, which is an aquatic plant, reproduces almost entirely by this
method. Observe the plantlets on demonstration (Figure 14).
68
Figure 14: Plant reproduction by the use of plantlets.
5) Bulbs Onions, chives, and lilies over-winter in the form of a bulb. Each bulb has a
very short stem which is surrounded by fleshy leaves. In the spring, the shoot apex
begins to grow using the nutrients stored in the leaves. Examine the onion bulb and find
the fleshy storage leaves and stem (Figure 15).
Figure 15: Plant reproduction by the use of bulbs.
6) Corms These are similar to bulbs except that there are no storage leaves. The
nutrients are, instead, stored in the swollen stem. Examine the demonstration of a corm
and compare it with a bulb. Gladiolus sp. and Crocus sp. produce corms (Figure 16).
69
Figure 16: Plant reproduction by the use of corms.
Sexual Reproduction
Although asexual reproduction is common in seed plants, sexual reproduction is the

major method of reproduction. It is essential to provide the degree of variability required
to exist in a diverse and changing terrestrial environment. It is impossible to examine all
the diversity found in these sexually reproducing plants. In this lab, we will deal with the
reproductive cycle itself and attempt to compare it with sexual reproduction in the mosses
and ferns. Much of the success of these plants over mosses and ferns in the terrestrial
environment is due to their reproductive methods. You should make note of features
found in these plants that are not found in the other groups. Also note how the basic life
cycle does not alter significantly from the other land plant groups.
The Flower
We will start the examination of the flowering plant life cycle by taking a detailed
look at the reproductive organ of the flower, which is part of the sporophyte generation.
Flowers are generally composed of the male and female reproductive structures
surrounded by attractive or protective leaf like structures collectively known as the
perianth. The flower functions to protect the developing gametes and to ensure gamete
transfer (pollination) and fertilization. Since plants are rooted structures, but still rely on
cross-fertilization to maintain genetic diversity, they must use other transfer agents to
complete fertilization. Physical factors such as wind and water and biological agents such
as insects, birds, and mammals transfer pollen (the male gametes) from plant to plant.
Flowering plants have not abandoned the use of sporangia as the final structure in
the sporophyte generation. The male reproductive structure, the stamen, and the female
reproductive structure, the carpel (pistil), contain the sporangia. There are two main
points you should note about the difference between these sporangia and those of the
mosses and ferns. First, there are two different types of sporangia in the flowering plants:
the male or microsporangium; and the female or megasporangium. Second, the
megasporangium is protected within the carpel.
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Dissect a preserved lily flower and find the sepals, petals, stamens, and carpels. Note
their relative position in the flower. They were produced in this order (from outside to
inside). Refer to Figure 17. You may need to use a dissecting microscope for this
exercise.
Remove and examine the fresh flowers of the plants provided on display. Find all of the
highlighted structures mentioned in the following paragraph.
The sepals develop first and are usually green in colour and leaf-like in structure.
They provide protection for the delicate internal structures and a small amount of food
through photosynthesis for the remainder of the floral parts. Inside the sepals are the
petals. These are also leaf-like in structure but are frequently brightly coloured. The
petals are also designed for protection and their arrangement is often an attractant to the
various animals such as insects that are necessary for the transfer of gametes or
pollination. Inside the petals are the stamens. The stamens are leaves modified for the
production of microsporangia. Each stamen consists of an anther suspended on a
filament. The anther is made up of one or more microsporangia and is the site of the
production of pollen grains, the multicellular or multinuclear haploid structures which
will eventually form male gametes by mitosis. The carpel is the most central structure
within the flower. A carpel is a leaf which has been modified to produce ovules. A
carpel is usually composed of a stigma, a sticky surface designed to receive pollen
grains; a style, the slender stalk through which pollen tubes grow; and an ovary
containing ovules. The ovules, in turn, contain the megasporangium which produces the
actual female gametes or egg cell nuclei. During fertilization, the pollen tube will
eventually transfer the male gamete to the egg in the ovule.
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Figure 17: Longitudinal section of a typical flower.
Development of the Gametophyte Plants
In the mosses and ferns, the only means of dispersal is the release of the mature
spores from the sporangia. The spores fall to the ground and must develop into
independent gametophyte plants. The early part of this development is particularly
susceptible to damage or desiccation in the environment. In contrast, seed plants do not
produce independent gametophyte plants nor does dispersal occur at the spore stage of
the life cycle. The gametophytes of seed plants are very small and are retained within the
sporangia throughout their development. In fact, the female gametophyte plant (or
embryo sac) is never released from the parent sporophyte. The male gametophyte plant,
the pollen grain, is only released from the microsporangium during pollination.
Development of Male Gametophyte Within each microsporangium there are many

cells called microsporocytes destined to undergo meiosis to produce four haploid
microspores. Each microspore then divides mitotically once, producing two nuclei that
become encased in a thick wall forming the pollen grain (Figures 18 & 19). One of
these nuclei is referred to as the generative nucleus and the other is the tube nucleus.
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At this stage, the pollen grains are released from the anthers and are transferred, via the
wind or animals, to the stigma of a flower. This process is known as pollination. The
stigma is specially designed to receive the pollen grain which then germinates. The tube
nucleus produces the pollen tube that penetrates the stigma and grows through the style
until it reaches the ovary. By this time, the generative nucleus has divided once to
produce two sperm nuclei. This three-nucleated structure is the mature male
gametophyte.
Observe slide #43, which illustrates male gametophyte development. On your slide, find
the following: anther, microsporangium, microsporocytes, microspores (at different
stages of meiosis), and immature pollen grains.
Figure 18: Development of the male gametophyte (pollen grain).
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Figure 19: The stages of the development of the male gametophyte (pollen grain),
a microscopic view.
Make a wet mount of the pollen grains from the flowers provided on display and note the
sculpted walls of the pollen grains.
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Development of the Female Gametophyte The development of the female
gametophyte (embryo sac) occurs within the ovule, inside the ovary. Each ovule
contains a megasporangium which produces one megasporocyte. The megasporocyte
undergoes meiosis producing four haploid cells or megaspores. Three of these
disintegrate and the remaining megaspore divides mitotically three times. The eight-
nucleate, seven-celled structure that is produced is the mature female gametophyte or
embryo sac (Figures 20 & 21). Observe slide #44, illustrating the female gametophyte
development. On your slide, find the following: ovary, ovule, megasporocyte,
antipodal cells, synergids, and polar nuclei.
Figure 20: Development of the female gametophyte (embryo sac).
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Figure 21: Development of the female gametophyte (embryo sac), a microscopic view.
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There are several important aspects of the gametophyte development in seed plants which
you should particularly note:
no release of spores from the sporophyte.

small size of the gametophyte the small size of pollen grains aids in
transfer to the stigma; the small size of the embryo sac allows it to be
better protected by the sporophyte.
neither antheridia nor archegonia are produced.
sperm nuclei, rather than sperm cells, are produced which eliminates the
dependence on water.
Figure 22 (below) summarizes the development of the male and female gametophytes.
Its a useful study tool for your exam.
Figure 22: Summary of male and female gametophyte development in flowering plants.
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Pollination and Fertilization
Pollination occurs when pollen is transported to the surface of the flowers stigma.
After pollination takes place, the pollen grain produces a tube, which grows down the
style and into the ovary to the ovule. It is only capable of growing through the opening in
the integument of the ovule to the embryo sac. The pollen tube fuses with the embryo sac
membrane and releases the two sperm nuclei into the sac. At this point, another
difference from the mosses and ferns occurs. One sperm nucleus fuses with the egg
nucleus to form a zygote. The other sperm nucleus fuses with the two polar nuclei to
form a triploid (3n) endosperm nucleus. This process is termed double fertilization
(Figure 23). The triploid endosperm nucleus is capable of dividing quickly and
governing the formation of food material to nourish the developing zygote. The ovule
with a zygote is now termed a seed. During the whole time from gametophyte
production through pollination to fertilization, the gametes have never been exposed to
the environment. The sperm nuclei were protected by the extremely resistant pollen grain
wall and the egg cell was protected by the ovary and the integument of the ovule.
Figure 23: Pollination and fertilization. Pollen is transferred from the anther to the stigma
of the carpel. Two sperm resulting from division of the generative nucleus move from the
pollen grain through the pollen tube to an opening in the ovule. One sperm fertilizes the
egg, and the other fertilizes the polar nuclei.
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Seed Development
Following pollination and fertilization, two major developmental steps occur in the
life cycle of flowering plants which do not occur in the mosses or ferns. One is the
development of the seed and the other is the development of the fruit. A seed is a mature
ovule that includes a seed coat, a food supply, and an embryo. The seed development
consists of a conversion of the integument of the ovule into a resistant seed coat, the
development of the endosperm, and the development of the embryo. The developing
embryo grows, absorbs endosperm, and stores those nutrients in seed leaves called
cotyledons. All these events take place within the original ovary. You will examine the
slides of the ovary of the plant Capsella bursa-pastoris. This plant is a common
Manitoba weed known as Shepherd's purse. The development of the embryo in the seeds
of this plant includes the following stages (Figures 24 & 25):
Pro-embryo stage During development, the zygote divides to form a mass of

cells called the embryo. Initially the embryo consists of a basal cell, suspensor, and
a two-celled pro-embryo. The suspensor is the column of cells that pushes the
embryo into the endosperm.
Globular stage Cell division of the pro-embryo soon leads to the globular stage
that is radially symmetrical and has little internal cellular organization.
Heart-shaped stage Division of the globular stage produces bilateral symmetry

and two cotyledons forming the heart-shaped embryo. The enlarging cotyledons
store digested food from the endosperm.
Torpedo stage The cotyledons elongate.
Mature embryo The mature embryo has large bent cotyledons attached to it.
The endosperm is depleted and food is stored in the cotyledons.
Examine the slides of the developing seeds (#45-48). The cross section most likely
passes through an entire fruit of Capsella and shows a number of sectioned, developing
seeds, each in a slightly different stage. Find examples of the globular, heart, torpedo,
and mature stages of the embryo. A section through a nearly mature seed will reveal an
embryo with two large cotyledons (Slide #46). Most or all of the endosperm has been
absorbed by the cotyledons and the integuments of the ovule have grown into a seed
coat. The basal portion of the embryo is termed the radicle, the middle portion is termed
the hypocotyl, while the part attached to the cotyledons is termed the epicotyl. The
epicotyl will eventually develop into the above ground structures of the plant (stem,
leaves, and flowers). The radicle will develop into the true root system, and the
hypocotyl will develop into the transition zone between the root and stem.
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Figure 24: Stages of development in a Capsella embryo within a seed.
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Figure 25: Stages of development in a Capsella embryo within a seed, a microscopic
view.
Obtain a bean seed. Section the seed along the plane indicated in Figure 26. Locate the
seed coat, cotyledons, and embryo. On the embryo, identify the epicotyl, hypocotyl,
and radicle.
Figure 26: Internal Anatomy of a Bean Seed.
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Fruit Development
They begin to develop after pollination and usually develop from the ovary, but
sometimes other parts of the flower are involved. The cells of the ovary wall expand and
the whole ovary enlarges to make room for the developing seeds. The extent of this
enlargement and the final differentiation of the cells depend on the type of fruit. Large
fleshy fruits develop a water resistant outer layer and several inner layers of large water
storage cells. These fruits are designed for seed dispersal by animals (the fruits, along
with the seeds inside them, are eaten). The seeds cannot be digested so they are spread
with the animal's feces (a great fertilizer!). These fruits, if they are not eaten, may also
provide some heterotrophic nourishment to the germinating seeds. In contrast, dry fruits
provide a covering that adheres tightly to the seed coat and will prevent desiccation.
Various modifications in dry fruits help disperse the seeds. In some plants, such as
dandelions and maples, the fruit functions like a kite or propeller, enhancing wind
dispersal. Other plants use animals to carry seeds by modifying the fruits to have burrs
that cling to animal fur or human clothing. Still others are designed to open violently and
thus disperse the seeds.
If available, examine the fruits on the bean plant. Note how the petals have withered.
Open one of these fruits and note the developing seeds.
Examine the selection of fruits available on the side bench and consider the probable
method of dispersal. (for example wind, water, ingestion by animals, adhesion to fur,
etc.)
Figure 27: Fleshy fruits
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Figure 28: Dry fruits.
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84
PRACTICAL LABORATORY 3 ANIMALS I:
INTRODUCTION TO THE ANIMAL KINGDOM

PHYLA NEMATODA, ANNELIDA, & ARTHROPODA
OBJECTIVES:
1. To review the concepts of phylogeny and classification.
2. Introduce body plans in higher animals.
3. To examine the basic body plan and the feeding,

respiration, circulation, and reproductive systems of the
Phyla Nematoda, Annelida, & Arthropoda.
INTRODUCTION
The animal kingdom includes multicellular organisms having eukaryotic cells that
lack cell walls, plastids, and photosynthetic pigments. Most members of this kingdom
take in nutrients by ingestion, and digestion of those nutrients takes place in an internal
cavity. Some take in nutrients by absorption and lack an internal digestive system. The
higher forms in the animal kingdom have evolved sophisticated levels of organization
and tissue differentiation. Reproduction for most of the organisms in the kingdom is
sexual, and except for some of the lowest phyla, haploid cells occur only in gametes.
Over the next several weeks, we are going to survey the following phyla of Kingdom
Animalia: Cnidaria, Plathyhelminthes, Nematoda, Annelida, Arthropoda, & Chordata.
For each phylum examined, you should consider:
- the basic body plan of the animals
- the system employed to obtain nutrients, and
- the reproductive systems of the animals.
I BODY PLANS IN HIGHER ANIMALS
Early in the development of many animals, the embryo becomes triple layered. As
a general rule, these layers, termed the germ layers (ectoderm, endoderm, and
mesoderm) form the various tissues and organs of the body as development progresses.
Ectoderm, covering the surface of the embryo, gives rise to the outer covering of the
animal and, in some phyla, to the central nervous system. Endoderm, the innermost
germ layer, lines the primitive gut, and gives rise to the lining of the digestive tract and
its outpocketings such as the liver and lungs of vertebrates. Between ectoderm and
endoderm is the mesoderm, the germ layer that forms the muscles and most of the other
organs between the gut and outer covering of the animal.
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There are three ways bodies can be organized around the cellular layers. The
simplest organization (seen in the flatworms - Platyhelminthes) is the acoelomate body
plan. In this body plan, there is no cavity between the digestive tract and outer body wall.
The space between the digestive tract and outer body wall is loosely filled with
mesodermal cells called parenchyma.
The pseudocoelomate body plan (seen in roundworms Nematoda) has a fluid

filled cavity called the pseudocoelom, located between the digestive tract and outer body
wall. There is a layer of mesoderm located underneath the outer body wall. There is no
mesoderm, however, surrounding the organs. Thus, the cavity is not completely lined by
mesoderm. The internal organs of pseudocoelomate animals are actually found free
within this fluid-filled pseudocoelom. This arrangement is known as a tube-within-a-
tube body plan.
The final organizational level (seen in the annelids and all other phyla above) is
the coelomate body plan. Coelomates have the body cavity (the coelom) completely
lined by mesoderm. There is a layer of mesoderm underneath the outer body wall and
there is another layer surrounding the organs (Figure 1). The two layers of mesoderm
connect dorsally and ventrally to form mesenteries that suspend the internal organs in the
coelom.
A body cavity has many functions. Its fluid cushions the suspended organs,
helping to prevent internal injury. The cavity also allows the internal organs to grow and
move independently of the outer body wall. If it were not for your coelom, every beat of
your heart or ripple of your intestine could deform your body surface, and exercise would
distort the shapes of the internal organs.
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Figure 1: Body plans.
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II PHYLUM NEMATODA
Members of Phylum Nematoda are worms with long, cylindrical bodies and for
this reason are commonly referred to as roundworms. The Phylum Nematoda is among
the largest of the animal phyla. Although only approximately 15,000 species have been
named; biologists estimate that if all the species were scientifically described, the number
would be closer to 500,000. Most nematodes are found free-living within the soil, but a
great many are important parasites of plants and animals.
Roundworms have two morphological advances absent in the flatworms (which

you will study in the tutorial lab). First, in addition to their digestive cavity, roundworms
have a body cavity called a pseudocoelom consisting of a fluid-filled space between the
body wall and digestive tract. Second, nematodes have a complete digestive tract with a
mouth and an anus.
External Anatomy of Ascaris lumbricoides
You will examine Ascaris lumbricoides, which is a large nematode that infects the
intestinal tract of humans and other vertebrates. It is one of the best-known parasitic
nematodes.
Use forceps to obtain one Ascaris for your group and place it in a dissecting pan. You
will also need a box of dissecting pins. Ascaris are dioecious animals (i.e. have separate
sexes). Compare the external characteristics of females and males. The larger animals
with straight tails are females. The thinner animals with hooked tails are males (see
Figure 2). At the anterior end, locate the mouth surrounded by three lobes (lips) of tissue
(you will need to use a dissecting microscope for this exercise). At the posterior end,
locate the anus (Figure 2). The three lobes (lips) will help you distinguish the anterior
from the posterior end of the animal.
Figure 2: Ascaris sp. external view of female and male.
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Dissection of Ascaris sp.
You will dissect Ascaris sp. to observe the internal features of the digestive and
reproductive systems. Before beginning your dissection, add a little bit of water into the
dissecting pan. This allows the organs to float and makes internal structures easier to see.
Pin the anterior and posterior ends of the animal to the bottom of the pan and carefully
slit the body wall longitudinally using the tip of a dissecting needle. Pin the sides of the
wall to expose the internal contents. Place pins at a 45 angle to the pan. Be very careful
not to remove or disturb any organs you will need to identify. Locate the muscular
pharynx, just behind the mouth. The intestine will appear as a thin ribbon-like structure
that extends from the mouth to the anus (see Figure 3). All other internal structures are
related to reproduction!
When you are finished with your dissection, be sure to trade with a group that dissected
an animal of the opposite sex.
**CAUTION: Ascaris eggs are extremely resistant to chemical treatment. Although it

is unlikely, some eggs may survive immersion in preservatives for short periods. For this
reason you should keep your hands away from your mouth and nose while performing
this dissection and wash your hands afterward.**
i) Feeding and Digestion in Nematoda
The specific feeding method utilized by a particular species of nematode is a

reflection of its habitat and way of life. Therefore, nematodes exhibit a wide range of
feeding habits: carnivorous, herbivorous, and parasitic. Although the diet may be
extremely varied, the nematode digestive system is relatively uniform throughout the
phylum.
Observe your dissected specimen as you read through the following information.
The mouth opens into a muscular pharynx, which acts as a pump to bring food
from the mouth into the intestine. The intestine is dorso-ventrally flattened. From the
pharynx, a long tubular intestine travels the length of the body to the anus. Digestive
enzymes are produced by the single layer of epithelial cells lining the intestine.
Digestion begins extracellularly within the intestinal lumen but is then completed
intracellularly.
ii) Respiration and Circulation in Nematoda
Nematodes lack respiratory and circulatory systems. The majority of free-living

nematodes are less than 2.5mm in length and are often microscopic. These animals are
largely the inhabitants of the interstitial spaces of aquatic sediments and soils. In the
soil, free-living nematodes live in the film of water surrounding each soil particle, so they
are essentially always in a moist environment. Free-living species obtain their oxygen
requirements by diffusion from the environment.
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iii) Reproduction in Nematoda
Males and females live in the intestines of humans where they graze on the
intestinal contents. Eggs pass out with the feces and if they contaminate food, are
introduced to another host. The larvae hatch in the intestine of the new host and then
burrow through the walls to be carried by the bloodstream to the lungs. At the lungs,
they burrow through the alveoli (air sacs) and crawl up the trachea and down the
esophagus. Occasionally larvae get lost and crawl up the esophagus and exit the nose. In
some areas of the world, Ascaris is so common that a child is not considered part of the
tribe until a larva is sneezed out and found in the bed!
Female ()
Locate the genital pore (a slit-like opening about one third of the way down from
the anterior end). The genital pore is connected to a short vagina. The vagina divides
into two uteri. At the end of each uterine segment, a thin oviduct can be seen, followed
by the thread-like ovaries. Eggs are produced by meiosis within the ovaries, and move
along the oviducts to the uterus. Fertilization of the eggs takes place in the uterus.
Male ()
The anus serves both an excretory and a reproductive function. The copulatory
spicule (a hook-like appendage used to hold the females genital pore open) should be
seen at the edge of the anus. At the posterior end of the intestine is a region known as the
cloaca (which is connected to the anus on the outside). This region is not visible on your
specimen, but you should be aware of its function. The cloaca serves as a common
collecting area for fecal material from the intestine and spermatozoa from the seminal
vesicle (the thick organ connected to the intestine at the cloaca). The relatively thick
tubes associated with the seminal vesicle are the sperm ducts or vas deferens. The
finest of the threads are the testes. Sperm cells are produced in the testes, and they
mature as they move along the vas deferens to the seminal vesicle. During copulation,
the sperm enter the cloaca before being deposited in the female. Nematode sperm are
unusual because they are amoeboid, not flagellated.
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Figure 3: Internal anatomy of Ascaris sp. female (a) and male (b).
At the end of your dissection:
- place dissected specimens in the special discard container provided (not

the regular garbage container)
- place gloves in the regular garbage container
- wash your hands
- clean your bench with paper towels and cleaning solution provided
III PHYLUM ANNELIDA
Annelids include a variety of marine, freshwater, and terrestrial worms including

the familiar leeches and earthworms. The most distinctive feature of the members of the
phylum Annelida is the division of their bodies into rings or segments (Latin annullus =
little ring). This type of body segmentation is called metamerism. In the annelids,
metamerism is evident in the external features of the worms, which accounts for their
common name, the segmented worms. As well, most annelids also show internal
metamerism with the division of the body into compartments by regularly repeated septa
(walls) and by the repetitive arrangement of organs and organ systems. Annelids show
several evolutionary advances over the Nematodes. They have a true coelom, fully lined
with a layer of mesodermal epithelium, the peritoneum. They have a more centralized
nervous system and a closed circulatory system. Most annelids also have setae, which
are small, bristle-like appendages.
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The Phylum Annelida is subdivided into four classes: Oligochaeta, Polychaeta,
Hirudinea, and Archiannelida.
Observe the specimen jars on display for the first three classes and note their
characteristic segmentation.
We will study the body plan, digestive, excretory, respiratory, circulatory, and
reproductive systems of the earthworm Lumbricus terrestris (a common oligochaete), and
the marine clamworm Nereis sp. (a polychaete).
A. THE EARTHWORM
External Anatomy of the Earthworm (Lumbricus terrestris)
Remove a preserved earthworm from the pail and transfer it to a dissecting pan.
Add a little bit of water to your dissecting pan.
a) Locate the conspicuous clitellum, a saddle-like swelling on the dorsal surface.

The clitellum produces a mucus sheath used to surround the worms during mating
and is responsible for making the cocoon within which fertilized eggs are
deposited. The anterior of the animal is more cylindrical than the flattened
posterior and is the closest to the clitellum. The ventral surface of the
earthworm is usually a lighter colour than the dorsal surface. The mouth is
located on the ventral surface of the first segment while the anus is found at the
end of the last segment (Figure 4).
Run your fingers over the ventral surface of the earthworms body. You should
be able to feel bristle-like setae used for locomotion.
Figure 4: External Features of the Earthworm

(Note: the dashed lines represent several segments).
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Internal Anatomy of the Earthworm
i) Digestive System
The earthworm is an example of a foraging herbivorous annelid, obtaining food by eating

its way through the soil and extracting nutrients from the soil as it passes through the
digestive tract.
a) Position your preserved earthworm dorsal side up and pin it down through the
first segment and then again further back behind the clitellum. Cut a slit in the
dorsal surface near the posterior pin. Using fine scissors extend the cut forward to
the first segment. Be careful not to cut too deep. Starting at the first segment, cut
the septa (thin membranes) that internally divide the segments, so the skin can be
laid flat. Use additional pins to hold the integument open and expose the organs.
Continue to fold the skin back until you have uncovered a centimetre or so of the
intestine.
b) Starting at the anterior end, locate the muscular pharynx (food ingestion). This is
followed by a tube-like esophagus which terminates in a crop (the wider organ)
which serves as a storage stomach. Posterior to the crop you will find the
gizzard. Gently press on the crop and gizzard to test their firmness. While the
crop is soft and thin, the gizzard is muscular (soil is ground up and churned within
the gizzard). The gizzard is followed by a long intestine in which both digestion
and absorption occur (Figure 5). On the dorsal surface of the intestine is a
longitudinal groove known as the typhlosole (Figure 5). The typhlosole increases
surface area for absorption. Undigested material is voided through the anus.
(a)
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(b)
(c)
Figure 5: Pictures of the internal features of the earthworm (a) & (b); and a diagram of
the internal features of the earthworm (c).
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ii) Excretory System
Annelids are considerably larger and more complex than flatworms. In addition,
they possess a well-developed circulatory system, which can function in conjunction with
other systems in the body. Increased size has made it impossible even for marine
members of the group to rely on simple diffusion for the removal of nitrogenous wastes
from their site of production. Thus, the excretory system functions both in
osmoregulation and in the removal of nitrogenous wastes. The excretory organs,
nephridia, are capable of removing fluids and wastes from both tissue fluids and blood.
Each nephridium (there is usually a pair of nephridia in all but the most anterior
segments) is a complex organ consisting of several parts (Figure 6). A ciliated funnel,
the nephrostome, opens into the coelom just anterior to the septum separating two
segments. Beating of the cilia drives fluids into the nephrostome from the coelom and
then through a long tubule which passes through the septum into the next most posterior
segment. A network of blood capillaries surrounds the tubule and works in the exchange
of materials, removal of wastes, and re-absorption of water and other useful materials.
This exchange takes place between the tubule contents and the blood. The tubule leads to
an enlarged bladder and materials finally exit the body through a small nephridiopore.
Examine the demonstration specimen of Lumbricus sp., which has been dissected to
illustrate the position of the nephridia in each segment. Use Figure 5 (above) to aid you
in orienting and locating the nephridia.
Figure 6: Diagram of one nephridium of Lumbricus terrestris.
iii) Respiratory and Circulatory Systems
All annelids live in moist environments; in fact, most of them are actually aquatic.
Even the earthworm can survive only in damp soil. This means that the body surface, or
parts of it, can still serve as the site of gas exchange. However, because annelids are
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considerably larger, more active, and more complex than the Nematodes, some means of
transporting gases, wastes, and nutrients through the body other than simple diffusion is
required.
The function of internal transport in annelids is performed by the coelomic fluid

which is moved around by contractions of the body wall musculature and, in the majority
of segmented worms, by a closed circulatory system. To enhance the capacity of the
blood or coelomic fluid to carry oxygen, these fluids may contain respiratory pigments.
These pigments easily combine with oxygen and increase the capacity of blood to
transport oxygen. When oxygen concentration is low the pigments release oxygen into
the respiratory system. In the annelids the respiratory pigment is hemoglobin which may
either be contained within cells or be in solution in the blood.
Locate the dorsal blood vessel on your dissected specimen and trace it toward the mouth.
There are five pairs of pseudohearts surrounding the esophagus (Figure 5). The
pseudohearts look like black rings on the dissected specimen. The blood in the dorsal
vessel flows anteriorly. It is pumped downward by five pairs of the pseudohearts to the
ventral blood vessel. The blood in the ventral blood vessel flows posteriorly. Small
vessels in each segment carry blood from the ventral vessel back to the dorsal vessel
(Figure 7). There are also blood vessels going to the major organs including the digestive
tract and excretory organs.
Figure 7: Diagram of circulation in the earthworm.
iv) Reproductive System
Earthworms are hermaphrodites with complex male and female reproductive

organs present in each individual, but they cross fertilize. Two earthworms mate by
attaching at their clitella and exchanging sperm, and then they separate. The received
sperm cells are temporarily stored in sperm receptacles while the clitellum secretes a
mucous cocoon. The cocoon slides along the worm, picking up the eggs, which are
produced in ovaries, and then the stored sperm. Finally, the cocoon slips off the worm's
head. The embryos develop within the cocoon.
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The reproductive structures of the earthworm start at segment nine. Identify (on
your dissected specimen) the seminal receptacles (bulb-like organs in segments nine and
ten) where sperm cells received from another worm are stored, and the three pairs of
whitish seminal vesicles (segments 9-12). Sperm are produced within testes located
inside the seminal vesicles and transferred to the male genital pore or gonophores via
the vas deferens. The female reproductive structures consist of a pair of ovaries
(segment 13) connected to the female genital pore or gonophores via a series of small
passageways. Many of the reproductive structures are very difficult to identify on the
preserved specimens so use the earthworm models on display as well as Figures 5 and 8.
Figure 8: Reproductive structures of the earthworm.
B. THE CLAMWORM
External Anatomy of the Clamworm (Nereis sp.)
The clamworm is a marine organism, living in sediment.
Obtain a preserved clamworm and place it in your dissecting pan. Add a little bit of water
to your dissecting pan. Note the well developed head region (Figure 9) with tentacles and
other sensory structures. You may also see jaws. If you do not see the jaws at this point,
you will see them later when you dissect your specimen. Also, note that the body of the
clamworm is distinctly segmented and that each segment bears a pair of fleshy
appendages, called parapodia. You will study the specific function of these appendages
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later, when you observe the respiratory system. Protruding from the fleshy parapodia are
many setae from which the class derives its name (poly-many, chaeta-setae). You will
also observe the setae later when you examine a demonstration slide of the parapodia.
Figure 9: External features of a clamworm (A); & a close-up view of the head region (B).
Internal Anatomy of the Clamworm
i) Digestive System
Place the specimen dorsal side up in a dissecting pan. Pin the anterior and
posterior ends of the animal and carefully cut through the skin to expose the internal
organs as you did for the earthworm. The marine clamworm, Nereis, is carnivorous and
therefore its digestive system differs from that of the herbivorous earthworm, Lumbricus.
The mouth and jaws are retractable (Figures 10, 11). If the jaws are retracted, cut along
the midline of the pharynx and pull the jaws out with a pair of forceps. The jaws are used
to capture small animals. The mouth opens into the muscular pharynx, which leads to
the esophagus. Lateral to the esophagus are two esophageal caeca (singular, caecum).
Posterior to the esophagus is the stomach-intestine.
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Figure 10: Internal anatomy of the clamworm.
(a)
(b)
Figure 11: A photograph of a dissected anterior region of the clamworm (a); and exposed
jaw (b).
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ii) Respiratory and Circulatory Systems
As you have observed earlier each body segment of the clamworm bears a pair of
lateral parapodia, fleshy paddle like appendages that may be used as legs in crawling or
as oars for swimming. The parapodia have a large surface area, are well vascularized
with blood vessels, and are the major sites of gas exchange (Figure 12). In addition, the
movement of the parapodia keeps fresh water with a high oxygen concentration
constantly moving over the respiratory surface. Parapodia can be considered primitive
gills.
Examine a demonstration slide of an intact parapodium and note the bristle-like setae.
Also, observe that the parapodium contains many blood vessels.
Figure 12: Cross-section of Nereis sp. showing the arrangement of blood vessels to the
internal organs (left); and showing a parapodium (right).
iii) Reproductive System
The reproductive organs in Nereis are difficult to see and will not be examined in
this dissection
IV PHYLUM ARTHROPODA
The arthropods have attained the greatest biological success of any group in the
animal kingdom. The phylum Arthropoda is the largest phylum of animals and includes
spiders, ticks, mites, scorpions, centipedes, millipedes, shrimp, crabs, and insects. Over a
million arthropod species are known, making up more than three quarters of all known
animal species. This vast assemblage of animals has representatives in virtually every
conceivable habitat. There are arthropods which fly, swim, burrow, and crawl. Members
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of the phylum can be found in fresh and salt waters, deserts, the Artic and Antarctic,
tropical rainforests and hot springs. Their tremendous success is mainly a result of a rigid
external skeleton and jointed appendages (arthro = jointed; poda = foot) that are used
for locomotion, feeding, reproduction, defence, and sensing the environment.
In this section, you will be examining the various systems in the Phylum
Arthropoda by looking at two representatives: the Crayfish and the Grasshopper. The
crayfish is an aquatic arthropod, which shows adaptations to this underwater
environment. The Grasshopper is a terrestrial arthropod, and therefore shows terrestrial
adaptations. You should be able to compare these two animals and their adaptations.
A. THE CRAYFISH
External Anatomy of the Crayfish (Cambarus sp.)
Crayfish are found in streams, rivers, lakes and ponds where the water contains
adequate amounts of calcium salts. They are primarily nocturnal, hiding in crevices and
under rocks during the day and emerging to feed at night. They will eat almost anything
organic plant or animal, living or dead.
Obtain a preserved crayfish (either a female or a male) and place it in your dissecting
pan. The body is divided into an anterior cephalothorax (fused head and thorax) and a
posterior abdomen (Figure 13). The chitinous exoskeleton protects the crayfish from
predators. Because the exoskeleton surrounds the body, it must be molted periodically to
allow for growth. Until the new skeleton hardens, the animal is helpless.
Observe a demonstration of a crayfish exoskeleton.
The carapace is a saddle-like covering over the cephalothorax. A transverse

groove separates the fused head from the thoracic region. The rostrum is an anterior,
pointed extension of the head. A pair of stalked eyes is located on the lateral side of the
rostrum. One pair of antennae is located on the head as well. At this point, look for a
mouth, which is located on the ventral side of the body, between the mandibles (Figure
14). The abdomen consists of several segments and is terminated by the telson (Figure
13).
Examine the appendages and note that the appendages are modified to serve a variety of
functions: feeding, walking, and swimming.
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Figure 13: External features of the crayfish (lateral view).
The male and female crayfish have an equal number of appendages. However, in
male crayfish the first two pairs of swimmerets have been modified. They are elongated
and can be brought together to form a trough-like channel, used for the transfer of sperm
from the male to the seminal receptacles of the female. In female crayfish, all of the five
pairs of swimmerets are of the same size and are used for swimming. Use Figure 14 to
assist you in determining if your specimen is a male or a female.
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Figure 14: Male (A); and female (B) crayfish (ventral view).
Internal Anatomy of the Crayfish
i) Digestive System
The dissection of the crayfish requires a great deal of care. Be certain to follow
instructions carefully and to ask for assistance if you are in doubt about any part of the
procedure.
Position the crayfish dorsal side up. Using a pair of scissors, remove the
carapace from the cephalothorax by cutting along the mid-dorsal line. Make the incision
at the posterior end of the carapace and cut up to the rostrum. Carefully cut the carapace
away along the side; do not disturb any of the underlying organs. Locate the large
yellowish digestive gland, which occupies much of the posterior part of the
cephalothorax (Figures 15, 16). It secretes enzymes and stores food. With a probe, move
it aside and find the remainder of the digestive system. The mouth (not visible at this
time) leads via a short esophagus to the large stomach which occupies most of the
anterior region of the cephalothorax. Open the stomach and locate the chitinous teeth
that form the gastric mill which is used to grind food. The intestine runs from the
stomach through the abdomen ending at the anus.
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Figure 15: Internal organs of the crayfish (lateral view). Note that the gills have been
removed.
Figure 16: Pictures of the internal organs of the crayfish.
ii) Osmoregulatory and Excretory Systems
Because of their diversity of form and life style, arthropods show considerable
variety in many of their organs including the excretory system. We will briefly examine
two common arthropod excretory systems, which function in both osmoregulation and
waste removal: green glands found in aquatic arthropods, and Malpighian tubules found
in terrestrial groups.
Locate the green glands on the dissected crayfish. Despite their name, they appear pink
and are located anterior to the stomach.
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They open to the exterior at the bases of the antenna (ventral surface). The green
glands consist of several parts, which will not be studied in this laboratory. At this point,
also locate the large mandibular muscles, which operate the jaw-like mandibles (Figure
14).
Aquatic arthropods generally have gills (Figure 17) for respiration except for a
few extremely small species in which there are no special respiratory structures.
Observe the "feathery" gills on your dissected specimen.
Note that the carapace covers the gills except at the anterior and posterior ends of
the gill chamber. Water containing oxygen is driven through the chamber over the gills
by the second pair of maxillae gill bailers. The gills are well vascularized.
Figure 17: Picture of the lateral view of the crayfish showing gills.
Arthropods have an open circulatory system with a prominent heart receiving

blood from the hemocoel and pumping it into vessels for distribution to the body.
Depending on the type of respiratory organ the arthropod possesses, the circulatory
system may or may not be important in the transport of oxygen to the body tissues. In
those animals where the blood is not important for the circulation of respiratory gases, no
respiratory pigment may be present. (e.g. insects)
Carefully remove the remainder of the carapace. Locate the pinkish heart lying on the
dorsal surface of the thorax of the crayfish (Figure 19).
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The crayfish, like any other aquatic arthropod, has an abundance of blood vessels
supplying the blood to the tissues. Find the ostia (small holes in the heart, shown in
Figure 15) through which blood re-enters the heart from the hemocoel. Note the location
and extent of the hemocoel (Figure 18).
Figure 18: Circulatory System of a Large Crustacean (a Lobster).

Arrows indicate the direction of blood flow. Note the large blood sinus (hemocoel).
Figure 19: Picture illustrating some of the internal features of the crayfish.
The gonads (testes or ovaries) are lateral to the anterior portion of the intestine,
as seen in Figure 15. Note that Figure 15 only illustrates the position of the testes. The
ovary, however, is located in the same area and looks like a thin-membraned sac filled
with eggs. Testes are generally white and ovaries are orange. The testes are difficult to
distinguish from the digestive glands, but the ovaries are coarser in texture and darker in
color. Sperm from the testes exit the body through a pore at the base of the fifth pair of
walking legs, and eggs are released at the base of the third pair of walking legs. If you
have problems finding the gonads on your specimen, observe the dissected specimens on
demonstration. During reproduction, males and females copulate and sperm cells are
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passed to the genital openings of the female. Fertilization is internal. Mating usually
takes place in autumn. The eggs remain "glued" to the female abdominal appendages
until spring while the young go through three molts and develop their appendages. At
this stage, the young leave the female to continue development to the adult form.
B. THE GRASSHOPPER (Romalea sp.)
External Anatomy of the Grasshopper
Romalea sp. is a large grasshopper common in the south-eastern United States.

Its main habitat is open grassland, weed patches, and roadside growth where it eats
grasses and other vegetation.
Obtain a preserved grasshopper (either a female or a male) and place it in your dissection
pan. The grasshopper belongs to the Class Insecta which is characterized by having three
body regions (head, thorax, and abdomen), one pair of antennae, and six legs (Figure
20).
Romalea has 10 abdominal segments and the terminal abdominal segment bears
the reproductive genitalia. The terminal segment of males is blunt, whereas that of
females is modified to lay eggs and is called an ovipositor (Figure 21). Determine the
sex of your grasshopper.
Figure 20: External anatomy of a grasshopper.
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Figure 21: Difference in the terminal segment between male and female grasshoppers.
Internal Anatomy of the Grasshopper
i) Digestive System
Position your grasshopper dorsal side upward and cut along the dorsal surface of
the thorax and abdomen and pull the exoskeleton aside. Be sure to keep your scissors
close to the body wall to avoid damage to the internal structures. The digestive system
fills most of the internal cavity of the grasshopper (except in mature females which may
be filled with eggs). A muscular tube, the esophagus, conveys food from the pharynx
into a large storage organ, the crop. Chewed food is stored in the crop. Next is the
stomach to which are attached six double finger-shaped digestive glands, the gastric
caecae, which produce enzymes that are secreted into the stomach to aid in digestion
(Figure 22). Because most of the digestive tract is lined with chitin (except the stomach
and crop) digestion and absorption take place mainly in the stomach. The digestive tract
continues as the intestine, a thin tube without accessory structures. It leads to the short
rectum which opens to the exterior via the anus. The hair-like tubules lying over the
intestine are Malpighian tubules, the excretory organs, which are discussed next.
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Figure 22: Internal anatomy of the female grasshopper (lateral view).
ii) Osmoregulatory and Excretory Systems
The grasshopper has Malpighian tubules. These organs are evaginations of the
gut, which serve as excretory organs in such terrestrial arthropods as insects, centipedes,
millipedes and arachnids. These blind pouches are bathed in blood in the sinuses of the
open circulatory system. Wastes enter the tubules and are removed via the hindgut. In
contrast to aquatic organisms in which nitrogenous wastes often take the form of
ammonia, in these organisms nitrogenous wastes are removed in the form of uric acid,
which is less toxic and needs little or no water for its removal.
Examine your dissected grasshopper and note the location of the Malpighian tubules.
Please see this section in the Tutorial Laboratories portion of the lab manual.
The sexes of the grasshopper are separate, and their reproductive organs are in the
terminal abdominal segments (Figure 22). The gonads are dorsal to the intestine in the
region of the Malpighian tubules. In the male, each of the two testes is composed of a
series of slender tubules, follicles, and is located above the intestine. Each testis is joined
to a longitudinal vas deferens. The vas deferens is joined to a single ejaculatory duct to
which accessory glands are attached. In the female, each ovary is composed of several
tapering egg tubes which produce the ova. Each ovary is joined to an oviduct leading to
the vagina, to which a pair of accessory glands and a single spermatheca is attached. The
latter organ is used to store sperm received at copulation.
Examine your dissected grasshopper and find either its ovaries or testes. Be sure to
observe another groups dissection to see the other sex.
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PRACTICAL LABORATORY 4 ANIMALS II:
PHYLUM CHORDATA
OBJECTIVES OF THE LABORATORY:
1. To examine the external features, the muscular, digestive,

urinary, reproductive, and respiratory systems, and the heart
of the perch and rat. (Phylum Chordata)
INTRODUCTION
Organisms belonging to Phylum Chordata are defined by the presence of five

major features:
1) A dorsal, hollow nerve cord. In mammals this becomes the brain and spinal
cord.
2) The notochord. This is a cartilaginous rod that develops dorsal to the
primitive gut in the early embryo. The notochord acts a cellular
hydroskeleton, which prevents distortion of the body when the body wall
musculature contracts during locomotion. In the lower chordates, the
notochord persists throughout life. However, in the vertebrates it is
surrounded and later persists as the soft centre of the intervertebral discs.
3) The presence, during some stage in the life cycle, of paired pharyngeal
pouches in the pharynx or throat. In most aquatic chordates, the pharyngeal
pouches perforate to form pharyngeal gill slits, but in higher chordates the
pharyngeal pouches are present only during embryonic development.
4) A gland (or tissue) which concentrates iodine.
5) A post anal tail. That is, a tail that extends past the exit of the anus.
Phylum Chordata is divided into three subphyla, of which Subphylum

Vertebrata is of interest to us. We will dissect and examine two animals belonging to
different classes within this subphylum: the perch (class Osteichthyes) and the rat (class
Mammalia). When doing a dissection be sure to follow all instructions carefully. If you
cannot find a structure, be sure to ask immediately. Many structures will vary from their
portrayal in the manual. The diagrams represent only the typical situation but no two
animals are identical. At various points throughout the lab, you will be asked to observe
both male and female specimens. Be certain that you observe a specimen of the opposite
sex to that which you are dissecting.
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A. THE PERCH
External Anatomy of the Perch (Perca flavenscens)
i) Introduction
The most numerous of all vertebrates are the bony fishes. They occupy every
marine environment and comprise the vast majority of the freshwater fish populations.
The perch belongs to this group of fish and is a good example of an aquatic vertebrate.
ii) General Features
Place a preserved perch in a dissecting pan.
Observe the fusiform body shape of the perch. It is laterally flattened and
streamlined to minimize resistance as the fish moves through its aquatic environment.
The body is covered by small overlapping scales. Note the direction in which the scales
overlap another mechanism to minimize resistance to movement. A layer of mucous
covers the scales externally to form a protective layer.
Remove a scale and observe it under a dissecting microscope. Scales of modern fish are
the remnants of bony armor, which covered the bodies of ancient vertebrates.
On the ventral surface of the fish, locate two openings. The larger more anterior
opening is the anus (Figure 1). The smaller more posterior opening is the urogenital
opening. On each lateral surface of the perch is a large crescent-shaped opening to the
gill chamber. Each gill chamber is covered laterally by a bony operculum.
Locate the pigmented line extending from the dorsal border of the operculum
posteriorly to the tail. This is the lateral line system and is formed of modified scales
containing sensory pits. The function of the lateral line system is thought to be detection
of vibrations of low frequency or movements in the water.
The body of the perch is divided into three parts: head, trunk and tail, which are
discussed in more detail in the following paragraphs (Figure 1).
Head: Note the terminal mouth of the perch (i.e. at the anterior, leading tip of the
head). The position of the mouth on the head is a modification reflecting the mode of
feeding. The perch feeds by overtaking prey while swimming. Pull down the lower jaw
and note the many sharp uniform teeth of the perch. Just above the mouth are four nasal
apertures (nares), two on each side. Push a fine probe through one of the anterior nasal
apertures and note that it emerges through a posterior nasal aperture. These nares are
not connected with the mouth.
The large eyes have no eyelids, but are covered by a layer of transparent
integument. The lateral location of the eyes prevents binocular vision.
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Bend the operculum backward and note four gill arches. Note the gills which are
attached to each of the the gill arches.
Trunk: The two sets of paired appendages (fins) located on the trunk of the perch are
homologous to the paired appendages of tetrapods. Just posterior to the opercula are the
two pectoral fins. They are attached to the pectoral girdle, which can be felt just
beneath the skin. Posterior and ventral to the pectoral fins are the pelvic fins.
Tail: Tapering from the trunk, the tail ends as a laterally compressed paddle. The
symmetrical caudal fin surrounds the fleshy end of the tail. This type of symmetrical
caudal fin is correlated with the presence of a swim bladder.
The Median Fins: There are four median, unpaired fins. These are: the caudal fin, just
seen; the anal fin, located posterior to the anus; the anterior dorsal fin, beginning at the
anterior end of the trunk; and the posterior dorsal fin, located posterior to the anterior
dorsal fin.
Figure 1: External Anatomy of the Perch (Lateral View).
Internal Anatomy of the Perch
i) Muscular System
Muscles are very important to an animal, for without them the animal could
neither move or eat or bear young. Most of the skeletal muscles of vertebrates can be
divided into two groups the axial muscles and appendicular muscles. The axial
muscles are in the axis of the body (associated with the axial skeleton) while the
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appendicular muscles are those of the limbs and girdles (associated with the
appendicular skeleton). The remainder of the skeletal muscles are associated with the
eyes, jaws, and in fish, the gills and opercula.
The musculature of the perch is not as complex as that of tetrapod vertebrates. Most
of the muscles of the perch are axial muscles and, considering relative body sizes, the
perch has approximately twice the muscle mass of most tetrapods.
The perch moves via lateral undulation of the body produced by waves of contraction
of the axial muscle mass. These contractions are relatively inefficient, thus a larger
muscle mass is required for locomotion in the perch.
Remove a 2 to 3 cm strip of skin from the left side of the perch approximately midway
between the pelvic and anal fins (see Figure 2). The strip should extend from the anterior
dorsal fin to the midventral line. To remove the skin, cut on the dorsal side of the
specimen. Exercise care in removing the skin, as some of the muscle fibers are anchored
in the dermis.
The axial musculature is composed of a series of muscle segments called myomeres.

Myomeres are separated by bands of connective tissue called myosepta (Figure 2). Each
myomere is zig-zag in shape and looks like a large W tilted on its side. A horizontal
septum divides the myomeres into a dorsal and a ventral portion. The main function of
the musculature is not locomotory but is to support the body viscera. The appendicular
muscles of most fish are small and simple because the paired fins are used primarily to
increase stability and maneuverability during locomotion. The remainder of the perch
musculature is associated with the jaws, gills, opercula, and skull. This musculature is
extremely complex because of the large number of separate moveable bones of the perch
skull. This head musculature has two main functions producing the movements
necessary for feeding and for respiration.
Skin removed on
dissection
Figure 2: Lateral View of Perch Musculature.
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ii) Digestive System
The gastrointestinal (GI) tract is a tube starting at the mouth and emptying either
into a cloaca or to the external environment via an anus. The ingestion, digestion and
absorption of food plus the elimination of undigested wastes are the functions of the GI
tract. The tube of the tract is, in chordates, subdivided into the oral cavity (mouth),
pharynx (throat), esophagus, stomach, small intestine, and large intestine. Accessory
organs are associated with the tract. Examples of accessory organs are teeth, tongue,
salivary glands, liver, and gallbladder.
Beyond the esophagus, the digestive organs are located in the coelom. Within the
coelom, most of these organs are suspended by a mesentery extending from the median
dorsal wall. Blood vessels and nerves extend to these organs via the mesentery. The
lumen of the digestive tract can be thought of as continuous with the external
environment and even though it contains fluids and enzymes that are released into the
cavity, small molecules, which result from digestion, must diffuse across the epithelial
cells, which line the GI tract to enter the bloodstream and tissue fluid. The major
products of digestion are monosaccharides, glycerol, fatty acids, and amino acids. These
and other small molecules are taken into the internal environment where further
processing occurs.
Caudal to the pharynx, differences in the anatomy of the digestive tract are
correlated primarily with the nature and abundance of food, regardless of the animal's
environment (i.e. whether aquatic or terrestrial). Because the walls of plant cells are not
digestible by animal enzymes and because they are mixed with the digestible matter,
processing takes considerably longer in animals whose diets include large amounts of
plant material. Consequently, the intestine of herbivores is usually longer than that of
omnivores and especially carnivores. Herbivores depend on bacteria and protozoa to
break down cellulose and many have a large blind (closed at one end) sac or tube called a
caecum at the junction of the small and large intestines where food is held and processed
by these microorganisms. Caeca increase the surface area available and can be found
almost anywhere in the digestive tracts of animals. Since most digestion and absorption
occur in the small intestine, a caecum at the end of the small intestine is not the best place
for the breakdown of cellulose to occur. Rabbits have solved this problem of poor
location by producing two kinds of feces, one type just from the caecum, which is eaten
immediately for additional processing.
In fish, the mouth opens into the oropharyngeal cavity, which extends to the
esophagus. Both respiratory water and food are taken in through the mouth. Fish have a
primary tongue that is an elevation in the floor of the oropharynx, which is immoveable.
Fish do not have any salivary glands. In carnivorous fish, such as the perch, the teeth are
used to capture and hold the prey so it can be swallowed.
In mammals, the mouth opens into the oral cavity, which extends to the pharynx.
The palate separates the respiratory and oral cavities enabling breathing and processing of
food in the oral cavity to occur simultaneously. Most mammals have a moveable,
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protrusive tongue, which is used primarily to manipulate materials in the oral cavity but
may also be used in procuring food or water. Virtually all mammals have salivary
glands. Saliva functions as a lubricant and may contain enzymes, which begin the
digestion of carbohydrates.
Except for baleen whales and anteaters, all mammals have teeth. The type of
teeth a mammal has reflects its diet. Molars have flat surfaces for crushing and milling
and are characteristic of herbivores. Premolars are used for shearing, cutting, and slicing
except in advanced herbivores where they function like molars. Incisors, chisel-shaped
teeth in the front, are useful for biting. Premolars and incisors are found in herbivores,
carnivores and omnivores. Pointed teeth (canine teeth in mammals) are characteristic of
carnivores and are useful for tearing flesh as well as catching and holding prey.
Because specialization in feeding reduces competition among mammals,

differences in dentition are important in identifying and classifying different groups. In
addition, with rare exceptions, each mammalian species has a set number of teeth.
The digestive tract of the perch (a carnivore) includes the gut tube and the
accessory digestive glands such as the liver and pancreas.
To expose the internal organs, hold the fish with the ventral side up and the head pointing
away from you. Insert the point of your scissors through the body wall in front of the
anus and cut up the midline of the body to the space between the opercula (Figure 3).
Now lay the fish on its right side in the dissecting pan, and remove the skin and the
muscles of its left side. To do that, continue to cut up around the back edge of the gill
chamber to the top of the body cavity. Make another incision from the starting point of
the ventral incision close to the anus, and cut upward to the top of the body cavity (Figure
3). Be careful not to disturb the internal organs. Remove the lateral body wall by cutting
along the top of the body cavity. This procedure will expose the body organs in their
normal position.
Figure 3: Incision Lines for the Dissection of the Perch.
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Find the following internal organs on your dissected specimen:
a) Esophagus located at the extreme anterior end of the body cavity, the esophagus is
a short, straight tube leading from the oropharynx to the stomach.
b) Stomach the stomach is a larger, thick-walled U-shaped tube. In the perch,
breakdown of the food by mechanical and chemical means begins in the stomach. The
size of the stomach varies according to how much food it contains.
c) Pyloric caeca (singular, caecum) the junction of the stomach and the intestine is
marked by the presence of three pyloric caeca. These are blind-ended tubes extending
from the gut that serve secretory and absorptive functions.
d) Intestine originating at the stomach, the intestine forms an S-shaped loop. At the
end of the loop, the intestine constricts and straightens. The intestine extends directly to
the anus. The perch's intestine is less than the length of its body. This correlates with
the animal's carnivorous life style. Herbivorous fish have a longer intestine (2 to 15
times the body length), which is required to provide greater digestive and absorptive
surfaces for the herbivores.
e) Liver situated just anterior to the stomach. On the undersurface of the liver is the
gall bladder. The gall bladder drains bile from the liver, and opens by a number of ducts
into the intestine. Bile is necessary for the proper digestion of fats.
f) Pancreas the pancreas is a digestive gland, usually found along the ventral border
of the intestine. In some fish, the pancreas is embedded in the liver. The pancreas
secretes digestive enzymes into the intestine and hormones (insulin and glucagon) into
the blood.
g) Spleen lying on the posterior dorsal surface of the stomach, the spleen is a football
shaped organ. It functions in the production and maintenance of blood cells. It might be
hard to find on some specimens.
Figure 4: Perch internal anatomy.
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iii) Osmoregulatory System
In first term, we examined the effects of the external environment on individual

cells and saw that changes in the external environment could have devastating effects on
cells. However, most organisms can survive fluctuations in their external environment
that are more extreme than even the hardiest of cells could tolerate. Animals survive
changes in their external environment by implementing physiological systems designed
to maintain a relatively constant internal environment -- a concept referred to as
homeostasis. A variety of regulatory systems and feedback mechanisms operate in order
to maintain homeostasis. Osmoregulation and excretion refer to the maintenance of
solute and water balance and the elimination of the nitrogen-containing waste products of
metabolism respectively.
To maintain water balance, the amount of water entering and leaving the body
must be equal. We acquire water through the ingestion of foods and liquids, as well as
water that is produced by cellular respiration. We lose water in urine and feces, as well
as the evaporative loss due to sweating and breathing. For all animals, individual cells
cannot tolerate a net gain or loss of water. There are two basic approaches to maintaining
water balance.
1. Osmoconformers are animals that do not adjust their internal osmolarity

and are isosmotic with their environment (e.g. most marine invertebrates).
2. Osmoregulators are animals that are not isosmotic with their environment
and have developed mechanisms to regulate their internal solute and water
concentrations (e.g. most saltwater vertebrates, freshwater and terrestrial
animals).
In Phylum Chordata, the kidneys eliminate the nitrogenous wastes of cellular

metabolism as well as a variety of other materials that may be present in the blood in
excess of the body's need. In addition, they conserve materials not in excess. Thus, the
kidneys have a vital function both in excretion and in maintaining an internal
environment that is nearly constant in the water and salt content of body fluids, in pH,
and in the content of sugar and other substances in the blood.
The perch urinary system consists of the kidneys, which lie dorsal to the swim
bladder. At the posterior ends of the kidneys, two small ducts unite to form one
common ureter. The ureter and the gonoduct (which passes genital products from the
gonad to the exterior) empty via the common urogenital opening (Figure 5). The urinary
bladder is a small sac at the most posterior, ventral end of the body cavity. It is a
separate structure in the male, but in the female, it is incorporated into the oviducts to
form the urogenital sinus. Try to find the above bolded structures on your dissected
specimen.
In freshwater fish, such as the perch, the elimination of nitrogenous waste occurs
primarily across the gill epithelium. In these animals, the kidneys functions are
osmoregulatory. The body fluids of freshwater fish are hyperosmotic (hypertonic) to
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their environment. Although the skin of the perch (with its mucous coating) is virtually
impermeable, the lining of the oropharyngeal cavity and the gill epithelium are permeable
to water and salts. Thus, there is a constant osmotic entry of water into the body fluids
and diffusional loss of salts (e.g. NaC1) from body fluids, across these surfaces.
The kidneys of freshwater fish continually produce large volumes of dilute urine.
In these animals, the kidneys are quite efficient in reabsorbing ions (salts) from the urine
being produced. Freshwater fish also actively take up ions at the gills to replace those
lost in urine and feces. The urinary bladder in the perch is small as urine is voided
almost as fast as it is produced.
Figure 5: Diagram of the internal anatomy of the perch (female).
Figure 6: Pictures of the internal anatomy of the perch.
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A brief introduction and overview of vertebrate reproductive systems can be

found in the Lab 4 Tutorial section of this manual.
NOTE: You are dissecting only one perch; therefore, you will only see one sex during
the actual dissection. You are, however, responsible for knowing the
reproductive organs of both the male and female. Be sure to prepare your
specimen so that it will be clear enough for another student to study. Also,
make certain you examine a specimen of the sex that you did not dissect.
Male ()
In the male, the testes are a pair of white elongated bodies lying just below the air
bladder to which they are joined by a thin sheet of tissue, the mesentery. They fuse
together toward the posterior end, and the sperm are passed to the outside through the
urogenital opening.
Examine the male perch and locate the testes.
Female ()
In the female, the ovary lies between the intestine and the air bladder (Figures 5
& 6). It is an epithelial sac filled with eggs. The posterior end of the ovary is tapered,
and the eggs pass to the outside through the urogenital opening just behind the anus.
The ovaries are paired in early stages, as they are in other vertebrates, but during
development, they fuse into a single organ.
Examine the female perch and find the ovaries.
v) Respiratory System
A detailed introduction to respiratory systems of vertebrates can be found in the

Lab 4 Tutorial section of this manual. Please be sure to read over it. In this lab, we will
examine the anatomy of the respiratory systems of vertebrates.
On your dissected specimen, remove the right operculum to expose the gills
(Figure 7). The gills are attached to each of the four pairs of branchial arches (Figure
8).
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Figure 7: Picture of the gills of the perch.
On the anterior surface of the arches are the gill rakers. Each gill has two sets of
filaments, one extending anteriorly, and the other posteriorly. Gills with double sets of
filaments like these are known as holobranchs (Figure 8). Each gill filament is made of
many small thin walled folds called lamellae. The lamellae contain the gill capillaries
and exchange of oxygen and carbon dioxide between water and blood occurs across the
thin walls of the lamellae. In fish, water is constantly being passed over the gills. Water
is drawn into the mouth and then into the pharynx while the opercula are closed. Valves
in the mouth close, the oral cavity contracts and water is forced past the gill rakers,
through the gill slits, over the gill filaments and out behind the posterior free ends of the
opercula which are now open. As water passes over the gill filaments the exchange of
oxygen and carbon dioxide occurs (Figure 8). The arrangement of gill capillaries within
the lamellae is such that the blood flow is opposite to the flow of water over the gills.
This is an example of a counter current system. It is an extremely efficient way to
maximize movement of oxygen into the blood.
In addition to their respiratory function, gills also perform important excretory

functions. Without exception, all fish primarily excrete their nitrogenous wastes via the
gills rather than via kidneys. Marine fish also excrete excess salts via the gills.
Freshwater fish absorb needed salts via the gills.
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Figure 8: Structure of a fish gill.
iv) Circulatory System
The circulatory system functions primarily as the major transport system of the
vertebrate body. Oxygen and food are carried from sites of absorption to all tissues and
carbon dioxide and other wastes are carried from the tissues to sites of removal.
Hormones are also transported by the circulatory system.
The closed circulatory system of vertebrates consists of the heart, arteries, veins,
capillaries, and blood. Arteries carry blood away from the heart to the sites of exchange
in the tissues, in the capillaries. Veins carry blood from tissue capillaries back to the
heart.
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The circulatory system of the perch is a typical low pressure single type system in
which the heart is a single pump and there is a single circuit of blood flow. Venous
(deoxygenated) blood from the body is pumped through the heart forward to the gills.
From the gills, where it is oxygenated, the blood goes directly to the body. Thus the
blood makes a single circuit during which it is pumped, oxygenated, and distributed to
the body, before it returns to the heart. In this pattern of circulation the heart pumps only
deoxygenated blood.
The Heart
To expose the pericardial (heart) cavity extend the ventral incision

approximately 2 cm anterior to the pelvic fins. Extend this incision dorsally under the
gills to the lateral line. The pericardial membrane should adhere to this flap of body
wall you have cut. Gently free the membrane as you raise the cut flap.
You will notice a heavy membrane separating the pericardial and abdominal
cavities. It is termed the transverse septum. The fish heart consists of four distinct
parts. Posterior to anterior (in the same direction as blood flow) these are the sinus
venosus, atrium, ventricle, and bulbus arteriosus (Figures 9 and 10).
Figure 9: Heart of the perch (ventral view).
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Figure 10: Heart of the Perch (right lateral view).
These parts are not arranged in a straight line, but have folded over one another to
produce a S-shaped organ. The top of the S, the sinus venosus, receives blood from two
common cardinal veins and the hepatic sinus. The sinus venosus is thin walled and opens
directly into the atrium. The atrium is equivalent to the paired atria of higher
vertebrates. It is thicker walled and larger than the sinus venosus. The ventricle itself is
a thick muscular structure. Like the atrium, it has a single internal chamber. Backflow of
blood from the ventricle during contraction is prevented by a valve. The last portion of
the perch heart is the bulbus arteriosus. The bulbus arteriosus is really an enlarged, very
muscular portion of the ventral aorta, the vessel in which blood flows away from the
heart and toward the gills. Backflow of blood from the bulbus arteriosus is also
prevented by the presence of valves. Figure 11 traces the flow of blood through the perch
heart.
Figure 11: Longitudinal section through the heart of the perch (lateral view). Arrows
indicate the direction of the blood flow.
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B. THE RAT
External Anatomy of the Rat (Rattus rattus)
i) Introduction
In general, orders of mammals are easily recognized and differentiated on the

basis of external appearance. For example, the external anatomy of a rat, a cat, a whale,
and a BIOL-1116 student are all very different. Features such as the appearance of the
limbs, the method of locomotion and characteristics of the teeth enable us to differentiate
these mammals. All mammals, though, have two unique external characteristics (as well
as other internal characteristics), which distinguish them from all other vertebrate classes:
a) all mammals have hair at some time during their development.

b) all female mammals possess mammary glands with external openings for
nourishing their young.
The major mammalian characteristics can be observed from the study of any
mammal. For this purpose, the white rat is used. It is an albino strain of the European
house rat. The rat is a rodent (Order Rodentia). Rodents have enlarged front teeth and
other adaptations for a gnawing, herbivorous mode of life. The rodents are the most
numerous mammals in both number of genera and species, and number of individuals.
There are no flying or marine rodents but they do occupy almost every known terrestrial
habitat. Other familiar rodents are squirrels, mice, guinea pigs, porcupines, and beavers.
ii) General Features
Place a preserved rat in a dissecting pan.
The rat is a tetrapod with digitigrade locomotion, meaning the animal walks on
the digits only, with the remainder of the foot elevated. The principle covering of the
body is hair, which is derived from the epidermis of the skin. The scales covering the
tail in the rat are also epidermal in origin as are the rats claws. The body of the rat is
divided into four general regions: head, neck, trunk, and tail, which are discussed in
more detail in the following paragraphs.
Head and Neck: The head is large and separated from the trunk by a distinct, moveable
neck. Examine the head. The mouth is bounded by well-developed upper and lower
lips. The upper lip is deeply cleft in the rat. A pair of large upper incisor teeth and a
pair of lower incisor teeth should be visible. External nares are situated on a more or
less flexible nose. The eyes are protected by upper and lower eyelids. The ear
possesses an external fold called the pinna, which directs sound waves into the ear canal
(external auditory meatus). A tympanic membrane lies out of sight at the base of this
canal. The rat, as well as many other mammals, possesses special long sensory hairs
called vibrissae (Figures 12 & 13). Vibrissae grow out from the snout, upper eyelid,
cheek, and chin of the rat.
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Trunk: The trunk is divided into an anterior thorax and a posterior abdomen.
Carefully feel along the ventral surface of the trunk. The thorax contains ribs but the
abdomen does not. Teats or nipples (the external openings of the mammary glands) can
also be located in two rows on the ventral surface of the trunk (Figure 13). There are
usually twelve teats in the rat, evenly distributed between the thoracic and abdominal
regions.
Most mammals have separate urogenital and anal openings. In female rats, the
urinary and genital openings are also separate. In females, the urethral opening is the
most anterior with the vaginal orifice just posterior to it. The anus is located at the base
of the tail, posterior to the vaginal orifice (Figure 13). In males, the urethra is a common
urogenital duct and opens at the tip of the penis. Associated with this, at the base of the
tail, is the sac-shaped scrotum containing the testes (Figure 12).
Tail: The tail of the rat has only a few scattered hairs, which are distributed between
reptile-like scales of epidermal origin.
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Figure 12: External Features of Male Rat (Ventral View).
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Figure 13: External Features of Female Rat (Ventral View).
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Internal Anatomy of the Rat
i) Muscular System
During the evolution from fish to tetrapod mammals, many changes occurred in
the muscular system. Some of these changes are correlated with changes in the mode of
support and locomotion that occurred with the move from an aquatic to a terrestrial
environment. Still other changes parallel changes in the mode of respiration, and the
development of a moveable tongue. Most of the axial musculature of tetrapod mammals
has lost its segmentation. The dorsal musculature supports and moves the vertebral
column and head while most of the ventral musculature has differentiated into thin broad
layers of muscle. These layers support the thoracic and abdominal walls and are
important in breathing movements. Some ventral muscles of tetrapods function to
transfer body weight to the pectoral girdle and appendages. With the evolution of the
appendages as organs of locomotion and support, the appendicular musculature
differentiated into numerous separate muscles.
Place your rat with its dorsal surface up. Using coarse scissors, make a middorsal
incision through the skin extending from the back of the head to the tail. From this
incision, cut through the skin down the lateral surface of the left limbs to the wrist and
ankle. Remove the skin from the left side of your rat (skin at least to the midventral line)
and from the left forelimb. Use a blunt probe to tease the skin away from the muscles.
Also remove any fat and connective tissue covering the muscles. With the aid of Figure
14 and the following instruction, locate the following muscles:
a) External oblique this is the outer layer of the body wall. It is visible on the ventral
and lateral surfaces of the abdomen.
b) Internal oblique using small scissors, make a short, very shallow cut in the
external oblique on the ventrolateral surface, as shown in the picture below. Beneath the
external oblique you will see muscle fibers running in a different direction. These are
fibers of the internal oblique muscle. Remove a square of external oblique to expose the
internal oblique.
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c) Transverse oblique cut the internal oblique as you did the external oblique.
Beneath it you will find a third layer. This is the transverse oblique. Thus, the body wall
is composed of three layers: external oblique, internal oblique, and transverse oblique.
d) Rectus abdominis using scissors, make an incision through the body wall along the
midventral line. Examine the inside surface of the body wall just lateral to the incision.
You will see a long, thin muscle running anterior-posteriorly. This is the rectus
abdominis.
e) Latissimus dorsi remove connective tissue and fat from behind the left arm and
you will see a fan-shaped muscle running from under the arm to the middorsal line and
over the ribs. This is the latissimus dorsi.
f) Appendicular muscles on the left arm locate the triceps brachii and the biceps
brachii. The biceps brachii functions to flex the forearm, the triceps brachii acts to
extend the forearm. These two muscles perform opposite functions, hence, they are
antagonists.
Figure 14: Lateral View of the Rat Musculature.
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ii) Digestive System
Salivary glands: To identify the salivary glands carefully remove the skin from one
side of the face and neck to expose an area as shown in Figure 15. When you are
removing the tissues to expose the glands, be certain not to cut too deep. There are three
pairs of salivary glands. The first is the parotid gland, which lies just beneath the ear
and extends over the ventrolateral surface of the neck to the shoulder. The submaxillary
glands are just ventral to the slightly larger parotids and are inseparable from the more
anterior overlying sublingual glands. Also locate a fourth gland, the extraorbital
lacrimal gland, which forms tears and other eye secretions. Examine Figure 15 to find
the relative location of the aforementioned glands.
Figure 15: Diagram (top); and picture (bottom) of the salivary glands of the rat.
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Oral cavity: With a pair of heavy scissors or bone cutters, cut the muscles, skin and
mandible at the angle of the mouth (only on one side) and depress the lower jaw with
your fingers. This cut should be done on the opposite side of the salivary glands
dissection. Locate the following structures: large incisor teeth, molar teeth, the
tongue, and the bony roof of the mouth or hard palate (Figure 16). The palate separates
the nasal and oral cavities. Unlike carnivorous mammals (and yourself) whose teeth stop
growing shortly after they emerge, the teeth of many herbivorous mammals (including
the rat) continue to grow throughout life. Note that the rat does not have canines or
premolars.
Figure 16: Picture of the oral cavity of the rat.
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Abdominal Cavity and Internal Organs: Follow the incision lines indicated in Figure
17 and the description below to expose the abdominal and thoracic cavities of the rat. A
pair of scissors (as opposed to a scalpel) should be used to make the incisions so that
damage to the visceral organs is avoided.
Make a small incision with your scissors just caudal to the posterior end of the
sternum. Do not cut too deeply into the body cavity. Snip through the abdominal wall
toward the pelvic area. If your specimen is a male, cut posteriorly and to the right of the
penis until you reach the scrotum. To expose the thoracic cavity, cut in an anterior
direction through the ribs, just to the right of the mid-ventral line. Cut through the body
wall laterally, just anterior to the diaphragm (Figure 17) and make two more lateral
incisions in the posterior region of the abdomen. Gently rinse the thoracic and
abdominal cavities with running water.
Figure 17: Incision lines for dissection of the rat.
The coelom is the body cavity within which the viscera (internal organs, such as
liver, stomach, intestines, etc.) are suspended. The abdominal cavity and viscera are
covered by a membranous tissue called the peritoneum, which is formed from
mesoderm. Recall that the coelomic cavity of evolutionarily advanced invertebrates and
vertebrates is always lined by mesodermal tissue. The peritoneum is extensive and forms
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the following delicate membranes. It is crucial to observe the membranes at this point
because they are very delicate and usually get damaged during the examination of the
internal organs.
a) Mesentery Proper A double layer of peritoneal membrane extending from

the dorsal body wall to the viscera (internal organs).
b) Falciform Ligament A layer of tissue extending from the ventral body wall
and diaphragm to the liver.
c) Parietal Peritoneum This is the shiny membrane which lines the body wall
of the abdominal cavity.
d) Visceral Peritoneum This is the shiny membrane which covers the
internal organs (viscera) within the abdominal cavity.
e) Greater Omentum This is a double walled peritoneal sac which extends
from the greater curvature of the stomach to the spleen.
f) Lesser Omentum This membrane joins the lesser curvature of the stomach
to the liver.
After finding all of the above membranes, locate the following organs and glands
associated with digestion (Figures 18 a and b):
a) Stomach As in other animals, this sac-like structure serves as a storage site for
ingested foods. The stomach is comprised of three areas, which are morphologically
similar but histologically different. These are the cardiac portion (entrance from the
esophagus), the fundic portion (large middle area) and the pyloric portion (constricted
posterior portion) (Figure 18). The stomach opens to the duodenum through the pyloric
sphincter. Cut open the stomach with a median incision and wash out its contents. Note
the folds (rugae) on the walls. Observe the pyloric sphincter at the point of junction
between the stomach and duodenum.
b) Small Intestine The small intestine is comprised of three regions, which can be
differentiated histologically. The anterior portion (the duodenum) receives the ducts
from the digestive glands, pancreas, and liver. The second portion is the jejunum and
the most posterior is the ileum. The small intestine is the site of most chemical digestion
and the absorption of nutrients. Digestive enzymes from the pancreas and the intestine
itself are secreted into the lumen of the small intestine where the chemical breakdown of
food occurs. Note the length of the small intestine. Cut a piece (1-2 cm long) of the
small intestine. Open it and wash it out. Examine it under the dissecting microscope.
The velvet texture of the lining of the small intestine is created by numerous minute
projections, the villi. The absorptive surface is greatly increased by the presence of villi.
c) Large Intestine The ileum opens to the large intestine via the ileocolic valve. The
large intestine consists of four areas: a large caecum or blind sac near the ileocolic valve;
an ascending colon on the right side; a short transverse colon; and a descending colon
heading posteriorly.
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The caecum is quite large in herbivores, like the rat, and may contain microorganisms,
which further breakdown the plant material not already digested by the enzymes of the
small intestine.
The major function of the large intestine itself is the re-absorption of the large
quantities of water secreted into the gut during digestion. Thus, as undigested material
moves along the colon, water is removed from it, resulting in a mass of waste material,
the feces. Feces are stored in the rectum (the terminal portion of the colon) until
eliminated through the anus.
d) Rectum The rectum is the continuation of the descending colon through the pelvic
region. It terminates with the anus, which opens externally.
e) Liver It is divided into several lobes. The bile produced in the liver passes directly
to the duodenum, via the bile ducts. The gall bladder, which stores the bile in most
mammals, is not present in the rat. Bile is needed for proper digestion of fats. The liver
has many functions such as detoxification of certain chemicals and production of
glycogen (a carbohydrate storage material).
f) Spleen This is an elongate flattened organ, which is attached to the greater

curvature of the stomach through the greater omentum. The spleen and the liver remove
old red blood cells from circulation and break them down. The products of the
breakdown are secreted into the stomach by the spleen and into the duodenum by the
liver.
g) Pancreas The pancreas is a diffused gland, which is embedded in the mesentery

proper and greater omentum. It is found along the anterior edge of the duodenum, just
after the stomach. The pancreas secretes many digestive enzymes into the small intestine
as well as hormones (insulin and glucagon) into the blood.
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Figure 18 (a): Diagram of the internal organs of the rat.
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Figure 18 (b): Close-up views of the stomach and caecum of the rat.
iii) Osmoregulatory System
Examine your dissection specimen and locate the paired kidneys, which are
embedded in fat on the dorsal body wall (Figures 19 & 20). Note the fat, which remains
around the kidneys. Locate the ureter, which is found exiting the kidney (along with the
renal blood vessels) at a depression called the hilus (Figure 21). The ureter is the duct
which carries urine from its site of formation to the urinary bladder for storage. The
urinary bladder is usually quite small and firm in its empty state and its muscles are
contracted in these preserved specimens. The duct leading from the bladder to the
exterior is the urethra. In the male, the urethra travels through the penis and is the
passage for sperm during reproduction as well as the tract for urine at other times. In the
female, the reproductive products do not exit through the urethra. It is also very difficult
to find in the female.
Examine the demonstration of a pigs kidney that has been cut in half with a median
longitudinal section. You will notice that there may be large amounts of red and/or blue
dye in some parts of the kidney; this is indicative of the highly vascularized nature of the
kidneys. In addition, the kidney is not homogeneous; the outer portion is called the
cortex and the inner region is called the medulla (Figure 21).
137
Figure 19: Female rat urogenital system. Figure 20: Male rat urogenital system.
Figure 21: Longitudinal section through kidney: diagram (left); and picture (right).
138
In this lab you will study the macroscopic features of the reproductive system. In
the tutorial lab you will study the microscopic features. As mentioned in the perch
section above, a brief introduction and overview of vertebrate reproductive systems can
be found in the Lab 4 Tutorial section of this manual.
NOTE: You are dissecting only one rat; therefore, you will only see one sex during the
actual dissection. You are, however, responsible for knowing the reproductive
organs of both the male and female. Be sure to prepare your specimen so that it
will be clear enough for another student to study. Also, make certain you
examine a specimen of the sex that you did not dissect.
Male ()
Locate the scrotum, a large sac of skin, muscle and connective tissue containing
the testes on the exterior of the body just ventral to the anus. During non-breeding
periods, the testes may be retracted into the abdominal cavity and the scrotum will not be
enlarged. However, sperm cannot develop completely within the high temperatures of
the abdominal cavity; the slightly lower temperature of the scrotum is necessary for the
final stages of sperm formation.
Carefully cut open the scrotum and locate the paired testes, which are oval. Around the
outside of each testis is a C-shaped structure known as the epididymis, which is a very
long, highly coiled tubule. Three regions of the epididymis can be recognized: the caput
(head) epididymis covering the anterior end of the testes, the corpus epididymis lying
along the lateral surface of the testes, and the cauda (tail) epididymis on the posterior
end of the testes. Sperm cells produced in the seminiferous tubules of the testes pass into
the caput epididymis via the vasa efferentia (very small tubules that are not visible).
Then, the sperm cells move through the corpus and cauda epididymis into the vas
deferens, which is a moderately large tube leading from the epididymis to the urethra
which is a tube located inside the penis. Notice that the penis is enclosed in an epithelial
sheath and held along the ventral wall of the abdomen (Figures 20 & 22).
The prostate glands are found on either side of the urinary bladder. Locate the
large, lobed vesicular glands, and the coagulating glands, which are closely applied to
the inner curve of the vesicular glands. The above glands and some other glands in this
region comprise the accessory sex glands. The secretions of these glands form the
seminal fluid, which carries the sperm during ejaculation, activates and provides certain
nutrients for them, and contains substances which neutralize the somewhat acidic
environment in the vagina.
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Figure 22: Pictures of the male rat reproductive system.
Female ()
Locate the ovaries, which appear as a mass of follicles just posterior to the
kidneys. They are often buried in a mass of fat, which should be carefully removed. The
adult non-pregnant rat is usually in breeding condition with mature or well-developed ova
in the ovaries. The oviducts are small, highly coiled tubes leading from the ovaries to the
uterus. Locate the right and left cornua (horns) of the uterus. The cornua of the uterus
unite to form the vagina (Figures 19 & 23).
Open one horn of the uterus and examine the interior for embryos, which might be
present. The vagina leads to the exterior separately from the urethra. The vagina opens
to the exterior via the external vaginal orifice, which opens just posterior to the urethral
opening. The clitoris, which is the female homolog of the male penis, is associated with
the urethra and therefore is not connected with the vagina in the rat.
B
A
Figure 23: a) Female rat reproductive system with developing fetuses in the uterine horns.
b) Female rat reproductive system in the non-pregnant state.
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v) Respiratory System
Earlier in this laboratory, you made an incision along the midline on the ventral
side of the rat. If you have not already cut through the rib cage, do so at this time. Use
your scissors and start at the base of the rib cage near the diaphgragm and progress
cranially to the lower jaw. Be careful, many glands, blood vessels, and organs lie just
under the skin and may be damaged if you cut too deeply. Spread the rib cage and
expose the organs of the thoracic cavity and the organs in the neck region. When this is
completed, the heart will be visible in the center of the cavity. Locate the lungs located
on each side of the heart. The membrane lining the wall of the thoracic cavity is the
parietal pleura; the membrane covering the lungs is termed the visceral pleura. There
is one lobe to the left lung and four lobes making up the right lung (cranial lobe, medial
lobe, caudal lobe, and accesory lobe) (Figure 24). Just caudal to the lungs, you should be
able to see a diaphragm, a thin muscular sheet.
Locate the trachea, which is a long tube supported with cartilaginous rings to
prevent collapse as the organism inhails. Follow the trachea posteriorly to the point at
which it branches into a right and left bronchus. These bronchi lead to the lungs where
they branch further into bronchioles (Figure 24).
Follow the trachea anteriorly and locate the larynx (the larynx is visible in Figure
27). It should appear as an enlarged, square-shaped box. The larynx allows mammals
to have a vast repertoire of sounds ranging from ultrasonic squeaks and chirps (in bats) to
the highly complex sounds of human speech.
Figure 24: Diagram of the rat respiratoy system.
141
vi) Circulatory System
The circulatory system of the rat typifies the high-pressure double circulatory
system of mammals in which the heart is a double pump and there are two circuits of
blood flow, systemic and pulmonary. In this type of system, blood is pumped through
the heart twice. Deoxygenated blood from the body is pumped to the lungs where it is
oxygenated (pulmonary circulation). Oxygenated blood from the lungs returns to the
heart whereupon it is pumped to the rest of the body (systemic circulation). Thus the
heart pumps both oxygenated and deoxygenated blood. In this lab we will examine only
the heart and its associated vessels. The vessels comprising the rest of the circulatory
system will be studied in the Tutorial lab.
The Heart and associated vessels
Be extremely careful during this portion of the dissection as the arteries and veins
are extremely fragile. You will examine the heart and locate only the major blood vessels
coming out of and going into it. Locate the thymus gland, which overlies the anterior
portion of the heart. Carefully remove the thymus gland.
The heart of the rat consists of four chambers. There is a right ventricle, and a
left ventricle, which are not easy to distinguish externally. Also, locate the easily distin-
guishable right atrium and left atrium which are dark ear-shaped structures on each
side of the anterior portion of the heart (Figure 25).
Figure 25: Ventral view of the heart of the rat.
142
Use forceps and carefully dissect the muscle and fatty tissue away from the major
arteries and veins in the neck region. This is a tedious process and will take some time.
Your rat has been double-injected with latex. Blue latex was injected into the veins
and red latex was injected into the arteries. Entering the right atrium are three main
blood vessels which bring the deoxygenated blood back to the heart from all regions of
the body. These blood vessels are the right superior vena cava, the left superior vena
cava, and the inferior vena cava. The left superior vena cava may be seen running
across the dorsal surface of the thoracic cavity to enter the right atrium close to the point
of entry of the inferior vena cava and the right superior vena cava. The right and left
superior venae cavae return deoxygenated blood to the heart from the right and left side
of the head and neck. The inferior vena cava may be located by lifting the heart and
carefully separating the lobes of the lung. It returns deoxygenated blood to the heart from
the lower part of the body. The inferior vena cava is a large vein running from the
diaphragm to the right atrium. The thoracic cavity is drained by the azygous vein which
empties into the left superior vena cava near its entry into the atrium. It is found only on
the left side of the thoracic cavity (Figures 26 & 29).
The deoxygenated blood in the right atrium is pumped to the right ventricle
through an opening guarded by the tricuspid valve. The blood is pumped from the right
ventricle through the pulmonary semilunar valve into the pulmonary trunk (usually
colourless) which divides into right and left pulmonary arteries going to the lungs.
The oxygenated blood returns from the lungs to the left atrium via the right and
left pulmonary veins. These can be found on the concave surface of the lungs but are
difficult to trace to the atrium (do not attempt).
From the left atrium the blood enters the large muscular left ventricle through the
bicuspid valve.
Figure 26: Dorsal view of the heart of the rat.
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144
TUTORIAL / THEORETICAL
LABORATORIES
145
146
TUTORIAL LABORATORY 1 BACTERIA, PROTISTS, & FUNGI:
INTRODUCTION TO THE LIVING WORLD

DOMAIN BACTERIA
DOMAIN EUKARYA: THE PROTISTS
DOMAIN EUKARYA: THE FUNGI
OBJECTIVES:
1. To introduce the organization of the living world.
2. To examine the ecological importance of the Domain

Bacteria.
3. To examine the evolutionary development of Eukaryotes.
4. To examine sexual reproduction in some representative

protists and the three main groups of Fungi.
5. To look at the ecological impact of some protists and

fungi.
In this weeks lab you will continue your exploration of the Bacteria, Protists, and
Fungi. Last week you examined asexual reproduction as well as many slides which
illustrated the morphology of these organisms. This week we will examine some of the
evolutionary and ecological aspects of these organisms including sexual reproduction.
I AN INTRODUCTION TO THE CLASSIFICATION OF ORGANISMS
The biological system of classification catalogs, organizes, characterizes, and

names the millions of organisms that constitute life on Earth. In order to systematically
study the diverse organisms on earth, scientists had to organize the vast amount of
different species of organisms. Taxonomy (Greek, organizing rules) is the science of
naming, describing, and classifying the organisms into similar groups. The classical
classification scheme as proposed by Carlos Linnaeus, and which is still in use today,
consists of a series of 8 major categories or taxa (singular, taxon). Actually, Linnaeus
only recognized 7 taxa. The level of Domain is a relatively recent addition. Biologists
place organisms in these taxa based on their similarities, then group similar taxa into
larger and larger categories. The basic structure of this hierarchical classification is as
follows:
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DOMAIN (largest, most inclusive taxon)
KINGDOM (subdivisions of domains)
PHYLUM (subdivisions of kingdoms)
CLASS (subdivisions of phyla or divisions)
ORDER (subdivisions of classes)
FAMILY (subdivisions of orders)
GENUS (subdivisions of families)
SPECIES (distinct type of life)
Consider, for example, the classification of humans:
Domain: Eukarya
Kingdom: Animalia
Phylum: Chordata
Class: Mammalia
Order: Primate
Family: Hominidae
Genus: Homo
Species: Homo sapiens
Except for the species, which we can define, the taxa are not subject to precise
definitions and they are somewhat subjective in nature. This often leads to some lively
disagreements on classification among biologists depending upon their own subjective
interpretations of the available evidence. In recent years the study of taxonomy and
systematics, has become a very dynamic and exciting area of biology. With the advent of
many new molecular techniques, it is now possible to compare the genetic structure of
different organisms and this has provided new insights into the evolutionary histories of
many groups. These new techniques have given us new insights into many of the major
macroevolutionary events which occurred early in the history of life, such as the origin of
the eukaryotes, or the origin of multicellularity, and the ultimate origin of the major
groups such as animals, plants and fungi. As we begin to better understand these events,
we reorganize the taxonomy to better reflect the actual evolutionary history of life on
Earth. This is an ongoing process which will undoubtedly continue for many years. In
this lab we will briefly examine how one of these macroevolutionary episodes (serial
endosymbiosis) has shaped the taxonomy of the protists.
One of the practical values of the classical taxa comes at the genus and species
levels. Each organisms needs to be uniquely identified. That is, everyone needs their
own unique name.
We use a binomial system of nomenclature based upon the scheme originally

introduced by Carolus Linnaeus, in the 18th century. In this system, each organism is
identified by a universally understood two-part Latin or Latinized name consisting of the
name of the genus to which the organism belongs and the specific epithet, which is
unique for each species within the genus. In this system, the rat is always Rattus rattus
and the earthworm is always Lumbricus terrestris whether it is being studied in Japan,
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Egypt, or Canada. The genus name is always capitalized while the specific epithet is not.
Both are underlined or italicized. The two words together are called the scientific name
of an organism.
This begs the very important question What make one species unique from
other species? There are several ways to think of a species. That is, there are several
species concepts. One species concept that has proven very useful, particularly when
thinking about evolutionary questions such as speciation, is the concept of a Biological
Species. A biological species is said to be: a population or group of populations whose
members have the potential to interbreed in nature and produce viable, fertile offspring,
but do not produce viable, fertile offspring with members of other such groups
(Campbell and Reece, 2011). So two organisms who can not interbreed and produce
viable, fertile offspring are said to be different species. This concept helps us to
understand the mechanisms that would work to reproductively isolate populations and
therefore helps us to understand the evolutionary processes of speciation. However this
biological species concept is of little value when considering organisms who do not
reproduce sexually, or for organisms only represented by fossils. We will not have the
opportunity to discuss the biological species concept in detail in the lab, however it is a
major topic that you will deal with in lecture.
In this course, you will also note that the spelling of certain biology words
changes depending on whether they are used in a singular or plural form, for example:
bacterium (singular) and bacteria (plural). Some other examples are: spirillum (spirilla),
ovum (ova), larva (larvae), alga (algae), coccus (cocci), bacillus (bacilli).
II IMPORTANCE OF PROKARYOTES
The process of nitrogen fixation.
Bacteria from the Rhizobium Genus can transform atmospheric nitrogen (N2) into
compounds that plants, and eventually other organisms, can use. These compounds
include ammonia, nitrate, and nitrite. This process is called nitrogen fixation and the
only organisms on earth that can carry out this biochemistry are certain prokaryotes. One
major group of nitrogen fixing bacteria are found in the soil. Bacteria like the genus
Rhizobium are a major source of useable nitrogen in the terrestrial environment. Many
species of Cyanobacteria are also important nitrogen fixers in the freshwater and marine
environments. The enzymes involved in this nitrogen fixation process are collectively
referred to as nitogenases. These enzymes are very sensitive to oxygen. Oxygen appears
to inactivate these enzymes and shut down the nitrogen fixation processes. As a result of
this oxygen sensitivity, many cyanobacteria have evolved special cells called
heterocysts that have adaptations to exclude and reduce the oxygen inside them, so as
to maximize the efficiency of the nitrogenase enzymes.
Certain plants, especially legumes (peas, beans, alfalfa, and clover) have formed
symbiotic relationships with these nitrogen-fixing bacteria. Legumes supply the bacteria
with carbohydrates and other organic compounds. Bacteria, on the other hand, supply the
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plant with fixed nitrogen. The nitrogen-fixing bacteria are commonly known as
root living because they live in swellings called nodules along the roots. Legume-
Rhizobium symbiosis generates more useful nitrogen for plants than all industrial
fertilizers and the symbiosis provides the right amount of nitrogen at the right time at no
cost to the farmer.
Observe the nodules on the roots of the plant on demonstration. Also, note Figure 1
shown below.
Fig. 37-10a
Nodules
Roots
(a) Pea plant root
Figure 1: Nodules on the roots of a pea plant.
III EVOLUTIONARY HISTORY OF EARLY EUKARYOTES
It appears from an examination of the fossil record that the first eukaryotic cells
arose about 2.5 billion years ago. The origin of the eukaryotic cell design was a
monumental step forward in the history of life on earth. For the first 2 billion years of the
history of life there were only prokaryotes. These prokaryotes were very successful, and
evolved all of the major biochemical processes of life that we see in modern cells
today. However the morphological evolution of diverse multicellular organisms was
not possible with the simple prokaryotic cell structure. The best the prokaryotes could do
was simple colonies and chains of cells. The more complex eukaryotic cell design
allowed for the internal specialization of cells, different internal compartments for special
cellular functions, and for specialization of entire cells for specific functions, and
therefore the development of multicellular organisms. But before the multicellular
organisms appeared there was a host of unicellular organisms that diverged and radiated
out as what we call the protists. The story of how these unicellular protists evolved is
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one of fusion and symbiosis, as well as natural selection.
Problem of the protists
In last weeks Practical lab we examined some of the protists by looking at

organisms, which we called algae, that where kind of like plants, in that they where
primary producers and carried out photosynthesis. Another group of protists the
protozoa, where more like animals, in that they were heterotrophs and tended to move
about. Finally we saw the fungal-like protists which looked a lot like fungi and often
fed saprophytically, like many fungi. As we said last week, this grouping was purely
for convenience. The actual evolutionary history of these organisms turns out to be far
more complex. As described in chapter 28 (page 617) of your textbook, the origin and
diversity of the protists can best be explained by the process of serial endosymbiosis.
The actual evolutionary history of this group is best represented by phylogenetic

hypotheses and the supergroups as presented in your textbook on page 620. Basically,
endosymbiosis involves a host cell (organism) engulfing a symbiont cell (organism) by
phagocytosis. Normally this type of feeding would result in the symbiont cell being
digested by the host cell. However according to the endosymbiotic hypothesis, these
endosymbionts, upon occasion, are not digested but remain within the cytoplasm of the
host and carry on performing the abilities it has, but now to the benefit of the host cell as
well. Figure 2 illustrates how this hypothesis explains the origin of mitochondria and
chloroplasts in early eukaryotes.
Figure 2: The endosymbiotic hypothesis for the origin of mitochondria and chloroplasts.
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IV SEXUAL REPRODUCTION AND LIFE CYCLES
One of the most important concepts in biology is the idea of reproduction and life
cycles. If you recall the list of characteristics of life described in chapter 1 of the text,
reproduction was a major activity of living things. All your discussions of cell division,
(mitosis and meiosis) last term in BIOL-1115 were leading up to this idea of
reproduction.
In the beginning there was simple cell division. That is, a cell copied its DNA,
then separated those two copies of DNA, and then divided up the cytoplasm into two
new daughter cells. When the prokaryotes do this we call it binary fission because of
the relatively simple mechanics involved. When it happens in more complex eukaryotic
cells, it is called mitosis. Although there are several variations on the mechanisms of
mitosis seen in several groups of protists, the process always involves; copying the
chromosomes, getting those copies of DNA apart, then dividing up the cytoplasm and all
that cellular machinery, and ending up with two genetically identical daughter cells.
Creating genetically identical offspring has its advantages and its disadvantages. One
advantage is, if the environment in which you live is very stable and you are well
adapted to that environment, then producing offspring that are genetically identical to
you means they will all be well adapted to that environment too. Another advantage is
that no other organism is needed to divide in two.
However, in a rapidly changing environment chances are that the combination of

genes you copy and pass on to your genetically identical offspring wont be as good in a
new environment as they were in the old environment. It would be a very good thing, in
a rapidly changing environment, if all your offspring were slightly different, genetically.
This would mean that there might be some offspring with genes that would give them
characteristics that might better adapt them to the new environment. Of course many of
the offspring would not have very good sets of genes and they would likely die very
quickly, but genetic variation in the offspring should allow at least some to survive in the
changing environment.
As you saw in BIOL-1115, the type of cell division involved in producing

genetically different offspring is called meiosis. You should go back and review the
processes of both mitosis and meiosis, but recall that eukaryotic cells can have one copy
of each chromosome (haploid) or they can have two copies of each chromosome
(homologous chromosomes) and be diploid. The process of meiosis takes a diploid cell
and divides it into 4 haploid cells. Its important to remember that these haploid cells
will all be genetically different from each other and genetically different from the parent
cell. If two of these haploid cells then fuse together (fertilization) they combine the two
sets of chromosomes to form a new diploid cell which is also genetically different than
any other diploid cell. You should go back and review what processes of meiosis create
this variation. This is a life cycle. The processes of meiosis and fertilization are the basis
of sex, and therefore these life cycles are often referred to as sexual life cycles.
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It is this genetic reshuffling that is the chief advantage of sex, and a very
important component of evolution and natural selection. What you will see as we work
through the algae and fungi (and eventually the plants) are variations on this basic sexual
life cycle.
In Practical Lab 1, we looked at asexual reproduction in the Fungi and downy

mildews. We also looked at sexual reproduction in the water molds. In this lab, we will
examine reproduction in the algae, as well as sexual reproduction in the plasmodial slime
molds and Fungi.
A. REPRODUCTION IN ALGAE
Like everything else involving algae, methods of reproduction are quite varied. In
Practical Lab 1 we introduced the algae and examined the characteristics of some
representative groups. In this lab we will continue our look at the algae by examining
some of the different ways in which reproduction is accomplished.
i) Asexual Reproduction
For aquatic organisms such as algae, the environment for the most part is fairly
stable. The temperature of the water can fluctuate, but this is usually over an entire year
rather than in a matter of hours. Nutrient levels can also change but usually do so
relatively slowly. Only light fluctuates in a rapid manner, and even this is in a rather
predictable manner, i.e. dark at night, light during the day. Since the stable environment
reduces the requirements for genetic variability, methods of asexual reproduction are very
common in the aquatic algae. Many algae never reproduce sexually. However, sexual
reproduction (which we will discuss shortly) is very important in many algae during the
production of cells or structures designed to withstand periods of seasonal dryness or
frozen winter conditions.
Two methods of asexual reproduction utilized by algae will be examined. These are:
a) Daughter colony formation

b) Sporulation
a) Daughter Colony Formation
A limited number of colonial algae produce miniature replicates of the colonies.

These are termed daughter colonies. These may be produced inside the hollow,
spherical colonies, or inside the actual cells of the parent colony. Eventually, the parent
colony will rupture and release the new daughters.
Examine the prepared slide of Volvox sp. (slide #73) and note the daughter colonies.
Also, refer to Figure 3.
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Volvox sp. consists of many Chlamydomonas-like cells bound in a common
spherical matrix. Each cell in the sphere has two flagella extending outward from the
surface of the colony. Synchronized beating of the flagella spin the colony through water
like a globe on its axis.
Observe a demonstration of a live culture of Volvox sp. and note the characteristic
movement of the colonies.
Figure 3: Volvox sp.
b) Sporulation
This is the most common form of asexual reproduction in the algae. Sporulation
refers to the process in which any cell of an organism produces one or more reproductive
cells inside its cell walls. The original cell is termed a sporangium and the new cells are
termed spores. Spores are often produced in large numbers for a rapid increase in
population size.
Examine the prepared slide of Ulothrix sp. (Slide #28) and locate developing spores,
known as zoospores (motile spores) located within zoosporangia. These zoospores
swim away from the parent, settle down and develop directly into new filaments (Figure
4).
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Figure 4: Sporulation in Ulothrix sp.
ii) Sexual Reproduction
Although most algae reproduce asexually, sexual reproduction is often associated

with the production of resistant stages or over-wintering structures. The algae have
evolved many variations on the basic theme of sexual reproduction, such as different
types of gametes, different means of gamete transfer, and different locations of
fertilization.
A gamete is a cell that fuses with another gamete in the process of fertilization
(also known as syngamy). By definition, gametes must be haploid since the result of
fertilization is a diploid cell (Zygote). The process of gamete formation is called
gametogenesis. The relative form of the two fusing gametes defines two categories of
sexual reproduction: isogamy and heterogamy.
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Isogamy
Isogamy is the form of sexual reproduction in which the gametes produced are
identical in shape, size, and motility. There is no structural distinction between "male"
and "female" gametes, which are known as isogametes.
A specific example exhibiting non-motile isogametes is the reproductive process

known as conjugation, which occurs in the filamentous green alga, Spirogyra sp.
Examine Slide #85 of conjugating Spirogyra sp.. Identify the four stages of the process
as outlined in Figure 5.
Figure 5: Isogamy in Spirogyra sp. (a) Resting filaments of alga cells. (b) Formation of
conjugation tubes between two adjacent filaments. (c) Cytoplasmic contents of each
cell form a compact mass, representing an isogamete. The isogametes from one filament
migrate through the conjugation tubes into the adjacent filament. (d) The two isogametes
unite to form a zygote. Each zygote eventually undergoes meiosis to form four haploid
cells. One haploid cell will form a new filament by mitosis, the other three degenerate.
Heterogamy
In heterogamy, two different types of gametes are produced. The male gamete,
the sperm cell, is typically very small, highly motile, and is produced in very large
numbers. The female gamete, the egg cell, is much larger and non-motile. Fewer female
gametes are produced but each is usually afforded some protection. Heterogametes are
also produced by higher plants and animals.
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Oedogonium sp. is a green alga that produces heterogametes. Figure 6 illustrates
the life cycle of this alga.
Examine Slide #90 and locate a mature egg cell and the small male filaments, which are
the site of sperm production. In the species you are examining, the egg cells and male
filaments are usually adjacent to one another on the same algal strand.
Figure 6: Heterogamy in Oedogonium sp.
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B. SEXUAL REPRODUCTION IN PLASMODIAL SLIME MOLDS
The plasmodium of Physarum that was on demonstration in the Practical Lab was
in its vegetative (non-reproductive) stage. It would continue to grow for as long as there
is adequate food and moisture. When either of these factors is in short supply, one of two
scenarios can occur. One scenario sees the plasmodium dry into a hard resistant structure
called a sclerotium and remain dormant until conditions improve. Alternatively, the
plasmodium will migrate to an exposed area and enter its reproductive stage. In this
stage, the plasmodium will produce fructifications (fruiting bodies) which are
characteristic of the species. There are two major types of fruiting bodies: sporangia
(singular; sporangium) and aethalia (singular; aethalium) (Figures 8 & 9). Sporangia are
usually borne at the tip of a stalk and can be extremely ornate. Aethalia are relatively
massive, single-chambered structures. They may have been formed by complete fusion
of large numbers of sporangia during their evolutionary development. No trace of
individual sporangial walls are found in most aethalia. It is within these fructifications
that meiosis occurs to produce the haploid spores. The spores of slime molds are very
small and easily dispersed by the wind. The spores will germinate as amoeboid or
flagellated organisms. These haploid stages may later fuse as gametes and grow into a
new plasmodium. Figure 7 illustrates the life cycle just described.
Figure 7: The life cycle of a plasmodial slime mold that produces sporangia.
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Figure 8: Sporangial forms of slime molds.
Figure 9: Aethalia.
Examine the display of different sporangial and aethalial forms and be able to
differentiate these two types of fruiting bodies.
Examine a demonstration slide of a longitudinal section through a sporangium of

Stremanitis sp.. Also, examine a demonstration slide of a cross section through an
aethalium of Lycogala sp.. Be sure to notice the spores in both slides.
Examine the demonstration posters that show some of the images of the Irish Potato
Famine.
C. SEXUAL REPRODUCTION IN THE KINGDOM FUNGI
Life cycles are a very important component of the survival and success of any
organism. Understanding why certain basic life cycle types have evolved helps us to
understand the diversity of life on earth. In Practical Lab 1 you examined many of the
basic structural features that separate the three main groups of fungi, as well as how they
reproduce asexually. In this weeks lab, we will look at the sexual lifecycles of these
fungi.
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a) Phylum Zygomycota
The Zygomycota have haploid, aseptate (coenocytic) feeding hyphae. This

haploid stage is the most dominant stage of the life cycle. By dominant we mean, most
obvious, longest lived, etc. (not dominant like in genetics). These organisms spend most
of their life cycle as haploid hyphae growing and feeding (saprophytically). As in
Practical Lab 1, we will use Rhizopus stolonifera as a representative of this phylum.
Sexual reproduction in R. stolonifera occurs only between different mating

strains, which have been traditionally labeled as + and types (or Strain 1 and Strain 2,
as seen in Figure 10). Although the mating strains are morphologically indistinguishable,
they are often shown in life cycle diagrams as different colours. When the two strains are
in close proximity, hormones are produced that cause their hyphal tips to come together
and develop into gametangia (Figures 10 & 11), which become separated from the rest
of the fungal body by the formation of septa. The gametangia are swellings in the two
hyphae which fill with haploid nuclei. Eventually the wall between the two gametangia
will break down and the two sets of haploid hyphae will mix together in the same
chamber (plasmogamy). This chamber becomes the zygospore. The zygospore will
form a thick outer cell wall which is so strong it can allow the zygospore to withstand
very bad environmental conditions such as drying out or freezing. These resistant
zygospores remain dormant until the environmental conditions improve. Each zygospore
has two sets of separate haploid nuclei and is therefore referred to as dikaryotic. (Recall
this from Practical Lab 1) Eventually the two sets of haploid nuclei fuse (karyogamy)
forming many diploid nuclei. It is these diploid nuclei that eventually undergo meiosis at
the time of germination. The zygospore then cracks open and produces a sporangium
that is similar to the asexually produced sporangium, and the life cycle begins again.
This life cycle is illustrated in Figure 10 below.
Examine the prepared slide #26 of Rhizopus stolonifera sexual reproduction. Note the
gametangia and the early and late stages of zygospore formation.
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Figure 10: Life cycle of Rhizopus stolonifera.
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(a) R. stolonifera, gametangia.
(b) R. stolonifera, young zygospore (c) R. stolonifera, a thick-walled zygospore.
Figure 11: Pictures illustrating the stages of sexual reproduction of R. stolonifera.
b) Phylum Ascomycota
It should be noted that sexual reproduction in the Ascomycota always involves the
formation of an ascus (plural, asci). An ascus is a sac-like structure that is characteristic
of this phylum and distinguishes the Ascomycota from all other fungi. Ascus formation
is usually within a complex structure composed of a tightly interwoven hyphal network
known as an ascocarp. Many ascocarps are macroscopic, and are the only part of these
fungi that most people ever see. An ascocarp may be open and more or less cup-shaped
(called an apothecium), closed and spherical in shape (called a cleistothecium), or flask
shaped with a small pore through which the ascospores escape (called a perithecium).
The layer of asci is called the hymenium, or hymeneal layer, and lines the interior of the
ascocarp. Figure 13 shows examples of these different types of ascocarps. In this lab
you will observe only an apothecium.
The life cycle of the Ascomycota starts with the feeding haploid hyphae. The
Ascomycota spend much of their lives as these haploid hyphae, growing and feeding
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saprophytically through the soil. As in the Zygomycota, the haploid hyphae of the
Ascomycota can fuse to form a dikaryotic stage (i.e. containing two haploid nuclei, one
from each strain). When two compatible mating strains of an Ascomycota species grow
close together they stimulate the production of structures that will facilitate the fusion of
the hyphae. These structures are often called gametangia as well. However, in the
Ascomycota the gametangia produced by the male hyphae are called antheridia
(singular, antheridium) and the gametangia produced by the female hyphae are called
ascogonia (singular, ascogonium). The male nuclei of the antheridium pass into the
ascogonium via a tubular outgrowth of the ascogonium known as the trichogyne.
Plasmogamy, or the fusion of the two cytoplasms, occurs when the trichogyne fuses with
the ascogonium. The male nuclei then pair with the genetically different female nuclei
within the common cytoplasm but do not fuse. Hyphae now begin to grow out of the
ascogonium. As the hyphae develop, pairs of nuclei migrate into them and
simultaneously mitotic divisions occur in the hyphae and ascogonium. Cell division in
the developing hyphae occurs in such a way that the resulting cells are dikaryotic. The
dikaryotic hyphae grow together to form the ascocarp.
The ascus first forms at the tip of the developing dikaryotic hypha. The two
nuclei in the terminal cell (ascus) of the dikaryotic hyphae then fuse into a single diploid
nucleus (karyogamy). This is the only diploid cell (structure) in the life cycle. The
diploid ascus then elongates and the diploid nucleus divides by meiosis, forming 4
haploid nuclei. Each haploid nucleus usually divides again by mitosis, resulting in a total
of 8 haploid nuclei. These haploid nuclei are then cut off in segments of the cytoplasm to
form ascospores. In most Ascomycota, the ascus becomes turgid at maturity and finally
bursts, sending its ascospores explosively into the air. Figure 12 below illustrates the
basic life cycle of the Ascomycota.
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Figure 12: Life Cycle of a Typical Ascomycota.
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(a)
(b) (sectioned)
(c)
Figure 13: Some typical ascocarps found in the phylum Ascomycota.
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Other examples of sexual reproduction in the Ascomycota
i) Yeast
Recall from Practical Lab 1 that yeast are predominantly unicellular and
reproduce asexually by fission or by budding (pinching-off of small buds). Sexual
reproduction in yeasts occurs when either two cells or two ascospores unite and from a
diploid zygote. The zygote may produce asexual buds, or may undergo meiosis to
produce four haploid nuclei. In some species there may also be a subsequent mitotic
division producing eight haploid nuclei. The single cell in these unicellular yeasts is
acting as an ascus, and therefore the whole organism becomes the reproductive structure.
Within the ascus/zygote wall, walls are laid down around the nuclei so that eight
ascospores are formed. These are liberated when the ascus wall breaks down. The
ascospores either bud asexually or fuse with another cell to repeat the sexual process.
Make a wet mount of the culture of yeast and find budding cells.
Two genera of yeasts, Saccharomyces sp. and Schizosaccharomyces sp. are

commonly used in the baking and brewing industry. In both these genera the asci are
formed by the fusion of two haploid cells.
Examine the prepared slide #84 of Saccharomyces sp. "sporulating", and note the
presence of 4 ascospores in each ascus. Also note that in these unicellular yeasts, there is
no ascocarp. Figure 14 shows yeast cells in various stages of asexual and sexual
reproduction.
Figure 14: Yeast cells budding and sporulating.
ii) Cup Fungi
The cup fungi are the most advanced group of Ascomycota. They produce an
ascocarp called an apothecium, with the asci arranged in an exposed layer. Although
many apothecia are disc or cup-shaped, other forms also exist.
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Peziza sp.
The apothecia of Peziza sp. often exceed 10 cm in diameter. These apothecia are
usually bowl-shaped when young but will become flattened and distorted with age. At
maturity, the thousands of asci on the surface of the cup develop hydrostatic pressure. If
the cup is disturbed, the asci rupture releasing thousands of ascospores in a visible "puff".
Wind currents then transport the spores to a new environment.
Examine the prepared slide #94 of the apothecium and note the asci and ascospores.
Also, examine the preserved material of different species of Peziza sp. on demonstration
(Figure 17).
(a)
(b)
Figure 15: (a) Diagram of the apothecium of Peziza sp.. (b) Microscopic views of asci.
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Morchella sp. (morels)
These are some of the most highly prized edible fungi. The apothecium of
Morchella sp. has a stalk, or stipe, and a fertile portion called the pileus (Figure 16). The
pileus is essentially discoid, but it is folded over the stipe apex and is highly contorted.
These distortions greatly increase the surface area of the pileus. The asci line the large
pits, which are separated by sterile ridges.
Examine the preserved material on the side bench and note the large, folded pileus.
(a) (b)
Figure 16: (a) Diagram of the apothecia of Morchella sp.. (b) Picture of morels
c) Phylum Basidiomycota
Sexual reproduction in the Basidiomycota always involves the formation of a

basidium (plural, basidia). A basidium is a sac-like structure that is characteristic of this
phylum and distinguishes the Basidiomycota from all other fungi. The basidia produced
have club-like extensions which give this group of fungi its common name: Club fungi.
Basidium formation is usually within a complex structure composed of a tightly
interwoven hyphal network known as a basidiocarp. Basidiocarps are large fruiting
structures, which are the most visible stage of the fungus. A typical mushroom is a
familiar example of a basidiocarp. The basidia are borne in a layer on the surface of gills
which, in turn, are produced on the underside of fleshy umbrella-like basidiocarps. The
basidiospores are forcibly ejected form each basidium. The basidiocarp consists of a
stout stalk (stipe) bearing a circular cap (pileus) from which the lamellae (gills) hang
down (Figure 17). It should be noted that the basidiocarp is an ephemeral structure
usually lasting only a few days, whereas the mycelium, living on organic matter in the
soil, may last for years.
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Figure 17A: Diagram of a typical basidiocarp.
Figure 17B: Basidiocarp of Coprinus comatus (inky cap mushroom).
Examine the mushrooms on the side bench, and note the basidiocarps in various stages of
development. In these basidiocarps you should note the stipe, pileus, ring (annulus), and
gills.
The Basidiomycota life cycle begins, as before, with the haploid feeding hyphae
which grow through the substrate and feed saprophytically. When two sexually
compatible mating strains (+ and -) grow close to one another, the hyphae simply fuse
(plasmogamy) and the two haploid nuclei move into the resulting cell. This forms a
dikaryotic cell. That cell continues to divide. As it does, each haploid nucleus divides
too, and the resulting daughter cells will end up with one of each kind of haploid nucleus.
This forms a chain of cells (a hyphae) in which all cells are dikaryotic, a dikaryotic
hyphae. As with the Ascomycota, the cells at the ends of the dikaryotic hyphae will be
the ones to form the sexual reproductive structure. In the Basidiomycota the terminal
cells of the dikaryotic hyphae form the basidium. When the basidium forms, the
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terminal cell of the dikaryotic hyphae swells and the two haploid nuclei finally fuse
(karyogamy). This diploid basidium now undergoes meiosis and the 4 resulting haploid
nuclei migrate through small stalks at the end of the basidium. The four basidiospores
wall off at the ends of these stalks. In many Basidiomycota, the mature basidiospores are
forcibly ejected off the ends of these stalks and carried away in the wind.
Mycologists, those who study fungi, often refer to the stages of the Basidiomycota
life cycle as passing through three distinct phases: primary, secondary, and tertiary.
When it germinates, a basidiospore produces the primary mycelium. Initially the
mycelium may be multinucleate, but septa soon form and the mycelium is divided into
haploid monokaryotic (uninucleate) cells. This septate mycelium grows by division of
the terminal cell. Branches do occur, and the mycelial mass can become very complex.
The dikaryotic hyphae (mycelium) that eventually forms is commonly called the
secondary mycelium. The tertiary mycelium, which is also dikaryotic, arises directly
from the secondary mycelium, and forms the basidiocarp. Figure 18 below illustrates
the life cycle of the Basidiomycota.
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Figure 18: Life Cycle of a Typical Basidiomycota fungus.
Examine the prepared slide #95 of a cross section of a pileus (cap). Note the gills, layers
of basidia, and basidiospores (Figure 19).
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(a) Gills x.s. (b) Gills x.s., close-up.
(c) Basidium and basidiospores, close-up.
Figure 19: Microscopic views of cross-sections through the gills of a basidiocarp.
Other examples of Basidiomycota fungi
i) Bracket Fungi
Members of this diverse group of fungi produce basidiocarps that are woody,
leathery, or papery, but never soft. The basidia are found covering the surface of gills (or
teeth), or lining the inside of pores. Many of the common bracket fungi (or shelf fungi)
found growing on the surface of living or dead tree trunks are members of this group
(Figure 20).
Examine the preserved material of members of this group.
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Figure 20: Some members of the bracket fungi.
ii) Puffballs and Earth Stars
In the puffballs, the mature basidiocarp consists of a papery outer covering with a
small opening or ostiole on the top. Inside is the mass of spores. When raindrops strike
the leathery covering, "puffs" of spores are ejected through the ostiole (Figure 21).
Examine the demonstration material of puffballs (Figure 22) and earthstars (Figure 23).
Remember the structure you see is a basidiocarp, similar to that of a mushroom.
Figure 21: Young and mature puffball basidiocarps.
Figure 22: Lycoperdon sp. (puffballs).
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Figure 23: Geaster sp. (earthstars).
V THE ECOLOGICAL IMPACT OF PROTISTS AND FUNGI
So far we have looked at the morphological characteristics and reproductive

cycles of some representatives of protists and fungi. It is important to understand,
however, that these organisms are integral parts of our living world and have important
roles in many natural systems. This section will provide you with some insight into the
ecological importance of these organisms. Keep in mind that their roles in any
environment can be positive or negative. Sometimes these organisms contribute to the
health of an ecosystem. However, sometimes these organisms can be detrimental, as can
be the case with human crops.
A. ALGAL ECOLOGY
We certainly dont have time to explore all the aspects of the ecology of the algae.
However you should be aware that as photosynthetic organisms in marine and freshwater
habitats, the algae are the primary producers of these aquatic food chains. They capture
the energy from sunlight and convert it into energy stored in the chemical bonds of
organic molecules. These molecules, that makeup their bodies, are then food for other
organisms.
B. PROTIST ECOLOGY
As with the algae, the ecology of the protists is too large a topic to tackle in this
lab. Therefore, we will briefly examine one specific example involving a downy mildew
that had a dramatic impact on humans.
As mentioned in Practical Lab 1, downy mildews generally live on land and are
parasites of plants. Many have significant impacts on economic plants and cause
tremendous losses of crops each year. The infamous Phytophthora infestans causes late
blight of potatoes, which turns the stem of potatoes into a black slime. "Phytophthora"
literally means "plant destroyer". This organism produced the great potato famines in
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Ireland. The famine of 1845-47 was responsible for over 1 million deaths from starvation
and initiated large-scale emigration from Ireland to the United States. Within a decade,
the population of Ireland dropped from 8 million to 4 million. Virtually the entire Irish
potato crop was wiped out in a single week in the summer of 1846. The disease remains
a major problem today and causes crop losses of about 15% in North America.
C. FUNGAL ECOLOGY
While there is no single fungal lifestyle, one thing common to all fungi is that
(unlike the green plants, for example) they do not make their own food. Fungi get their
nutrients from existing organic matter, and there are many sources of organic matter in
the world: leaf litter, dung, soil, animals, dead wood, and living plants are just a few
examples. If a fungus feeds on dead organic matter, its known a saprophyte. Fungi that
get their nutrients from living organisms do so in a variety of ways. These various
methods of nutrient acquisition can be categorized into one of two types of relationships.
If a fungus is in a relationship where there is no benefit to the other organism, the fungus
in question is a parasite. If, however, there is some benefit to both the fungus and the
other organism, the relationship is considered to be mutualistic. Note that in mutualistic
associations, the benefits need not be equally shared. You will often see the word
symbiotic used to describe associations between different organisms and that word
literally means "living together". Its a general term covering all types of associations,
with no implication about the nature of the association. Hence, parasitic and mutualistic
are two examples of symbiotic associations.
i) Saprophytes
Fungi are the primary decomposers in natural ecosystems and without their
activities significant amounts of dead material would accumulate and nutrient recycling
would be limited. In particular, one major component of plant cell walls known as lignin
(which is responsible for the hardness of wood) is only significantly broken down by
some fungi. Many free living soil fungi feed by secreting digestive enzymes through
their outer cell walls and the digestion of the dead organic matter in the substrate occurs
externally. They then absorb the digested nutrients through their cell walls.
Agaricus is a common genus of saprophytic fungi (Figure 24). The ordinary Field
Mushrooms belong to this genus and you will often see various Agaricus species growing
in gardens, parks, and paddocks. In such situations the mycelium is feeding on dead
organic matter in the ground. Commercially, Agaricus is grown on composted plant
material.
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Figure 24: Agaricus sp.
ii) Parasitic Fungi
Many fungi are parasites of plants and other organisms. In this role, fungi
sometimes act as crop pests and frequently have disastrous effects on our food supplies.
As an example, it is estimated that between 10% and 50% of the worlds fruit harvest is
lost each year on account of fungal pests (Figure 25). On the other hand, the antibiotic
revolution which has saved millions of lives first started with the discovery of the
antibacterial properties of penicillin. We also use fungi extensively in food
manufacturing to produce alcohol, bread, cheeses, and much more.
Figure 25: Rhizopus sp. on strawberries
iii) Mutualistic Fungi
An important type of mutualistic relationship is the mycorrhizal association

between fungi and plants. The word mycorrhiza is derived from the Classical Greek
words for 'mushroom' and 'root'. In a mycorrhizal association the fungal hyphae of an
underground mycelium are in contact with plant roots, but without the fungus parasitizing
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the plant. In fact, the association is commonly mutually beneficial. Through
photosynthesis, a chlorophyll-containing plant makes simple carbohydrates (using carbon
dioxide, water, and sunlight). About 90% of plant species form mycorrhizal associations.
In many of these relationships, 10% to 30% of the food produced by the plant moves
through to the fungi. The associated fungal mycelia are adept at extracting minerals
(especially nitrogen and phosphorus) from the soil and these pass through to the plants.
Mycorrhizal fungi can also protect plants against pathogenic fungi and micro-organisms.
It has also been shown that plants who have their fungal partner removed do not grow as
well as plants with intact mycorrhizae. All in all, mycorrhizal fungi are very important
for plant health.
(a) (b)
Figure 26: (a) Fungal mycorrhizae on plant roots. The fungus appears as small white
strings on the outside of the root. (b) Increase in root area of a plant with mycorrhizae.
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REVIEW/STUDY QUESTIONS
(These are questions about the material that has been covered in both the Practical AND
Tutorial labs. Use your lab manual and experiences in the labs to answer the questions.
These are meant to help you review the material and study for the lab exam)
Green Algae
Question 1: What particular morphological features do the following algae possess

which enhance their survival?
Chlamydomonas sp.:
_______________________________________________________________________
_______________________________________________________________________
Chlorella sp.:
_______________________________________________________________________
_______________________________________________________________________
Question 2: Is there any feature lacking in the above mentioned organisms, which
they need to survive?
_______________________________________________________________________
_______________________________________________________________________
Question 3: Suggest a reason as to why colonial algae, such as Gonium sp. and
Scenedesmus sp., would have evolved their particular shape.
_______________________________________________________________________
_______________________________________________________________________
Question 4: How many planes of cell division occur in each of the following types of
filamentous algae?
Ulothrix sp. _____________________________________
Cladophora sp. ___________________________________
Fritschiella sp. ____________________________________
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Question 5: Which genus (out of the ones mentioned in Question 4) would be most
successful in a damp soil environment? Why would this particular genus be most
successful in a damp soil environment?
_______________________________________________________________________
_______________________________________________________________________
______________________________________________________________________
_______________________________________________________________________
Question 6: How are green algae different from Cyanobacteria?
_______________________________________________________________________
_______________________________________________________________________
Question 7: What is the advantage of asexual reproduction to the algae?
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
Question 8: What is the main disadvantage of asexual reproduction to the algae?
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
Animal-like Protists
Question 9: How does the movement of Paramecium sp. compare with that of
Amoeba sp., Volvox sp., and Chlamydomonas sp.?
_______________________________________________________________________
_______________________________________________________________________
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Question 10: Why is the division of Paramecium sp. called fission (transverse or
binary) and why is it an example of asexual reproduction?
_______________________________________________________________________
_______________________________________________________________________
Fungal-like Protists
Question 11: What is the purpose of the grain of oat seen in the culture of Physarum
sp.?
_______________________________________________________________________
_______________________________________________________________________
Question 12: What is the specific function of haustoria (singular, haustorium)?
_______________________________________________________________________
_______________________________________________________________________
Kingdom Fungi
Question 13: Are conidia haploid or diploid?
_______________________________________________________________________
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TUTORIAL LABORATORY 2 KINGDOM PLANTAE:
PLANT HISTOLOGY
SEED PLANT ENVIRONMENTAL ADAPTATIONS
OBJECTIVES:
1. To introduce land plant evolution with regard to the non-

reproductive structures.
2. To examine land plant histology.
3. To examine some of the key adaptations seed plants have

developed in order to become so dominant in the biosphere.
I THE EVOLUTION OF VEGETATIVE CHARACTERISTICS
In last weeks lab you examined the evolution of the reproductive structures and
processes in land plants. This week you will examine the evolutionary history of the
vegetative (morphological) improvements. The story begins, as it did last week, with the
initial colonization of the land by the Charophytes (green algae). The story continues
even today, with natural selection constantly improving the abilities of land plants to
survive the hostile terrestrial environment. Recall from Practical Lab 2 that land plants
have three important things they need to do in order to survive in the harsh terrestrial
environment:
1) They must be able to absorb light for photosynthesis.

2) They need to absorb water from their surroundings and make sure it is
available to all the cells of the organism.
3) They must find a supply of carbon dioxide and nitrogen compounds
that are needed to build the cells organic molecules.
There are some basic differences between the aquatic and terrestrial habitats that
we need to examine. In the aquatic environment, the essential nutrients needed to survive
are generally available in the surrounding water. Light, another essential requirement, is
readily available as well (particularly in shallow waters). As such, less effort is required
by the aquatic plant to absorb and distribute these resources to the algal body. The
terrestrial environment, however, does not provide all the essential elements for survival
in one location and desiccation is a constant threat. Light is generally coming from
above, and carbon dioxide can be absorbed from the air. However, water and minerals
are found in the soil. Land plants must therefore have separate organs for absorbing light
from above, and water and minerals from the soil below. The aquatic environment also
provides significant support for the body of the plant underwater, via the surrounding
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water pressure. However the air is less dense and provides virtually no support against
the pull of gravity. So land plants must have an organ that is strong enough to hold up
the weight of the plant against the pull of gravity. As we work our way through the three
groups of land plants you will see how these organs (leaves, stems, and roots) have
evolved to better adapt land plants to this difficult environment.
Morphological & Anatomical Diversity in the Plant Kingdom
Plant morphology is the gross or entire structure of the plant. This includes size
and shape of leaves, type of root systems, etc. Plant anatomy is the arrangement of cells
and tissues within the organism.
Plants are obviously different than animals. Plants are relatively simple in design.
They have only four organs: leaves, stems, roots, and a reproductive organ (either cones
or flowers). Plants are also sedentary (they dont move around) except in the sense that
their spores or seeds may travel to new locations and germinate, eventually growing into
new plants. Animals are far more complex than plants. Animals have many complex
organ systems such as the nervous system, muscular system, and digestive system. Why
are plants and animals so different? Because plants can feed themselves, and animals
need to find food. That is, plants are photosynthetic. All they need to create their food
by photosynthesis is a source of water, carbon dioxide, and sunlight. Animals on the
other hand must move around and search for food. Then they have to catch, ingest, and
digest their food. All the complex animal body plans, structures, and even behaviours are
adaptations to find food.
There is much evidence that the first land plants invaded the swampy shorelines
of the Middle Silurian period, about 425 million years ago. The evolution of land plants
then followed a pattern of adaptation from an aquatic to a terrestrial existence. The early
primitive land plants would have lacked adaptations to the very dry terrestrial
environments and would have been restricted to swampy, boggy types of habitats. As
time progressed, natural selection would have selected for better adaptations to the drier
terrestrial habitats and so the more recently evolved plants would have continued to
invade drier terrestrial environments. The increase in complexity from algae to seed
plants reflects, to some extent, an increase in the ability to survive in drier climates.
Recall that most algae live in aquatic habitats. These habitats are very stable
environments because the daily temperature fluctuations are minimal, there is no threat of
desiccation, nutrients are usually readily available, and a dispersal medium is present.
The major problem that the algae face is the possibility of sinking to a depth where light
is not sufficient for photosynthesis. The terrestrial environment, on the other hand, does
not offer this stability. It is subject to climatic changes, water shortages, and nutrient
deficiencies. Perhaps the most significant feature of the terrestrial environment is the
spatial separation of basic plant requirements. As was mentioned above, light is above
the soil and water and inorganic nutrients are in the soil. This dictates that one portion of
the plant should be adapted for photosynthesis, and the remainder of the plant should be
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adapted for water and mineral absorption as well as anchorage. There must also be a
route of exchange for the products of each of these regions. The environment has,
therefore, determined that the plant, if it is to be successful, should have a division of
labour. Simply put, each plant organ has a job. Leaves are adapted to maximize the
processes of light absorption and photosynthesis. Stems, to a certain extent, help
photosynthesis by elevating the leaves high into the air to catch more sunlight. Roots
anchor the plant and also absorb water and minerals from the soil. Cones or flowers are
adapted to maximize reproduction as we will see later.
As you examine the morphologies of the different groups, notice the increasing
complexity of each of these organs. Also, be aware that these groups, while reflecting a
possible pattern of plant evolution, do not necessarily illustrate the actual way in which
plants evolved.
II PLANT HISTOLOGY
All multicellular organisms are made of cells. These cells have become specialized
to perform specific functions in the organisms body. Before we examine the morphological
adaptations of a few groups of land plants, we need to review the basic types of cells and
tissues that make up their bodies. Your text has a very good review of plant histology. You
should read Chapter 35 Plant structure, growth and development from Biology,
Canadian edition by Campbell, N.A. and Reece, J.B. (2013).
As cells differentiate, they become quite dissimilar from one another and must be
integrated structurally and functionally to form coordinated units. Multicellular organisms
are not random collections of different cells performing different functions. Rather, they
possess different levels of organization: cells tissues organs organ systems. A
tissue may be defined as a group of similar cells performing a common activity. Histology
is the study of cells and tissues. An organ may be defined as a group of tissues collectively
performing a specific function. An organ system is defined as a group of organs
performing a major body function.
In this weeks lab there are demonstrations of microscope slides and related
information set out on the side benches. As you work through these different plant cell and
tissue types, you should be able to recognize each type of cell and be able to describe its
primary function. We will start by looking at the basic plant tissue types and then move on
to examining the histology of some different plant groups.
i) Basic Plant Tissue Types
Plants derive all of their cells from rapidly dividing zones known as meristems.
Meristems are domes of rapidly dividing cells. The cells produced by these meristematic
regions then mature and differentiate in the permanent tissues that make up the plants body.
There are typically two apical meristems on any plant: one at the tip of each above ground
shoot which produce cells of the stems and leaves; and another at the tip of each
underground root which produce all the root tissues.
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The cells produced by the apical meristems develop into one of three basic
permanent tissues; fundamental tissue, epidermal tissue, or vascular tissue. These
permanent plant tissues consist of mature, differentiated cells that do not generally
undergo any further cell division.
Epidermal (dermal) Tissues
These tissues form the protective outer covering of the plant body. In young plants
the principle surface tissue of the roots and stems is epidermis. Epidermis is also the
surface tissue of all leaves. Usually the epidermal layer is only one cell thick, however it
may be thicker in certain species that live in very dry habitats and where protection against
water loss is critical. Most epidermal cells are relatively flat, have very large vacuoles, and
only small amounts of cytoplasm. Epidermal cells on the above ground parts of the plant
often secrete a waxy, water-resistant cuticle onto their outer surfaces.
Figure 1: The epidermis and cuticle of a generic land plant.
Vascular Tissues
Vascular, or conductive tissue, is a distinctive feature of the higher plants; one that
has made possible their extensive exploitation of the terrestrial environment. This tissue
includes cells that function as tubes or ducts through which water and numerous substances
in solution move from one part of the plant body to another. There are two principle types
of vascular tissue: xylem and phloem. Both of these are complex tissues (i.e. they consist
of more than one kind of cell).
Xylem is a vascular tissue that functions in the transport of water and dissolved
substances upward in the plant body. It forms a continuous pathway running through the
roots, stem, and appendages of the stem, such as leaves. The xylem tissue is always located
more internally than the phloem tissue. In the ferns, xylem consists of only one kind of cell
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known as tracheid cells. However in the flowering plants the xylem usually includes two
unique types of cells: tracheids and vessel elements. The xylem also includes numerous
parenchyma and sclerenchyma cells. The parenchyma cells are the only living cells in the
mature, functioning xylem. The cytoplasm and nuclei of tracheids, vessel elements, and
sclerenchyma cells disintegrate at maturity, leaving the thick cell walls as the functional
structures. Another function of the xylem is support, particularly of the aerial part of the
plant. The numerous fibres present in xylem tissue function almost exclusively in this way.
In addition, the thick-walled tracheids are also important as supportive elements.
Figure 2: Diagram of the constituent cells of xylem tissue.
Phloem is also a complex tissue which contains conducting cells as well as

supportive fibres and parenchyma. The function of the phloem tissue is to conduct water
and carbohydrates (used by plants for food) upward and downward in the plant. The
conducting cells in phloem are called sieve elements. The sieve elements are arranged in a
vertical series to form a structure called a sieve tube. Unlike the tracheids and vessel
elements of xylem tissue, the sieve tubes retain their cytoplasm at maturity, but their nuclei
disintegrate. The sieve elements are elongated cells with specialized porous areas in their
end walls called sieve plates. Food material located in the cytoplasm can move from one
cell to the next by means of cytoplasmic streaming. Usually one or more specialized,
elongated parenchyma-like cells, called companion cells are closely associated with the
sieve elements in most flowering plants. Mature companion cells retain both their
cytoplasm and their nuclei. It has been suggested that all of the cellular functions of sieve-
tube elements are controlled by companion cells.
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Figure 3: Diagram of the constituent cells of phloem tissue.
Fundamental (ground) Tissues
These tissues have functions other than protective or conductive. They may function
in storage, support, photosynthesis, etc. Parenchyma, sclerenchyma, and collenchyma
tissues are representatives of fundamental tissues and are discussed in more detail below.
Parenchyma cells are the most common type of plant cell. They are quite large and
possess thin walls, which remain thin and flexible even when the cells are mature. These
cells contain leucoplasts which function in the storage of starch produced in photosynthesis.
Leucoplasts appear as purple stained granules on your slide. Parenchyma cells of the stems
of some plants contain chloroplasts and have photosynthetic functions.
Figure 4: Parenchyma cells.
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Sclerenchyma cells are small and posses tough, thick cell walls. These walls are
often impregnated with lignin, a highly branched polymer that makes cell walls more rigid.
As such, sclerenchyma is used as a structural support tissue in land plants. Mature
sclerenchyma cells can not elongate and are found in regions of the plant that have stopped
growing in length. At maturity, sclerenchyma cells may actually be dead.
Figure 5: Sclerenchyma.
Collenchyma cells are also specialized for structural support and are located as
cylinders of tissue beneath the epidermis. Contrary to sclerenchyma, they support
elongating (still growing) parts of the plant. Collenchyma cells are relatively flexible and
allow the plant to bend without breaking. Their cell walls are thinner than those of
sclerenchyma and slightly thicker than those of parenchyma. The walls do not contain any
lignin.
Figure 6: Collenchyma cells.
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ii) Phylum Bryophyta (Mosses)
These plants normally live in damp, shady, terrestrial environments. There are a
few aquatic mosses and some that live in extremely dry environments, but these are the
exception. There is mounting evidence that the first plants that made the transition from
the aquatic to the terrestrial habitat were indeed vascular plants. The modern mosses
appear to have lost the use of the specialized internal water transport tissues. While the
ferns and other vascular plants retained the use of their xylem and phloem. Regardless,
the morphological and reproductive characteristics of the Bryophytes are thought to be
similar to those first land plants.
The Bryophytes have traditionally been classified as non-vascular plants. They

are the only land plants that lack functional vascular tissues for the transport of water and
organic and inorganic nutrients. They also require free water for reproduction. For these
reasons, mosses tend to be small in size and to grow in clumps (Figure 7). When you
examine representatives from this group, you should become aware of the features which
distinguish them from the algae.
Morphological and Anatomical Diversity in Mosses
Examine a whole moss plant under a dissecting microscope. It is best to separate one
plant from the whole culture and place it under the dissecting microscope. Note the
obvious differences in various areas of the plant. These differences reflect a division of
labour far more extensive than that illustrated in the alga, Fritschiella sp. Which you saw
in Practical Lab 1. You should be able to find leaves, the stem, and root-like rhizoids.
Note the spiral arrangement of the leaves on the stem.
Cluster of mosses Individual plants
Figure 7: Parts of the moss plant.
Examine the prepared slide of a moss stem (Slide #98). Note the epidermis, storage
cortex of parenchyma cells, and the central cylinder. This central cylinder in most
mosses is made up of cells that look like vascular tissue, but dont work. This means that
any transport of water and nutrients throughout the plant must occur by diffusion between
cells or up the outer surface of the moss plant through capillary action. Note Figure 8.
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Central cylinder
Cortex
Epidermis
Leaf
Figure 8: X.S. of a moss stem.
Make a wet mount of a leaf and examine it under a compound microscope. Note the
simplicity of the moss leaf. The blade portion of the leaf is only one cell layer thick.
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Figure 9: XS moss leaf.
iii) Phylum Pterophyta (Ferns)
Ferns, like mosses, are considered to be terrestrial plants. Most are located in
areas of damp soil, high humidity, and low light, although there are minor habitat
exceptions in this group. Some of the more common habitats are: moist woods, dry
woods, mossy boulders, and rock crevices. They are mainly found in moist tropical areas
with only a few adapted to temperate regions. While this dependency on an environment
which does not readily lead to desiccation is partially due to morphological and
anatomical (vegetative) deficiencies, it is also due to the reproductive system. The ferns
have advanced over the mosses primarily through the evolution of vascular tissue (which
allows rapid, long distance transport of materials) and true roots (which allow for
absorption of water and nutrients from the soil). This has allowed the ferns to increase in
size, to become more distinct in their division of labour, and for the absorbing portion of
the plant to become more remote from the photosynthetic sites.
True roots allow these terrestrial plants to access water from deeper in the soil.
Upper layers of soil often dry out through evaporation, but water is often more
consistently available in deeper layers of soil. The roots provide a large surface area
through which the water and minerals can be absorbed. Notice on the diagram below
(Figure 10) that the stems of many ferns grow horizontally underground. These types of
stems are called rhizomes. The roots of these ferns grow out of the horizontal
underground stems. This origin makes these roots adventitious. An adventitious root is
any root that arises from tissues on the plant other than an apical meristem.
A problem still offering some restrictions to the growth and survival of the ferns
is the primitive nature of the vascular system, which is not as efficient as the vascular
system of the seed plants. When you examine representatives of this group, note how
they have advanced over the mosses with regards to features promoting a terrestrial
existence.
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Morphological and Anatomical Diversity in the Ferns
Examine the whole fern plant on demonstration. Note the whole aerial leaf (frond), the
stipe, the underground stem (rhizome), and the small roots (Figure 10).
Figure 10: Mature fern sporophyte.
Examine the fiddleheads, which are young, coiled fronds (Figure 11). Fiddleheads are
considered a culinary delicacy in some parts of the world.
Figure 11: Fiddlehead.
Examine the prepared slide of the fern frond (Slide #56) and find structures shown in
Figure 12 (cuticle, epidermis, mesophyll, stomata). Ignore the reproductive structures
on the under side of the leaf.
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Figure 12: Fern frond X.S. diagram (above), and picture (below).
You should note a few important things about the fern leaf structure.
There are now a number of different cell types making up the structure of
the fern leaf that you did not see in the moss leaf. There is an upper and
lower epidermis. Epidermal cells secrete a waxy substance call cutin onto
the outer surface of the leaf. This forms an outer water-proof waxy layer
called the cuticle. This water-proof waxy cuticle prevents loss of water
from the leaf surface to the surrounding dry air.
There are large green cells inside the leaf parenchyma cells that contain
chloroplasts. Chloroplasts contain the photosynthetic pigments and other
components of the photosynthetic apparatus. These cells are rather
loosely arranged within the leaf, with lots of air space between them. This
loose arrangement allows gasses like CO2 to rapidly move through the leaf
to be absorbed by these cells so they can use it to make sugar in the Calvin
cycle of photosynthesis. Oxygen (O2), a waste product of photosynthesis,
also diffuses out of these cells and moves through the air spaces out of the
leaf.
Since the cuticle makes the outer surface of the leaf water proof, it also
makes it air tight. There are small holes in the epidermis called stomata
which allow CO2 in and O2 out of the leaf. However, these stomata would
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also allow water to evaporate out of the leaf. So in order to reduce the rate
of water loss through these stomata, they are surrounded by two special
epidermal cells called guard cells. These guard cells can close the
stomata at night or during very dry periods to help the ferns retain water,
but open the stomata when the photosynthetic cells need CO2 for
photosynthesis.
Finally, we also see bundles of vascular tissue throughout the fern leaf.
The xylem and phloem in these veins transport water and minerals to the
leaf cells, and transport the products of photosynthesis out of the leaf.
Examine the prepared slide of a cross section of a fern rhizome (Slide #99). Note the
cuticle, epidermis, cortex, and presence of vascular tissue (Figure 13).
(a)
(b) (c)
Figure 13: (a) Fern rhizome X.S. diagram.
(b) & (c) Fern rhizome X.S. pictures taken under the microscope.
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iv) The Flowering Plants
As mentioned above, a lot of the success of seed plants can be attributed to some
important morphological and anatomical features. These include a well developed
vascular system, a highly efficient absorptive and anchoring system, protected
photosynthetic tissues, and a sufficient support system. Cellular differentiation and
selective combinations of cell and tissue types have produced these features. Division of
labour is complete in this group. These higher plants possess only four organs, each
comprised of numerous tissues. Each plant organ conducts specific functions for the
plant, thus contributing to the success of the plant as a whole. These organs are the root,
stem, leaf (vegetative organ), and flower (reproductive organ). The major functions of
the root are anchorage, storage of photosynthetic products, and absorption. The stem's
functions are storage, support of leaves and flowers, and transport of water and dissolved
nutrients (both inorganic and organic). Photosynthesis is the major function of the leaf.
The function of the flower is reproduction.
We will begin our examination of the histology of flowering plants with a look at
the general features of a typical plant. Following that, well look at details of each of the
organs mentioned above (with the exception of the flower since it was studied in Practical
Lab 2).
Gross Morphology
For this section of the lab you will examine a bean plant (or other available plant) as
a representative of the flowering plants. Remember that plants are stationary organisms.
They must get all their water and nutritional requirements from the environment in their
immediate vicinity. The fact that these requirements are in two extremely different
environments (above and below ground) means that the plant must also be considerably
different in these two habitats. Also, because the materials acquired from the two habitats
must be utilized in all parts of the plant, there must be an adequate transport system to
provide links throughout the plant.
Observe the overall body structure of the bean plant (or another available plant) on
demonstration and identify the following parts (Figure 14).
Nodes regions of the stem from which leaves, buds, and branches arise.
Internodes the regions of the stem located between the nodes.
Terminal buds located at the tips of the stem and branches. They enclose a
growing region which continues to give rise to new tissues and organs.
Axillary or lateral buds located in the axis of the leaves. They may give rise to
lateral branches.
Blade and petiole specific parts of a leaf.
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Primary and secondary roots the primary root is the first root produced by the
plant. The secondary roots arise as root branches from the pericycle, deep within the
primary root tissue.
Root tips the last few millimetres of the root. This is the site of production of
new root length and the area where water and minerals are absorbed from the soil.
Figure 14: Morphology of a typical flowering plant.
The Root
The root constitutes the underground portion of the plant axis. The three main
functions of roots are absorption, anchorage, and food storage. The tissues of the root
are arranged in a fashion that best facilitates these functions. The anchoring ability of the
root depends on the substrate in which they are found, the size of the roots, and the mode
of root branching. The absorptive portion of the root lies immediately behind the tip (i.e.
the area containing the root hairs). In this area, the plant cells are not fully differentiated
and there is, as yet, no cuticle. This facilitates the rapid absorption of water and nutrients.
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The older portions of the root have a cuticle that has been produced by the epidermal
cells. These older regions have a large area called the cortex which is filled with
parenchyma cells. This cortex region of the root is where the plant often stores the
products of photosynthesis. They also have a well developed conducting system for the
transport of water, nutrients and photosynthetic products.
All the tissues of the root originate from the rapid cell division going on in the
root apical meristem. This is a region of rapidly dividing cells at the tip of each root. It
is covered by several layers of cells called the root cap. This root cap protects the
delicate apical meristem as the root is pushed downward through the soil.
The roots of flowering plants show a couple of different patterns of growth and
development. The three basic types of roots are tap, fibrous, and adventitious (Figure
15).
Tap typical taproots include a prominent main (primary) root directed vertically
downward with numerous smaller lateral roots.
Fibrous have many long slender roots of about equal size with many secondary
and
tertiary roots off the main roots.
Adventitious arise from an organ in the plant other than the primary root (apical
meristem) or its branches.
Observe the demonstration material showing different types of roots.
Figure 15: Three typical flowering plant root systems.
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Examine the prepared slide (demo) of a cross section through the root of Ranunculus sp.,
(Slide #8) and identify the following, highlighted structures (Use Figure 16(a)-(c) as a
guide):
Epidermis a single layer of cells which covers the root. Certain

epidermal cells differentiate as root hairs. These will not be seen in your
section because the section was taken past the point at which root hairs
arise. In older sections, a cuticle is visible on the epidermal cells.
Cortex a wide multi-cellular region of parenchyma cells. The purple

stained organelles are leucoplasts which function in the storage of starch.
The inner cell layer of the cortex is comprised of smaller endodermal cells.
The heavy wall between adjacent endodermal cells is the Casparian strip
which functions to prevent lateral flow of materials. This structure ensures
that all materials moving laterally must pass through endodermal cell
membranes which control the movement of various materials into and out of
the stele.
Stele the central portion of the root, whose outer extremity is termed the
pericycle. In the Ranunculus sp. root, it is comprised of parenchyma cells.
The pericycle is responsible for forming root branches. The inner portion of
the stele contains two types of vascular tissue; xylem and phloem. The star-
shaped arrangement of cells in the centre is the xylem tissue. Some
sclerenchyma cells are present outside the xylem. The cells between the
points of the star form the phloem tissue.
Figure 16 (a): Diagram of a root X.S.
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Figure 16 (b): Microscopic view of a X.S. of Ranunculus sp. root. An overview of a
mature root is seen on the left. A higher magnification view of the cortex (on the right)
shows parenchyma cells with leucoplasts.
Figure 16 (c): Microscopic view of a X.S. through the centre of a root of Ranunculus sp.
The Stem
The stem is usually an above ground part of the plant. It typically serves as a
mechanical support for the leaves, flowers, and fruits. It also furnishes a path of
conduction between these organs and the roots. Depending on the plant, it may also act
as a storage organ for water and products of photosynthesis, to a limited extent. Stems
are characterized by having a highly developed vascular system which is critical in their
transport and support functions. As in the case of the root, the internal anatomy of the
stem is well suited to its function. The single layer of epidermal cells have thin inner
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walls, while the outer walls, which are exposed to air, are thicker and often covered with
a cuticle (a waxy substance to prevent water loss). The epidermis may also develop
spines for protection, glandular extensions to attract insects, or hairs to reduce the rate of
transpiration. In addition, stems possessing photosynthetic tissue (i.e. they are green)
have stomata in their epidermis. These are openings which facilitate gaseous exchange
and limit water loss. Stems also possess varying amounts of fundamental tissues
(parenchyma, sclerenchyma, etc.) to perform the storage and support functions.
Examine the prepared slide of the cross section through the stem of Medicago sp., (Slide
#
11) and identify the following, highlighted structures (Use Figure 17(a)-(c) as a guide):
Epidermis a single layer of cells which covers the stem. Observe the
cuticle, which is present on the outer extremity of each epidermal cell.
Cortex and Pith the main storage part of the stem of Medicago sp.
(alfalfa) is divided into two distinct regions by the ring of vascular tissue.
The region between the epidermis and the vascular tissue is called the
cortex. It is composed of several layers of parenchyma cells, some of
which may contain chloroplasts and thick-walled collenchyma cells.
Collenchyma cells are specialized for structural support and are located as
cylinders of tissue beneath the epidermis. The region internal to the
vascular tissue is called the pith and is composed of parenchyma.
Vascular Bundle an area of conducting tissue. Each vascular bundle is

surrounded by sclerenchyma cells for support. Each bundle possesses both
xylem and phloem. The xylem tissue is always located more internally than
the phloem tissue. Observe the several large xylem cells which are known
as xylem vessels. These cells are dead when mature, lose their end walls,
and form lengthy continuous tubes for conducting water and minerals
upward in the plant. The phloem tissue, on the other hand, conducts water
and organic nutrients upward and downward in the plant.
Figure 17 (a): Schematic X.S. of the stem of Medicago sp. (alfalfa).
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Figure 17 (b): Microscopic views of cross sections of the stem of Medicago sp. (alfalfa).
Figure 17 (c): Close up view of the vascular bundle of the stem of Medicago sp. (alfalfa).
The Leaf
Leaves are perhaps the most conspicuous parts of plants and also the most
important. Being rich in chloroplasts, they function in photosynthesis. In general, they
are characterized by their thin, expanded form and are able to maintain their shape by an
internal skeleton of vascular tissue which is continuous with the vascular tissue of the
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stem. Thus, the vascular tissue carries food to all parts of the plant, gives support to the
leaf, and brings water to the sites of photosynthesis.
The internal arrangement of the tissues of the leaf can be seen in cross sections cut
at right angles to the broad surface of the blade (Figure 18 a). The outermost layer of
cells, which extends over the surface of the leaf, is the epidermis. The interior of the leaf,
between the upper and lower epidermis, is called the mesophyll. This corresponds to the
cortex of the root and stem. The mesophyll is further differentiated into an upper region
(the palisade mesophyll) and a lower region (the spongy mesophyll). The vascular
tissue extends through the spongy mesophyll.
Examine the prepared slide of a cross section through the leaf of Lilium sp. (Slide #14)
and identify the following, highlighted structures in Figure 18 (a)-(b).
Epidermis a continuous layer of cells extending over the upper and

lower surfaces of the leaf. Locate both the upper epidermis and the lower
epidermis, each with a cuticle. The cuticle is a waxy substance, which
prevents water loss through epidermal cells and protects the leaf against
mechanical injury and foreign invasion. The lower epidermis contains a
number of stomata, each of which is located between two highly
specialized guard cells. The guard cells regulate the opening and closing of
the stomata by changes in turgor pressure. The function of the guard cell is
to regulate the exchange of gases for the photosynthetic process.
Mesophyll an area corresponding to the cortex of the root and stem. It is

composed of a palisade mesophyll and a spongy mesophyll. The palisade
mesophyll is a region of packed cells located beneath the epidermis. This
region is composed of chlorenchyma cells and is the primary
photosynthetic area of the plant. The spongy mesophyll is composed of a
loosely knit network of chlorenchyma cells with numerous intercellular
spaces between the cells.
Vascular Bundle an area of conducting tissue composed of xylem tissue

and phloem tissue. The vascular bundles in the leaves are the terminal ends
of the continuous vascular system. In the more prominent vascular bundles,
a bundle sheath of parenchyma or sclerenchyma cells is present.
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Figure 18 (a): Diagram of a leaf X.S.
Figure 18 (b): Microscopic view of a leaf X.S.
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III SEED PLANT ENVIRONMENTAL ADAPTATIONS
Seed plants do not only dominate the terrestrial environment, they are found in
basically all environments, including both desert and aquatic habitats. Morphological
features of the roots, stems, and leaves have allowed seed plants to adapt to xeric (arid)
and hydrophytic (aquatic) areas. In the demonstrations in this lab you will explore some
of these morphological adaptations.
A. ADAPTATIONS OF XEROPHYTIC PLANTS
Aridity is the major (and almost the only) environmental factor that creates a
desert, and it is this functional water deficit that serves as the primary limitation to which
desert organisms must adapt. Desert plants survive the long rainless periods with three
main adaptive strategies: succulence, drought tolerance, and drought evasion. Each of
these is a different, but effective suite of adaptations for prospering under conditions that
would kill plants from other regions.
Succulent plants store water in fleshy leaves, stems, or roots in compounds or

cells from which it is not easily lost. All cacti are succulents, as are such non-cactus
desert dwellers as agaves, aloes, elephant trees, and many euphorbias. (Figure 19)
Several other adaptations are essential for this water-storing habit to be effective.
Succulents must be able to absorb large quantities of water in short periods, and they
must do so under unfavorable conditions. Because roots take up water by passive
diffusion, succulents can absorb water only from soil that is wetter than their own moist
interiors. Desert soils seldom get this wet and dont retain surplus moisture for long.
Desert rains are often light and brief, barely wetting the top few centimeters of soil,
which may dry out after just a day or two of summer heat. To cope with these conditions,
nearly all succulents have extensive, shallow root systems. A succulent must be able to
guard its water hoard in a desiccating environment and use it as efficiently as possible.
The stems and leaves of most species have waxy cuticles that render them nearly
waterproof when the stomata are closed. Water is further conserved by reduced surface
areas. As an example, most succulents have either few leaves (agaves), no leaves (most
cacti), or leaves that are deciduous in dry seasons (Elephant trees & Boojum trees). The
water can also be bound in extracellular mucilages or inulins (plant compounds that
readily absorb water).
Many succulents possess a water-efficient variant of photosynthesis called CAM,

an acronym for Crassulacean Acid Metabolism. The first word refers to the plant family
(Crassulaceae) in which the phenomenon was first discovered. CAM plants open their
stomata for gas exchange at night and store carbon dioxide in the form of an organic acid.
During the day, the stomata are closed and the plants are nearly completely sealed against
water loss; photosynthesis is conducted using the stored carbon dioxide. At night the
temperatures are lower and humidity higher than during the day, so less water is lost
through transpiration. Transpiration is the evaporative loss of water from a plant. Plants
using CAM lose about one-tenth as much water per unit of carbohydrate synthesized as
do those using standard photosynthesis. However, there is a trade-off: the overall rate of
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photosynthesis is slower, so CAM plants grow more slowly than their ordinary
counterparts. (An additional limitation is the reduced photosynthetic surface area of most
succulents compared with ordinary plants.)
Stored water in an arid environment requires protection from thirsty animals.

Most succulent plants are spiny, bitter, or toxic, and often all three. Some unarmed, non-
toxic species are restricted to inaccessible locations.
Water scarcity is the most importantbut not the onlyenvironmental challenge

to desert organisms. The aridity allows the sun to shine unfiltered through the clear
atmosphere continuously from sunrise to sunset. This intense solar radiation produces
very high summer temperatures which are lethal to non-adapted plants. At night, much
of the accumulated heat radiates through the same clear atmosphere and the temperature
drops dramatically. Daily fluctuations of 22C are not uncommon when the humidity is
very low. Microphylly (the trait of having small leaves) is primarily an adaptation to
avoid overheating but it also reduces water loss. A broader surface has a deeper
boundary layer of stagnant air at its surface, which impedes convective heat exchange. A
leaf up to 10 mm across can stay below the lethal tissue temperature of about 46C on a
calm day with its stomata closed. (because it has less stagnant air above it impeding heat
loss) A larger leaf requires transpiration through open stomata for evaporative cooling.
Since the hottest time of year is also the driest, water is not available for transpiration.
Non-succulent, large-leafed plants in the desert environment would overheat and be
killed. Desert gardeners know that tomatoes will burn in full desert sun even if well
watered; their leaves are just too big to stay cool. Desert plants that do have large leaves
produce them only during the cool or rainy season or else live in shaded microhabitats.
There are a few mysterious exceptions, such as jimson weed (Datura wrightii) and desert
milkweed (Asclepias erosa). (Figure 20) Perhaps their large tuberous roots provide
enough water for transpiration even when the soil is dry.
Leaf or stem color, orientation, and self-shading are still more ways to adapt to
intense light and heat. Desert foliage comes in many shades, but rarely in typical leaf-
green. More often leaves are grey-green, blue-green, grey, or even white. The light color
is usually due to a dense covering of trichomes (hair-like scales), but is sometimes from a
waxy secretion on the leaf or stem surface. Lighter colors reflect more light (= heat) and
thus remain cooler than dark green leaves. Brittlebush and white bursage leaves show no
green through their trichomes during the dry season, while desert agave (Agave deserti) is
light grey due to its thick, waxy cuticle. Other plants have leaves or stems with vertical
orientations; two common examples are jojoba and prickly pear cactus. This orientation
results in the photosynthetic surface facing the sun most directly in morning and late
afternoon. Photosynthesis is more efficient during these cooler times of day. Prickly
pear pads will burn in summer if their flat surfaces face upward. Some cacti create their
own shade with a dense armament of spines; teddy bear cholla (Opuntia bigelovii) is one
of the most striking examples. (Figure 19)
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(a) (b)
(c) (d)
Figure 19: Examples of xerophytic plants.
(a) Teddy bear cholla cactus; (b) Queen Victoria agave; (c) aloe plant;
(d) Elephant tree (B. microphylla)
(a) (b)
Figure 20: (a) Jimson weed (Datura wrightii); (b)
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B. ADAPTATIONS OF HYDROPHYTIC PLANTS
Plants such as the water lily are hydrophytic, having stems and leaves which are
either partially or completely submerged in water. Totally submerged plants are the true
water plants or hydrophytes. Because they are truly aquatic, they have the greatest
number of adaptations to life in water. Some of these adaptations are:
The presence of little or no mechanical strengthening tissue in stems and

leaf petioles. If these plants are removed from the water, they hang limply.
They are normally supported by water all around them and so have no need
of mechanical strengthening. Indeed, this would be a distinct disadvantage
as it would limit flexibility in the event of changes in water level or water
movements. (Figure 21)
Submerged plants lack the external protective tissues required by land plants
to limit water loss. The epidermal (outermost) layer shows very little, if
any, sign of cuticle formation. All the surface cells appear to be able to
absorb water, nutrients, and dissolved gases directly from the surrounding
water. As a result, the internal system of tubes (xylem) which normally
transports water from the roots to all parts of the plant, is often greatly
reduced or even absent. Thus, if these plants are removed from the water,
they wilt very quickly, even if the cut stems are placed in water. This is
because the normal water transport system is poorly developed. As might
be expected, there are also no stomata (breathing pores) on the leaves.
Roots, which normally play a very important role in the absorption of

nutrients and water from the substrate, are often also reduced and their main
function is anchorage. The root hairs which function in absorption are often
absent and roots themselves may be entirely dispensed with. (e.g. Bladderwort)
Many species have very specialized leaf shapes. The submerged leaves are
often highly divided. This has the advantage of creating a very large surface
area for absorption and photosynthesis. It also minimizes water resistance
and hence potential damage to the leaves. Heterophylly, where leaves of
different shapes are produced depending on where on the plant they are, is
common. This can create great problems for identification! In many cases,
the submerged leaves are totally different to floating or emergent leaves on
the same plant. The emergent leaves are usually much less divided and have
a more similar internal structure to those of land plants.
Air-filled cavities often extend throughout the leaves and stems of aquatic
plants, providing an internal atmosphere. Certain aquatic fly and beetle
larvae have a novel adaptation which allows them to take unusual advantage
of this. A sharp appendage on the end of their abdomen is used to pierce
into submerged plants, giving them access to the internal air-filled cavities
as their own personal oxygen source.
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(a) (b)
Figure 21: Examples of submerged leaves.
(a) A bladderwort. (b) Thin flexible leaves of Eel Grass (Vallisneria spiralis).
Floating plants are of two types: those which are rooted with floating leaves (e.g.
Water Lily); and those which are not rooted in the sediment, but just float on the surface
(e.g. Duckweed). See Figure 22 for examples of these plants. Floating leaves are
generally tough because they have to withstand the weather and water movement. The
green pigment-containing chloroplasts important for photosynthesis are restricted to the
upper surface of the leaves which are the only surface to be well lit. Stomata, through
which gas exchange takes place in the leaf, are also found only on the upper surface of
the leaf. This upper surface often has a thick waxy cuticle to repel water and help to keep
the stomata open and clear. Air-filled internal cavities are also often present.
Terrestrial plants such as trees have to develop an enormous quantity of structural

material in order to rise above all the other plants and collect the lion's share of the light
available. Water lilies provide a neat example of a plant which has managed to do
exactly the same thing, but with the minimum of structural material. Weak stems
produce a massive floating canopy of leaves which dominate the local aquatic plant
community just as effectively as trees dominate a forest. The difference lies in their
external medium; water provides all the necessary support whereas air does not.
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(a) (b)
Figure 22: Examples of floating plants.

(a) Water lilies have floating leaves which are rooted in a substrate.
(b) Duckweed is not rooted and the whole plant floats on the surface.
Observe the xerophytic and hydrophytic plants on display. Try to determine what
morphological changes to their roots, stems, and/or leaves have allowed them to
successfully survive in their environment.
Table 1: Comparison of the Morphological Adaptations Observed in the Roots, Stems,

and Leaves of Xerophytic and Hydrophytic Seed Plants.
XEROPHYTIC PLANTS HYDROPHYTIC PLANTS
ROOTS
STEMS
LEAVES
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Phylum Bryophyta (Mosses)
Question 1: What is the purpose of the spiral leaf arrangement in mosses?
_______________________________________________________________________
_______________________________________________________________________
Question 2: What is the function of each area of the moss plant?
Rhizoids: ______________________________________________________________
Stem: __________________________________________________________________
Leaves: ________________________________________________________________
Question 3: Why do mosses grow in clumps?
_______________________________________________________________________
_______________________________________________________________________
Question 4: Why is the growth in height of mosses severely restricted?
_______________________________________________________________________
_______________________________________________________________________
Question 5: What types of support allow mosses to remain upright and tall?
_______________________________________________________________________
_______________________________________________________________________
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Question 6: Is there a cuticle present on the surface of the leaves in mosses?
_______________________________________________________________________
_______________________________________________________________________
Question 7: Do the leaves of mosses have stomata?
_______________________________________________________________________
_______________________________________________________________________
Question 8: Why is fragmentation an inefficient means of reproduction?
_______________________________________________________________________
_______________________________________________________________________
Question 9: What type of cell division occurs in the moss capsule?
________________________________________________________________________
Question 10: How does the moss sporophyte obtain its organic nutrients and water?
________________________________________________________________________
Question 11: What does the presence of flagellated male sperm cells suggest about
the means of gamete transfer?
________________________________________________________________________
Question 12: What advantage(s) does the embryo have by being enclosed by the
female gametophyte?
________________________________________________________________________
________________________________________________________________________
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Question 13: What is the main method of spore dispersal in mosses?
________________________________________________________________________
Phylum Pterophyta (Ferns)
Question 14: Why is it advantageous for the plant to possess vascular tissue?
_______________________________________________________________________
_______________________________________________________________________
Question 15: Fern plants are taller in comparison to mosses. What is the advantage
of the increased size to the plant?
_______________________________________________________________________
_______________________________________________________________________
Question 16: Explain why it is more beneficial for ferns to have only their leaves
above ground.
_______________________________________________________________________
_______________________________________________________________________
Question 17: What is the function of the epidermis?
_______________________________________________________________________
Question 18: What is the function of the mesophyll in the frond?
_______________________________________________________________________
Question 19: What is the function of the stomata on the frond epidermis?
_______________________________________________________________________
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Question 20: Are prothalli haploid or diploid?
_______________________________________________________________________
Question 21: What is the main method of spore dispersal in ferns?
_____________________________________________________________________
Question 22: Name three features of fern reproduction which could be considered
detrimental (harmful) to a terrestrial existence.
i) ___________________________________________________________________
ii) __________________________________________________________________
iii) __________________________________________________________________
Question 23: Why is it advantageous for ferns to reproduce almost entirely by

sexual reproduction?
___________________________________________________________________
__________________________________________________________________
Question 24: Ferns and mosses must still be considered marginal terrestrial plants.
Suggest two reproductive features that both possess which forces this marginal
existence.
i) ___________________________________________________________________
ii) __________________________________________________________________
Question 25: What structure in the moss life cycle is comparable to the sporangium
in the fern life cycle?
_____________________________________________________________________
_____________________________________________________________________
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Question 26: What structure in the fern life cycle is comparable to the mature moss
gametophyte?
_____________________________________________________________________
The Flowering Plants
Question 27: How does the function of xylem tissue differ from that of phloem tissue?
_____________________________________________________________________
_____________________________________________________________________
Question 28: What is the function of sclerenchyma tissue?
_____________________________________________________________________
_____________________________________________________________________
Question 29: What is the function of collenchyma tissue?
_____________________________________________________________________
_____________________________________________________________________
Question 30: What is the function of parenchyma tissue?
_____________________________________________________________________
_____________________________________________________________________
Question 31: Give the specific function(s) of:
Root:
_____________________________________________________________________
_____________________________________________________________________
215
Stem:
________________________________________________________________
_____________________________________________________________________
Leaf:
_____________________________________________________________________
_____________________________________________________________________
Question 32: What is double fertilization? Support your answer with sketches.
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
Question 33: Give the function of each part of the embryo.
Epicotyl: _____________________________________________________________
_____________________________________________________________________
Hypocotyl: ____________________________________________________________
_____________________________________________________________________
Radicle: ______________________________________________________________
_____________________________________________________________________
Question 34: What is the specific function of the fruit?
_____________________________________________________________________
_____________________________________________________________________
216
TUTORIAL LABORATORY 3 ANIMALS I:
PHYLA CNIDARIA, PLATYHELMINTHES, NEMATODA, ANNELIDA, &

ARTHROPODA
OBJECTIVES:
1. To examine the basic body plan and the feeding,

respiration, circulation, and reproductive systems of the
Phyla Cnidaria & Platyhelminthes.
2. To examine certain aspects of the anatomy and

morphology of the Phyla Nematoda, Annelida, &
Arthropoda.
I PHYLUM CNIDARIA
The phylum Cnidaria includes a diverse group of about 10,000 living species.
The name Cnidaria refers to the specialized cells called cnidocytes (Greek knide =
nettle), which are unique to this phylum. Cnidocytes contain stinging structures,
nematocysts that are used to immobilize prey. The cnidarians are exclusively aquatic
animals with the majority of species found in marine habitats. The phylum includes
many brightly coloured and beautiful animals such as jellyfish, sea anemones, sea fans,
corals, and hydras.
Observe various cnidarians on demonstration.
Members of the phylum Cnidaria are radially symmetrical organisms

characterized by a digestive body cavity known as a gastrovascular cavity (not a coelom),
some muscle fibers and many nematocysts. Mature cnidarians are composed of only two
cellular layers, an outer epidermis (developed from embryonic ectoderm) and an inner
gastrodermis (developed from embryonic endoderm). Thus, they are known as
diploblastic organisms. In cnidarians, these cell layers are thin and the animals are
therefore very fragile. Between these two cellular layers is a gelatinous (non-cellular)
layer of mesoglea. Contrary to epidermis and gastrodermis, the mesoglea is poorly
developed (Figure 1).
Animals above the cnidarians on the evolutionary tree have three germ layers of
cells. Thus, they are known as triploblastic organisms (the third layer lies between the
ectoderm and endoderm and is called the mesoderm). The mesoderm allows animals to
become more organized in their structure. Cnidarians, for example, are organized mainly
at the level of tissues, while those with three layers possess organs and elaborate organ
systems.
217
The members of Phylum Cnidaria have one of two forms: a polyp or medusa. A
polyp is a tubular organism closed at one end with a mouth surrounded by tentacles at the
other end. A medusa is an umbrella-shaped, jellylike, free-swimming organism with a
mouth at the end of a central projection called a manubrium (Figure 1). We will examine
one representative member for each body form.
Figure 1: Body forms in Cnidaria.
Hydra sp.
The Hydra is an excellent organism for the study of the polyp form of the Phylum
Cnidaria. Although a very simple organism, it illustrates a definite level of organization
into tissues.
Examine slide #59 of stained cross sections through the body wall of Hydra. Note that
the body wall surrounding the gastrovascular cavity consists of two distinct layers of
cells: an outer layer called the epidermis and an inner layer called the gastrodermis.
These two layers are held together by a thin, non-cellular layer, the mesoglea (Figure 2).
218
Figure 2: Diagram of the cross-section through the body wall of Hydra sp. (top) and the
microscopic view of Hydra sp. (bottom).
Feeding and Digestion in Cnidaria
Place a living Hydra in a small dish of water and examine with the dissecting
microscope. At the upper end of the animal, observe several elongate, actively moving
tentacles, which are used to capture food (Figure 3). At the centre of this circle of
tentacles, is the mouth. Tap the edge of your dish and observe the extreme contractility
of this organism.
To observe your specimen ingesting food, add several Daphnia (or other available
food organisms) to your dish and observe with the dissecting microscope. When the
tentacles of the Hydra come into contact with its prey, the prey jerk for several minutes
before becoming still. The food organism reacts in this way because it is being stung by
numerous wart-like nematocysts on the tentacles. Once the food organism has been
ingested, it will be degraded within the gastrovascular cavity and the small particles will
be engulfed by the gastrodermal cells by phagocytosis.
219
Osmoregulation in Cnidaria
Being small, exclusively aquatic organisms cnidarians require neither a special

respiratory nor circulatory system. The entire body surface can participate in gas
exchange since all cells in these diploblastic (two-layered) animals are constantly moist
and exposed to the gas-containing medium. In addition, no cell is ever far enough from
the source of either nutrient materials or gases to necessitate any more elaborate transport
than either direct diffusion from cell to cell or diffusion through the very watery
mesoglea that exists between the two cell layers (the epidermis and the gastrodermis).
Reproduction in Cnidaria
Hydra reproduce asexually by simple budding and sexually by the production of

sperm and ova. Budding consists of a simple outgrowth from the body wall. The
gastrovascular cavity of the bud is initially continuous with that of the parent. The bud
will separate from the parent at maturity.
Examine slide #58 and observe Hydra budding (Figure 3).
The gametes for sexual reproduction are produced in organs called spermaries
(testes) and ovaries, which are protuberances along the body wall (Figure 3). Both sex
organs may be present in a single organism or in two separate organisms.
Examine slide #60 to see spermaries and slide #61 to see ovaries. Also, find other
structures labeled in Figure 3. Note, that on most of the slides, you will not see the bud
shown in Figure 3.
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Figure 3: Longitudinal section through Hydra.
Obelia sp.
Obelia is a colonial marine organism that is found attached to seaweeds, rocks,

shells or pilings in the shallow waters off seacoasts. This animal alternates between a
polyp stage (which is the asexual generation) and a medusa stage (which is the sexual
generation).
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Examine Figure 4 that illustrates the life cycle of Obelia. The plantlike colony consists
of numerous branches, which terminate in two kinds of polyps. The feeding, or nutritive
polyps possess tentacles and resemble Hydra. The reproductive polyps are club-shaped
and lack tentacles. The branches of the colony are covered by a transparent sheath - the
perisarc (Figure 5), which extends around the nutritive (feeding) and reproductive polyps
as the hydrothecae and the gonothecae, respectively (Figure 5).
Examine slide #63 of a hydroid colony of Obelia and slide #62 of a medusa of Obelia sp.
Figure 4: Life cycle of Obelia sp.
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Figure 5: Different microscopic views of Obelia sp.
The medusa is umbrella-shaped and swims with its convex surface upward so that
the tentacles are facing downward. The tentacles have numerous cnidocytes (with
nematocysts inside them) to assist in capturing prey. Extending into the cavity of the
medusa is the manubrium, which contains a mouth at its tip surrounded by four oral
lobes. Food organisms enter the mouth and into a gastrovascular cavity. The gonads
(testes or ovaries) can also be seen on the undersurface of the medusa.
Examine the medusae on demonstration of the common jellyfish, Gonionemus sp. (Figure
6). Gonionemus sp. is dioecious; that is it has the male and female organs in separate
organisms. The eggs and sperm are released into the sea where fertilization takes place.
The zygote develops into a planula larva, which swims about, then settles, and is
transformed into a polyp.
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Figure 6: Medusa of Gonionemus sp. (a); & diagram of the structures of a medusa (b).
II PHYLUM PLATYHELMINTHES
Members of the phylum Platyhelminthes (Greek platy = flat, and helmins =

worms) are dorso-ventrally flattened, which accounts for their common name, the
flatworms. In contrast to the cnidarians, flatworms are bilaterally symmetrical animals,
with distinct anterior and posterior ends and right and left sides. As well, the flatworms
are triploblastic, with a third cellular layer (parenchyma tissue developed from
embryonic mesoderm) located between the epidermis and the gastrodermis. Flatworms
are simple organisms having no body cavity other than the digestive cavity. Thus, they
are acoelomate. This group has advanced considerably beyond the cnidarians in that
they possess well-developed nervous, excretory, muscular, digestive and reproductive
systems.
Dugesia sp.
In this laboratory, you will study Dugesia sp., often called a planarian, which is a
common freshwater organism. It is often found in ponds and streams. Parasitic
flatworms, such as flukes and tapeworms also belong to the Phylum Platyhelminthes.
Place a living planarian in a small glass dish containing water and examine with the
dissecting microscope. Observe its general shape and size and follow its movements for
a few minutes. From its movements, determine its anterior, posterior, dorsal and ventral
sides.
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Place a small piece of liver in with the live planarian and watch it feeding. Note the
extrusion of the pharynx when it is feeding. Also, note the position of the pharynx and
mouth, which is located at the end of the pharynx (Figure 7).
Figure 7: Microscopic view of Dugesia sp. and diagram of a planarian with extended
pharynx.
Examine the whole mount slide (Slide #64) of Dugesia as well as the prepared slide of a
cross section (Slide #65) through this planarian and locate the structures shown in Figure
8. Note the extensive nature of the gastrovascular cavity.
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Figure 8: Longitudinal (top) and cross section (bottom) of a planarian - Dugesia sp.
Feeding and Digestion in Platyhelminthes
Free-living platyhelminthes are mainly carnivorous and prey on small

invertebrates that can easily be captured. As well, they may feed on the remains of dead
animals. Dugesia sp. has a blind digestive tract (i.e. there is only one opening) but it is
extensively branched and thus considered more complex in structure than that of the
cnidarians. The digestive process in planaria is similar to that in Hydra except that food
is brought to the gastrovascular cavity through a muscular protrusible pharynx (Figure
7). There are three main branches of the digestive tract each with numerous lateral
branches, which greatly increase the surface area for absorption (Figure 8, top).
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Parasitic platyhelminthes, which possess a digestive tract, feed on the tissues and
tissue fluids of the host. For example, Clonorchis sinensis is a flatworm that lives in the
bile ducts of the human liver. Examine Slide #66 of Clonorchis sinensis, the human liver
fluke. Like Dugesia sp., Clonorchis sinensis has a muscular pharynx, which leads into
a blind digestive tract (Figure 9). However, the pharynx of Clonorchis sinensis is not
protrusible. Instead, Clonorchis sinensis has an anterior mouth located at the bottom of
a muscular sucker. This oral sucker is used to suck in tissue fluids.
Figure 9: Longitudinal section of a flatworm - Clonorchis sinensis.
Other parasitic platyhelminthes do not posses a digestive system and absorb

simple nutrients directly through the body wall. This is possible because many internal
parasites are often bathed in partly or completely digested food -- especially if they live
in the digestive tract or blood of the host.
Examine a demonstration of preserved tapeworms (Taenia sp.), which live in the
intestines of humans.
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Osmoregulation and Excretion in Platyhelminthes
Free-living, freshwater flatworms, planarians, have a discrete excretory system

(protonephridia), which is primarily osmoregulatory but may have an excretory function
as well. Marine members of the group often lack this system, supporting the hypothesis
that its main function is osmoregulation. The system consists of a network of tubules
within the animal's body tissues. One end of the tubule opens through a small pore to the
exterior. The other end of the tube ends blindly within the body in a spherical structure
containing long cilia - these are called flame cells (Figure 10). Excess water (and
possibly wastes) enters the flame cell system and is propelled through the tubules toward
the outside by the beating of the cilia (the "flame").
Figure 10: Planarian Excretory System (top) and One Flame Cell (bottom).
(Note the long cilia or "flame" which move material toward the excretory canal)
Unfortunately, the flame cell system of planarians is difficult to locate, even in

live animals and will not be observed during this lab.
Respiration and Circulation in Platyhelminthes
Platyhelminthes lack respiratory and circulatory systems. These animals are

small and are always found in a moist environment whether they are free-living or
parasitic. When present (as in Dugesia sp.), the gastrovascular system (meaning gut-
transport) serves to transport nutrient materials. Otherwise, all internal transport occurs
by simple diffusion through and between cells of the small body.
Reproduction in Platyhelminthes
Planarians are hermaphroditic organisms that have both male and female sex
organs in the same individual. Copulation in flatworms is usually mutual, each partner
inseminating the other. The reproductive system of planaria is difficult to locate, even in
live animals, and will not be observed during this lab.
Note that the body plan, digestive systems, and methods of reproduction of tapeworms
and liver flukes are quite different compared to planarians.
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III PHYLUM NEMATODA
Cross Section of Ascaris lumbricoides
Examine slide #88, which is a cross section through the anterior end of Ascaris sp.
and identify the cuticle, epidermis, pseudocoelom, muscle cells, and pharynx (Figure
11).
Figure 11: Diagram of Ascaris sp. cross section.
As seen on the cross section, the musculature of Ascaris sp. consists of only longitudinal
muscles. Groups of these muscles contract simultaneously working against the fluid in
the pseudocoelom.
IV PHYLUM ANNELIDA
External Anatomy of the Earthworm
As you saw in the practical lab, the earthworm has bristle-like setae on its ventral
surface which are used for locomotion. In the earthworms natural habitat (the soil),
setae work extremely well to provide the grip required for movement. To demonstrate
their effectiveness, obtain a living earthworm. Place the worm on a piece of moist paper
towel and observe its movement. Then, observe the animals movement on a smooth
surface (e.g. the bottom of an enamel pan). You should see a marked difference in the
ability of the animal to move on the two different surfaces.
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Cross Section of the Earthworm
Examine slide #71 of the cross section of Lumbricus sp. and note the following
highlighted structures (Figure 12):
Cuticle thin external layer.
Epidermis outer cellular layer composed of epithelial cells. Try to locate Goblet cells
in the epidermis of the earthworm. They appear empty.
Circular muscle layer located just beneath the epidermis, with the fibres cut
longitudinally in the section.
Longitudinal muscle layer these fibres are arranged in blocks of feather-like bundles
extending toward the center; they are cut transversely.
Peritoneum a thin epithelial lining which separates the body cavity from the body
wall. (epidermis and muscles)
Coelom the body cavity (fluid-filled in the living worm).
Intestine note the infolding of the dorsal surface of the intestine. This is called the
typhlosole.
The cells surrounding the exterior surface of the intestine are called chloragogue cells.
These function as: an accessory digestive gland; the site of glycogen synthesis,
deamination of proteins, and production of ammonia and urea.
Dorsal blood vessel just above the intestine.
Ventral blood vessel just below the intestine.
Nephridia within the coelom, between the intestine and the body wall. In this section,
only incomplete portions of nephridia can be seen, usually appearing as wavy lines.
Setae two pairs ventrally and two pairs ventrolaterally projecting from the body wall.
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Figure 12: Cross-section of Lumbricus sp
V PHYLUM ARTHROPODA
Respiratory and Circulatory Systems of the Grasshopper
Terrestrial arthropods use several different respiratory organs, the most unique of
which is the tracheal system. Air enters small openings in the body wall called
spiracles which lead to a system of branching tubules, tracheae. The tubes branch
repeatedly so that extremely fine tubules, tracheoles, reach the individual cells or small
groups of cells inside the body. As you can imagine, the surface they provide for
respiratory exchange is very large (Figures 13, 14).
Figure 13: Generalized Diagram of an Insect Tracheal System.
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Figure 14: Schematic diagram and actual picture of one part of the tracheal
system showing relationships of spiracle, trachea and tracheoles to the body wall and
internal muscles.
Examine a demonstration slide of the spiracle and tracheae of an insect. Note the spiral
thickening of the tracheal tubes.
Like crayfish, the grasshopper also has an open circulatory system. However, it
does not preserve well and will be impossible to locate.
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Phylum Nematoda
Question 1: During the dissection of Ascaris, you observed the digestive tract. On
the basis of the structure of the digestive tract, what conclusions can you draw about
this organisms diet?
_____________________________________________________________________
_____________________________________________________________________
Phylum Annelida
Question 2: In the tutorial lab, you were asked to observe the earthworms
movements on both smooth and rough surfaces. Compare the worms movements
on both surfaces and explain any differences you noticed.
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_________________________________________________________
_________________________________________________________
Question 3: What is the specific function of the Goblet cells in the epidermis of the
earthworm?
_____________________________________________________________________
_____________________________________________________________________
Question 4: What happens to the body of the earthworm when circular and
longitudinal muscles contract?
Circular: ______________________________________________
Longitudinal: ____________________________________________
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Question 5: What is the specific function of the typhlosole in the earthworm?
_____________________________________________________________________
_____________________________________________________________________
Question 6: In the clamworm, what is the specific function of esophageal caeca?
_____________________________________________________________________
_____________________________________________________________________
Question 7: Suggest reasons for the differences in the organization between the
digestive tract of Lumbricus and Nereis.
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
Question 8: With respect to diet and behaviour, why does Nereis have a much more
elaborate sensory system than does Lumbricus?
_______________________________________________________________________
___________________________________________________________
Question 9: In what ways are the parapodia of Nereis sp. morphologically suited to
the function of gas exchange?
_____________________________________________________________________
_____________________________________________________________________
Phylum Arthropoda
Question 10: Where are the gills of the crayfish attached to the body? What is the
advantage of this attachment?
_____________________________________________________________________
_____________________________________________________________________
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Question 11: Why is it important that the gills of the crayfish are well vascularised?
_____________________________________________________________________
_____________________________________________________________________
Question 12: Explain the difference between closed (seen in earthworm) and open
(seen in crayfish) vascular systems.
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
Question 13: When referring to the excretory systems of arthropods, why is it

advantageous in a terrestrial environment to excrete uric acid instead of ammonia?
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
Question 14: In the tutorial lab you were asked to examine a demonstration slide of
the spiracle and tracheae of an insect. What is the function of the spiral thickenings
of the tracheal tubes?
_____________________________________________________________________
_____________________________________________________________________
Question 15: What is the adaptive significance of the difference in respiratory organ
structure between organisms living in terrestrial versus aquatic environments?
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
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For study purposes, fill out the following table. This table is not to be handed in, and
you should add info to it as we continue our survey through the Kingdom Animalia.
Phylum Symmetry Tissue layers Coelom type Digestive system
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Respiratory Circulatory Excretory Reproductive Special
system system system system features
237
238
TUTORIAL LABORATORY 4 ANIMALS II:
PHYLUM CHORDATA
OBJECTIVES OF THE LABORATORY:
1. To examine the skeletal and skin (integumentary) systems of

the perch and the rat. (Phylum Chordata)
2. To examine the microscopic features of the reproductive

system of the rat.
3. To examine the vessels comprising the circulatory system of

the rat.
4. To provide background information on the respiratory

systems of vertebrates.
A. THE PERCH
i) Skeletal System
The skeletal system of vertebrates is composed of bone and/or cartilage. Bone

tissue is found only in the Subphylum Vertebrata. Some of the lower vertebrates do not
possess bone, but all the higher vertebrates do. As such, bone is often thought of as being
typical of vertebrates. In vertebrates, bone functions as a supporting tissue, a calcium
reserve, and as a hemopoetic (blood forming) tissue.
The skeleton is the basis of form and support of the vertebrate body. Muscles
attach to the skeleton and vital organs are surrounded and protected by skeletal elements.
As you examine the skeletons of the perch and the rat, you should note a number of basic
changes that have occurred in the evolution of the vertebrate skeleton. Some of the
changes involve a reduction in the number of bones in the skull and a reduction in the
number of ribs. Correlated with the move from an aquatic to a terrestrial environment are
the increase in the complexity of the limbs and limb joints, the development of the
pectoral and pelvic girdles and the strengthening of individual bones to support the
weight of the organism on land. The central structure of support in the lower vertebrates,
the notochord, is progressively replaced functionally by elements of the vertebrae.
Although the notochord runs the length of the vertebral column in fish, in many it has
been greatly restricted by the vertebrae. In adult tetrapods, the only remnant of the
notochord is the gelatinous material found in the intervertebral discs between successive
vertebrae of the vertebral column.
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In the study of the skeletal system in the perch and a more advanced vertebrate,
the rat, you should try to determine which skeletal features are signs of typical
evolutionary advancement and which may be specializations due to the animals way of
life.
The skeleton of vertebrates is broadly divided into two parts:

i) the axial skeleton consists of the skull, vertebrae and ribs.
ii) the appendicular skeleton consists of the pectoral and pelvic
girdles and the bones of the appendages.
NOTE: The bones of the prepared skeletons are delicate and easily broken.
HANDLE THESE SKELETONS WITH EXTREME CARE!
1. The Axial Skeleton
a) Skull
The skull of the perch is actually a double structure consisting of two boxes of
bone, one enclosed by the other. The outer skull is an armor of dermal bone. Primitive
extinct bony fish had dermal bony armor covering most of their bodies. In the modern
fish, the outer skull is virtually all that remains of this armor. Dermal bone forms, as its
name implies, in the dermis of the skin, and is not preceded by a cartilage structure. The
inner skull is composed of endochondral bone. Endochondral bone develops under the
dermis and replaces existing cartilaginous structures. Hence the name endochondral
denotes the bony tissue develops within existing cartilage structures. Elements of the
inner skull form the cranium or brain case.
The perch skull consists of many small bones. You are not responsible for knowing the
identity of these bones, but look closely at the skull to see the inner endochondral skull
encased by the outer dermal skull.
b) The Vertebral Column
A series of endochondral bones called vertebrae form the vertebral column.

Vertebrae have several common features. The large spool-shaped central portion of each
is the centrum. Extending through the middle of each centrum is a canal for the passage
of the notochord. As mentioned previously, many fish retain a notochord throughout life.
Above the centrum, an arch of bone surrounds and protects the spinal cord. This is
the neural arch. A dorsal projection, the neural spine, extends outward from the
vertebral column (Figures 1 and 2).
The fish vertebral column is divided into two subdivisions: the trunk and the tail
(caudal). Although fish do not have a neck, the first two trunk vertebrae are modified.
These vertebrae lack ribs. The rest of the trunk vertebrae possess ribs. Caudal (tail)
vertebrae do not have ribs. They possess a ventral portion that forms a hemal arch,
which surrounds blood vessels (Figures 1 and 2). The neural arch with the neural spine is
present in both trunk and tail vertebrae
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Figure 1: Representative Vertebrae of the Perch.
Figure 2: Skeleton of the Perch.
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2. The Appendicular Skeleton and Fins
Using the preserved perch (on demonstration) and the skeleton, locate the
following structures:
The Median Fins: The dorsal fins of the perch have fin rays for support. The anterior
dorsal fin has ossified fin rays, which provide stiff support, while the fin rays of the
posterior dorsal fin are not ossified and are flexible. Only the first two fin rays of the
anal fin are ossified. The caudal fin is composed entirely of soft, unossified fin rays.
The Pectoral Girdle and Fins: The pectoral fins are attached to a bony girdle, the
pectoral girdle. The pectoral girdle is composed of a number of fused elements. The
girdle is also fused to the skull (the head and trunk of the perch move as a unit). The fins
are supported by soft fin rays.
The Pelvic Girdle and Fins: The pelvic fins are attached to the pelvic girdle, which is
composed of two bony pelvic plates. The plates may be fused along the midline. The
pelvic girdle is not attached to the vertebral column or to the pectoral girdle, but is free-
floating (embedded in muscle only). Only the medial fin rays are bony, the rest are the
typical soft unossified type.
ii) The Skin (Integumentary System)
The skin is an extensive organ that forms the interface between an animals
internal environment and the outside world. Vertebrates maintain an internal
environment different from their external environment. Thus, the skin in vertebrates is
well developed and has many important functions. It is primarily a protective covering
but it does not isolate the body. The skin typically contains many sensory receptors as it
is an organ which is in constant contact with the outside world. Skin structure is
correlated closely with an organisms mode of life and environment. Thus, among the
vertebrates skin varies considerably, but its basic structure is the same.
Skin is composed of two main layers; the outer epidermis overlaying the thicker
dermis. The epidermal layer is composed of stratified squamous epithelial tissue. This
type of epithelium is usually associated with structures, such as the skin, which are
primarily concerned with the protection of underlying tissues from abrasion.
The dermis consists of dense fibrous connective tissue containing a number of

blood vessels. This tissue has an arrangement of protein fibres, cells, and non-living
intercellular substance.
The aquatic environment is less stressful and more stable than the terrestrial
environment. Since all fish are aquatic, fish skin is relatively simple in structure. The
epidermis in fish is relatively thin. Unicellular mucous glands are abundant in the
epidermis. The dermis is composed of dense fibrous connective tissue and is attached to
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the underlying muscles by a layer of loose connective tissue. The scales of fish are
located within the dermis.
Examine Slide #35 (a section of fish skin) noting the features above. Refer to Figure 3.
Figure 3: Cross-section through perch skin.
B. THE RAT
i) Skeletal system
1. The Axial Skeleton
a) Skull
Mammals have, when compared with lower vertebrates, higher levels of

metabolism, increased levels of activity, larger and more complex arrays of behavioral
responses, and endothermy.
Along with the evolution of these features were structural changes in all organ
systems. The structure of the mammalian skull has been affected most by the increase in
the size and complexity of the brain and changes in breathing and feeding mechanisms.
In the evolution of the mammalian skull, elements of the outer dermal skull and
inner endochondral skull (seen in the perch) have fused to form a single large skull to
house the enlarged brain. There has also been a reduction in the number of bones
forming the skull and fusion of these bones in the adult skull. These changes have
243
increased the strength of the skull necessary to resist the forces produced by the large
and powerful jaw muscles.
Higher levels of activity and metabolism require mammals to consume large

quantities of food and exchange large volumes of air in their lungs. Mammals use their
powerful jaw muscles and teeth to cut up and chew food before it is swallowed.
Mammals can breathe and chew food simultaneously because the nasal passages and the
oral cavity (mouth) have been separated by a shelf of bone and soft tissue called the
palate. Locate the following bones on a rat skeleton (Figure 4A):
i) the tooth-bearing bones of the upper jaw: the maxilla and premaxilla,
ii) the mandible,
iii) the occipital.
Also note the location and different types of teeth (incisors and molars).
b) The Vertebral Column
In an aquatic environment, the weight of an organisms body is supported by the

water. In a terrestrial environment, the vertebral column must, instead, support the body
and also transfer the weight of the body to the girdles and appendages. In mammals, the
vertebral column shows a considerable variation in structure due to the different functions
and stresses along its length. Vertebrae are divided into five groups (cervical, thoracic,
lumbar, sacral and caudal) although no two vertebrae are exactly alike. Virtually all
mammals have seven neck or cervical vertebrae. Unlike the fish, a mammalian tetrapod
has a neck and can freely move its head independently from movement of its trunk. This
is due to modifications of the first two cervical vertebrae and their articulations. The first
cervical vertebra is the atlas and it articulates with the occipital condyles of the skull.
Movement at this joint produces vertical movement of the head (the yes or nodding
motion). The second vertebra is the axis (Figure 4). Movement at the joint between the
atlas and axis produces rotational movement of the head (shaking the head no).
Thoracic vertebrae bear ribs while lumbar vertebrae do not. Differentiation of trunk
vertebrae into thoracic and lumbar regions is correlated with the evolution of a
diaphragm. Most mammals have 1215 thoracic and 4 7 lumbar vertebrae. The
sacrum is formed by the fusion of 3 5 sacral vertebrae. In terrestrial mammals, body
weight is transferred to the pelvic girdle through the sacrum.
Mammalian tails vary greatly in length and function and, therefore, the number
and size of the caudal vertebrae varies between species.
Examine the skeleton of the rat and distinguish the five vertebral regions described above
(Figure 4A).
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c) Sternum and Ribs
The sternum and ribs encase and protect the thoracic organs and help in
ventilation of the lungs. As mentioned, differentiation of the trunk vertebrae into thoracic
and lumbar regions is correlated with the evolution of a diaphragm. The diaphragm is a
sheet of muscle that separates the thoracic cavity containing the lungs from the
abdominal cavity. You observed this muscle when you dissected your specimen in the
practical lab. The ribs help prevent the collapse of the pleural cavities when the
diaphragm contracts and movement of the ribs occurs during deep or laboured breathing.
All ribs attach dorsally to the thoracic vertebrae and curve ventrally. True ribs (7
pairs) attach directly to the sternum, while false ribs (3 pairs) attach to the rib(s) anterior
to themselves. Floating ribs (3 pairs) do not have any ventral attachments. The sternum
is formed of a number of elements. The clavicle of the pectoral girdle articulates with the
first of these elements. A xiphoid process composed of cartilage is attached to the last
element - the xiphisternum. On the rat skeleton, locate the sternum, xiphisternum and
all rib types (Figure 4B).
2. The Appendicular Skeleton
The pentadactyl (five digit) limbs of terrestrial vertebrates evolved from the fins
of fishes. In the evolution of the tetrapod mammals, the limbs became more and more
important in locomotion and in supporting the body. Thus, through evolution the
position of the limbs changed such that they are located essentially beneath the body of
the mammal and move back and forth in the vertical plane.
In the transition from an aquatic to a terrestrial environment and the development

of appendages, the pectoral and pelvic girdles became larger and stronger. In addition,
with the evolution of a distinct neck, the pectoral girdle lost its connection to the skull.
The basic terrestrial vertebrate limb is composed of three segments. The proximal
segment contains a single bone, the middle segment contains two bones and the distal
segment contains multiple bones and typically has five digits (pentadactyl). All
vertebrate limbs have evolved from this basic plan.
a) Pectoral Girdle and Appendages
Examine the rat skeleton and locate the bones forming the pectoral girdle: the scapulae
(singular, scapula) and the clavicles. Locate the bones of the pectoral limb the
humerus, radius, ulna, carpals, metacarpals and phalanges.
b) Pelvic Girdle and Appendages
Examine the pelvic girdle of the rat skeleton. On each side, three bones have fused to
form a single strong element. These elements are also fused to each other and to the
sacrum. Locate the bones of the pelvic appendages the femur, tibia, fibula, tarsals,
metatarsals and phalanges.
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Figure 4A: Skeleton of the Rat (lateral view).
Figure 4B: Sternum and Ribs of the Rat (lateral view).
246
ii) The Skin (Integumentary System)
The epidermis in mammals is thick and has many layers of dead cells at the
surface. The epidermis is effective in protecting the underlying tissues from water loss,
invasion by microorganisms and mechanical abrasion. The epidermis on the soles of the
feet and toes is very thick and forms horny pads.
The dermis is composed of dense fibrous connective tissue and contains blood
vessels, nerve fibres and sensory endings, arrector pili muscles, glands, and hair
follicles. Sebaceous glands and sweat glands are multicellular glands of epidermal
origin. Sebaceous glands produce oily and waxy secretions that are usually secreted into
hair follicles. Sweat glands occur in most, but not all mammals. Mammary glands,
characteristic of all mammals, probably evolved from sweat glands.
The presence of hair is a distinct mammalian characteristic and hair is the most
obvious derivative of mammalian skin. In most mammals, hair forms a dense protective
and insulating fur over the body. Eyelashes and vibrissae are modified hairs. Claws are
present in most mammals but have been modified as nails in primates and hooves in
ungulates.
Observe your own finger nails. To see your nails clearly, place your hand under the
dissection microscope on your desk.
Examine slide #51 (a section of mammalian skin) noting the features described above
(Figure 5). If your slide is of poor quality, examine the slide of mammalian skin on
demonstration.
Figure 5: Cross section through the skin of a mammal.
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iii) Reproductive System
In the practical lab you studied the macroscopic features of the reproductive
system of the rat. In this lab, you will study the microscopic features. A brief overview
of vertebrate reproductive systems follows.
The gonads, ovaries and testes, of all vertebrates are similar in structure and
function. The gonads are usually paired (although they may be fused or single in some
groups). The testes in all vertebrates produce millions of sperm at a time. As we shall
see, the major differences in male reproductive systems involve the mechanisms for
transferring those millions of sperm to the ova.
Ovaries produce ova surrounded by fluid filled sacs called follicles. These
follicles burst releasing ova into the coelom and then the oviduct. The number of ova
produced at any one time varies considerably depending upon the habitat of the animal
and the amount of parental care the young will receive. Thus, fish, like the cod, which
release unprotected eggs into the ocean and do not care for their young may produce and
release more than a million eggs at once. Other fish and amphibians which do not
broadcast their ova as far afield or which give some measure of parental protection to the
eggs may produce thousands of eggs at once. Birds and mammals, which invest
considerable parental energy in protecting their offspring both before and after birth,
produce only a few eggs (ova) at one time.
Once ova are released from the ovary, they are transported through the oviduct
toward the exterior. In the case of fish and amphibians, which release shell-less eggs, the
oviduct is essentially an unspecialized tube leading indirectly to the exterior. Birds and
reptiles produce shelled eggs; the shell is critical in protecting the egg from desiccation in
the terrestrial environment. The oviducts in these animals are modified, being in close
association with a shell-forming gland and capable of considerable distension to
accommodate the enormous, inflexible eggs that result.
Mammals, which bear their young live, have developed a different modification
of the oviduct system. The distal portion of the duct, involved in shell deposition in birds
and reptiles, is enlarged as the uterus. This thick walled muscular structure, the lining of
which will form an intimate relationship with the membranes around the embryo to form
the placenta, will participate in nourishing the young as they develop within the female
reproductive tract. In primitive mammals, (the rat is a fine example), the two uterine
horns remain separate; in higher mammals (primates) which produce fewer, more
developed young, the two horns of the uterus are fused to form a single large uterus.
A critical difference between vertebrates, which release unprotected eggs, and

those which release shelled eggs or in which the young develop within the female's body,
is the timing and location of fertilization. In fish and amphibians, which release
unshelled eggs into an aquatic environment, fertilization is external, occurring in the
water in which the males and females release their gametes. Embryonic development, as
well as fertilization, will take place in the aquatic environment. No special organs are
248
required for transfer of sperm. However, in some cases the males may have structures,
such as thumb pads of frogs, for holding the female in place so sperm and eggs are more
likely to be in the same place at the same time.
In animals with shelled eggs or internal development, fertilization must be

internal and these animals need a mechanism for getting the sperm to the egg while the
egg is still in the appropriate region in the female's body. Interestingly, birds lack
copulatory organs and rely on close apposition of males and female genital openings for
sperm transfer. Male reptiles and mammals have accessory structures (the penis) which
can be inserted into the female genital opening releasing the sperm well within the
female's body.
In many ways the problems of reproduction in aquatic and terrestrial

environments are similar for plants and animals. As a result, many of the solutions are
similar.
Examine slide #41, which is a cross section through the testes of a rat. Note the circular
outlines of the sections of seminiferous tubules, which are the site of sperm production.
Locate mature sperm in the centers of some of these tubules (Figure 6).
Figure 6: Diagram of rat testis (cross section).
Examine slide #40 of a cross section of a rat ovary. Note the large sac-like follicles in
which the ova (eggs) mature. Some of the follicles may have ova visible within them
(Figure 7). When a follicle is ripe (Graafian follicle) and its ovum is mature, it bursts,
releasing the ovum, which is then picked up by the oviduct. The oviduct is ciliated
internally. The cilia beat to create currents to draw the ovum inside and propel it towards
the uterus.
Also, examine the demonstration slide of a mature Graafian follicle.
249
Figure 7: Microscopic view (top) and diagram of a rat ovary (cross section).
iv) Circulatory system
In the practical lab we studied the heart and its associated vessels. In this lab we
will examine some of the other vessels that make up the circulatory system of the rat.
You will not perform any dissections in order to study these vessels. Instead, you will
use the information in your lab manual along with the demonstration materials on display
in the lab.
1. The Arteries and Veins Of The Head
Find the heart on the demonstration dissection (you may need to review the
section on the heart from the Practical lab). Oxygenated blood leaves the left ventricle
through the aortic semilunar valve and enters the aorta. The aorta immediately divides
250
into the innominate artery and the aortic arch, which sends off two branches; the left
common carotid artery and the left subclavian artery (Figure 8). The aortic arch
continues as the dorsal aorta.
Locate the innominate artery and carefully trace its path to the point at which it
branches. (you can use the probes provided) The right common carotid artery is the
branch which carries the blood to the right anterior portion of the head and the right
subclavian artery carries blood to the right front leg. The right and left subclavian
arteries give off a number of branches which carry oxygenated blood to the thoracic
cavity, the vertebral column, the brain, the muscles of the neck and shoulder, and
continue on as the right and left axillary arteries.
The venous system in the head region carries deoxygenated blood to the heart.
The right and left superior venae cavae, which you located in the Practical lab, are
parts of this system.
Figure 8: Arteries and veins in the head region of the rat.
251
2. The Visceral Arteries and Veins
Locate the dorsal aorta again. Notice the numerous intercostal arteries that arise
from the dorsal aorta and go to the ribs. Just posterior to the diaphragm locate the aorta.
The first large unpaired abdominal artery that branches from the aorta is the coeliac
artery (Figure 9). After a short distance, the coeliac artery gives rise to three smaller
arteries - the splenic, hepatic, and the gastric arteries. The splenic artery supplies the
pancreas and spleen. The hepatic artery supplies the liver with oxygenated blood and the
gastric artery serves the stomach.
The second unpaired branch of the dorsal aorta is the superior mesenteric
artery. Carefully trace the superior mesenteric artery. It gives rise to a number of blood
vessels, which lead to the pancreas, duodenum, and small intestine.
Locate the paired renal arteries, which come off the aorta after the superior
mesenteric artery and carry blood to the right and left kidneys.
Just posterior to the renal arteries are the small paired ovarian arteries (female)
or the testicular arteries (male) which supply blood to the ovaries or testes. Locate the
two iliolumbar arteries, which carry blood to the dorsal body wall. In the region of the
iliolumbar arteries the dorsal aorta gives off the inferior mesenteric artery, which
carries blood to the colon and rectum.
The dorsal aorta splits into two branches; the left and right common iliac
arteries, which divide into a number of branches supplying blood to the reproductive
organs and hind limbs.
Follow one of the common iliac arteries as it goes out towards the leg. It will first
give off a small artery ventrally which runs to the uterus or to the accessory glands of the
male. Look deep under this vessel - here you will find the internal iliac artery. The main
artery continues out to the leg as the external iliac where it will further subdivide to serve
the pelvic region and the leg itself.
From the branching of the common iliac arteries, the dorsal aorta is known as the
caudal artery which supplies blood to the tail (Figure 9).
In many instances, the venous system has comparable blood vessels to that of the
arterial system (Figure 10). Locate the left and right common iliac veins, which flow
into the inferior vena cava. Trace the inferior vena cava anteriorly and locate the paired
iliolumbar veins, testicular veins (male), ovarian veins (female), and the renal veins.
The inferior vena cava enters the liver where the hepatic vein enters it. (This is usually
buried deep within the liver tissue so do not attempt to locate the hepatic vein). The
inferior vena cava continues on to enter the right atrium.
252
Figure 9: Visceral arteries of the rat.
253
Figure 10: Visceral veins of the rat.
254
VERTEBRATE RESPIRATORY SYSTEMS
The following information is meant as an introduction to general vertebrate

respiratory physiology. It should be used to supplement the knowledge you gained from
the dissections you performed in the practical lab.
Oxygen is used as the terminal acceptor of the electron transport system which is
found in mitochondria. This means oxygen must diffuse into all cells, even in large
multicellular organisms. In addition, carbon dioxide is produced during cellular
respiration by decarboxylation reactions which are the exact reverse of carboxylation
reactions occurring in photosynthesis. This gas must be removed because if it
accumulates it will change the internal environment of the organism which must remain
relatively stable for proper function. Carbon dioxide combines with water to form
carbonic acid which will change the pH of blood and tissue fluids interfering with
enzyme reactions and causing a multitude of difficulties. Consequently, highly
developed structures have evolved for gas exchange.
Oxygen cannot diffuse directly to the mitochondria, it must become dissolved in

the fluid environment of the cell. Regardless of the size or living conditions of the
animal, all gas exchange requires the gas to go into solution. Because the volume of a
cell increases as a cube function and the surface area as a square function, the size of cells
is limited. This is necessary as eventually not enough surface area would be available for
exchange between the cell and its environment. By becoming multicellular, surface area
is increased. With division of labour, some cells become gas exchange surfaces. As the
organism increases in size, more respiratory surface is needed. This has resulted in the
development of infolded (invaginated) and outfolded (evaginated) respiratory organs.
In general, lungs have developed as invaginations and gills as evaginations.
Because oxygen is about 20 times less soluble in water than air (210 cm3 of
O2/litre of air, 4 to ll cm3 of O2/litre of water) aquatic animals have developed a wide
variety of mechanisms for extracting it from water. Most of these depend on keeping the
concentration of oxygen as high as possible on the outside of the respiratory membrane
and as low as possible on the inside, so inward diffusion occurs as rapidly as possible.
Respiratory pigments combine with oxygen so that the concentration is kept low in the
surrounding fluid. The flow of oxygen-rich water is often in the opposite direction of
oxygen-poor blood to keep the concentration gradient as high as possible. To facilitate
diffusion, respiratory membranes are very thin and, consequently, fragile. Gills are often
protected by a covering which sometimes becomes involved in pumping water across
them. Terrestrial animals also require oxygen to be dissolved in water in order to be
absorbed, and organisms with lungs, therefore, must keep them moist. Because lungs are
the result of invagination, they are internal and well protected.
255
256
Question 1: In the rat, what is the name of the single bone of the proximal segment
of the pectoral limb?
_____________________________________________________________________
Question 2: How many digits does the rat have?
_____________________________________________________________________
Question 3: Is the pectoral girdle of the rat attached to the skull?
______________________________________________________________________
Question 4: In the rat, which two bones make up the middle segment of the pelvic
limb?
_______________________________________________________________________
Question 5: What is the reason for the absence of canines and premolars in the rat?
_____________________________________________________________________
_____________________________________________________________________
Question 6: Are the sperm cells of the rat produced by mitosis or meiosis?
_____________________________________________________________________
257
258
APPENDIX
THE MICROSCOPE
This is a review of how to use one of the most important tools in Biology -- the light
compound microscope. You will be putting this instrument to a great deal of use in the
weeks and months to follow. Knowledge of its potential, its limitations, and the
techniques involved in using it to the best of its advantage is an asset which no student
can afford to be without.
I The use of the light compound microscope (OLYMPUS CX21)
Handle your microscope with care and learn to use it properly. It is one of the best
education microscopes available!
Many parts of the microscope, such as lenses, condensers, mechanical stages, etc., can
be removed by unscrewing them or pulling them off. Never, under any circumstances,
remove part of the microscope unless you are specifically instructed to do so. Loose
parts may be lost, or put on a microscope to which they do not belong, and on which they
may not work as well as the parts that do belong. Removal of parts may also permit dust
to enter the microscope and thus impair its effectiveness.
The successful use of the microscope is largely dependent on good microscope

technique. It is a precision instrument and should be treated as such. Some points of
good micro-technique are outlined below:
1. Carefully lift the microscope from its cupboard onto the desk. When carrying the
microscope, hold both sides around the hole of the arm as shown in the diagram below.
2. Remove the dust cover, fold it and place it in your cupboard. This ensures that the
dust cover will not be torn or lost during the laboratory period.
259
3. Cleanliness
A great deal of the difficulty which students encounter is a result of dirt -- on
microscope lenses, on the condenser, or on the slide. It should be a matter of routine
to clean the top ocular lens (eyepiece), the bottom objective lenses, the top lens of the
condenser, and both surfaces of the slide before starting to use the microscope. Use
only lens tissue to do so -- never paper towels, kleenex or handkerchiefs. The latter
may contain particles of dust or grit which could scratch the lenses. If you are unable
to get a lens clean, or if the microscope lenses are dirty inside, cover your microscope
with the plastic dust cover and report the difficulty to the Instructor or Demonstrator.
4. Resolution
Carefully following the steps outlined below will result in the best resolution possible
with the microscope, and the sharpest image. Resolution is defined as the ability to
see fine detail, and may be measured as the minimum distance between two points at
which they are discernable as two distinct points, rather than a single point -- the
shorter this distance, the greater the resolving power. Resolution depends, in part, on
your own visual power and on the quality of the microscope you are using -- factors
which cannot be altered. However, it also depends, to a very large degree, on the way
in which you use the microscope. Cleanliness and the correct use of the condenser,
iris diaphragm, and light all contribute to improving resolution.
5. Learn to Use the Microscope with Both Eyes Open

Remember that the use of the microscope with one eye closed results in eye strain,
fatigue, and very often, severe headaches.
6. Start with Low Power and, if necessary, work up through Medium to High Power.
In looking at any microscope slide, start with low power and, if necessary, work up
through medium to high power. Locate and centre the object you are looking for at
the lowest possible magnification. If you can clearly distinguish the details described
on medium power (10X objective) no advantage is gained by using high power (40X
objective).
7. Some slides need to be examined at the lower powers.

Some slides are to be examined at the lower powers, and will be pointed out to you as
they are to be examined. Under no circumstances are these slides to be examined
under the 40X objective, as slide breakages will usually result.
8. Switch the light off when you are not using the microscope.
9. At the end of your observations:

- make sure that you remove the slide from the stage and place it in the appropriate slot
of the slide box,
- rotate the revolving nosepiece to bring the 4X objective into position,
- wrap the cord around the frame and base of the microscope,
- cover the microscope with the plastic cover and place it in the cupboard.
260
10. Using the eye shades:
When Wearing Eyeglasses

Use with the eye shades in the normal, folded-down position. This will prevent
the eyeglasses from being scratched.
When Not Wearing Eyeglasses
Extend the folded eye shades in the direction of the arrow (see diagram below) to
prevent extraneous light from entering between the eyepieces and eyes.
II The parts of the light compound microscope
The microscope has two lens systems which are for magnifying the objects under
observation. The lenses through which you look are termed the oculars or eyepieces
(Figure 1). On the Olympus CX21 these are marked 10X, which tells you that this lens
can magnify an image ten times its size. On the left ocular, there is the diopter adjustment
ring. It compensates for the difference in eyesight between your eyes. The lower set of
lenses (there are three on the revolving nosepiece) are termed the objective lenses.
These are marked with the following numbers: 4.0X; 10X; and 40X. These numbers
signify the magnification given by each of these lenses, i.e. 4.0 times, 10 times, and 40
times. The different objectives are also colour coded. There are also other numbers on
each of the objectives. These numbers are related to the aperture size of these lenses and
glass thickness. A third set of lenses is found in the condenser. Their role is to gather the
light from the microscope lamp and concentrate it so that it passes through the slide to the
objective lens. Carefully turn the microscope on its side so that you can see the bottom
surface of the condenser (Figure 1). Locate the iris diaphragm, a structure made of thin
overlapping metal plates with a hole in its centre. Movement of the iris diaphragm ring
results in the altering of the diameter of the hole, thus controlling the amount of light that
reaches the condenser. The large round knob on either side of the main body of the
microscope is the coarse adjustment knob. It s responsible for coarse focusing and
should only be used with the lower power objective. The smaller knob is the fine
adjustment knob and is used to sharpen the image you are viewing. It can be used with
all of the objective lenses.
On the large object stage, you will find a clipping device known as specimen holder,
which holds the microscopic slide in place. Note the specimen holder scales, which
identify the specific position being observed on the specimen. The specimen holder can
be moved to change the area of the slide under observation. The specimen holder knobs
are located on the left-hand side of the microscope, below the stage. They allow for the
261
rapid and systematic scanning of slides. The upper knob moves the slide along Y axis
while the lower one moves the slide along the X axis. On the left-hand side of the base of
the microscope, you will find main switch and the light intensity adjustment knob.
Figure 1: A Light Compound Microscope.
III Steps in focusing the light compound microscope
DO NOT FORCE ANY OF THE KNOBS. IF YOU FEEL RESISTANCE, ASK

FOR HELP!
1. Set the microscope in front of you at a comfortable working distance for observing
and reading.
2. Plug the microscope in and switch it on. You may need to adjust the light intensity.
Rotating the light intensity adjustment knob towards you increases brightness and
rotating it in the opposite direction decreases brightness. It is best to set the knob mid
way and later adjust, if necessary.
262
3. Turn the revolving nosepiece until the low power objective lens (4X) is in position.
4. Rotate the coarse adjustment knob to lower the stage.
5. Place a slide on the stage. Set it in position with the specimen holder.
6. Use the specimen holder knobs to centre the slide over the area you want to look at (it
works best if you watch from the side). Rotating the upper knob moves the slide in
the vertical direction (Y axis). Rotating the lower knob moves it in the horizontal
direction (X axis).
Do not move the specimen holder directly by hand, for this will damage the rotary
mechanisms of the above knobs.
When the specimen holder reaches the stopper position, the rotation force of the above
knobs become heavy. Stop rotating the knob at this time.
7. Turn the coarse adjustment knob to focus. The safest way to bring the object into
focus is to bring the stage up as far as it will go while looking from the side. Then
move the stage down slowly while looking into the oculars until the object comes into
focus. This procedure will prevent the breakage of slides.
8. Adjust the condenser position and iris diaphragm aperture. The condenser is usually
used in the highest position. If the entire observed field of view is not bright enough,
brightness may be improved by lowering the condenser slightly. To move the
condenser, rotate the condenser height adjustment knob (1), seen in the diagram
below. The iris diaphragm ring (2), seen in the diagram below, has an objective
magnification scale (4X, 10X, 40X). Rotate the ring so that the magnification of the
objective in use faces frontward.
Once you have found the object youre looking for on low power (objective 4X), try
finding it on higher powers (objectives 10X and 40X) by doing the following:
263
9. While looking into the microscope, centre the object of interest on the slide. Then
switch to the 10X objective. A fine adjustment to focus may be needed. If so, use the
fine adjustment knob. Your microscope is parfocal -- once the image has been
brought into focus under low power, it is not necessary to adjust the coarse focus
when switching to a higher power.
Also, adjust the iris diaphragm to match the 10X objective.
10. Now focus on the letter object of interest with the 40X objective. Before doing so,
make sure that the letter object is centred, moving the slide if necessary. Watching
from the side, slowly rotate the revolving nosepiece to bring the 40X objective into
position. Look through the lens and alter the fine adjustment, if necessary, to
sharpen the image. The coarse adjustment should not be used while looking into the
microscope. Otherwise you may destroy the microscopic lens, break the slide or
coverslip (although the spring-loaded lenses help to reduce this possibility), and
scratch the lens.
Note that the working distance (the distance from the objective to the slide) is very
short for this objective, usually less than 1 mm. If the high power objective will not
swing into position without touching the slide, it means that the coverslip and mount
on the slide are too thick. You should not attempt to use the 40X objective on this
type of slide.
11. Adjust the interpupillary distance of the two oculars so that you can observe a single
microscopic image through the two oculars, as seen in diagram below. While
looking through the eyepieces, move both oculars until the left and right fields of
view coincide completely. The position of index dot () indicates the interpupillary
distance value.
12. If necessary, adjust the diopter. The diopter adjustment compensates for the
difference in eyesight between your eyes. While looking through the right ocular
with your right eye, rotate the coarse and fine focus adjustment knobs to bring the
specimen into focus. While looking through the left ocular with your left eye, rotate
only the diopter adjustment ring to focus on the specimen.
264

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LAB MANUAL 2017

LABORATORY SCHEDULE WINTER 2017

Week # Practical room (0RC046) Demo/Tutorial room (0RC044)

THE LAB IS WORTH 40% OF YOUR MARK IN THIS COURSE:

Midterm Lab Exam (Exam I) = 12%; week of February 13, 2017

The laboratory course in Biology is designed to serve several purposes:

1. Illustration of some of the lecture material.

2. Acquisition of information first hand.

3. Providing training in careful observation and deduction, the accurate

Laboratory work is extremely important in Biology. Textbook, lecture

The role of the laboratory Instructor and student demonstrators is to provide

MATERIALS SUPPLIED BY THE STUDENT:

1. Textbook: Campbell Biology in Focus, 2nd Ed. by Campbell et al.,

2. Laboratory Manual for BIOL-1116.

3. a) Dissecting Kit including:

4. A lab coat is mandatory. Please be sure to bring a lab coat to every

NOTE: THE ABOVE ARE THE MINIMUM REQUIREMENTS.

MATERIALS SUPPLIED BY THE UNIVERSITY:

Lab attendance in BOTH practical and theoretical labs is compulsory not

Missed Quizzes and Exams:

NO SPECIAL CONSIDERATIONS WILL BE MADE IF THE INSTRUCTOR (OR

In this course, laboratories are subdivided into practical and demo/tutorial

Depending on the nature of the exercises, students will work individually or in

Most laboratory exercises include a number of demonstrations, specimens, or

Basic Dissection Techniques

1. Practice safe hygiene when dissecting. Wear appropriate protective clothing,

5. A good strategy to use if working in groups is to read aloud the directions

4. Slide Box Policy

i) Cranial, caudal and rostral

ii) Dorsal and ventral

iii) Lateral and medial

iv) Proximal and distal

vi) Right and left

INTRODUCTION TO THE LIVING WORLD

1. To briefly introduce the organization of the living world.

2. To describe the basic characteristics of the Domain

3. To describe the basic characteristics of some

4. To examine asexual reproduction in the three main groups

I THREE DOMAINS OF LIFE

The diversity of the Bacteria Domain should not be surprising. Representatives of

Binary fission is a form of asexual reproduction. However, simple sexual

Figure 2: Binary fission in bacteria.

Figure 3: Bacterial types.

Figure 4: Example of Gram-positive and Gram-negative bacteria.

Cyanobacteria are commonly known as blue-green algae. They are

A number of cyanobacteria produce specialized, nitrogen-fixing cells called

Figure 7: Gloeocapsa sp.

III DOMAIN EUKARYA

The Domain Eukarya includes three very diverse Kingdoms of eukaryotic

i) the algae (plant-like protists)

In this lab, we will examine some characteristics of a few representatives of each

i) Algae (Plant-like protists)

Figure 8: The freshwater dinoflagellate - Ceratium sp.

Almost all euglenids are unicellular flagellates which contain a spiral or

Examine slide #57 of Euglena sp.

Figure 9: The unicellular flagellate - Euglena sp.

Diatoms are a major component of plankton in oceans and lakes. Their

Examine slide #72 of diatoms and note many different shapes.

Figure 10: A freshwater diatom.

d) Chlorophytes (Green Algae)

Make a wet mount of Chlamydomonas sp. and Chlorella sp..

Observe the similarities and differences between these two organisms.