BIO1AP Prac Manual 2023 - v2

School of Agriculture,
Biomedicine and
Environment
Animal & Plant Life

(BIO1AP)
2023
PRACTICAL MANUAL
La Trobe University, Bundoora & Albury-Wodonga
Department of Environment and Genetics, Department of Animal, Plant and Soil Science
BIO1AP Timetable 2023
Theory
Topic content Day Time Venue
Monday 10 am – 11 am Check in your timetable (Allocate+)
LECTURES
(2 x 1hr per week) Monday 11 am – 12 pm Check in your timetable (Allocate+)
(not a repeat of 10 – 11 am)
ONLINE MODULE
Open Tuesdays at 9 am BIO1APM LMS in relevant week section
(weekly)
Practical
• For the location of your f2f laboratory and workshop sessions see Allocate+. If you have not
already allocated yourself to a lab session (LC01), go to the BIO1AP LMS for further information.
• To the right of the timetable below the due weeks for in-semester assessment are indicated. This
includes Topic quizzes (theory). Check the Subject Learning Guide and BIO1AP LMS for further
details about assessments.
Week Week Topic Lecturer Lab (LC01) / Workshop (WS01) Obs (in Topic Report
Commencing lab) Quiz due
Refln due Mon
due Fri Mon
Online Workshop (via LMS): Lab safety
1 Jul 31 Lab drop-in: familiarisation for students
new to La Trobe
F2F Lab 1 skills re microscope/drawing & Obs
2 Aug 7 Ass. Prof. set up pea expt. for Report 1. Demo expts set
Tony up.
Plant Life
Gendall F2F Workshop – Report writing Refln


3 Aug 14
F2F Lab 2 skills re microscope/drawing & Obs
4 Aug 21 collect pea data for Report 1, Examine demo
expts.
5 Aug 28 F2F Workshop – review pea experiment Refln

Ass. Prof. results / questions about Report 1.
Monika F2F Lab 3 skills re microscope/drawing - Obs
6 Sept 4 Doblin plant tissues

MID SEMESTER BREAK – NO CLASSES (Mon 11 Sept – Fri 15 Sept)
7 Sept 18 F2F Workshop – invertebrate movement Refln

Dr Pam (slides/displays) / vertebrate locomotion
Sept 25 Hurst F2F Lab 4 Report 1 general feedback. Obs
8 (Fri 29th public Invert behaviour / daphnia expt. for Report 2.
holiday TBC)
Animal Life
9 Oct 2 F2F Workshop review daphnia data 

collected in lab 4 class > prep for Report 2.
10 Oct 9 F2f Lab 5 animal adaptations for feeding – Obs

Ass. Prof. skulls / insect mouthparts
Kylie Online Workshop (model organisms, Refln
11 Oct 16 Robert e.g., hydra, (plus vertebrates?)
 
F2F Lab 6 – Thermoregulation simulation Obs
12 Oct 23
Oct 30 Swot vac – no classes 
Name:_____________________________________
Lab/Workshop Day:_________________________ Time:______________
Demonstrator Name:________________________
Animal and Plant Life (BIO1AP)

2023
If you have not already allocated yourself to a lab session, go to the BIO1AP
LMS for further information.
f2f classes begin in week 2 with online activities in week 1

see the timetable on the previous page.
Signposts used in this manual:
Safety note: Carefully read the

associated safety instructions
Reading: List of reading to

support the lab/workshop and
associated lectures.
To do: Complete a table,

drawing, graph, etc.
Watch: Your demonstrator will

show you a particular technique
you will need to use or there will
be group discussion.
LMS: There is an LMS activity

to support the lab/workshop
session. It will usually need to
be done prior to the class.
CONTENTS
CONTENTS ....................................................................................................................... 2
LABS AND WORKSHOPS - GENERAL INFORMATION .................................................. 3
Timetable and Venue ................................................................................................................. 3
Laboratory equipment for class .................................................................................................. 3
General Safety Procedures in the Laboratory ............................................................................. 4
COVID-19 (& flu etc.) Safety in the Laboratory ........................................................................... 4
Practical assessment (38% towards final grade) ........................................................................ 5
WORKSHOP 1 Lab Safety online / teaching lab drop-in ................................................. 10
LAB 1 PLANT ROOTS AND PLANT GROWTH REGULATION...................................... 11
Part 1 - PLANT ROOTS ........................................................................................................... 11
Part 2 - PLANT GROWTH REGULATION EXPERIMENTS – set up ........................................ 17
WORKSHOP 2 Scientific report writing ........................................................................... 23
LAB 2 PLANT GROWTH REGULATION – Data collection ............................................. 29
Part 1 – Plant Growth Regulation data collection...................................................................... 29
Part 2 - PLANT AND MICROBIAL SYMBIOSES ...................................................................... 36
WORKSHOP 3 Effect of GA (Report 1) data analysis/presentation ................................ 41
LAB 3 Plant leaves and stems ........................................................................................ 45
Introduction ...................................................................................................................... 45
PART 1: Primary Growth in Stems ........................................................................................... 48
PART 2. Secondary Growth in Stems ...................................................................................... 50
PART 3. Leaf epidermal peel ................................................................................................... 53
WORKSHOP 4 Animal locomotion.................................................................................. 55
Earthworm................................................................................................................................ 55
Animal locomotion – comparing running, flying and swimming ................................................. 57
LAB 4 Effect of temperature on heart rate (Report 2) / Report 1 feedback ..................... 61
Background: Effect of temperature change on physiology ........................................................ 61
Specimen preparation .............................................................................................................. 64
Recording heartbeat and experimental set up. ......................................................................... 65
Report 1 feedback .................................................................................................................... 70
WORKSHOP 5 Effect of temperature on heart rate (Report 2) results............................ 71
LAB 5 Animal Adaptations for Feeding ........................................................................... 77
Part 1 What can I learn from a skull? ........................................................................................ 77
Part 2 Insect mouthparts .......................................................................................................... 82
WORKSHOP 6 Asynchronous activity via LMS .............................................................. 84
LAB 6 Thermoregulation in simulated animals in cold conditions ................................... 85
Part 1: Freezing point depression in an ectotherm ................................................................... 85
Part 2: Effectiveness of insulation on thermoregulation in an endotherm .................................. 90
LAB AND WORKSHOP CLASSES - GENERAL INFORMATION
Timetable and Venue
The lab and workshop class schedule is on the inside cover of this manual.
You should attend the same session (i.e., day, start time) each week and if you do not currently have
a BIO1AP LC01 (lab class) activity timetabled in Allocate+ go to the BIO1AP LMS for further
information. (information on using Allocate+ can be found on this webpage:
http://www.latrobe.edu.au/students/timetables.
Laboratory equipment for class

Bring the following equipment/items to every lab class:
• A bound copy of this Practical Manual For some classes you will also need:
• Laboratory coat • Calculator
• Safety glasses (can be borrowed if • Small paintbrush (may be in a
needed) dissecting kit)
• Pens, pencils (including B for drawing), • Sharp/blunt ended probes (as in a
eraser, ruler dissecting kit)
• Permanent marker pen
At all times in the laboratory, you must wear fully enclosed shoes to protect your feet from spills and
sharps. Students wearing thongs, open toed shoes or ballet flat style shoes will not be permitted to
remain in the laboratory. Disposable gloves will be provided when required – if you are allergic to
latex please tell your demonstrator and alternative gloves can be provided.
For workshops, make sure you bring this manual and wear closed shoes in the lab.
Repeat students
It is highly recommended that you attend all learning activities to give yourself the best chance of
passing. If you feel your performance in labs/workshops and associated assessments was adequate
in a previous attempt, then you will need to contact the subject coordinator, Pam Hurst at
BIO1AP@latrobe.edu.au.
Practical Coordinators
Bundoora - Dr Pam Hurst. She is also the subject coordinator and will be in all practical sessions.
Please feel free to see Pam during class with any questions. For out of class time, Pam’s contact
details and weekly consultation time can be found on LMS.
Albury-Wodonga - Dr Michael Shackleton. He is also your instance coordinator and will be in all
practical classes. Please feel free to see Michael during class with any questions. For out of class
time, Michael’s contact details can be found on LMS.
3
General Safety Procedures in the Laboratory

All students need to complete an online lab safety induction quiz scoring 10/10 before attending
your first laboratory class, which is in week 2. Even if you have completed a lab induction in semester
one for BIO1MGC or have done the quiz for BIO1EEB, you need to complete a lab safety induction
for each subject, even if the same labs are used. Go to BIO1AP LMS for further information.
When working in a science laboratory there are rules that have been established to ensure the safety
and welfare of both students and staff. Important areas covered include:
• Personal Protective Equipment (PPE). FULLY enclosed footwear is to be worn at all times
when working in the laboratory during labs or workshops. Laboratory coats are to be worn at
all times during lab classes. Safety glasses will be needed for some activities, and when will
be indicated in the manual. Hair tied back during lab classes.
Students not wearing the appropriate PPE will need to leave the class (see below re COVID-19).
• Laboratory manual. This contains instructions related to activities and safe procedures and for
this reason a hardcopy is to be brought to all classes. You will not be able to have a
laptop/device with you for all activities.
• Waste disposal. Sharp items such as glass slides, cover slips and razor blades are hazardous
and must be disposed of in the special yellow 'Sharps' bins or containers provided on lab
benches. Biological waste must be disposed of in the appropriate bins. Ask your demonstrator
if unsure of correct disposal.
• Correct movement/storage of equipment such as microscopes and also of bench stools.
• Report any accidents broken equipment or near misses to your demonstrator immediately.
Take care with the use of chemicals, razor blades, thermometers, and glassware.
• Tidy and clean bench. Equipment to be returned to bench station and your bench area to be
wiped with cleaning fluid and paper towel before leaving.
COVID-19 (& flu etc.) Safety in the Laboratory

During face-to-face classes we all need to provide the safest possible environment for everyone
present.
Face masks are strongly recommended whenever in the lab as maintaining

1.5m distance from others will not be possible. Wear the highest quality one
that you can afford.
Face masks are available from the library and will also be available in the lab. If
you would prefer higher quality, e.g., N95, you will need to bring your own.
If you test positive for COVID-19 you are required to inform the university using an online form.
See this webpage for student information regarding COVID-19 and a link to the form: Student
FAQs for COVID-19, About La Trobe, La Trobe University
Steps you can take to keep yourself and others safe this winter and minimise the spread of COVID:
• Get a COVID-19 test if you have symptoms (even people who have had COVID-19
recently should get tested if they experience symptoms).
• Do not come to campus if you have symptoms, even if your COVID-19 test is negative
• Tell us if you test positive to COVID-19 – go here for a link to the form: Student FAQs for
COVID-19, About La Trobe, La Trobe University
• Get a flu shot. These are widely available from pharmacies and GPs across the state.
4
Practical assessment (38% towards final grade)
1. Observations/Reflections 12% of final grade See timetable inside front

(best 8 of 10) cover for which will occur in
which week
2. Scientific Reports x 2 26% of final grade See timetable inside front
cover for due dates for each
For an outline of all assessments see the BIO1AP Subject Learning Guide in the BIO1AP LMS Start
here block. Further information on all assessments is in the Assessment block on LMS. Below are
details relevant to practical assessments, some of which is also on LMS.
1 A. Observations
During lab classes (LC01 in Allocate+) you will show your demonstrator a biological observation
made during the session. At the start of each lab class there will be more information about the
observation for that week. For example, for a particular lab, the lab notes in this manual may indicate
three drawings to do and you will select the one that you wish to show to your demonstrator as your
observation for the week.
Biological observations will be marked out of 5. Criteria are below and are the same as on BIO1AP
LMS.
5 Drawing accurately represents required features together with all the required details. Drawing
very clear and well sized.
Table/Figure very clear and accurately contains required information/detail.
4 Drawing generally accurately represents the required features together with all/most of the
required details. Drawing clear.
Table/Figure clear and accurately contains required information/detail. Only minor errors.
3 Drawing represents structures/features but could be clearer. Most required details present.
Table/Figure clear with mostly accurate required information/detail.
2 Drawing unclear and does not clearly represent structures/features and/or missing most
required details.
Table/Figure with omissions/errors of important required information/detail.
1 Drawing shown but does not represent structures/features as required and/or is missing
required details.
Table/Figure with many omissions/errors of important required information/detail.
0 Not shown.
5
1.A.1 Recording your observations through drawings
For some labs you will need to make drawings of the plant or animal structures
you observe. The purpose of drawing, rather than taking a photograph, is to
record the relevant structures and their relationship to one another. You do not
need to be able to produce a drawing that is like a photograph. Producing a
labelled drawing is also an excellent way to ensure you have correctly observed
and understood the plant or animal anatomy.
General instructions for drawing
1. Drawings should be made directly when viewing the specimen and completed in the
laboratory. It is unscientific and inaccurate to make rough sketches to be completed later,
when no specimen is available.
2. Use a sharp B pencil for drawing.
3. Drawings are to be line drawings only, i.e., only the outline of structures should be drawn.
Shading should not be used. Lines must be smooth and clean.
4. Low power drawings do not require individual cells to be drawn, only the boundaries of each
cell type/structure.
5. See that the proportions of the drawings are correct by roughly estimating the relative sizes
of the main structures and outlining them faintly first. Drawings should be well-spaced on the
page and take up at least half an A4 page.
6. Drawings should contain all features seen. If you cannot see a feature, you have been
directed to look at, use the lab copies of Campbell Biology and/or ask your demonstrator for
assistance.
7. All drawings should be completely labelled, regardless of whether the same structure has
been labelled in a preceding drawing. The labels should be neatly written and placed
horizontally. The labelling lines should be ruled (no arrows) and label lines should not cross
each other.
8. Every drawing should have the appropriate heading, containing relevant information such as:
a) the name of the organism (e.g., Genus species – when handwriting underline the two names
to indicate this is the scientific name. When typing a scientific name, italics are used and no
underlining);
b) the nature of the specimen or preparation being drawn, e.g, whole specimen, longitudinal
section (L.S.), transverse section (T.S.), smear;
c) the part of the organism, e.g., root tip;
d) in the case of magnification, state the amount, e.g., X40 (low power), X100, X400 (high
power).
6
Example of a low power magnification drawing

• A low power diagram is a plan representation only (Figure 1)
• No detail of individual cells is drawn.
• The arrangement of different cell types/tissues/structures in a specimen is illustrated,
indicating relative positions/sizes.
Ranunculus repens
Transverse section, root
10x
Figure 1: Example of a low power drawing in relation to how the specimen would appear under a
microscope (Figure 2)
Figure 2: Black and white depiction of how the specimen would appear under a microscope
7
1.A.2 Recording your observations through graphs/figures
For some labs you will need to record data collected and may be required to produce a
graph/figure to represent these biological observations. These graphs may need be produced on
your laptop in class or sketched in the appropriate place in this lab manual.
To produce graphs on your laptop you should use Excel. If you are not familiar with Excel, or wish
to have a refresher, you can do free online training at LinkedIn Learning (previously Lynda.com).
See this uni webpage for more information:
https://www.latrobe.edu.au/students/opportunities/careers/linkedin-learning
There are a large number of Excel courses but not all are relevant. Suggested beginner courses to
look at: Learning Excel Online (beginner level), Excel 2016: Introduction to formulas and functions
(beginner level).
1 B. Reflections
Following four of your six workshop classes, you will write a short reflection related to the workshop
activity. See the Assessment section of BIO1AP LMS for more information on what a reflection will
entail. Reflections will need to be completed online before 11:59 pm on Friday of the week of your
workshop. See the timetable inside the manual cover for due weeks. A link for online completion
of your reflection will be on LMS but note that the reflection ‘portal’ is not part of LMS and cannot
be set to accept late submissions.
FAQ: If I miss a f2f class (lab or workshop), how do I catch up?

Carefully read the relevant lab or workshop information in the manual. Check to see if there was
any online preparation for the missed class in the relevant week on LMS and do this if not already
completed. Answer any questions in the manual as best you can by looking at a textbook/your
notes on lectures.
Depending on your timetable and when the missed session was, you may be able to attend
another session, but you need to contact Pam Hurst (BIO1AP@latrobe.edu.au) to see if a seat is
available.
If you have any questions about missed class activities, contact a Subject Support Tutor (see the
Student Resources block on LMS for Subject Support Tutor contact details and info about any
sessions they may be running). Depending on your question, they may be able to answer via
email or you can ask to meet up (f2f or via Zoom) if you would like to talk through anything.
FAQ: I missed completing a biological observation/giving a reflection, what happens?
As the best eight of 10 marks will go towards your final mark, missing one (or two) need not impact
your final mark.
Missed observation: Later in semester there will be the opportunity, if you would like to, to come to
the lab and catch up on any missed observation mark. There will be a time during the mid-
semester break, week 11 and swot vac. Details of the time/date will be given closer to the mid-
semester break. Note, this will not be a repeat of the missed lab session, just the opportunity to do
the observation. You will need to contact a Subject Support Tutor to make sure you have caught
up on anything missed during the lab session.
Missed reflection: Poor time management is not a valid reason missing submitting a reflection. For
example, if you wait until just before the deadline and internet problems mean you cannot leave
your reflection, that is a risk you took in waiting until the last minute.
If an extended period of illness has impacted your study, then contact Pam Hurst, subject
coordinator - BIO1AP@latrobe.edu.au
8
2. Scientific reports (based on lab class experiments)
Biologists communicate the results of their experiments or studies both in written form and orally.
Communicating results in a scientific report (or article) is the most common written method and you
will be introduced to the format of written scientific reports and what is expected in written reports
for BIO1AP. In preparation for your week three workshop on report writing, you will complete an
online activity (‘lesson’) in LMS that explains the structure of scientific reports and information in
relation to those that you will write for BIO1AP. Once the LMS activity is completed, you will be
able to download a pdf to refer to when writing your reports.
In the week 3 workshop on report writing, you will discuss with other students the marks you
awarded a sample report, which is part of the online preparation for the f2f workshop. Discussion
with other students and your demonstrator and referring to the marking criteria will develop your
understanding of what is expected.
Writing at university requires you to cite the sources of information that you use in written
assessments, that is, display this aspect of academic integrity. Everyone starting at La Trobe has
automatic enrolment in LTU0AIM (Academic Integrity Module). This module, via LMS, explains all
aspects of academic integrity and, if not already completed, should be completed as soon as
possible so that you understand what constitutes plagiarism (e. g., not citing your sources of
information) and collusion. The two scientific reports you will write are to be submitted via the LMS
and a plagiarism detection software tool called Turnitin will be used. Information for students about
Turnitin is provided here http://www.latrobe.edu.au/students/academic-integrity/explanation/turnitin
Report 1 will be based on a plant experiment that is set up in Lab 1 (week 2), with data collected in
Lab 2 (week 4). The report will be due in week 6 (see timetable inside front cover of manual).
Report 2 will be based on an animal experiment in Lab 4 (week 8), where you will set up and collect
data in the same class. The report will be due in week 11 (see timetable inside front cover of manual).
For each report there will be a separate submission link, available in the ‘Assessment’ section of
BIO1AP LMS.
Allow sufficient time to upload your report to Turnitin and if you wish to view the originality report
(more information about originality reports is here http://www.latrobe.edu.au/students/academic-
integrity/explanation/turnitin) the initial report typically takes ten to fifteen minutes to be generated
following successful submission. If subsequent submissions are allowed, then the second (or later)
originality report takes up to 24 hours to be generated.
Late Submission of Reports

There are University wide policies and procedures to guarantee fair, consistent, and transparent
treatment of late submission of assessment tasks which provide equity around extensions of
submission dates and penalties associated with not submitting assessment tasks by the due date
and time.
Please refer to the information/links given in the Assessment section of BIO1AP LMS.
Report FAQ’s – see BIO1AP LMS

For each report there will be a specific FAQ page in the Assessment section of LMS.
9
WORKSHOP 1 Lab Safety online / teaching lab drop-in
Lab Safety – online induction on LMS

Go to BIO1AP LMS Week 1 section to view the induction pdfs and
complete the associated quiz before your timetabled lab session next
week. Make sure to allow time to complete before the quiz deadline.
Use page 4 of this manual as a reminder of the Personal Protective

Equipment you need for labs.
Microscopy – online module on LMS
If you want a refresher or have not used a compound microscope before,

work through this module before week 2 as during your first lab in week
2 you will need to use a compound microscope to view prepared slides.
Teaching lab drop-in
Bundoora: LIMS 1 teaching laboratories (Rooms 203/204)
There are no formal f2f workshop classes this week, but Pam Hurst (BIO1AP Subject
Coordinator) will be present in the labs to answer any questions. Students starting at La
Trobe in semester two or students who did not do first semester biology are strongly
encouraged to drop in to become familiar with the labs before the first lab in week 2.
For this week, you do not need to stick to your Allocate+ listed session - attend at any of
the times below:
Tuesday 1 August: 10 am
Tuesday 1 August: 2 pm
Wednesday 2 August: 10 am
Albury-Wodonga: Your instance coordinator will be in touch with you via email prior to
Week 1.
10
LAB 1 PLANT ROOTS and PLANT GROWTH REGULATION
Online safety quiz

Campbell Biology 12e (Aus. & Microscopy
New Zealand version)
module on LMS
Concepts 35.1 - 35.3, 800-812
Concept 39.2, 894-900
(both in Week 1)
AIMS
- To be able to use a compound (light) microscope correctly to view prepared specimens.
- To observe and record the structural features/tissues of monocot and dicot roots.
- To set up and understand the design of the plant growth regulation experiment examining the
effect of GA..
Part 1 - PLANT ROOTS

The Structure and Function of Roots
Introduction
A single ryegrass plant occupying less than 0.6 m3 of soil may have as many as 15 million individual
roots and branch roots with a combined length of 644 km and a total surface area larger than a
volleyball court. Significant additional surface area is provided by root hairs, which for the ryegrass
plant may number more than 14 billion with a total surface area almost the size of a football field
(Stern, 1997, Introductory Plant Biology).
Given these calculations, it is not difficult to identify the primary function of roots. Such an
extensively branched network and substantial surface area facilitates the absorption of water and
dissolved mineral nutrients from soil. It also anchors the plant firmly in the soil. The vascular tissues
of the root and the stem conduct water and dissolved mineral nutrients from the roots to the aerial
parts of the plant and sugar from the leaves to the roots where it is used for root growth,
development and maintenance. The structure of roots relates directly to their function. The tissues
of developing roots are predominantly primary, that is, they derive from a primary meristem within
the root apex which allows the roots to grow through the soil. Only in perennial woody plants such
as woody dicots and gymnosperms does secondary thickening occur in roots.
11
1A The root tip

Background
As discussed in Plant Life lecture 1, the apex in developing young roots can be divided into four
major regions or zones. The first, the root cap, is clearly defined and envelops the tip of the
developing root. The other three are not always as clearly defined as they represent a
developmental sequence. These are the region of cell division (meristematic zone), the region of
elongation (elongation zone) and the region of maturation (specialisation zone). The region of cell
division comprises the apical meristem of the root and is the region from which cells and tissues of
the root are derived, giving rise to growth of roots through the soil.
In the region of cell division, some cells will be observed in various stages of cell division (i.e.,
mitosis; prophase, prometaphase, metaphase, anaphase, and telophase (see Figure 2) and the
condensed chromosomes can be observed as stained a dark red-purple colour.
The region of maturation is often not included in the specimens provided but is sometimes evident
where root hairs are beginning to expand.
Figure 2: The condensed chromosomes of cells in mitosis (stages 1-5) may be visible in the root tip
section and are useful indicators of the region of cell division. In a dividing cell the short mitotic phase
alternates with the longer interphase (Figure from Campbell Biology 11th edition (Australian and New
Zealand version), 0245. 12.11 Elisabeth Pierson, FNWI-Radboud University Nijmegen, Pearson
Education).
12
Examine the longitudinal section of a root (prepared slide #1 or #5). Start at 4x objective lens
and then go to the 10x.
In the space below, draw a low power (10x objective lens) diagram of the root
tip and label the root cap, apical meristem and zone of cell elongation. Can you
see where cell division is taking place? Depending on the slide you have, cell
division may not be visible – check with your demonstrator. Cell division does
not need to be included in your diagram.
13
1B Root anatomy
The vascular tissues of primary roots are aggregated into a cylinder in the centre of the root. This
cylinder of vascular tissue, which is referred to as the stele, may or may not contain a central region
of pith. Most monocot roots have a central pith, whilst most dicot roots do not. The outermost layer
of the stele comprises a relatively undifferentiated tissue called the pericycle that is usually only cell
layer wide (e.g. as in Ranunculus sp.). The pericycle is particularly important in lateral root
formation. Endodermal cells are characterized by the presence on the radial walls of waxy (suberin)
bands called Casparian strips that ensure that the water and mineral nutrients taken up from the
soil pass through the cytoplasm of the endodermis. This allows a degree of control over the uptake
of water and salts into the stele.
Examine the prepared slide labelled # 30 (Ranunculus root c.s). or # 39 (monocot and dicot
root c.s.). Start at 4x objective lens to find and focus on the monocot c.s., then go to 10x.
In the space below, draw a low power (10x objective lens) diagram of the
monocot root and label the epidermis, endodermis, phloem, xylem, cortex
and pericycle.
Compare this cross section to that of the dicot root on the same slide. A
diagram of this cross section is in the drawing instructions at the front of the
manual.
14
1C Root hairs
Mount the tip (approximately 20 mm) of the root of a cucumber (Cucumis sativus) seedling in a
drop of water on a microscope slide and examine it under low power using a dissecting
microscope. You may need to cut the root tip off if the seedling is large or difficult to mount flat on
the slide.
Identify the root hairs. If you cannot see hairs on the root tip, check with your
demonstrator. In the space below, draw the root, showing the root hairs and
the root apex.
15
Questions
1. What is the function of the root cap?
2. What is the function of the region of cell division? How does this assist the
functioning of the root?
3. What are the functions of the phloem and xylem tissues in a root?
4. How do the root hairs contribute to the functioning of the root?
16
Part 2 - PLANT GROWTH REGULATION EXPERIMENTS – set up
Campbell Biology 12e (Aus. & NZ version)

Concept 39.2, 894-900
• Set up the experiment that you will write up for Report 1. You may also be involved in setting
up demonstration experiments. All experiments involve investigating the effects of interfering
with the normal levels of growth regulators in plants.
• Make notes on the methods for Experiment 1 - this will be written up and submitted for
assessment (Report 1 due week 6 – see Timetable at the front of this manual)
• Report 1 will be based on Experiment 1 and in Lab 2 the class results will be collected
Summarising the data will be discussed the following week in Workshop 3 (week 5).
Introduction
Plant development involves both growth and differentiation. The term growth is applied to
quantitative changes occurring during development and it may be defined as an irreversible
increase in the size of a cell, organ, or whole organism. However, during development there appear
not only quantitative differences in the numbers and arrangement of cells within different organs,
but also qualitative differences between cells, tissues and organs, to which the term differentiation
is applied. Differentiation at the cell and tissue level is well known and is the primary object of study
in plant anatomy. However, we may also speak of differentiation of the plant body into shoot and
root. Similarly, the change from the vegetative to the reproductive phase may also be regarded as
another example of differentiation.
Thus, we may say that growth and differentiation are the two major developmental processes.
Usually, growth and differentiation take place concurrently during development.
One of the most fundamental of all biological principles is that every organism is the product of the
interaction of its genetic material and its environment. The form and function of plants are
dependent upon their genetic makeup and the ability of the genes to be expressed through
regulation of protein synthesis. The environment influences growth, differentiation, and metabolism
of a plant by controlling, through the effects on protein synthesis and enzyme activity, expression
of its genetic makeup.
The internal environment of a cell within the plant is equally important. This environment may
include the proximity and type of neighbouring cells, and products, especially plant hormones,
secreted by other cells and tissues. For various reasons the term ‘growth regulators’ is preferred to
‘plant hormones’. Growth regulators, which occur in five classes (viz. auxins, cytokinins,
gibberellins, abscisic acid and ethylene), are known to control cellular and tissue development in
plants. Their action is usually tissue specific and dependent on their relative concentrations. It has
been shown that growth regulators have specific effects on gene expression.
17
Experiment 1: The Role of Gibberellic Acid in Plant Growth (Report 1)
Aim: To study the effect of gibberellic acid application on plant growth. To do this you will set up
control and treatment pea plants. Control plants will be sprayed with water and there will be two
other groups of plants, each with a different concentration of gibberellic acid (GA) applied. In Lab
2 you will have your plants returned and then measure their height. Data from your whole lab class
will be made available to you and the average heights of the three groups of plants will be
compared. This is the data you will use for Report 1.
Plants: You and your partner will set up a plant each. Your demonstrator will tell you whether your
plants will be control plants or are to be sprayed with GA and at which concentration. Record your
treatment in the table below. Collect a plant containing a seedling (young plant) of Pisum sativum
variety ‘William Massey’, which is a dwarf pea variety. The dwarf pea plants are deficient in
gibberellic acid (GA) due to mutation of a gene in the pathway for biosynthesis of GA.
Before setting up your plants, look carefully at them and record the number of
internodes present between the scale leaf and the node below the apical shoot. Do
not be confused between a compound leaf with tendrils and the apical shoot, which
may be enclosed by stipules.
Seat# Treatment Number of internodes
Procedure:
1. Label each plant and pot with the appropriate information.
Plants: Use a pencil for writing labels (pencil does not run/wash off). Using
jeweler’s tags write either C, TA or TB on one side and on the other side, your lab
day & time and seat no. Attach the label carefully to the plant.
Pots: Add a label as directed by your demonstrator.
SAFETY NOTE: Re #2 below, disposable gloves and protective glasses are required
when spraying GA. The GA solution is in the fume cupboards at one end of the
laboratory. Dispose of gloves after you have finished handling the plant pots and then
wash your hands.
2. Treatment application
a) Treatment plants will be sprayed with a 15 mg / litre (mg L-1) solution of gibberellic acid
(TA) or a 30 mg / litre (mg L-1) solution of gibberellic acid (TB). The plants must be sprayed
in a fume cupboard with the provided hand atomizer (preset to a fine spray), taking extreme
care to avoid drifting of the spray. The leaves and shoot apex should be sprayed only until
fine droplets begin to form (avoid over-spraying). Rotate the plant during spraying to ensure
uniform coverage of the plant. Each fume hood will have one of the two GA concentrations.
Do not move sprays between fume hoods.
b) Control plants (C) will receive a fine spray of water. Take your plants to the designated area
(bench below dividing window between labs) to spray with water. The leaves and shoot
apex should be sprayed only until fine droplets begin to form (avoid over-spraying). Rotate
the plant during spraying to ensure uniform coverage of the plant.
3. Put your pot in the location as directed by your demonstrator, check plants are clearly labelled.
Plants will be returned for Lab 2 (in Week 4).
18
Experiment 2 (demonstration): Apical Dominance in Phaseolus vulgaris
Your demonstrator will select students to set up control and treatment plants that everyone will
look at during Lab 2 in week 4.
Aim: To study the effect of the apical bud on the growth of axillary buds.
Plants: Three pots, each containing a bean seedling (Phaseolus vulgaris).
Procedure:
1. Label, plants.
Using jeweller’s tags, label each of the three plants with the treatment it will receive
(see below), your bench number and lab day & time. Use a pencil for writing labels
(pencil does not run/wash off). Label pots as directed by your demonstrator.
Label Treatment
C Control (no decapitation)
D+L Decapitated + Lanolin
D + IAA Decapitated + IAA (indole acetic acid) in Lanolin
2. Control (C) plant: set aside as it will remain an intact plant.

3. Decapitated + Lanolin (D + L) plant: use a clean sharp razor blade or fine scissors to cut off
the main shoot about 1 cm above the point of attachment of the first pair of leaves (see
Figure 3 on the next page). Next using a clean swab stick cover the cut with plain lanolin
paste (no IAA added). Discard the used swab stick into the container provided.
SAFETY NOTE: Re #4 below, take care using the lanolin paste containing IAA – wear
gloves, avoid skin/eye contact and wash immediately if this does occur. When
procedure completed, discard gloves and wash hands thoroughly.
4. Decapitated + IAA (D + IAA) plant: use a clean sharp razor blade to cut off the main shoot
about 1 cm above the point of attachment of the first pair of leaves (see Figure 3 below).
Next using a clean swab stick cover the cut with lanolin paste containing 2% (114 mM) IAA
(indole acetic acid – an auxin). Discard the used swab stick into the container provided.
5. Place the three pots in the large white tray provided, insert a plastic label in one of the pots
with your group seat #, check plants are clearly labelled and take the tray to the glasshouse
when directed to by your demonstrator.
19
Figure 3. Diagram of an immature (seedling) bean plant indicating where to decapitate and
location of axillary buds.
20
Experiment 3 (demonstration): Auxin and Leaf Development
Your demonstrator will select students to set up control and treatment plants that everyone will
look at during Lab 2 in week 4.
Auxin affects the patterning of vascular tissues (i.e. xylem and phloem) throughout the plant. The
correct synthesis, transport and response to auxin are fundamentally important if the plant is to
develop normally. In this experiment, you will examine the effects of exogenous (introduced) auxin
on developing and mature tissues of Phaseolus vulgaris.
Aim: To study the effect of auxin on the development of leaf tissue.
Plants: You will be supplied with two or more Phaseolus vulgaris plants (bean plants). If more
than two plants, identify the two plants that are closest in developmental stage. Label pots as
directed by your demonstrator.
Procedure:
1. Using a pencil and jeweller’s tags label one plant as “Expt 4-C”, and one plant as
“Expt 4-T”. Also, write your bench number and lab day/time.
2. On each of the two plants, using a waterproof marker, carefully draw a series of lines as
indicated by the dotted lines in Figure. 4. These lines will guide you in applying either lanolin
(control) or the lanolin + auxin (treatment), and assist with the observation of the effects of
auxin in the next lab class in two weeks,
A. Along the margin of a

mature, fully expanded
simple leaf.
B. Along the adaxial (upper
surface) midvein (the large
central vein) of a second
mature, fully expanded
simple leaf.
C. Along the margins of a
single leaflet of an
expanding tri-foliate
compound leaf.
D. Along the adaxial midvein
of a single leaflet of an
expanding tri-foliate
compound leaf.
E. Directly on the youngest,
developing leaves nearest
the shoot apex. (depending
on plant development, you
may not have a leaf E)
Figure 4. Diagram of a Phaseolus vulgaris seedling indicating where lines are to be drawn (see
instructions above)
21
3. Control (C) plant: With the applicator provided, smear a small amount of lanolin paste (no
auxin) onto the plant, tracing over the lines you made at Step 2.
SAFETY NOTE: Re the procedure below at #4, take care using the lanolin paste
containing IAA – wear gloves, avoid skin/eye contact and wash immediately if this does
occur. When procedure completed discard gloves and wash hands thoroughly.
4. Treatment (T) plant: With the applicator provided, smear a small amount of lanolin paste
containing 2% (114 mM) IAA (indole acetic acid – an auxin) onto the plant, tracing over the
lines you made at Step 2.
5. Stake plants use bamboo stakes provided to keep the plants upright and the treatment plant
from cross contaminating the control plant during the next two weeks in the greenhouse.
A Subject Support Tutor will stay in the lab after the Tue am and Wed am
classes and be available to assist anyone with questions about the class or LMS or just about
anything! For the Tue pm class, see the Student Resources block in LMS for SST support after
class.
Last check before you leave class:
Has your demonstrator seen your labelled drawings for the biological observation assessment?
Clean and tidy if you haven’t already done so. Lab coat off and wash hands before leaving.
22
WORKSHOP 2 Scientific report writing for biology
Before coming to your workshop, you need to complete the

‘Introduction to scientific report writing’ lesson in the Week 3 section of
BIO1AP LMS. One of the activities is to mark an example report using
the BIO1AP Report Assessment Criteria and Standards (two copies
provided on the following pages). You will need to bring the marks you
awarded to the workshop.
AIMS
- Discuss the example report from ‘Introduction to scientific report writing’ lesson in the Week
3 section of BIO1AP LMS.
> To understand the structure of scientific reports and requirements for BIO1AP reports.
> To understand of the marking criteria and standards for BIO1AP reports.
In this workshop, you will be introduced to the aims of scientific report writing for biology,
in particular for BIO1AP. Before you write your first report for BIO1AP, it is important to
know the general structure, format, and type of content of a formal scientific report as
well as the criteria that will be used for marking your submission for assessment.
Workshop activity
Understanding how you will be assessed will assist you to submit an appropriate
scientific report for assessment. The best way to gain this knowledge is to mark sample
scientific reports using the same assessment criteria and description of standards that
your demonstrator will use to mark your reports.
In small groups (max. 4 students), you will discuss the good and
bad qualities of the marked example report and come with a
consensus mark for each section. You will compare marks that
each student group awarded and discuss the interpretation of the
criteria and standards with your demonstrator.
There will be time to ask questions about report writing, either with your demonstrator
or at the end of the workshop with the Subject Support Tutor.
23
Space for any additional notes about report writing:
24
BIO1AP Report Assessment Criteria and Standards

Marks More than Satisfactory Satisfactory Unsatisfactory
(>70%) (50-70%) (<50%)
Title is concise, relevant, and informative and Title is relevant, informative and includes correct Title is uninformative and/or lacks
TITLE
includes correct information. Is different from lab information. Is different from lab manual. correct information or is copied from
/5 manual. lab manual.
Clear and well written flow beginning with There is an attempt to provide a context for the Introduction is limited in scope and/or
relevant general concepts. An integration of investigation through presentation of concepts unclear and/or unstructured and/or
INTRODUCTION
previous research that explains the context for the and previous research, however, this may not be irrelevant. Fewer than three sources
(250 words)
/ 20 investigation. Clear link from background to clearly presented. There may be some irrelevant cited, or sources cited are not scholarly.
general aim and then to specific hypothesis/es. information included. Aims and hypothesis/es
Hypothesis/es are correctly structured and clearly are present but may be unclearly worded. At least
articulated. At least three scholarly sources cited. three sources cited, and most are scholarly.
A concise but clear, well written and logical Procedures are described in a logical order using Procedures incorrectly described and/or
presentation of the procedures used (including the text. Minor omissions and/or problems with not as text and/or contain major
METHODS
(150 words)
/ 15 species used) and is in an accepted scientific style. clarity/scientific style. May or may not state the omissions.
The statistical test used to analyse the data is statistical test used to analyse the data.
stated.
Comprehensively, clearly and accurately describes Accurately describes the data obtained, however, Written description lacking or
Written the data and, if required, analysis. All the limited in description of trends and notable incorrect/unclear/missing statements
(100 words) following included features of the data. Other requirements below regarding statistical tests (if required).
are present.
/ 15 • Description of the trends and notable features of the data
• Reference to tables and/or figures appropriately placed in the text.
• Correct statements regarding data and, if required, correct statistical test results/statements.
RESULTS
Clear and appropriate presentation of correct Presentation of correct summary data but Presentation of summary data unclear
Data summary data in sequentially numbered tables omissions and/or placement of information below or absent or only raw data presented.
presentation and/or figures. Includes: reduces clarity. Data presentation lacking identified
requirements below.
/ 15 • Tables and/or figures have captions positioned in the correct location and include appropriate information to understand the data presented.
• All graphs have labeled axes and axis labels include correct units.
• All tables have labeled columns/rows and include correct units.
• Appendix/ices contain appropriate data and correct calculations/statistical analysis.
25
(>70%) (50-70%) (<50%)
Clear, well written and correct biological Begins with reference to key findings but support A lack of understanding of the
interpretation of the results that begins with or not for hypothesis/es not clearly expressed. results, hypothesis/es and biological
reference to key findings (without repeating results) Correct biological interpretation may not be clear. interpretation is evident. Fewer than
DISCUSSION
and whether hypothesis/es supported. At least three At least three sources cited, and most are three scholarly sources or poor-
(300 words)
scholarly sources cited. Clearly and concisely scholarly. Logical flow of the points below may quality sources cited. Limited/no
/ 30 includes the following in a logical flow: be lacking, or a point not addressed. discussion of points below.
• Key findings discussed in the context of previous research.
• Methodological issues which may have affected results are discussed.
• Suggestions for future study on this topic are provided.
TOTAL
/ 100 Before any deductions (see below)
Deduction per
Not following instructions, e.g., formatting errors (examples given but not limited to these):
error (marks)
Reference formatting
In-text - not as author(s) surname, year 2
(use Harvard style – see
Academic Referencing Tool
DEDUCTIONS
Reference list - not alphabetical by surname, inconsistencies in formatting 2

via Library website)
̶ /10 General formatting
Incorrect line spacing of section text 2
(see Week 3 LMS lesson on
Scientific Report writing)
Section heading errors (including omission) 2
Font size/style inconsistencies 2
Final Total / 100 (less any late deductions - for info on late deductions, see Assessment section of BIO1AP LMS)
26
BIO1AP Report Assessment Criteria and Standards

(>70%) (50-70%) (<50%)
Title is concise, relevant, and informative and Title is relevant, informative and includes correct Title is uninformative and/or lacks
TITLE
includes correct information. Is different from lab information. Is different from lab manual. correct information or is copied from
/5 manual. lab manual.
Clear and well written flow beginning with There is an attempt to provide a context for the Introduction is limited in scope and/or
relevant general concepts. An integration of investigation through presentation of concepts unclear and/or unstructured and/or
INTRODUCTION
previous research that explains the context for the and previous research, however, this may not be irrelevant. Fewer than three sources
(250 words)
/ 20 investigation. Clear link from background to clearly presented. There may be some irrelevant cited, or sources cited are not scholarly.
general aim and then to specific hypothesis/es. information included. Aims and hypothesis/es
Hypothesis/es are correctly structured and clearly are present but may be unclearly worded. At least
articulated. At least three scholarly sources cited. three sources cited, and most are scholarly.
A concise but clear, well written and logical Procedures are described in a logical order using Procedures incorrectly described and/or
presentation of the procedures used (including the text. Minor omissions and/or problems with not as text and/or contain major
METHODS
(150 words)
/ 15 species used) and is in an accepted scientific style. clarity/scientific style. May or may not state the omissions.
The statistical test used to analyse the data is statistical test used to analyse the data.
stated.
Comprehensively, clearly and accurately describes Accurately describes the data obtained, however, Written description lacking or
Written the data and, if required, analysis. All the limited in description of trends and notable incorrect/unclear/missing statements
(100 words) following included features of the data. Other requirements below regarding statistical tests (if required).
are present.
/ 15 • Description of the trends and notable features of the data
• Reference to tables and/or figures appropriately placed in the text.
• Correct statements regarding data and, if required, correct statistical test results/statements.
RESULTS
Clear and appropriate presentation of correct Presentation of correct summary data but Presentation of summary data unclear
Data summary data in sequentially numbered tables omissions and/or placement of information below or absent or only raw data presented.
presentation and/or figures. Includes: reduces clarity. Data presentation lacking identified
requirements below.
/ 15 • Tables and/or figures have captions positioned in the correct location and include appropriate information to understand the data presented.
• All graphs have labeled axes and axis labels include correct units.
• All tables have labeled columns/rows and include correct units.
• Appendix/ices contain appropriate data and correct calculations/statistical analysis.
27
(>70%) (50-70%) (<50%)
Clear, well written and correct biological Begins with reference to key findings but support A lack of understanding of the
interpretation of the results that begins with or not of hypothesis/es not clearly expressed. results, hypothesis/es and biological
reference to key findings (without repeating results) Correct biological interpretation may not be clear. interpretation is evident. Fewer than
DISCUSSION
and whether hypothesis/es supported. At least four At least three sources cited, and most are three scholarly sources or poor-
(300 words)
scholarly sources cited. Clearly and concisely scholarly. Logical flow of the points below may quality sources cited. Limited/no
/ 30 includes the following in a logical flow: be lacking, or a point not addressed. discussion of points below.
• Key findings discussed in the context of previous research.
• Methodological issues which may have affected results are discussed.
• Suggestions for future study on this topic are provided.
TOTAL
/ 100 Before any deductions (see below)
Deduction per
Not following instructions, e.g., formatting errors (examples given but not limited to these):
error (marks)
Reference formatting
In-text - not as author(s) surname, year 2
(use Harvard style – see
Academic Referencing Tool
DEDUCTIONS
Reference list - not alphabetical by surname, inconsistencies in formatting 2

via Library website)
̶ /10 General formatting
Incorrect line spacing of section text 2
(see Week 3 LMS lesson on
Scientific Report writing)
Section heading errors (including omission) 2
Font size/style inconsistencies 2
Final Total / 100 (less any late deductions - for info on late deductions, see Assessment section of BIO1AP LMS)
28
LAB 2 PLANT GROWTH REGULATION (DATA COLLECTION) /
PLANT SYMBIOTIC RELATIONSHIPS

Concept 39.2, 894-900
Concept 37.3, 861-867
Concept 36.3, 833-834
AIMS
- To correctly measure and record pea plant height for the effect of GA experiment (Report 1).
- To observe the demonstration experiments set up in Lab 1 (apical dominance, auxin and leaf
development)
- To observe and record the differences in plants with/without root nodules.
Part 1 – Plant Growth Regulation data collection
Effect of GA experiment – Report 1

Experiment 1 (Effect of GA) measurements need to be done first so that class data sheets can be
checked before plants are disposed of. Collated data will be available for you to work with in next
week’s workshop.
Remember, you will be individually writing up Experiment 1 as a scientific report and submitting it
for assessment (Report 1). Last week’s workshop (Workshop 2) you became familiar with the
report marking criteria and the section content required in a scientific report for BIO1AP.
Day 14 measurements:
1. Locate your plant, and if necessary, carefully disentangle it from nearby plants. Then take it to
your bench.
2. Using a clear plastic ruler, carefully measure your plant’s height in millimetres, measuring from
the scale leaf to the node below the apical shoot. Record your measurement below
and also record the number of internodes present.
3. Also, using pen or a dark pencil, clearly/neatly write your plant’s height on
the appropriate class data sheet (one for your lab by the lab dividing window).
Treatment Height (mm) Number of internodes
3. Using pen or a dark pencil, clearly write your plant’s height on the appropriate class data sheet
(one for each lab – see prac coordinator)
4. Leave your plant on your bench until given the ok to dispose of it.
Hypothesis
The aim of this experiment was to investigate the role of gibberellic acid in plant
growth. In addition to a general aim, we also need a more specific hypothesis,
a statement that can be tested to see if the data collected supported or did not
support the hypothesis.
29
Questions
These questions will help to direct your learning and assist with writing your report. However, this
is not the only information to include in your report! Go to BIO1AP Reading List (link on LMS) for
relevant scientific articles on this topic.
Which question is relevant for the Introduction? Which for the Methods?
1. What does the plant hormone, GA, do in stems?
2. Why was it necessary to spray one pot with distilled water?
Example data presentation
Using just your bench’s data, calculate the average pea plant height for each treatment and using
the graph paper on the next page, sketch how this summary data could be presented. Include
axis titles and measurement units.
In Workshop 3, calculating and presenting the class data needed for Report 1 will be discussed.
30
This graph paper is provided for you to sketch ways of presenting the summary data on pea plant
height for each treatment just for your bench’s plants.
31
Space for any additional notes about the Effect of GA experiment.
32
Experiment 2: Apical Dominance in Phaseolus vulgaris
For full details of the experiment see the Labl 1 set up notes. Below is a summary of the
treatments.
Label Treatment
C Control (no decapitation)
D+L Decapitated + Lanolin
D + IAA Decapitated + IAA (indole
acetic acid) in Lanolin
Day 14 observations:
Look at the axillary buds of all three plants.

Which bean plant displays the most growth
of axillary buds?
Questions
1. What can you now conclude about the effect of IAA on axillary bud growth?
2. Why is it necessary to have a treatment Decapitated + Lanolin (no IAA)?
3. IAA is an auxin. List at least four functions of auxins in plants
4. What is apical dominance? (one of Tony’s lectures and a topic module cover this
concept)
33
Experiment 3: Auxin and Leaf Development
For full details of the experiment see the Labl 1 set up notes. Below shows where either plain
lanolin or IAA in lanolin was applied to bean plants (Phaseolus vulgaris).
Day 14 observations:
Examine and compare the control and auxin (IAA)
treated leaves for visible changes. Pay particular
attention to:
a. size
b. overall shape
c. orientation (i.e. the angle at which the
leaf is held)
d. surface topology (curvature)
e. petiole (e.g. straight, twisted)
Record your observations in the table

below.
Leaf Control Treatment
34
Questions
Answering these questions will help you understand the differing effects of IAA on leaf growth and
that differences are related to leaf age.
1. What does the plant hormone, auxin, do in leaves?
2. What was the effect of auxin on mature leaves (leaves A & B)?
3. What was the effect of auxin on developing leaves (leaves C, D & E)?
4. What is the effect of auxin on the petiole?
35
Part 2 - PLANT AND MICROBIAL SYMBIOSES
B1. Macroscopic analysis of a plant - bacterial interaction
Introduction
In this experiment, the mutualistic symbiosis of legumes and Rhizobium sp. bacteria and the effect
of inorganic Nitrogen (N) nutrition on this relationship are examined. Legume roots infected with
the bacterium Rhizobium produce outgrowths known as 'nodules' within which the bacteria are able
to chemically reduce gaseous N2 (dinitrogen, an inert form unavailable for biological processes) to
NH4+ (ammonium ions, a form of N plants can use to satisfy their N requirements). This process is
known as Nitrogen fixation (N-fixation). Legumes, like all other plants, can use various forms of
inorganic Nitrogen (e.g. NO3–, NH4+) which are taken up by the roots and dispersed throughout the
plant body via the transpiration stream (in xylem). Therefore, legumes can satisfy their Nitrogen
requirements in two ways, by Nitrogen fixation and by uptake of inorganic Nitrogen by the roots.
Aim
The aim is to demonstrate the role of the Rhizobium–legume symbiosis and examine the effect of
inorganic N nutrition on both nodulation and growth in this system.
Materials and Methods

Seeds of Glycine max (soybean) were moistened with water. Some of the seeds were dusted with
an inoculum containing the bacterium Rhizobium japonicum. All the seeds were planted into pots
containing the inert supporting material Perlite and watered with different nutrient solutions as
shown in the Table below. One set of plants will be available as a demonstration for each group.
Treatments for Rhizobium-inorganic N-legume interactions in Glycine max

Treatment no.: 1 2 3 4
NH4NO3 NH4NO3
CaSO4 CaSO4 CaSO4 CaSO4
MgSO4 MgSO4 MgSO4 MgSO4
Solution for K2SO4 K2SO4 K2SO4 K2SO4
watering
KH2PO4 KH2PO4 KH2PO4 KH2PO4
contains:
K2HPO4 K2HPO4 K2HPO4 K2HPO4
CaCl2 CaCl2 CaCl2 CaCl2
Trace elements Trace elements Trace elements Trace elements
Nitrogen (N) in
solution - + - +
Inoculated
with
Rhizobium - - + +
japonicum
36
Examine the soybean plants, carefully noting any differences in growth and
condition (colour, size, overall health, etc.) and the degree of nodule formation.
Do not confuse the nodules, normally a pale pinkish colour, with pieces of white
Perlite that may be adhering to the roots. Record your data in the table on the
next page. Next answer the questions below.
1. Are there differences in growth between plants in Treatments 1 and 2? Suggest an

explanation for any differences.
2. Are there differences in growth between plants in Treatments 1 and 3? Suggest an

explanation for any differences.
3. Are there differences in growth and nodulation between plants in Treatments 2 and
4? If so, suggest an explanation for the differences.
4. Are there differences in the growth and degree of nodulation between plants in
Treatments 3 and 4? If so, suggest an explanation for the differences.
5. How might the presence of root nodules containing Rhizobium sp. bacteria benefit
the growth of legumes?
37
38
Rhizobium-inorganic N-legume interactions in Glycine max
Results
NH4NO3 in R. japonicum
Treatment Nodules
Growth Medium Inoculum Growth
(present / absent, Other observations
(1 [min] to 10 [max])
location, etc.)
1 - -
2 + -
3 - +
4 + +
Optional activity. Observation of a plant – fungal interaction
Many species of pant form symbiotic associations with soil fungi. The kingdom Fungi includes a
great diversity of organisms that are all heterotrophic eukaryotes and are (along with bacteria) the
major group of organisms that form symbioses with plants. They mainly form colourless filamentous
structures (hyphae), although some types (the yeasts) are unicellular. The fruiting bodies of some
fungi form large macroscopic structures that are commonly called mushrooms.
Examine the Pinus radiata seedling (2-5 plants per lab). Identify roots with and
without a covering of fine white growths - the ectomycorrhizal fungi.
What are the main differences between mycorrhizal roots and uninfected roots, in terms of
branching, thickness and colour?
A Subject Support Tutor will stay in the lab after the Tue am and Wed am
classes and be available to assist anyone with questions about the class or LMS or just about
anything! For the Tue pm class, see the Student Resources block in LMS for SST support after
class.
Last check before you leave class:

Has your demonstrator seen your biological observation for assessment?
Clean and tidy if you haven’t already done so. Lab coat off and wash hands before leaving.
39
Space for any extra notes about Lab 2
40
WORKSHOP 3 Effect of GA (Report 1) data
analysis/presentation
Check the Week 5 LMS area for any addition information related to this workshop.
AIM
To use Excel for class data entry for the Effect of GA on growth experiment and to produce an
appropriate graph suitable for inclusion in Report 1.
Bring to class: a laptop (set up for free wifi access via eduroam), with a desktop version of Excel
installed. See this Student IT support webpage for how to download a free version of Office,
which includes Excel. The desktop version is preferable to the online version as there is more
functionality.
If you are not familiar with Excel, or wish to have a refresher, you can do free online training at
LinkedIn Learning. See this uni webpage for more information:
https://www.latrobe.edu.au/students/opportunities/careers/linkedin-learning There are a large
number of Excel courses, however, note that many use financial data examples. Suggested
beginner courses to look at: Learning Excel Desktop (beginner level), Excel: Introduction to
charts and graphs (beginner – intermediate level).
Data analysis - background
1. Number of replicates in an experiment
Because different pea plants will not grow in exactly the same way, for each treatment group
(control and two different concentrations of GA applied) we use measurements from more than one
individual pea plant and compare the average values for the variable of interest. The different
individuals can be referred to as experimental replicates as each individual in a treatment receives
the same conditions at the same time. Repeating the experiment at a later date is not increasing
the number of replicates (i.e., sample size).
2. Raw data
When there are a number of replicates with a measurement for a numeric variable this raw data
can be graphed as a histogram of the frequency distribution (see the example in the figure
below). A frequency distribution is just an arrangement of observations with the number of
individuals with a particular value (values indicated on the x-axis) shown on the y axis.
Frequency distributions are not usually presented in scientific reports, but there are statistics that
can describe frequency distributions. Getting an average value is one of these descriptive
statistics.
Descriptive statistics provide information about the frequency distribution and are a way of
describing the distribution without having to create or present a frequency histogram. There are
three common descriptive statistics; mean (or average), standard deviation and standard error
of the mean.
Example histogram showing the variation in shell height of the gastropod Bembicium nanum.
𝑋𝑋� indicates the mean (average) shell height.
41
3. Summarising raw data
For scientific reports, the Results section contains summarised data, not raw data. For a numeric
measurement, summarised data usually includes the mean value and a measure of the spread of
the values, usually the standard error of the mean. Summarised data can be presented as a
table or a figure. Figures are usually better as trends in the data are more easily seen.
Calculation of mean, standard deviation and standard error
Mean
The mean (𝑋𝑋�) for a sample is given by:
∑ 𝑥𝑥𝑖𝑖
𝑋𝑋� =
𝑛𝑛
where ∑ = sum of
xi = individual measurements
n = number of measurements
Standard Deviation
The sample standard deviation provides a measure of dispersion of the measurements

about the sample mean. For a normal distribution, a range of one standard deviation each
side of the mean includes about 68% of the total number of measurements. The standard
deviation is used to calculate the standard error of the mean.
The sample standard deviation (s) of the sample measurements is given by:
∑(𝑋𝑋 − 𝑋𝑋�)2
𝑠𝑠 = �
𝑛𝑛 − 1
where ∑ = sum of
X = individual measurement
𝑋𝑋� = sample mean
Standard Error
The standard error of the mean (𝑆𝑆𝑆𝑆𝑋𝑋� ) is a measure of the precision of the mean. The smaller
this value, the more reliable is the estimate of the mean.
The standard error of the mean (𝑆𝑆𝑆𝑆𝑋𝑋� ) is given by:
𝑠𝑠
𝑆𝑆𝑆𝑆𝑋𝑋� =
√𝑛𝑛
where s = standard deviation of the sample
42
In biological experiments, only a sample of the population is studied/used in an experiment. The
values for the mean and standard deviation of an infinitely large population cannot be determined.
For the samples that are used, there will be variation between them, and this sampling variation
becomes larger as the size of the sample decreases. The standard error of the sample mean gives
a measure of how good an estimate the sample mean is of the population mean.
Note, the descriptive statistics explained above assume that the measurement x is continuously
variable, and that the frequency distribution of x is described by a normal distribution.
Data analysis for the Effect of GA on growth experiment (Report 1)
1. Number of replicates (sample)
You will use class data, and this will be 20-25 replicates per treatment group, depending on your
session. You will compare the height of pea plants between control and GA groups. The data you
will use will be from multiple benches. The raw class data was collected in Lab 2 last week.
2. Raw data
You will download the raw data from LMS and enter the values into an Excel file. The mean
height for each treatment can be easily calculated in Excel.
If you are not familiar with Excel, or wish to have a refresher, you can do online training at
LinkedIn Learning (previously Lynda.com). See this uni webpage for more information:
https://www.latrobe.edu.au/students/opportunities/careers/linkedin-learning There are a large
number of Excel courses but not all are relevant. Suggested beginner courses to look at:
Learning Excel Online (beginner level), Excel 2016: Introduction to formulas and functions
(beginner level).
3. Summarising raw data
Once raw data is in Excel, you will be able to present the raw data graphically to indicate the
spread of values and descriptive statistics for each group. The Excel chart type ‘box plots’ will be
used.
4. Describing the results
In the Results section of your report, you will need to write about the results, you do not just
present a figure (graphs are called figures). At the beginning of the Results section, you use text
to describe important features of the data presented. For example, state which treatment had the
tallest plants.
5. Hypothesis – supported?
To test if the data supports or rejects the hypothesis, ideally, a statistical comparison of the mean
growth of the treatments is made. The appropriate statistical analysis is more complex than you
are required to understand at this stage (first year), so you will not be required to perform
statistical analysis for the report. However, in the Results section you are required to present
summarised data and to describe in words the data trend(s) and include in the Discussion section
whether or not the results support the hypothesis proposed.
43
Space for any extra notes about Workshop 3
44
LAB 3 PLANT STEMS and LEAVES

Figures 35.10, 35.11
Concept 35.4, 813-816
Concept 36.4, 837-838
LAB 3 Aims
- To examine the cells that constitute the stems and leaves of plants.
- To understand the distribution and function of these cells.
- To prepare a leaf epidermal peel to view stomata under the microscope.
PART 1: Primary Growth in Stems

Examine the prepared slide of a stem of a Ranunculus (an herbaceous dicot).
PART 2: Secondary Growth in Stems

A. Examine the prepared slide showing secondary growth in the stem of Tilia (a woody dicot).
B. Examine the stem structure of Eucalyptus/Pinus.
C. Prepare and stain a slide of wood pulp and examine its composition.
PART 3: Epidermal peel
Introduction
The stems of plants serve several important functions which are reflected in their anatomy. First,
stems support the leaf canopy which is essential for absorbing light and assimilating CO2 for
photosynthesis. The strength and flexibility of stems, which often support huge canopies against
extreme winds, is reflected in the anatomy of certain cells and the way they are arranged in the
stem. The strength of stems is most evident in the trunks of large trees, in which secondary, lateral
growth of the stems results in massive, woody and yet flexible structures. Secondly, the organic
products of CO2 assimilation formed in the leaves are transported via stems to other tissues,
especially non-photosynthetic tissues such as roots. This process involves the movement of
products such as sucrose, via sieve tube cells associated with phloem in the stem. Thirdly, stems
provide the route for the movement of water and inorganic ions from the roots to the leaf canopy.
This involves another set of specialized cells (tracheids and vessels) which comprise the xylem.
Clearly, the anatomy of stems is important in understanding many aspects of the functioning of
plants.
The anatomy of stems not only reflects their function but also patterns of stem growth. Tissues
comprising the plant body are designated as primary or secondary, depending on their origin, that
is, from what meristem or growth centre (centre of mitotic cell division) they derive. If they originate
from a terminal or apical meristem (e.g. the shoot apex) they are called primary tissues, but if they
originate from a lateral meristem (e.g. vascular cambium) they are called secondary tissues. Growth
resulting from an increase in the production of primary tissues is known as primary growth and
results mainly in increased height of the plant. Growth resulting from the production of secondary
tissues is known as secondary growth, which results in increased stem diameter. Primary growth
is universal in vascular plants; secondary growth occurs generally in woody dicotyledons and
gymnosperms, resulting in the formation of shrubs and trees with larger stems.
In Part 1 and 2 of this lab class you will investigate the anatomy of an herbaceous dicot (Ranunculus
– Buttercup) that has only primary growth and of a woody dicot (Tilia sp.) in which secondary growth
occurs giving rise to a small tree. You will also study the wood of Eucalyptus or Pinus and the
composition of wood pulp used for making paper.
45
Main type of cells and tissues in plants

Plants, like animals, consist of a system of cells arranged into tissues that form a highly organised
body. The component tissues area derived from the major growth centres or meristems where
cell division (mitosis) occurs. The meristem associated with the shoot and root apices are referred
to as primary meristems. The assembly of plant organs consisting of cells derived from these
meristems is termed the primary body of the plant.
Cells derived from meristems develop in various ways by a process called differentiation to
produce cell types that have specialised structures and functions. Differentiated cells are
arranged in characteristic ways in plants to form tissues. In broad terms, a tissue can be defined
as a group of cells that perform a specific function. Some tissues are composed of only one type
of cell (simple tissues) while others are composed of two or more types (complex tissues). During
the lab class you will observe the above cell types and tissues
Cell types seen in plant stems

Parenchyma cells Parenchyma cells occur in tissues throughout the whole plant body. Apart
and tissues from meristematic cells, parenchyma cells are the least differentiated cells
of the plant body. They are large, have thin walls, are approximately
spherical in shape and retain a living protoplast, often with a large
vacuole.
Sclerenchyma cells Sclerenchyma cells have thick walls that are often highly lignified and lack
and tissues a living protoplast. They function to strengthen and support. The most
common type of sclerenchyma cells are known as fibres, Fibres are
elongated and usually pointed at the ends and often occur in association
with tracheids and vessel elements in xylem (e.g. in wood). Sclereids
have very thick, lignified cell walls and are found in the cells of fruits such
as pears.
Tracheids and Tracheids and vessel elements are water-conducting cells comprising the
vessel elements in tissue called xylem which extends throughout the plant body from the
xylem tissue roots to the leaves. Mature tracheids and vessel elements are elongate
cells with thick lignified walls and lack living protoplasts. Tracheids are
generally much longer than vessel elements and have pointed ends. Their
walls are often pitted and vary in thickness. Vessel elements are generally
larger in cross-section and have oblique, pointed, or transverse end walls
with perforations.
Sieve tube Sieve tube elements and companion cells are components of the tissue
elements and called phloem, which is responsible for the translocation of organic
companion cells in solutes, mainly sucrose, throughout the plant. Sieve tube elements are
phloem tissue elongate cells with structures called sieve plates in their end walls or side
walls. The protoplast persists at maturity but lacks a nucleus. Companion
cells are specialized parenchyma-like cells which are closely associated
with sieve tube elements. They are thought to be intimately involved in
supporting the function of the sieve tube elements and there are
numerous cytoplasmic connections between the two. Companion cells
have persistent nucleate protoplasts.
46
Using a biology textbook and the above notes name the different cell types below:
Parenchyma cells, sieve tube elements and companions cells of the phloem
tissue, fibres, vessel elements, sclereids, and tracheids.
3 7&8
1
50 µm
50 µm
2
8
7
4 5a 50 µm
6a 6b
20 µm
Name the cell types

1. 100 µm
2. Collenchyma
5b
3.
Sclerenchyma
4.
5. a+b
6. a+b
7. & 8.
47
Principal tissues of plant stems.
Tissue types Location and characteristics

Cork cambium A secondary lateral meristem which produces a protective tissue (cork) which
or phellogen replaces the epidermis in woody stems and roots.
Cortex The tissue, usually of parenchyma cells, located between the epidermis and the
vascular cylinder of stems and roots; often for food storage; can be photosynthetic
in young stems.
Epidermis The outermost cell layer of the plant body; usually of parenchyma cells; often
covered by a protective waxy cuticle.
Hypodermis A layer or layers of distinctive cells immediately inside the epidermis of a stem;
supportive and protective function.
Medullary ray Radial clumps of parenchyma which link the pith and the cortex in stems, especially
in wood; often give wood its distinctive “spoke-like” appearance when cut and
dressed. Conducts fluids horizontally
Periderm A complex secondary tissue commonly called bark derived from the cork cambium
or phellogen and replacing the epidermis; typically in woody stems and roots;
provides protection.
Phloem Tissue principally responsible for translocation (transport of organic products of
CO2 assimilation) in vascular plants; outermost component of vascular tissue of
most stems and roots; comprises sieve elements, parenchyma and sclerenchyma
cells and companion cells (in angiosperms) and albuminous cells (in
gymnosperms).
Phloem or A group of sclerenchyma fibres associated with a vascular bundle and appearing
bundle cap like a cap on the phloem; provides support.
Phloem ray Radial clump of parenchyma tissue located in secondary phloem.
Pith Central tissue, usually of parenchyma cells, of stems and roots; often a storage
tissue.
Vascular A secondary lateral meristem from which the secondary vascular tissues
cambium (secondary xylem and phloem) develop; comprises meristematic cells which
develop (i) between the primary xylem and phloem of primary vascular bundles to
produce a fascicular component, and (ii) from parenchyma cells of the medullary
rays between primary vascular bundles to produce an interfascicular component.
Xylem Tissue mainly responsible for conduction of water and inorganic salts in vascular
plants; innermost component of vascular tissue of most stems and roots; comprises
vessel elements, tracheids, parenchyma cells and sclerenchyma fibres; also aids as
a structural or supportive tissue, especially secondary xylem.
Xylem ray Radial clump of parenchyma tissue located in secondary xylem – also known as a
medullary ray; give a distinctive appearance or ‘grain’ to dressed wood.
PART 1: Primary Growth in Stems
Stem anatomy of Ranunculus sp.
Buttercup (Ranunculus sp.) is a herbaceous dicot which has little or no secondary growth and
never develops wood (i.e. the stem shows no further lateral expansion once the primary tissues
have matured). However, it has a degree of strengthening of the stem tissues sufficient to support
a small herbaceous plant.
Examine the transverse section of Ranunculus stem. Note that the lignified xylem cells and fibres
are stained red. Cells with mainly cellulose walls tend to stain blue
Prepare a LP diagram and label each tissue listed on the next page
Indicate the type of cells (e.g. parenchyma cells) comprising the tissue and note
the likely function of the tissue (e.g. outer layer of cells with protective coating).
48
Diagram below to include:
• epidermis (the distinctive outermost layer of cells);

• cortex (the ground tissue between the epidermis and vascular bundles and between the
vascular bundles);
• pith (the central tissue);
• primary xylem;
• primary phloem;
• and bundle sheath
49
Question: What tissues provide a degree of strengthening of the buttercup stem? What cell
types comprise these tissues and what chemical constituents contribute to the strength of
these cells?
PART 2. Secondary Growth in Stems
Stem anatomy of the woody dicot, Tilia sp.
Tilia sp. (common name ‘linden’ or ‘basswood’) is a woody dicot tree which exhibits extensive
secondary growth giving rise to a woody plant. Examine the prepared slide showing transverse
sections of stems with one, two-, or three-years’ lateral growth of tissues from the vascular
cambium. Locate the position of the vascular cambium in each section and identify the secondary
tissues.
Tilia sp. also forms a second lateral meristem called the cork cambium from which a protective bark
develops to replace the epidermis. This may be evident in the three-year-old stem. The cork
cambium also serves to compensate for expansion of the stem caused by extensive secondary
growth. The multilayered surface tissue (including the cork cambium) is called a periderm.
Examine the cross section of Tilia sp. stem

Prepare a LP diagram that includes all tissues (does not need to include whole
specimen). If multiple years growth present, label each year, with Year 1 being the
oldest. Label these tissues: cortex, pith, primary xylem, secondary xylem, xylem
ray, secondary phloem, phloem ray, and vascular cambium, epidermis
50
Questions
1. What tissues strengthen the Tilia stem?
2. What cell types comprise these tissues?
3. What is the main difference between the mature stem of Tilia and that of the
buttercup, and how does this relate to the plant growth forms of the two species?
4. What is an “annual ring”?
5. Compared to some species it is relatively easy to discern one annual ring from
another in the Tilia stem. Can you explain why this is so?
Stem structure of Eucalyptus.
Observe with the unaided eye the cross section of the Eucalyptus sp./Pinus sp stem provided or
the large cross section of a woody stem.
Identify the wood (secondary xylem), bark (secondary phloem and periderm) and vascular
cambium (secondary meristem). Note the thickness of the bark.
Question: To what environmental stress, characteristic of southern Australia, is a thick

bark in the eucalypts likely to be an adaptation?
51
The composition of wood and wood pulp
Wood has long been one of the most important materials in human societies, especially for use
as a fuel for heating and cooking, building material for houses and boats, and, more recently, pulp
for making paper. Most of the world’s paper is made by matting and pressing together the fibrous
cells derived by maceration of wood from trees (e.g., Eucalyptus pulp is one of the world’s major
sources; the ‘bright’ paper used in this manual is made from Eucalyptus pulp).
1. Place a drop of macerated wood on a clean glass slide.

2. Drain excess fluid from the preparation with the edge of a tissue/paper towel
3. Add a small drop of Toluidine Blue which stains cell walls.
4. Place a coverslip over the preparation and use a tissue to remove excess liquid.
5. Examine the preparation under a compound microscope, starting at low magnification
6. Identify tracheids, vessel elements and sclerenchyma fibres (refer to the Figure you labelled
earlier or a textbook).
Draw at least one of the above three (tracheid, vessel element or sclerenchyma
fibre) that you can see in your preparation.
Question: What characteristic of the constituents (tracheids, vessel elements,

sclerenchyma fibres) of macerated wood (known as ‘pulp’) makes them suitable for
making paper?
52
PART 3. Leaf epidermal peel
In some of Tony’s lectures he discussed leaf structure, and this technique allows you to view the
leaf stomata. If you continue to second year botany (BOT2ILP) you will be setting up an
experiment with leaf epidermal peels to examine stomata size changes.
For this activity you will work in pairs, one student preparing a leaf epidermal peel from a monocot
and the other student one from a dicot plant. You can then swap preparations to compare
stomata and their arrangement.
Procedure:
1. Have a microscope slide and coverslip ready.

2. Add a drop of water to the slide.
3. Collect either a leaf from a corn plant (monocot) or from a pea plant (dicot).
4. Determine the abaxial (lower) surface of your leaf.
5. Bend the leaf to break the abaxial surface or tear the leaf from the edge.
6. Tear off some epidermis, the transparent thin layer of cells.
7. Holding the part of the leaf with the detached epidermis in the drop of water on the
microscope slide, cut of the epidermis.
8. Place a coverslip on the sample.
9. View under the compound microscope, starting with low power.
If it is difficult to see the cell walls, it may help to close the iris diaphragm a little to increase the
contrast. This should make the cell walls clearer.
Examine the preparation and identify guard cells. These form the stomata and are bean shaped
and joined at the ends. You should see green chloroplasts within the guard cells.
If you are having difficulty in seeing the guard cells, your preparation may be too thick. Do a
second epidermal peel to see if you can get a thinner preparation.
Compare the preparations at the same magnification and make notes/sketches

below in relation the arrangement and number seen.
Monocot (corn) Dicot (pea)
53
Space for extra notes
54
WORKSHOP 4 ANIMAL LOCOMOTION

Chapter 33 Annelids, 742-743
Concept 40.1, 923
Concept 50.6, 1184-1188
AIMS
• To identify muscles involved in earthworm movement from a prepared slide.
• Compare the costs of swimming, running, flying and hopping.
Earthworm
How earthworms use two different types of muscle to move their bodies was explained during
lectures this week. Below is a 3-D schematic diagram of the internal anatomy of an earthworm
Not all these features may be visible in a prepared slide.
Part of Figure 33.25 in Campbell Biology. Ventral surface of the worm is at the bottom
55
Look at a prepared slide of an earthworm cross section and identify the
longitudinal muscle, the circular muscle and chaetae (setae). Indicate these
on a low power magnification plan drawing (see pages 6-7 for what should be
included/not included in a low power drawing). Also label any other
structures/tissues you can identify. On your drawing indicate the dorsal and
ventral sides of the worm.
Movement of a simple animal - demonstration (optional)
On the bench below the window dividing the two labs will be microscopes set up to view a
live simple animal (flatworm or cnidarian). Note what animal is available to view.
Can you see the animal moving?
Is it moving location?
If it is not moving location, can you think of how it might?
56
Animal locomotion – comparing running, flying and swimming

Running, flying and swimming require more energy than sitting still, but how do they compare?
The greatest differences between moving on land, in the air, and in water result from the
differences in support and resistance to movement provided by water and air. The weight of
swimming animals is fully supported by the surrounding water, and no effort goes into supporting
the body, while running and flying animals must support their full weight. On the other hand, water
presents considerable resistance to movement, air much less, so that flying and running require
less energy to push through the medium.
How do we compare the ‘cost’ of particular modes of locomotion between animals?
A simple way to compare the costs of moving for different animals is to determine how much
energy it takes to move. Energy cost can be measured as the amount of energy required to move
one unit of body mass over one unit of distance by that mode of locomotion. The graph below
depicts differences in energy cost, as measured by the number of calories (cal) required to move
one kilogram (kg) over a one metre (m) distance for three modes of locomotion. The graph has a
logarithmic (log) scale for both x and y axes and an explanation of the difference between linear
and log scales is in the box on the next page.
Figure above is from Campbell Biology 11th edition (Aus & NZ version), p1164. Source: Adaptation of
figure 4 from “Locomotion: Energy Cost of Swimming, Flying, and Running” by Knut Schmidt-
Nielsen, from Science, July 1972, Volume 177(4045). Copyright © 1972 by AAAS. Reprinted
with permission.
57
Difference between linear and log scales

A log (or logarithmic) scale is used when there is a large range of values that would be difficult to
depict on on a graph using a linear scale. Divisions on a logarithmic scale are different from a
linear scale. Major divisions increase by a factor of 10. See below for an example of a
logarithmic scale grid. On the y-axis, the two major intervals, after 1, are 10 and then 100 (10 x
10). 10 x 10 = 100 can also be written as 101 x 101 = 102.
In the graph of the three modes of locomotion, the far-right x-axis label is 106, which equals
1,000,000 as shown in the grid on the previous page. To highlight the difference between log and
linear scales comparisons between the two are shown below. From 1 to 10 is the same distance
on both scales, but this changes dramatically after 10. The short brown line by the logarithmic
scale represents 10 to 20 on that scale.
58
From the energy cost graph, you can see that there are differences in energy cost for different
forms of locomotion and these differences are due to the different physical properties of the
medium through which travel occurs. There are two aspects to consider when answering the
questions below:
1. support provided by the medium
2. resistance to movement through the medium
Questions to discuss:
For any given mode of locomotion, what is the impact of body mass on the cost of moving?
Which mode of locomotion is least costly and why might this be so?
How do the other two modes of locomotion differ from the least costly mode?
Which is the costliest mode of locomotion? How does it differ from the next most costly?
Hopping
There is another mode of locomotion that is common in Australia - hopping. Large macropods
increase their speed by increasing the length of their stride, not the frequency of hops, and the
stride length is dependent on leg length. However, the smaller species attain their maximum
speed by having a higher hopping frequency than the larger species. The economy of locomotion
achieved by kangaroos is enabled by the elasticity of the tendons and the leg geometry, with the
maximum efficiency being achieved at speeds above 10 km/h.
How does hopping compare to running? (See the graph shown during the workshop)
59
60
LAB 4 THE EFFECT OF TEMPERAURE ON HEART RATE
(REPORT 2) / REPORT 1 FEEDBACK

Concepts 40.4, 938 - 940
Concept 42.1, p971-972
AIMS
• To conduct an experiment to observe the heartbeat of daphnia (water flea) at different
temperatures (written up for Report 2) and demonstrate your understanding of the
predicted relationship between temperature and heart rate.
• Demonstrate your understanding of cardiac muscle cells.
• Understand the feedback given on Report 1 / ask specific questions about your report.
Background: Effect of temperature change on physiology
Temperature change has striking effects on many physiological processes. For example, oxygen
consumption and heart rate of many animals (particularly aquatic invertebrates) often increase in a
regular fashion with increasing temperature.
In general, a rise of 10cC in temperature causes many biological rates to increase about two to
three-fold. The increase in a rate caused by a 10oC increase in temperature is called the Q10.
Therefore, if the rate doubles the Q10 is 2 and if the rate triples the Q10 is 3. To solve for Q10 when
the rates at two different temperatures have been observed the following equation can be used:
10
 R2 
Q =  T 2 − T 1
10  R1  Equation 1
Where: T2 = higher temperature R2 = rate at T2

T1 = lower temperature R1 = rate at T1
The relative body size of most animals means that they require some sort of circulatory system to
transport respiratory gases, nutrients and waste products sufficiently rapidly between exchange
surfaces (gills, lungs, gut lining) and cells of their body where these materials are produced or used
up. Many invertebrates possess an open circulatory system in which the circulating fluid is not
anatomically separated off from the spaces between cells (= interstitial spaces). This type of
circulatory system is in contrast to the situation in vertebrates and a few invertebrates (e.g. worm),
which possess a closed circulatory system. In a closed circulatory system, the circulating fluid is
kept separate from the spaces between the cells by a system of continuous vessels. In both types
of circulatory system some type of pump (usually called a heart) is needed to move the circulatory
fluid around the body. (see Campbell Biology Concept 42.1 for more information about open and
closed circulatory systems)
In this lab class we will examine the functioning of the open circulatory system of Daphnia a small,
freshwater crustacean. Daphnia is common in billabongs and other standing inland water bodies
and so has to cope with variations in water salinity and temperature. You will measure the heart
rate of Daphnia at two different temperatures that have a difference of 10 oC.
With class data the average Q10 of the animals observed can be calculated. In addition, class data
will be used to examine if there was a significant difference in heart rate at the two temperatures
by using a paired t-test.
61
Background: Daphnia (common name: water flea)

Classification:
Kingdom: Animalia
Phylum: Arthropoda
Class: Crustacea
Subclass Anchiopoda
Family: Daphniidae
Genus: Daphnia
Daphnia sp. are small aquatic crustaceans, related to crayfish and shrimp. Since they are
transparent animals, they are very useful in the laboratory as you can see the internal organs
functioning under a microscope. They are abundant in most standing freshwater. The animals you
will work with were collected from the La Trobe University moat freshwater system.
Left:Lateral view
diagram of Daphnia
sp. from Lankester
1909 A Treatise of
Zoology
62
Lateral view of Daphnia pulex. By K.Tapdıqova - Own work, CC BY-SA 4.0,

https://commons.wikimedia.org/w/index.php?curid=68869411
Internal anatomy can be observed through the carapace (see diagrams on previous page).
• The single black compound eye is the most conspicuous internal organ and is easily
recognised by its constant motion. Careful observation may reveal the nearly transparent
muscles that attach to the eye.
• The ocellus (or nauplius eye) appears as a small black speck, it can be difficult to locate.
• The gut is easily seen, particularly if the animal has been feeding. The gut runs from the
mouth, loops through the head and continues along the body through the thorax and
abdomen to end at the anus.
• The gut is divided into three distinct regions:
- foregut and hindgut, which are lined with cuticle and
- midgut that is lined with epithelium, adorned with microvilli (site of absorption).
• In the head region and attached to the anterior part of the midgut are two small sacs, the
hepatic cecae (or hepatic diverticulum).
• Lying along the gut in the thoracic region of adult females you should see one of the pair
of ovaries, which may contain maturing eggs that are slightly green in colour. The number
of eggs depends on the nutritional state and size of the female. The eggs are extruded into
the brood chamber formed by the new carapace following a moult. The eggs develop during
the between-moult period into embryos and then neonates that are released when the
female moults. Sometimes if the females are very well-fed fat globules are present closely
associated with the gut in the thoracic region.
• The heart is easily seen in living animals because of its rapid beating and is located dorsal
the gut and just behind the head.
63
Heart Anatomy and function in Daphnia
The central organ of the circulatory system of Daphnia is the heart. As seen in Figure P5.1 and in
the large model at the front of the lab, the heart is situated under the clear shield-like plates of the
carapace towards the dorsal (‘back’) surface of the animal.
When the heart contracts, the circulatory fluid (= haemolymph) is pumped forward in a series of
arteries that extend forward (= anteriorly) from the heart towards the head. The haemolymph then
travels through a series of sinuses (= cavities) scattered throughout the body before making its way
into a sinus (=pericardial sinus) which lies around the heart. From the pericardial sinus it enters the
heart through the ostrium (=’opening’). Entry to the heart occurs when the heart is relaxed (=
diastole). Valves that only open inwards prevent backflow of haemolymph through the ostium.
The heart of Daphnia is easily visible through the transparent carapace, making it an ideal organ
with which to observe the effect of environmental variables. To examine a live animal and prevent
it darting out of the microscope’s field of view the specimen can be anchored to a small dab of
grease.
Specimen preparation
Grease mounting a live specimen (each student):
1. Using the handle end of a paint brush, place a small dab (pin-head size) of grease on the
bottom of the lower half of a small petri dish.
2. Using a plastic pipette, with the narrow end cut off at an angle, collect a large active
individual from the beaker containing Daphnia.
3. Expel the water and daphniid on to the grease in the base of a small petri dish.
4. Place the petri dish under your stereomicroscope. Turn on light.
5. While looking down the stereomicroscope, check the position of the daphniid and if
necessary, use the brush end of a paintbrush to carefully position the daphniid on to its side.
If not on the grease, try to move the daphniid to the grease. If this is not easily done, then
mount another specimen. Avoid getting the thoracic limbs stuck. Water needs to be able to
pass over the thoracic appendages.
6. Once the daphniid is in place, add room temperature pond water to completely immerse the
daphniid (to approx. ¾ depth).
7. While looking down the stereomicroscope, locate the heart. It lies near the gut, or alimentary
tract, which may be pulsating. The heart is a clear, or almost clear sac and it is likely to be
beating very rapidly. You may need to adjust the light and/or turn the petri dish around to
locate the heart.
8. Record the sex of your specimen M / F / unknown and, if female, is there embryos
present in the brood chamber Y / N.
9. Record the temperature of the pond water in the petri dish to the nearest whole degree. Be
careful when placing the thermometer in the water not to dislodge the mounted daphnia.
Pond water in petri dish is o

C
10. Allow the animal 10 minutes to acclimatize to the chamber. Turn off the light when not
observing.
After 10 minutes the heartbeat and thoracic limb movement should slow to a more regular
pattern as the animal overcomes the shock of being handled.
11. While waiting, read the preparation for cooling the specimen after the practice heartbeat
measurements have been recorded (see next page).
64
Recording heartbeat and experimental set up.
To get an idea of what it will be like to view daphnia and observe their heartbeat, go
to the Week 8 block in LMS and look for the link under the Lab 4 heading.
Recording heartbeat (practice)
1. After 10 minutes of acclimation, turn the light on and record the heart rate at room
temperature by counting the heartbeat for 10 seconds. You may wish to download a ‘tap
counting’ app on your phone.
2. Record the count for 10 seconds in the table below and take a second count straight away
and record below.
3. Turn the microscope light off.
4. Calculate the heart rate for each count and then the average heart rate.
Heartbeats in 10 Heart rate Average
seconds (heart beats per minute) heart rate
Count 1 Count 2 Calculation 1 Calculation 2

Example results 20 21 120 126 123
Room temperature
= o
C
65
Experimental set up/procedure
Once you have practiced observing and recording the heartbeat at room temperature, follow the
instructions below to measure heart rate a further two times at two different temperatures.
18 oC
1. Using a plastic pipette, carefully remove a small amount of the room temperature water from
the small petri dish and replace with cooled pond water.
2. Wait 1 minute and check if the temperature is at 18 oC. Adjust as necessary by replacing some
of the petri dish water with room temperature/cooled pond water.
3. Place the petri dish under the microscope and position/focus the specimen so that you can see
the heartbeat.
4. Turn the light off and wait 10 minutes before recording heartbeats (see table below).
5. After recording heartbeats, cool the daphniid as per the instructions below.
8 oC
1. Prepare a large petri dish with a mix of mainly ice and a small amount of cold tap water. For
the experiment, the small petri dish needs to sit in the ice/water mix without the icy water getting
into it or the small petri dish floating.
2. Using a plastic pipette, carefully remove about 2/3 of the pond water from the small petri dish
and replace with cooled pond water.
3. Carefully place the small petri dish in the larger petri dish that contains the ice/water mix.
4. Wait 1 minute and check if the temperature in the small petri dish is at 8 oC. If a couple of
degrees above, wait a couple of minutes to see if it decreases. If necessary, replace some of
the small petri dish pond water with cooled pond water. If below, replace some of the small
petri dish pond water with room temperature pond water.
5. Place the petri dish set up under the microscope and position/focus the specimen so that you
can see the heartbeat.
6. Turn the light off and wait 10 minutes before recording heartbeats (see table below).
Heartrate data > class data sheet

After recording your heartbeats in the table below, calculate the heart rate and transfer the average
heart rate of your daphniid to the class data sheet (on bench below lab dividing window). Write
clearly and neatly with a dark pen/pencil.
Next, tidy/clean your bench area and put away the stereomicroscope.
Heart beats in 10 Heart rate Average

seconds (heart beats per minute) heart rate
Count 1 Count 2 Calculation 1 Calculation 2

18 oC
8 oC
66
Relationship between temperature and heart rate

Using the graph paper on the below, indicate the sort of relationship you think you would
see with the class data If heart beats were also counted at four temperatures in between
the two used (total of six temperatures). Label axis and give units of measurement.
67
Experimental Hypotheses
Write a hypothesis in relation to Daphnia sp heartrate as examined in this experiment.
You should also write a hypothesis regarding what is expected regarding Q10.(hint –
see introductory information for this lab)
Data summary/analysis
Class data will be available on LMS, in the Week 9 block. Next week’s workshop will go over how
to summarize and analyse the results.
Further background reading for Report 2

In addition to the Campbell Biology sections given at the beginning of these lab notes,
there is a selection of relevant journal articles in the BIO1AP Reading List. Access
these articles via the link given in Week 8. You can also access them via the whole
Reading List which you can access via the Library block on LMS, or via the Library
webpage.
68
HEART MUSCLE
Animals have four main tissue types, epithelial, connective, nervous and muscle. Tissue types
are explained in Campbell Biology near the start of Chapter 40. There are three types of muscle
– skeletal, smooth, and cardiac (or heart) and below are images of the three types.
Skeletal Smooth Cardiac
[Images above taken from Figure 40.5 in Campbell Biology 12e]
Each cardiac muscle cell has one nucleus, which is not the case with skeletal muscle, and are
connected by intercalated disks that are involved in signalling between cells.
View the prepared slide of vertebrate heart muscle, first at low power and then high
power (40x objective lens – do not use the 100x objective lens as this is only used
with an oil immersion procedure).
Question: Compared to the prepared plant slides seen in previous labs, how easy is it to
distinguish individual muscle cells? Why might this be?
69
Report 1 feedback
Individual feedback
Make sure you know how to view all possible types of feedback on your report (see the instruction
document under 'Other Assessment Resources' in the Assessment LMS section). Refer to the
Report Assessment Criteria and Standards when viewing your feedback (pdf available below the
Report 1 submission link if you completed the Introduction to Scientific Report Writing lesson in
Week 3). Note any questions you have - these may be addressed in the general feedback given
by demonstrators.
Group feedback
During this week’s lab class, your demonstrator will give general feedback to your whole group,
for example, in general, what aspects were done well and those that could be improved for the
next report. You will also have the opportunity to clarify with your demonstrator any questions
you have about their feedback on your first report. Feedback should help you improve your
scientific report writing skills and therefore, your performance on future reports.
To help you focus on what you can improve for Report 2, complete the table below
while the feedback (general and/or individual) is fresh in your mind.
Report section that I can improve on: ________________________________

Feedback about this section Improvement(s) to be implemented in Report 2
Report section that I can improve on: ________________________________

Feedback about this section Improvement(s) to be implemented in Report 2
Space for additional notes:
70
WORKSHOP 5 Effect of temperature on heart rate results
(Report 2)
Go to LMS, Week 10 for the data file to use and for any additional
resources for this week’s class
AIMS
• To produce the type of graph required for Report 2.
• Correctly calculate Q10.
• To conduct a paired t-test and understand how to interpret the result.
Summarizing your data > graph
Use all the data in the file you can see in LMS (Week 10), there may be data for more than your
lab. Some data may have been excluded (blacked out) and if so, this will be explained during the
workshop.
Enter data into an Excel file - one line for each daphnid's data. The first two columns of data do
not need to be entered.
You will need to summarise the heartrate class data as a box plot, similar to what was done with
the pea plant class data for Report 1, The daphnia data will need to be arranged differently to
produce the boxplot in the same way. The best way is to copy your data sheet first so that you
can still use the original data sheet for calculations. We will go over this in class.
When including the graph in your report, do not use a screen grab/snip. Use copy/paste but do
not link the graph to the excel file. Pasting as a picture is best.
Calculate Q10
You will need to calculate the average Q10 for the 10 oC interval using Equation 1 given in the
Introduction to Lab 4. In the results section of your Report, report the temperature range and
average Q10 value. You should have also included a hypothesis regarding the Q10 value from the
experiment in your Introduction.
For temperatures that are 10oC apart, T2 – T1 = 10, so the equation (see p 61) reduces to Q10 =
R2/R1. For the class data this is heartrate at 18 oC/heartrate at 8 oC and this can be calculated in
Excel for each individual.
Statistical test
With two sets of data, a simple statistical test can be performed to see if the mean heartrate at the
two different temperatures is significantly different. When comparing means for two groups, the t-
test can be used. There are two types of t-tests: one for independent samples and one for paired
samples. As each daphnia had its heartbeat counted at both temperatures, these are paired (or
repeated measures) results, so the paired t-test is the test that will be used with class data. How
to conduct the paired t-test is given on the next pages and will be gone through in the workshop,
together with interpreting the result.
71
(Report 2)
Statistical test (cont.)
Last week, during Lab 5, you were asked to write a hypothesis in relation to Daphnia heartrate.
State it again below.
Now write a hypothesis that states that no difference is occurring. This is called the ‘null
hypothesis’ and is important as this is what the statistical test is based on.
To test if the class data supports or rejects the null hypothesis, we need to statistically compare
the mean heartrate at each temperature. The appropriate statistical test to use for the class data
is called a t-test. A type of t-test called a paired t-test is used where you have two samples in
which observations in one sample can be paired with observations in the other sample. As the
same daphnid had their heart rate measured at the two different temperatures, the two
measurements are paired.
Paired t-test equation
The following formula is used to calculate the t-ratio (t-value):

∑ 𝑑𝑑
𝑁𝑁
𝑡𝑡 =
(∑ 𝑑𝑑)2
∑ 𝑑𝑑2 −
� 𝑁𝑁
𝑁𝑁(𝑁𝑁−1)
where,
d = difference between matched pairs
N = number of pairs
Σ = sum for all values
Options for a one tailed paired t-test calculation
• By hand – using Excel and the instructions (example data/calculations on the following
pages)
• Using a statistical program that you are familiar with, for example, Jamovi if you have
done/are doing first year statistics.
For either option, you will need to state the method of calculation of the t-test in your Methods
section. Also, for either option, you will need to have an Appendix - see the end of these notes
for what should be included in the Appendix
72
(Report 2)
Example one tailed paired t-test calculation
Below are example hypotheses for a study investigating respiration rate in a small marsupial,
Antechinus agilis, at two different temperatures.
Hypothesis: The respiration rate of Agile Antechinus (Antechinus agilis) at 12oC will be less
than that at 28oC.
Null There is no difference between respiration rate of Agile Antechinus (Antechinus
hypothesis: agilis) at two different temperatures (12oC and 28oC).
Below is example data for 12 individual Agile Antechinus (or replicates) and the last two columns
indicate columns that can be added in Excel to calculate d and d2 that are required for the paired t-
test equation (given on p1). The last row is where other values have been noted, such as N, and
where summing d and d2 using Excel’s summing function have occurred. This example will be
worked through during the workshop.
Respiration rate per minute at:

Replicate 28oC 12oC d d2
1 42 20 22 484
2 40 25 15 225
3 41 26 15 225
4 39 19 20 400
5 41 20 21 441
6 42 21 21 441
7 40 20 20 400
8 39 19 20 400
9 46 25 21 441
10 41 24 17 289
11 45 23 22 484
12 40 22 18 324
N = 12 Mean = 41.3 22 Σ 232 4554
Calculating the t value using the formula and values calculated above:
∑ 𝑑𝑑 232
𝑁𝑁 12 19.3 19.3
𝑡𝑡 = = = = = 26.80
2 4554−4485.3 √0.52 0.72
2 −(∑ 𝑑𝑑) �
�∑ 𝑑𝑑 𝑁𝑁 132
𝑁𝑁(𝑁𝑁−1)
To find out what this t-value of 26.80 means in relation to the null hypothesis we need to use a
table of the critical values of t for a one-tailed test (see last page). A one-tailed test is used when
in your hypothesis you predict that one of the matched values for an individual will be more/less,
faster/slower etc. than the other, rather than stating that there will be a difference.
To use the table of critical values, we also need to know the degrees of freedom (df), which are
calculated as N – 1. In the example above, this is 12-1 = 11.
73
(Report 2)
How to use the Critical values of t for one-tailed tests table (next page):
1. The first column is for degrees of freedom.

2. The headings above the other columns refer to the probability of the result occurring and
is expressed as a decimal. By convention, if the t-value has a probability of less than 0.05
then the null hypothesis is rejected and the biological/experimental hypothesis is
accepted.
3. Find the line for 11 degrees of freedom (df).
4. Go along that line in the table and find the t value of 26.80.
5. Along this line, 26.80 has a p value of less than 0.001. This means that the calculated t-
value has a less than 0.01% (p < 0.001) chance of occurring when the null hypothesis is
true. This is below the conventional significance value (α) of 0.05 (5%).
6. Therefore, in this example we would reject the null hypothesis and conclude from this
result that the increase in respiration at the higher temperatures is significant.
Now calculate the one-tailed paired t-test with the class data
The results of your one tailed paired t-test (but not the actual calculations) must be presented in
the Results section of your report. You must include the t-value, the degrees of freedom (df) and
p.
74
(Report 2)
75
(Report 2)
Appendix
For Report 2, one Appendix will be required, and you will need to refer to it in the appropriate
place/s in your report. Your Appendix must have a title and be divided into two parts. Part 1
includes relevant information on the paired t-test calculation and Part 2 includes Q10 details.
Part 1 [give it an appropriate heading]

It should include:
 hypotheses statements
If calculated by hand: If calculated using a
statistical program
 Data table as on p2 for example data showing  Appropriate output that
calculated values indicates test
 Calculated t-value as on p2 for example data conducted and
includes N, mean
 Critical t value at significance level of 0.05. For the
values and the
example data this would be reported as:
calculated t-value, df
 Critical t value one tailed paired test: tα (d.f.) = t.05(11) = and p
1.796.
 Statement indicating if null hypothesis rejected or supported
 Statement indicating if biological/experimental (alternate) hypothesis supported or
rejected.
Part 2 [give it an appropriate heading]

It should include:
 How individual Q10 values calculated
 Individual Q10 values used to calculate the average Q10
Questions about Report 2?
Have any questions ready to ask your demonstrator.
76
LAB 5 ANIMAL ADAPTATIONS FOR FEEDING

Concept 33.4 Insects, 749-750
Concept 41.4, 960
LMS Week 10 block

Online skulls
Insect adaptations for feeding
AIMS
• To use observations of mammal skulls to identify if the animal is a carnivore, herbivore or
omnivore.
• To identify the mouthparts of a hemipteran insect (or other insect) and match them to the
unmodified mouthparts of a generalised insect.
Part 1 What can I learn from a skull?

In this lab class you will consider functional aspects of mammalian skull morphology that provide
evidence for the animal’s diet and feeding mode. Observe differences between the three skulls
shown below.
Skulls of three Australian mammals; from left to right, a carnivore (Eastern quoll, Dasyurus
viverrinus), herbivore (Koala, Phascolarctos cinereus) and an omnivore (Bilby, Macrotis lagotis)
(Images: Museum Victoria)
Teeth, jaws, and other skull features can provide important clues about what an animal ate and
how it captured and consumed its food. Adaptations for feeding among mammals are clearly
demonstrated by different patterns of dentition both in numbers and shape of teeth.
Mammalian teeth can be differentiated into four basic types: incisors (I), canines (C), premolars
(PM) and molars (M) (see figure next page). Incisors are chisel-shaped and generally used for
holding, biting, gnawing, shearing, etc. Canines are sharp and conical in shape and are used for
griping, piercing and ripping during feeding or fighting. Premolars and molars are used to grind or
break up food. Premolars differ from molars in that they have fewer roots and are often simpler in
form. In some mammals it is not possible to distinguish premolars from molars and they are
combined as the cheek teeth (PM + M).
77
The dental formula describes the number of each kind of tooth that is present in the upper and
lower jaw on one side of the skull. A horizontal line separates the upper and lower jaw formula. To
calculate total teeth, add up those found in the upper jaw (x 2 for both sides of the skull) and the
lower jaw (x 2).
Features of a mammalian skull and tooth types
78
The location of eye sockets in the skull is one of the best indicators of whether an animal plays
the role of predator or prey. Prey animals, such as kangaroos and rabbits, tend to have eyes on
opposite sides of their heads. This provides a nearly 360-degree field of view at all times, a
tremendous advantage when surveying the landscape for movement and possible threats. In
contrast, predators, such as dingos and cats, tend to have forward-facing eyes (see figure below).
While this limits their field of view, it allows the view that each eye sees to overlap with the other
(more binocular vision) to perceive depth and distances more accurately, a huge advantage when
pursuing prey. The size of the eyes can also tell us something about activity patterns, nocturnal
animals have larger eyes than their diurnal counterparts.
Position of the eyes – on the side of the head vs. facing forwards, gives us clues to whether they
are predators or prey.
Herbivorous mammals
Herbivorous mammals tend to have flattened premolars and molars to grind up plant material,
and their incisors are specialised for biting / clipping or tearing vegetation, but they may only
occur on the lower jaw. Often, herbivores feature ridged molars and jaws capable of moving
sideways. Both of these traits help herbivores to grind their food more effectively. Notice that
many herbivorous animals reduce or lose the canine teeth and develop a diastema, or gap
between the incisors and the premolars (see figure below).
What function do you think loss of the canine and development of the diastema serves?
Carnivorous mammals
Carnivorous mammals have long, sharp canines for stabbing and tearing, and premolars and
molars specialised for shearing off bites of meat. The number of premolars and molars are
often reduced. The incisors are adapted for biting and holding with the help of large temporalis
muscles, which are responsible for pulling the lower jaw upwards and backwards towards the
skull. The temporalis muscles attach to the jaw at one end, and the top of the skull at the other
end. To help accommodate larger temporalis muscles, some predators have evolved to have an
enlarged ridge, termed the sagittal crest that acts as an attachment point or anchor for the
muscle (see image below).
Omnivorous mammals – including Insectivores, Granivores

Omnivorous mammals consume both plant and animal material; hence, they have dentition,
skulls, and teeth suitable for handling a variety of foods. Mammals that primarily eat insects
(insectivores) tend to have teeth with sharp points to pierce the exoskeleton. Gnawing mammals
(granivores) have large, ever-growing, chisel-shaped incisors and grinding cheek teeth.
79
Dentition of a carnivore, a herbivore and omnivore (from Campbell Biology 11th edition - Aus. &
NZ version, p 937).
Examine and discuss the three skulls for your group and complete the table on the next page
80
Observations of skull characteristics for three mammal skulls.
Skull A Skull B Skull C
Incisors (large/small)
Canines (long/short,
pointed/dull)
Molars/Premolars –
upper & lower surfaces
(surface contact/no
contact)
Molars/Premolars
(grinding
surface/shearing-
cutting
surface/multipurpose
surface)
Jaws can move side to
side (yes/no)
Sagittal crest
(present/absent)
Orbital socket –
diameter (small/large)
Orbital socket –
orientation
(forward/side)
Diastema
(present/absent)
Dental formula
(I.C.PM.M)
(I.C.PM.M)
Likely mode of
nutrition?
(Carnivore / Herbivore
/ Omnivore)
Predator or Prey?
Possible animal?
Actual species
81
Part 2 Insect mouthparts
Insects do not manipulate food

internally in a mouth but use
external appendages. Multiple
pairs of appendages near the
mouth are modified to process
food. A ‘generalised’ insect is
usually drawn with chewing mouth
parts. Mandibles are the main
chewing appendage.
Specialisation of mouthparts occurs

in different groups of insects so that
other food types can be consumed,
for example, free liquids or liquids
within other organisms. Different
insect feeding types was discussed
in lectures in week 8 and a
schematic diagram shown to
illustrate modification of mouthparts
in different insects.
Hemipteran insects, commonly called bugs, have modified mouthparts and the diagram below
shows how different they are from an insect with chewing mouthparts (a colour version of the
hemipteran diagram is in the Week 10 area of LMS.
Source: from Panfilio and Angelini (2018). By land, air, and sea: hemipteran diversity through the
genomic lens. Current Opinion in Insect Science 25: 106-115.
82
View the prepared slide of hemipteran (bug) mouthparts using the 4x objective
lens (40x magnification) and draw a plan diagram below indicating the
following structures: compound eye, antenna, labium, labrum and stylets. It
will not be possible to distinguish between mandibular and maxillary stylets.
As you will not be able to get the full length of the mouthparts in the field of
view, ensure the head is included.
83
WORKSHOP 6 ASYNCHRONOUS ACTIVITY VIA LMS
See the Week 11 area of LMS for details of this activity to be completed in
your own time but remember that the reflection is due 11:59pm Friday of this
week.
84
LAB 6 THERMOREGULATION IN SIMULATED ANIMALS IN
COLD CONDITIONS

Concept 40.3, 933-937
AIMS
• Determine the freezing point depression and plot the cooling curve for four simulated
ectothermic animals.
• Understand the description of the effectiveness of insulation experiment using simulated
endothermic animals and the data presented graphically.
Part 1: Freezing point depression in an ectotherm
Arctic animals that live in ambient temperatures below freezing have to avoid the fatal formation
of ice in their body tissue. These animals depend on physiological as well as biochemical
adaptations in order to live. High concentration of glycerol is found in the blood of the Antarctic
fish, Trematomus borchgrevinki. Glycerol increases the cold tolerance and lowers the freezing
point of the tissue fluid, preventing the deadly formation of ice crystals in the blood and tissues,
thus enabling the fish to live in seawater at a temperature of –1.8°C.
You will simulate an ectothermic animal’s internal environment and examine the effectiveness of
glycerol, antifreeze, and sodium chloride on freezing point depression.
Freezing-point
depression
describes the
phenomenon in
which the freezing
point of a liquid is
depressed when
another compound is
added, meaning that
the solution has a
lower freezing point
than before. The
normal freezing point
of tap water is 0oC.
85
COLD CONDITIONS
Procedure
Work in groups of four. Each student monitors one test tube ‘animal’ (see diagram below):
- glycerol (pre-dispensed)
- 1:1 mix of saline and antifreeze (pre-dispensed) [wear gloves for this ‘animal’]
- 0.9% saline
- tap water
1. Prepare an ice-salt bath using layers of crushed ice and thin layers of rock salt. Alternate
the layers of ice and salt until the Styrofoam container is full. Stir the ice-salt mixture using
a plastic rod. Firm mixture and top up with ice if necessary. Use the plastic rod to make
four spaces to place large test tubes.
2. Insert four large test tubes into the ice-salt mixture deep enough that when the test tube
‘animal’ is placed inside the large test tube it is fully below the level of the ice-salt mixture.
3. Place the test tube ‘animal’ inside the larger test tube. Placing the smaller tube in a larger
one ensures even distribution of temperature around the small tube and prevents cold
spots outside the small tube (see diagram below).
4. Insert the thermometer/thermocouple into the tube. Make sure the
thermometer/thermocouple is immersed in the solution, and the stirrer moves up and
down easily. If you have a solid thermoprobe, carefully use this as a stirrer as there will
be no space for a plastic stirrer. Be careful not to break the test tube.
5. For each ‘animal’ stir the solution with the stirrer approximately every 30 seconds and
allow the solution to cool to about 5oC. It is unlikely that all solutions will reach 5oC at the
same time.
Note- stirring the solution avoids the development of cold spots, ensuring the
temperature of the solution is even through the small test tube.
6. Once the temperature for an ‘animal’ is at about 5oC, measure the temperature every 2
minutes and record in the table on the next page. Each ‘animal’ needs to be monitored
and timed separately. Keep stirring regularly until your animal freezes.
7. Continue measuring the temperature for each ‘animal’ until six successive readings for
that animal are of the same temperature.
8. If your animal is still liquid at six consecutive temperatures, then the ice/salt mixture has
reached its cooling capacity.
Diagram of the set up for measuring the freezing point of fluid in the small test tube.
86
COLD CONDITIONS
Internal temperature over time for different test tube ‘animals’.

Temp o
C
Time
(1:1 Saline and
(Min.) Glycerol 0.9% Saline Tap water
antifreeze)
0
2
4
6
8
10
12
14
16
18
20
22
24
26
28
30
32
34
36
38
40
87
COLD CONDITIONS
Using Excel or on the graph paper below plot the cooling curve for the 3
solutions and tap water from the data in the table on the previous page. Include
a Figure caption.
88
COLD CONDITIONS
Calculate the freezing point depression for each of the test-tube ‘animals’ using the formula
below.
ΔTF =TF (pure liquid) - TF (solution)
where:
ΔTF = the freezing point depression
TF (pure liquid) = freezing point of water
TF (solution) = freezing point of solution
Glycerol =
1:1 mix saline and antifreeze =
0.9% saline =
Tap water =
Q4 How do glycerol, antifreeze and NaCl affect freezing point?
89
COLD CONDITIONS
Part 2: Effectiveness of insulation on thermoregulation in an endotherm
BACKGROUND
Endothermic animals can tolerate only a narrow range of body temperature, but they are able to
live in a wide range of environmental temperatures. The ambient temperature range within which
the metabolic heat production of an animal is unaffected is considered the animal’s thermoneutral
zone. It is advantageous for an animal to modify its behaviour and physiological adaptations, such
as thicker insulation, to remain in the thermoneutral zone. While in the thermoneutral range, the
animal’s resting metabolic rate remains fairly constant and effort for thermoregulation is reduced to
a minimum. Thermoregulatory heat production is enormously expensive and makes up the single
largest component of the energy budget in endotherms. When ambient temperature is below the
thermoneutral zone, the resting metabolic rate of an animal has to be increased by shivering to
replenish heat that was lost to the environment.
In situations where ambient temperature (Ta) is below the body temperature (Tb), heat is
transferred from the animal to the environment by conduction and radiation. Endothermic animals
adjust their rate of metabolic heat production (H) to equal the rate of heat loss (Q). The heat balance
of an animal can be expressed by the following equation:
H α Q α C (Tb-Ta)
The loss of heat is mediated by the gradient between the (Ta) and (Tb). The greater the difference
between the ambient and body temperatures, the faster and greater the heat loss is. For example,
wolves decrease their Tb to reduce the gradient between the Ta and Tb, thus reducing heat loss to
their environment. Conductance (C) represents heat flow from the body to the environment or vice
versa. To reduce their thermal conductance, animals rely on behavioural adaptations and the
efficiency of their insulation such as fat, blubber, and fur. As insulation is increased, conductance
decreases, heat loss is minimized, and the animals remain in their thermoneutral zones with
minimal expenditure of energy to regulate their Tb.
This part of the lab class examines the effectiveness of fat and/or fur as insulators on
thermoregulation at ambient temperatures of 22 oC and 8oC using data from an experiment
conducted using simulated endothermic animals.
'Animal' insulation types:
• No insulation
• Layer of fat only
• Layer of fur only
• Layer of fat and fur
The 'animals' were empty aluminium soft drink cans filled with 0.9% saline at 37oC, to replicate the
salinity of body fluids and an internal temperature appropriate for a mammal. The relevant
insulation type was then wrapped around the can.
The fat layer was created with lard (animal fat) at approx. 30oC enclosed in a zip-lock plastic
bag. the bag was folded lengthwise, wrapped around the can, and held in place with an elastic
band. The fur layer was created with a length of rabbit pelt wrapped around the can and held in
place with an elastic band. For fat and fur together, the fat was wrapped around the can first, then
the fur.
90
COLD CONDITIONS
The four simulated ‘animals’ with different insulation
Materials used to create the simulated animals. Aluminium can filled with filled with 0.9% saline
at 37oC, rabbit fur, animal fat.
91
COLD CONDITIONS
8oC environment
Before creating the four 'animals' the 8oC ambient temperature condition was set up. This was
achieved using icy water in a tub as no suitable cool room was available. Once the tub
temperature was 8oC, the 'animals' were then prepared as described on the previous page. As
each can was to be immersed in the tub of icy water, each 'animal' was placed in a plastic bag to
keep dry before adding to the 8oC environment. To monitor the internal temperature a
thermometer was placed in each can. The internal temperature for each 'animal' was measured
and recorded every five minutes for one hour.
22oC environment
This environment was the room temperature of the lab and ‘animals’ were placed in a tub on the
bench and the same measuring procedure occurred as for ‘animals’ in the 8oC condition.
Questions
Which of the four ‘animals’ do you predict to be best insulated in the 8oC environment? What
would be the next best? What would be the evidence to indicate this?
Do you predict the same pattern will occur at 22oC?
Consider the data you have been shown for the simulated animals at two different ambient
temperatures,
Which ‘animal’ had the greatest conductance of heat?
Which ‘animal’ had the lowest?
Does the data match your prediction?
Can you think of why the simulated animals with insulation didn't maintain an 'internal'
temperature as would a real animal? For example, in humans a temperature of 35oC or
below is dangerous and is considered as hypothermia.
92

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School of Agriculture,

Animal & Plant Life

Gendall F2F Workshop – Report writing Refln

9 Oct 2 F2F Workshop review daphnia data 

10 Oct 9 F2f Lab 5 animal adaptations for feeding – Obs

Lab/Workshop Day:_________________________ Time:______________

Animal and Plant Life (BIO1AP)

f2f classes begin in week 2 with online activities in week 1

Signposts used in this manual:

Safety note: Carefully read the

Reading: List of reading to

To do: Complete a table,

Watch: Your demonstrator will

LMS: There is an LMS activity

Timetable and Venue

Laboratory equipment for class

General Safety Procedures in the Laboratory

COVID-19 (& flu etc.) Safety in the Laboratory

Face masks are strongly recommended whenever in the lab as maintaining

Practical assessment (38% towards final grade)

1. Observations/Reflections 12% of final grade See timetable inside front

General instructions for drawing

Example of a low power magnification drawing

1.A.2 Recording your observations through graphs/figures

FAQ: If I miss a f2f class (lab or workshop), how do I catch up?

Late Submission of Reports

Report FAQ’s – see BIO1AP LMS

Lab Safety – online induction on LMS

Use page 4 of this manual as a reminder of the Personal Protective

Microscopy – online module on LMS

If you want a refresher or have not used a compound microscope before,

Teaching lab drop-in

Bundoora: LIMS 1 teaching laboratories (Rooms 203/204)

Online safety quiz

Part 1 - PLANT ROOTS

1A The root tip

4. How do the root hairs contribute to the functioning of the root?

Part 2 - PLANT GROWTH REGULATION EXPERIMENTS – set up

Campbell Biology 12e (Aus. & NZ version)

Experiment 1: The Role of Gibberellic Acid in Plant Growth (Report 1)

Experiment 2 (demonstration): Apical Dominance in Phaseolus vulgaris

Plants: Three pots, each containing a bean seedling (Phaseolus vulgaris).

2. Control (C) plant: set aside as it will remain an intact plant.

Experiment 3 (demonstration): Auxin and Leaf Development

Aim: To study the effect of auxin on the development of leaf tissue.

A. Along the margin of a

Before coming to your workshop, you need to complete the

BIO1AP Report Assessment Criteria and Standards

/ 100 Before any deductions (see below)

Reference list - not alphabetical by surname, inconsistencies in formatting 2

Font size/style inconsistencies 2

BIO1AP Report Assessment Criteria and Standards

/ 100 Before any deductions (see below)

Reference list - not alphabetical by surname, inconsistencies in formatting 2

Font size/style inconsistencies 2

Campbell Biology 12e (Aus. & NZ version)

Part 1 – Plant Growth Regulation data collection

Effect of GA experiment – Report 1

Treatment Height (mm) Number of internodes

1. What does the plant hormone, GA, do in stems?

2. Why was it necessary to spray one pot with distilled water?

Example data presentation

Space for any additional notes about the Effect of GA experiment.

Lab/Workshop Day:___________ Time: