Cell Structure

30/11/2009

CELL STRUCTURE
Objectives: To learn to use a compound microscope properly To review the main parts of a cell, using your textbook as a reference Before you leave at the end of the laboratory period you should know the parts of a microscope and how to determine total magnification. You should be able to recognize the organelles in a plant and an animal cell, and recall the functions of each. You should also know the stains that you used and what they stained. Introduction: As you will find in this course, there is no such thing as a "typical" cell. During this period you will look at four "representative" eukaryotic cells: two plant cells (one with chloroplasts that are easy to focus on, and one without chloroplasts that will be more challenging), one animal cell, and one example of a single-celled organism. In each case you will try to locate as many organelles as you can. You will also examine a prepared slide of some prokaryotic bacterial cells. Your textbook is an excellent reference, so you may want to refer to it during this exercise. I. MICROSCOPES A. Parts of a Compound Microscope Your goal in this section is to learn the names of all of the parts of the microscope. B. Magnification

The magnifying power of the ocular and objective lenses used on the microscope is usually engraved on the lens. Let's look at some typical lenses.

The magnification of the ocular lens is 10 X or ten times magnification.

There are three objective lenses shown on this microscope. You can magnify the image and move around to see the magnification engraved on each lens.

The lowest power lens on this microscope is 4 X

magnification. It is often referred to as the scanning lens and should be used first when viewing a new specimen. On this microscope the scanning lens has a red band around it to make it easy to identify.

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The next highest magnification is the 10 X lens also called the low power lens. It has a yellow band. The highest power dry lens is the 40 X lens. It has a blue band.

Some microscopes will have additional higher power objective lenses (for example 100 X). These lenses require that a drop of immersion oil be placed between the lens and the specimen.

Now that we know the magnifying power of the ocular and objective lenses, we can calculate the total magnification using each of the lens combinations.

To calculate the total magnification, multiply the power of the ocular lens times the power of the objective lens you are using.

C. Field of View

Sometimes it is necessary to determine the size of an object that you are viewing under the microscope. There is an easy way for you to estimate size. If you know the diameter of the field you are seeing in the microscope, you can estimate the size of the object you are viewing.

For example:

Here you see the same object with increasing magnification. At the lowest magnification the object occupies only a small portion of the field.

At the highest magnification the object nearly fills the field.

millimeters are too large of a unit of measure. Each box on the paper is a 1 mm square.• If you could fit a clear ruler under the microscope you could determine exactly how wide the field diameter is at the different magnifications and determine the approximate size of the object you are viewing. . You will do this more simply by placing a piece of graph paper on a microscope slide and viewing it under the microscope. In fact. you can buy a ruler mounted on a microscope slide that is especially designed for this. 1 mm = 1000 micrometers Let's try this at our lowest magnification using the scanning lens (40 X total magnification). Microscopic objects are measured in micrometers. but for microscopy.

.Now estimate the field diameter in micrometers.

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Now do the same using the 10 X ocular and the 10 X low power objective lens at a total magnification of 100 X (shown on the right). As you can see. the block on the graph paper gets larger. . each time the magnification is increased.

if not impossible. . the block will be so large that we will not be able see the lines clearly and it will be difficult. We can use a mathematical formula to estimate the field size at 40 X. the field diameter is inversely proportional to the magnification. Therefore. to estimate the field size.If we try to estimate field size using graph paper with the 40 X objective. how do we estimate a field size for the 40 X objective lens? We know that the higher the magnification is. the smaller the field diameter is. So.

Remember. . the field diameter with the 10 X objective lens was 1700 micrometers.Let's do this using the numbers we just calculated for the 10 X objective. Now fill in the numbers in the equation.

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For example. Image Formation When a microscope magnifies an image it shifts the orientation of the object you are viewing. D.Now solve the equation for the field diameter of the high magnification lens. if you cut out the letter "e" from a newspaper and put it under the microscope what happens to the orientation of the letter? . you get the .

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The "e" is now inverted and shifted from right to left. . Hold down the left mouse button and slide the cursor to the right. Now move the cursor to the left. place your cursor on the letter "e". What happens if we move the specimen stage to the right? To simulate this.

Cells A. Now move the cursor downward. Plant Cells . Place your cursor on the letter "e".What happened? Now try this. What happened? II. Hold down the left mouse button and slide the cursor upward.

Elodea: leaf cells .1.

If you examine the leaf using the scanning lens you will find the midrib running down the center of the leaf.Elodea is a decorative aquatic plant often found in fish tanks. The midrib contains the main vein for conducting materials to and from the leaf. A small leaf has been removed from the plant and placed with the lower surface down in a drop of water on a microscope slide. Can you identify the chloroplasts in the upper photo and the cell walls in the lower photo? . Now turn to higher magnification and examine a portion of the leaf away from the midrib.

.Click here to confirm your answers. .

Remember that a cell, such as this leaf cell, is a three-dimensional structure. The cell wall surrounds the cell on all six sides. The chloroplasts and other organelles are held against the sides of the cell by the large central vacuole. You can confirm the 3 dimensional nature of the cell by focusing up and down on while observing one cell.

Try this.

The cytoplasm of the cell is not static but "streams" around the perimeter of the cell. The chloroplasts are carried by the streaming cytoplasm.

In the biology laboratory you also will treat your specimens with different stains to enhance the contrast and make them easier to see under your microscope. This portion of the lab is not done in the virtual laboratory.

2. Onion: Epidermis A small piece of epidermis from the scale of an onion bulb has been removed and a a wet mount has been prepared for microscopic examination. If you are not experienced with the microscope, you may be able to see little besides the cell walls which appear as little boxes. If you reduce the amount of light by using the condenser diaphragm, you may be able to see the nucleus, cytoplasm, and vacuole.

. Now place the cursor over the image of the onion and close the diaphragm. What happens to the contrast? With the diaphragm closed you can easily see the onion epidermal cells. Now determine the size of a single cell using the estimate of the field diameter.Look at the onion preparation with the condenser diaphragm wide open.

An Animal Cell . B. the diameter of the field is about 560 micrometers. What is the approximate length of the cell? Now try another cell from the same field.In this example.

A small amount of material has been gently scraped from the inside of a mouth and mixed with a drop of water on a glass slide.You will use a cheek cell (simple squamous epithelium) as an example of an animal cell. A cover slip was then added. What do you see under the microscope? .

Most of the time. Now look at a higher magnification view of an epithelial cell. When the diaphragm is in the closed position. you have maximum resolution but the image has very low contrast. you will have maximum contrast. You can increase the contrast by closing down the diaphragm.The image has low contrast. When it is in the fully open position. . we compromise and close the diaphragm about half way to maximize resolution and contrast as much as possible.

Euglena a single-celled eukaryote .What cell structures can you identify? C.

What organelles can you see in the cell? . Look at the swimming Euglena cells.Euglena is a single-celled eukaryotic organism that is often called a "plant-like" organism since it is photosynthetic. Now look at a single Euglena cell.

Flagellum – Nucleus – Eyespot – Chloroplast – .

Bacteria single-celled prokaryotes .D.

. Notice that not all of the rods are of the same length. four or more cells. The round bacteria are often clustered in groups of two. The prepared slide of bacteria has three types of bacteria on it. Now find the rod-shaped bacteria.Bacteria are extremely small and often are not visible with the 10 X objective lens.

To estimate the sizes of these bacteria switch to a higher magnification image.Now find the spiral-shaped bacteria. At this magnification the field diameter is about 45 micrometers. .

Estimate the size of the rod-shaped bacteria.1. Estimate the size of the spiral-shaped bacteria. . 2.

. Estimate the size of the round bacteria.3.

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and this species has a relatively small number of chromosomes. into two equal parts. is one of the regions in the plant where cells are actively dividing and elongating. The root tip is responsible for the downward growth of the root and therefore. Part 1. Because of this. the root tip is an excellent system in which to study the process of cell division (cytokinesis)and nuclear division (mitosis) Furthermore. . At the right is a longitudinal section through an onion (Allium) root tip. by a process involving chromosomes. Mitosis is the sequence of events by which the nuclear material of one cell is distributed. a. Can you find dividing cells in the onion root tip? b.What differences can you see when you compare the nucleus of a dividing cell with that of a non-dividing cell? Part 2.Cell Reproduction 30/11/2009 How do cells reproduce? Introduction to the Lab: The emphasis of this laboratory period will be on mitosis. Click on the root tip to magnify the image. the chromosomes are fairly large and distinct.

Metaphase – During metaphase the chromosomes are distinct and line up near the center of the cell. Microscopy Lab Now that you have seen how nuclei divide. Review the diagrammatic summary of cell division in your textbook before you begin.Cell Structure 30/11/2009 a. the nucleolus is present and chromosomes are not distinct. View a video of mitosis in an animal cell Part 3. . b. the nucleolus is gone and chromosomes are distinct and wound throughout the nucleus. Identify the stage: Interphase .At interphase the nuclear envelope is still intact. Prophase .During prophase the nuclear envelope disappears. you can begin the microscope exercises.

By telophase the two groups of chromosomes have completely separated and are positioned at opposite sides of the cell. The nuclear envelope begins to reform. Telophase/Cytokinesis . .During anaphase the chromatids are separated and the two groups of chromosomes migrate towards opposite sides of the cell. Part 4. Click to begin Slides of whitefish blastulae will be used to show mitosis and cell division in animal cells. Mitosis in an Animal Cell.Cell Structure 30/11/2009 Anaphase . proceed to the next section. When you have identified at least one cell in each stage.

. Place the cursor over a dividing cell and click once. Identify the stage of division.Cell Structure 30/11/2009 Although the result of these processes and many of the events are the same or very similar to that of the plant cells. Identify at least one cell in each stage. Click on any of the slides at the right to magnify.During prophase the nuclear envelope disappears. the nucleolus is present and chromosomes are not distinct. the nucleolus is gone and chromosomes are distinct and wound throughout the nucleus. there are some differences. See what differences you can detect. Prophase . Use the scroll bars to move around the slide.At interphase the nuclear envelope is still intact. Identify the stage: Interphase .

.Cell Structure 30/11/2009 Metaphase – During metaphase the chromosomes are distinct and line up near the center of the cell.

During anaphase the chromatids are separated and the two groups of chromosomes migrate towards opposite sides of the cell.Cell Structure 30/11/2009 Anaphase .By telophase the two groups of chromosomes have completely separated and are positioned at opposite sides of the cell. Telophase/Cytokinesis . . The nuclear envelope begins to reform.

Cell Structure 30/11/2009 .

) • To understand how the relative position of meiosis and fertilization varies in different life cycles. You should know how to recognize the haploid and diploid (and dikaryotic) phases of each organism studied. located on two complete sets of chromosomes. noting the transition from single-celled or multi-cellular organisms. Alternately. [You may want to review the discussion of these terms in Lab 2.] . (The animal-like protists will be examined next semester. Before you leave at the end of the laboratory period you should be able to tell a plant-like protist (alga) from a fungus. meaning that they contain two complete sets of their genetic materials. containing only one complete set. as in the mushrooms. and what cells were formed by mitosis and by meiosis. some plant and animal cells are haploid (1n). • To appreciate the tremendous reproductive capacity of organisms reproducing asexually. and specify the kingdom of each.Reproduction: Protists and Fungi 30/11/2009 Reproduction: Protists and Fungi Objectives: • To examine some of the structural variation found in the simpler organisms on earth. Introduction: Most plant and animal cells are diploid (2n). This capacity may also be observed in some sexual cycles.

Cell Reproduction 30/11/2009 Dependent upon the organism.) The diploid number is restored when two haploid cells fuse during fertilization. In both cases. . a diploid cell undergoes a "reduction division" to form four haploid cells. (The reason that four cells are formed will be covered later in the semester. During a second type of cell division called meiosis. each "daughter cell" has the same amount of genetic material (chromosome number) as the "mother cell". both diploid and haploid cells can divide by mitosis.

g.g. a simplified cycle will be found at the upper right corner of each illustrated cycle. all of the other cells are formed by mitosis . On the right. as indicated in the sexual cycle to the right. Therefore. This zygote immediately undergoes meiosis upon "germination". The mature adult is then formed by a large number of mitotic divisions. Oedogoniium and Spyrogyra). Chlamydomonas). many protists have a sexual cycle where the only diploid cell is the zygote. the only haploid cells are the gametes. the relative number of haploid cells is small. the only diploid cell is the zygote.. forming the zygote.Cell Reproduction 30/11/2009 The actual location of meiosis and fertilization in a sexual cycle depends upon the organism. In some instances the "many haploid cells" remain separate (e. To help you keep track of which cycle goes with each organism you will study in this lab. on the left. egg and sperm. On the other hand. . For example. the only haploid cells produced by humans (and other animals as well) are the sex cells (gametes). they immediately combine to restore the diploid condition by fertilization. are formed by meiosis.. in other cases the cells are attached to form a filament (e. These two cycles can be abbreviated by simply showing meiosis and fertilization as gray arrows. Compare these sexual cycles. The gametes. and in still other cases the cells actually form a multicellular organism (not seen in lab.including the cells that are capable of fusion again. but described in your textbook).

Fucus is a macroscopic organism. The fungi that you will observe today are given their own kingdom. This can be an asexual reproduction of diploid organisms. The prokaryotic organisms. they are therefore called protists. Many organisms also have an asexual cycle where the offspring are simply produced by mitosis and therefore are genetically identical to the parent (in other words. We will study this cycle in the next two laboratories. Most biologists today subdivide all of life on earth into six kingdoms. Many if not all of the simpler plant-like organisms ("algae") are placed with simpler animal-like organisms into a Kingdom Protista. can produce both diploid and haploid cells asexually. such as yeast. It can be an asexual reproduction of the haploid organisms. resulting in an "alternation of generations". is associated with the sexual cycle.Cell Reproduction 30/11/2009 Plants (and some larger plant-like protists) have a third sexual cycle in which both diploid and haploid phases are multicellular. Some Plant-Like Protists Although the term "algae" is no longer used in classification. Chlamydomonas is a single-celled protist. such as the bacteria you observed in Laboratory 1. and the resultant recombination of genes. placed in the plant kingdom in some textbooks. the alternation of meiosis and fertilization. I. as the production of spores by many fungi. Spirogyra and Oedogonium are filamentous. are in Kingdom Eubacteria and Kingdom Archaebacteria. And some organisms. Finally. it is still a useful term to describe aquatic photosynthetic organisms with little tissue differentiation. as in the propagation of some commercial plants. they are "clones"). .

Single-celled protists .Cell Reproduction 30/11/2009 • A.

Euglena What characteristics do these cells have that are like plants? Are like animals? hints Note that these cells divide by mitosis. . Does Euglena have a sexual cycle? Yes No 2. Paramecium Although you will spend more time looking at animal-like protists ("protozoa") in Bio 102. one example is added here. and therefore asexually.Cell Reproduction 30/11/2009 1.

where plus and minus types "clump" together.Cell Reproduction 30/11/2009 Observe Paramecium under low and then high power. Since you cannot tell one from the other microscopically. two flagella. Chlamydomonas This small organism requires that you use a high-power objective for viewing. Click the thumbnail below to view a video of the first stage of mating. What characteristics do these cells have that are like animals? Are like plants? 3. If you look carefully. an eyespot (similar to Euglena). Chlamydomonas fusion This small organism has at least two "mating types". Do you see any of these specialized structures in the cell below? 3. . they are called "plus" and "minus". you may observe in each tiny cell: a cup-shaped chloroplast with a round starch-like product in the middle. and you may even discern the centrally-located nucleus.

fusion begins. On the right. but may also reproduce clonally. Watch the two cells at the top of the window. Chlamydomonas cells have a sexual cycle. click the thumbnail below to view a video clip of "pairing". each mating stage is shown in its correct temporal sequence. . by undergoing mitotic division. After pairing. where two Chlamydomonas cells come together to mate.Cell Reproduction 30/11/2009 Next. as shown below.

identify which cells are haploid and which are diploid. Click on the image at the right to begin. What compensating process must occur in the diploid (2n) cells? Mitosis Meiosis Chlamydomonas life cycle In the figure below. .Cell Reproduction 30/11/2009 Are the cells above haploid or diploid? haploid diploid How can you distinguish the haploid from the diploid phases in Chlamydomonas? In any organism? You have observed "fertilization" in this organism.

from a prepared slide of this alga. Oedogonium Note the top image on the right. or zygote. round structures within some of the cells. The lower image shows a fertilized egg. Some filamentous algae 1. eventually becoming attached to the lake bottom and differentiating into a new filament. these are the eggs. the zygote is the only diploid structure in the alga's life cycle. The Chlamydomonas zygote goes through meiosis to form four mobile "zoospores" that swim about.Cell Reproduction 30/11/2009 B. It shows the chain of cells that makes up one filamentous Oedogonium organism. note the clear. As in Chlamydomonas. This organism also reproduces asexually by simple fragmentation or by the formation and liberation of single zoospores from vegetative cells. In the middle image. .

In this image. notice that cells are connected end-to-end to form a long chain of cells.Cell Reproduction 30/11/2009 Oedogonium is a filamentous organism. Spirogyra . The bulb-like cells in the filament are egg cells. Also notice the smaller. Also note the small brown nucleus centrally located in each cell. cells below the egg on the top Zygotes are similar in size and shape to unfertilized eggs. sperm forming filament. 2. They are distinguished by their thickened cell walls. increased starch content. and darker color when stained.

Note the shape of the chloroplast(s) in the figure below. In the image below. From wet mounts. .Cell Reproduction 30/11/2009 This is another filamentous alga. Fusion in this genus begins as two filaments attach. you can see that it is also a long filament of cells. you can see both the front and back of the single spiral-shaped chloroplast that gives this genus its name. commonly found in ponds and puddles around New Jersey. The protoplast of the cell is mostly transparent. therefore. notice the small conjugation tube that joins adjacent cells. and the figure below.

The heavily stained ovoid structures in the figure below are zygotes. Spirogyra life cycle . Notice that the filament on the left is empty of cytoplasm. and though cell walls remain visible. you can no longer see the spring-like chloroplasts inside.Cell Reproduction 30/11/2009 Conjugation is complete after the cytoplasm from the adjacent cells fuse to form zygotes.

while other times it is exposed to air. air bladder. . Look at the images of Fucus below. Describe how these specializations are adaptations to an intertidal existence. a.Cell Reproduction 30/11/2009 C. Note the various specialized structures such as holdfast. and receptacles containing conceptacles. Specialization in a large alga The marine alga Fucus has adapted to life in the intertidal region: part of the day it is submerged.

Examine images of these conceptacles below.Cell Reproduction 30/11/2009 Fucus b. Inside the receptacles are small pit-like areas that contain the male and female reproductive structures. How are male and female structures similar? How do they differ? Why is one called "male" and the other "female"? What is the adaptive significance of this differentiation? .

b. bulb-shaped oogonia with egg or egg progenitor cells inside. The antheridia contain sperm or sperm progenitors. The lighter-stained thread-like hairs are non-sexual. they help retain moisture when the receptacle is exposed to dry conditions. darkly-stained. sterile hairs protect against dessication. Examine images of the antheridia and oogonia below. antheridia. How are these male and female structures similar? How do they differ? Why is one called "male" and the other "female"? What is the adaptive significance of this differentiation? .Cell Reproduction 30/11/2009 The male conceptacle (above) can be identified by the many small. The female conceptacle is characterized by its large. Male and female reproductive structures are located inside the conceptacles. As in the male conceptacle.

sterile hairs appear blue. Reproductive structures are stained red in the micrographs. Decide whether each structure is part of the haploid or diploid phase of the life cycle. slim antheridia (above) with the bigger. Some Fungi . compare the small. Study the life cycle of Fucus below. II.Cell Reproduction 30/11/2009 In these higher magnification images and diagrams. rounder oogonium (below).

with the zygote being the only diploid cell in the entire cycle.Cell Reproduction 30/11/2009 The sexual cycles of the fungi are basically the same as you have seen before. Fungal Life Cycle: In the diagram above. or karyogamy. The stage between plasmogamy and karyogamy is called dikaryotic. since each cell has two haploid nuclei: one from the plus strain and one from the minus strain. each arrowhead represents an event in the life cycle of many fungal cells. or (n + n). or plasmogamy. A Filamentous Fungus: Rhizopus (Black bread mold) . is not immediately followed by the fusion of the nuclei. in fungi the fusion of the cytoplasm. each having one nucleus from the plus strain and one from the minus strain. Each mitotic division therefore results in two pair of nuclei that are dikaryotic. A. However.

Sporangia can be seen in the image of Rhizopus at the right (top).the asexual reproductive cells of the fungus. appearing as a fuzzy white mat growing all over the surface. Note that each sporangium is composed of hundreds of smaller spores . one may note small. zygospores composed of numerous nuclei) in regions where the plus and minus join. • Through a microscope. These are the sporangia. sporangium through 16x Rhizopus zygosporangium as seen through 16x objective.Cell Reproduction 30/11/2009 • On an amenable medium. • After the organisms in a culture initiate the sexual cycle. And even over the top of the petri dish sometimes. or "body" of a fungus. . Each strand is called a hypha (plural. Rhizopus can be seen with the naked eye.) This is the mycelium. hyphae). (And down into the medium. black balls growing at the ends of certain hyphae. Rhizopus: Rhizopus as seen objective. one may find zygotes (actually.

Cell Reproduction 30/11/2009 Rhizopus life cycle In the figure below. identify which cells are haploid and which are diploid. .

Basidiospores are formed by meiosis. in Coprinus four basisiospores arise from each basidium. Can you find all four basidiospores attached to a basidium in Figure 4 on the right? (Why not?) . Mushrooms Examine the diagrams and images of the mushroom Coprinus below. Therefore. Remember that the product of a meiotic division is four daughter cells.Cell Reproduction 30/11/2009 B.

above. What is the ploidy of basidiospores? Haploid Coprinus or Diploid . Figure 4 above: Basidiocarp gills as seen through 40x objective. Figures 2 and 3: Higher magnification (as seen through 10x objective) through the basidiocarp.Cell Reproduction 30/11/2009 Figure 1 above: Diagram of cross section through mushroom cap. Notice the gills radiating from the central stalk. and Figure 4. Notice the different stages of development of basidiospores. Notice the basidiospores (stained red in Figure 2. below) attached to gills at basidia.

Make a diagram like the one below. diploid. C. showing how one dikaryotic cell can divide by mitosis to form two dikaryotic cells. karyogamy. Remember that plasmogamy refers to the fusion of the cytoplasm. or dikaryotic (n + n). A Cup Fungus . (Click here to review mitosis). of the nuclei.Cell Reproduction 30/11/2009 In the life cycle below determine if each tissue is haploid.

or asci. or "cup". holding the spores (each ascus contains 8 ascospores. In the lower image. numbered in one ascus in the lower image). look for the sac-like structures. The eight ascospores are formed by meiosis (to produce 4 cells) followed by a mitotic division (to double the number to 8).Cell Reproduction 30/11/2009 Examine the diagrams and images of Peziza (an ascomycete) that show longitudinal sections through the fruiting structure. Peziza .

Cell Reproduction 30/11/2009 The life cycle is similar to that for basidiomycetes (like Coprinus) except the dikaryotic stage is much shorter. Are the spores part of the sexual or asexual cycle of a basidiomycete's life history? Sexual or Asexual .

By the end of this exercise. a gametophyte from a sporophyte. To study the evolution of these vegetative structures in representative plants: a moss. a gymnosperm. and whether they are formed by mitosis. You should understand which cells are haploid and which diploid. a typical fern.Plant Evolution 30/11/2009 Plant Evolution Objectives: The objectives of this lab are as follows: • 1. • 2. You should be able to apply this information to the reproductive cycle in gymnosperms such as pine. and both a woody and a nonwoody angiosperm. you should be able to identify the various lignified cell types in cross sections of stem. and a gymnosperm. To examine the evolution of sexual cycles in a moss. meiosis. To look at cellular specialization in plants. a fern. You should know how to tell a moss from a fern. . Section 1: The evolution of multicellular plants involved the modification of different cells to perform distinct functions. • 3. with emphasis on lignified cells that function in water transport and/or support. or fertilization. Part of the success of land plants was based upon the evolution of a very strong molecule called lignin. In this laboratory you will learn how to identify three lignified cells.

Fibers:also evolved from tracheids. combined the functions of support and water transport.Tracheids: first to evolve. 2. 3.Reproduction: Protists and Fungi 30/11/2009 • • • 1. . specialized for water transport.Vessel elements: subsequently evolved from tracheids. cells specialized for support only.

stem-like. Evolution of Vegetative Structures A. Moss 1. You can see more of the details by magnifying the image. . and other parts of the plant. phloem.Reproduction: Protists and Fungi 30/11/2009 All three can be found in xylem. I. while fibers can be found in xylem. Observe the living green "leafy" gametophyte stage on the right. and root-like structures. Remember that these organisms have leaf-like.

c. d.Reproduction: Protists and Fungi 30/11/2009 2.The transport tissue is located in the middle of the stem. • Find: a. the "leaves" attached to the epidermis .The cortex is composed of all the tissue from the epidermal cells to the transport tissue. Now examine the cross section of a moss (Mnium) "stem". You can see the general structure of the "stem. the multicellular cortex region . the transport tissue .Epidermal cells are located on the outer wall of the moss stem. the outer epidermal cells . b." Magnify the image and identify the following structures. ."Leaves" are an extension of the epidermis.

stems. Ferns . they are not lignified so they are not called "vascular tissue. moss is a nonlignified plant.] B. While the central cells are specialized to transport water. and roots. The central portion of the "stem" is composed of much thinner cells. moss plants do not have leaves." [And therefore.Reproduction: Protists and Fungi 30/11/2009 (Although some of the cell walls in the cortex are stained red.

How tall are these sporophytes? What structures are specialized for photosynthesis? 2. Osmunda cinnamomea. . Examine the pictures of live ferns. each smaller bundle extends to a different leaf. In the fern. the leaves extend in clusters from an upright position of the stem. Study the cross section of a fern rhizome (an underground stem) of the Cinnamon fern.Reproduction: Protists and Fungi 30/11/2009 1. Thus. or frond. Magnify the image to identify the different structures. you will find a central vascular region surrounded by a ring of vascular bundles.

the vascular bundles .Reproduction: Protists and Fungi 30/11/2009 • Find: a. the central vascular region b.

xylem tracheid cells .Reproduction: Protists and Fungi 30/11/2009 Click here to enlarge the image yet again. Within a vascular bundle find: a.

Examine the picture of a live pine tree. the xylem in this fern is composed of tracheids only.Reproduction: Protists and Fungi 30/11/2009 As is the case with most ferns.) C. How tall is the sporophyte stage? Are there specialized photosynthetic structures. lignified cell walls are stained red -. Pine (a gymnosperm) 1. (Remember. and other portions of the plant to support them? Do these organisms have specific structures that penetrate deep into the soil to obtain water? .along with tannins and other materials with a net negative charge.

Examine the cross sections of a young twig on the right of a 3-year-old pine (Pinus) stem.Reproduction: Protists and Fungi 30/11/2009 2. .

Reproduction: Protists and Fungi 30/11/2009 .

the bark 1. . phloem Now click here to magnify the image even more.Reproduction: Protists and Fungi 30/11/2009 • Under low magnification find: a. the central pith b. three layers (annual rings) of xylem c. cork 2. cortex 3. You should see more of the cellular detail.

cells that function both for water transport and support. This resin makes the cut ends of twigs sticky. Under low magnification you can see them in both the woody part of the twig as well as in the bark. Remember that in pine wood.Reproduction: Protists and Fungi 30/11/2009 You should notice that all of the cells with red cell walls look alike -. . and are a defense mechanism in these plants. The cylindrical structures surrounded by green-staining cells are the resin canals. the only lignified cells are tracheids.other than slight differences in diameter in the early "spring" wood and the late "summer" wood found in each annual ring.

Click on the image to magnify it. the tracheids b. Note the bordered pits in the cell walls. Angiosperms I. the protective cell that surrounds the outside of the stem (What are they called?) Since it is difficult to determine the cell types using cross sections. D. A Woody Plant: Basswood (Tilia) 1. the thin-walled phloem cells that transport "food" throughout the tree d. an alternative method can be used called "maceration. the resin canals c. Examine the macerated pine wood on the right to see what the tracheids look like from the side.Reproduction: Protists and Fungi 30/11/2009 • Under high magnification find: a. . Examine the cross section through a basswood twig." The wood is treated with chemicals to make the cells come apart.

Reproduction: Protists and Fungi 30/11/2009 .

the central pith b.Reproduction: Protists and Fungi 30/11/2009 • Find: a. those in angiosperm wood are composed of at least two cell types. Click here. . phloem 2. three layers (annual rings) of xylem c. cortex 3. cork Notice that although the water conducting cells in pine wood were fairly uniform (Pinus stem). the bark 1.

Reproduction: Protists and Fungi 30/11/2009 .

Therefore. if you identify anything with red-stained cell walls as xylem. But note that there are lignified cells in the phloem region also! Click here. Therefore. examine the prepared slide of macerated basswood . You can click on the image to magnify it. It is easy to tell vessel elements in cross sections because of their size. The lignified cells with smaller diameters are either tracheids or fibers. you may be wrong! Lignified fiber cells can be found in the phloem as well. In the outer portion of the twig. the outermost layer is cork and the inner bark is phloem.Reproduction: Protists and Fungi 30/11/2009 The lignified cells with the largest diameters are the vessel elements. but more difficult to distinguish a tracheid from a fiber. . the difference can only be determined from a side view (see diagram of lignin cells in this lab's introductory page). Under higher magnification you will be able to see that the red-stained cell walls are very thick. and that the hollow cavity is very narrow.

Reproduction: Protists and Fungi 30/11/2009 .

the vascular bundles 2. the cortex Using higher magnification find in the vascular bundle: . • Under low magnification find: 1. a tracheid b.Reproduction: Protists and Fungi 30/11/2009 • Identify: a. the pith 3. a fiber c. A nonwoody plant: Buttercup (Ranunculus) Examine the Cross-Section of a Buttercup Ranunculus Stem. a vessel element II.

Reproduction: Protists and Fungi 30/11/2009 1. vessel elements 2. fibers .

In moss. While the fern sporophyte is well adapted to land. In ferns and all other vascular plants. In ferns (and other "seedless vascular plants") the gametophyte is a free-living. these are phloem fibers. autotrophic organism.Reproduction: Protists and Fungi 30/11/2009 There are lignified cells in the portion of each vascular bundle closest to the outside (cortex) and other lignified cells in each bundle closest to the pith. All plants have the sexual cycle that alternates a diploid sporophyte generation and a haploid gametophyte generation. These changes will be observed in the coniferous gymnosperm. And. you are seeing fibers. you are more likely to encounter this phase of the sexual cycle). the fern gametophyte is not. once again. since this is the phloem on the outside of each bundle. Once again. the gametophyte is "dominant" (or predominant. However. The large diameters of the lignified cells on the inside are indicative of vessel elements. Section 2 The evolution of plants also involved alterations in the sexual cycle. in the subsequent evolution of land plants. this haploid portion of the sexual cycle was modified in several ways that allow these plants to survive increasingly dry climates. and are therefore in the xylem. and the hollow cavity is very narrow. the cells on the outside have red-stained cell walls that are very thick. pine. the gametophyte forms gametes by mitosis. Changes in Sexual Cycles . The sporophyte forms spores by meiosis. the sporophyte is predominant. Therefore.

Reproduction: Protists and Fungi 30/11/2009 .

.Reproduction: Protists and Fungi 30/11/2009 Remember that in the last lab you learned two types of sexual cycles. • A third cycle (center) occurs in all plants and some multicellular protists. the diploid phase is usually multicellular. In this cycle. meiosis is immediately followed by fertilization. haploid spores are formed by meiosis. In this instance. the only diploid cell being the zygote (top). The zygote formed by the fusion of two gametes divides by mitosis to produce a multicellular diploid phase. fertilization is immediately followed by meiosis. • In the other. These spores divide by mitosis to produce a multicellular haploid phase that produces gametes by mitosis. the only haploid cells being the gametes (bottom). • In one. The rest of the cells in this cycle are haploid.

.Reproduction: Protists and Fungi 30/11/2009 In the alternation of haploid and diploid generations the diploid phase produces spores and is therefore called the sporophyte (“spore-plant”). The haploid phase produces gametes and is therefore called the gametophyte (“gamete-plant”). See the top diagram to the right.

the gametophyte generation is predominant ("dominant") and the sporophyte grows out of the gametophyte (right bottom). as in seed plants. the gametophyte may be free-living. Moss 1.or . as in ferns. Study the image of a moss antheridium.one haploid cell dividing to form two haploid cells Meiosis: one diploid cell dividing to form four haploid cells Fertilization: two haploid cells fusing to form one diploid cell (the zygote) A.Reproduction: Protists and Fungi 30/11/2009 NOTE: Both the gametophyte and the sporophyte stage may be duplicated by asexual reproduction .the resulting offspring being clones. . In the bryophytes such as moss. What type of cells are formed in these structures? Click on the image or the word "magnify" to get a better look at the cells. In all vascular plants the sporophyte is predominant (left bottom). This basic alternation of generations has two variations. or may be dependent upon the sporophyte. ALWAYS REMEMBER: Mitosis: one diploid cell dividing to form two diploid cells .

Reproduction: Protists and Fungi 30/11/2009 .

Reproduction: Protists and Fungi 30/11/2009 2. . The sperm must swim through liquid water to the opening at the top of the vase. then swim down the channel before the egg is fertilized. Click on the image or the word "magnify" to enlarge the image. forming the zygote. you will see a large cell (egg) within a vase-like structure. Now study the image of a moss archegonium. If it is a good section.

Reproduction: Protists and Fungi 30/11/2009 .

Examine the whole moss plant in the sporophyte stage. or is it dependent upon "food" from the gametophyte? .Reproduction: Protists and Fungi 30/11/2009 Is this method of gamete transfer well adapted to a terrestrial habitat? 3. Do you think this brown structure can carry on photosynthesis.

sporophyte or. gametophyte . b.Reproduction: Protists and Fungi 30/11/2009 Label each stage as either: a..

antheridium b. egg e. archegonium d. you can pass your hand across the top. The results will be a cloud of fine particles. sperm c. spore g. These are the spores formed by meiosis that will germinate to develop into the new gametophyte generation. zygote f.Reproduction: Protists and Fungi 30/11/2009 • Label: a. protonema NOTE:If you should discover living sporophytes outdoors. Ferns . B.

Some ferns. the spores (and gametophyte cells) are haploid. will germinate to form the gametophyte generation. under the correct conditions. The small brown or black cells are spores that.Reproduction: Protists and Fungi 30/11/2009 1. such as the cinnamon fern (previous section). Examine the live fern leaf ("frond") that has brown or black specks ("sori") on the underside. have all of the sporangia on a separate "fertile fond. Were the spores formed by mitosis or meiosis? ." The sporophyte cells are diploid.

. Examine the images of the fern antheridium and archegonium. Click on the images to get a closer look at the cells.Reproduction: Protists and Fungi 30/11/2009 2.

Reproduction: Protists and Fungi 30/11/2009 .

Reproduction: Protists and Fungi 30/11/2009 .

Reproduction: Protists and Fungi 30/11/2009 Answer the following questions: • • • • A. Are the egg and sperm haploid or diploid? C. Are the gametes formed by mitosis or meiosis? D. Do the gametophyte cells contain chloroplasts? How do you know? . Are the gametophyte plants haploid or diploid? B.

Reproduction: Protists and Fungi 30/11/2009 Examine the drawing of the fern life cycle to the right. .

these plants are heterosporous. or pollen grain. Pine In both of the plants you have observed so far. b)gametophyte Click here to reload the image. spore C. each forming four haploid microspores. sperm c. Some of the nuclei. All gymnosperms and angiosperms are heterosporous. egg e. the plants were homosporous. the sperm nuclei. zygote f. a) sporophyte or.Reproduction: Protists and Fungi 30/11/2009 • Label each stage as either. archegonium d. antheridium b.. . Each scale in the smaller male "staminate" cones contains numerous diploid microsporocytes or "microspore" mother cells that go through meiosis. Each microspore undergoes several mitotic divisions to form a microgametophyte. are capable of fusion with an egg nucleus.. the spores formed by meiosis all look the same. Now.... A variation on this theme is where some spores are larger than others. • Label: a.

• 1. forming a multicellular megagametophyte. each containing a single megasporocyte (megaspore) mother cell which goes through meiosis to form four haploid megaspores. each containing one egg. three megaspores degenerate while the fourth continues to divide mitotically. Observe the image of the smaller male pine cone.Reproduction: Protists and Fungi 30/11/2009 The scale in the larger female ("ovulate") cone has two ovules. . In this case. This megagametophyte has two archegonia.

.) You can see pollen being released from the male pine cones in the third image on the right. pollen is a MICROGAMETOPHYTE.Reproduction: Protists and Fungi 30/11/2009 What kinds of cells are found in these cones? Are the cells released from these cones haploid or diploid? (Remember.

.Reproduction: Protists and Fungi 30/11/2009 The forth image on the right is a magnified picture of actual pollen grains.

. Now observe the image of the larger female (ovulate) pine cone.Reproduction: Protists and Fungi 30/11/2009 • 2. the type you usually see in pine cone wreaths.

.Reproduction: Protists and Fungi 30/11/2009 What kinds of cells were formed WITHIN each of these leaf-like scales? The sixth image on the right depicts a pine seed found within the scales of the female pine cone. How do the gametes produced by the male cone get to the egg in the female cone? And THEN what happens? • 3. Examine a seed which is cut open. the seventh image on the right.

Reproduction: Protists and Fungi 30/11/2009 Look for the embryo with its numerous cotyledons. Look for the white storage tissues around the embryo. Remember. this storage tissue is the MEGAGAMETOPHYTE. .

surrounding nutrient tissues (MEGAGAMETOPHYTE) 3. outside seed coat (including a larger "wing"): Examine the drawing of a pine life cycle. .Reproduction: Protists and Fungi 30/11/2009 Where was this seed formed? What specialized structures help disperse these seeds away from the parent plant? (And why is this important?) Which of these cells are haploid and which diploid? • 1. embryo with cotyledons 2.

b) gametophyte Now click here to reload the image.. What other gymnosperms still live on earth? How are they grouped by taxonomists? Do any gymnosperms have a wider distribution than conifers? III Review: Putting It All Together .. seedling Pine is only one example of a gymnosperm. seed coat i. archegonium d. microspore e. • Label: a.Reproduction: Protists and Fungi 30/11/2009 • Label each stage as either. megagametophyte c. megaspores b. embryo h. pollen tube g. microgametophyte (= pollen) f. a) sporophyte or.

What are the functions for tracheids? How can you identify a tracheid in a cross section? What plants did you observe today that had tracheids? What do they have in common? • 2. What do they have in common? How do they differ? 6. What do they have in common? How do they differ? 5. What is the main function of a fiber? How can you identify a fiber in a cross section? In what tissues did you find fibers? What plants did you observe today that had fibers? What do they have in common? • • 4. with lots of parts. such as most ferns. Think about all of the figures that illustrate fibers. Change this life cycle to represent the situation for a heterosporous plant. What is the main function of a vessel element? How can you identify a vessel element in a cross section? What plants did you observe today that had vessel elements? What do they have in common? • 3. The "basic life cycle" illustrated in this lab is for a homosporous plant. Think about all of the figures that illustrate vessel elements. Compare the sexual cycles for moss and ferns. Think about all of the figures that illustrate tracheids.Reproduction: Protists and Fungi 30/11/2009 This is a complicated lab. • 1. so it is essential that you get an overview before try to sort out the details. • . such as pine. Compare the sexual cycles for ferns and pine.

Reproduction: Protists and Fungi 30/11/2009 .

• To practice forming three-dimensional images from two-dimensional observations. and relate these modifications to changes in function.Angiosperm Reproduction 30/11/2009 Angiosperm Reproduction Objectives: • To study the development of the different parts of the flower. • After completing the laboratory exercise you should know the parts of a flower and the functions of each. Again. Next. To observe some of the variation in different parts of sample fruits. Finally. Introduction: This lab is a continuation of the previous lab in which you studied some of the basic aspects of plant evolution. you should appreciate how plants have evolved different structures for pollen transfer and for fruit dispersal. review the generalized life cycle at the beginning of Part II of lab 7. and how these regions further develop to form the fruit with its seeds. Click here to briefly review the evolution of land plants. You should be able to recall which part of the flower develops into each portion of a fruit. you should be able to recognize which cells are haploid and which diploid. and whether they are formed by mitosis or meiosis. .

The megagametophyte was still large enough to provide the nutrients for the young embryo in the mature seed. I. starting as a free-living. The megagametophyte was maintained within the sporophyte cone.Plant Evolution 30/11/2009 Finally. you will observe several of the structures associated with the reproductive portion of the plant life cycle. This trend of reducing the gametophyte generation is continued in angiosperms. You will start by observing the development of the megagametophyte ("embryo sac") as well as that of the microgametophyte ("pollen"). Review the main parts of the flower below. The Flower You have now come to a major evolutionary advancement of the plant kingdom. In the diagram. There are six stamen in this flower. These and other trends in plant evolution are diagrammed on the next page. You will then study the subsequent development of the seed and fruit. and surrounded by the sporophyte layer called the integument (later to become the seed coat). windblown structure called "pollen". photosynthetic organism in ferns. review the evolution of the gametophyte generation. parts of the male reproductive structures. are labeled in green. or stamen. In this laboratory. In gymnosperms like pine. the microgametophyte was reduced to a small. multicellular. You will also examine examples of the different types of fruits to see how they have evolved different structures for protection and dispersion. .

On the right. at the base of the style. On the left. label the external parts of the pistil (sometimes also called the "carpel"). but is located in the center of the flower. the male reproductive structures. label the parts of the stamen. Note that the ovary is not visible in the diagram.Plant Evolution 30/11/2009 The female reproductive structures are labeled in blue. The diagram below illustrates the components associated with sexual reproduction. the female reproductive structure. Sexual Structures of a Flower . style and ovary make up the single pistil in this flower. The stigma.

• We can speculate that one of the functions of sepals is to hide developing flowers from pollinators. and may be one of the selective advantages for having inconspicuously colored sepals--they are often green! They may also protect fragile flower parts from damage.Anther 2. Ovary Although you did not label the sepals (outer whorl(s) of modified leaves) and petals (inner whorl(s) of modified leaves) in the previous illustration. than petals. you should know the function of each: • Sepal: To protect the developing flower bud. to provide a landing area for insect pollinators.Plant Evolution 30/11/2009 1. and so they are sometimes thicker. or fleshier. This prevents colored petals from attracting insect visitors before pollen is ready for dispersal. Stigma 4. Are there other ways to distinguish sepals from petals? . Filament 3. Petal: To attract pollinators with colors and/or odors. Style 5.

Gladeolus Flower Dissection If the petals and sepals are removed from a gladeolus flower. The Anther . Ovule 7. Stigma 3. Pollen grains 2. Label the parts and identify each as belonging to either the sporophyte (diploid) or gametophyte (haploid) generation. the remaining parts are the stamens and the pistil (sometimes called the carpel. Pollen tube 5.) When a longitudinal cut is made down the ovary. the line of ovules inside becomes visible. A. The following diagram shows the gladeolus flower dissected in this way. Style 4. Flower Pistil After Pollination 1.Plant Evolution 30/11/2009 The figure below depicts the internal features of the pistil after pollination. Ovary 6.

Anther 6. Each anther consists of two pollen sacs. Note the style in the center of the circle of anthers. one on each side of a vascular bundle. Stigma 2. Filament Cross Section through Flower Pistil and Anther . Style 3. Ovules 5. which illustrates a cross section through flower sexual structures.Plant Evolution 30/11/2009 Study the diagram below. 1. Ovary 4.

. to the right. An anther with mature pollen is drawn in the diagram below. anthers (and their pollen sacs). two pairs per anther. filament. This is a large structure-. continued.) 2. and vascular bundles.Plant Evolution 30/11/2009 1. A. The Anther. Style Compare the diagram above with the prepared slide of the Lilium anther. Anther (pollen sacs are within the anthers.this image was captured with the 6x scanning lens! Scroll to find the style.

and vascular bundles. (Look for grains where the section was right across the upper or lower surface of the pollen.edu/~gb101/lab8_angio_repro/lilyanther2. Examine a prepared anther using higher power by clicking on the image below to magnify. Count the number of grains and divide this into the known field of view. . Then estimate the size of the pollen grains.) Second. (You will have to look to find a pollen grain where the histological section was made in such a manner that both nuclei were in the same slice.rutgers.2mm. the cell wall has a definite structure. pollen sacs. Cross section through Lilium anther In the high magnification image to the right (http://bio.) Remember that pollen is a MICROGAMETOPHYTE formed from a MICROSPORE. If the field of view in the thumbnail image below is 1. find pollen.jpg). each pollen grain has two nuclei.Plant Evolution 30/11/2009 Two features of the pollen are worth noting: First. how large is one pollen grain? Find a cluster of grains that line up across the diameter of the field.

Microgametophyte Development 0. 3. divides to give rise to four haploid microspores.003 mm. You may review the procedures for estimating cell size using a known field of view in Laboratory 1.Plant Evolution 30/11/2009 1. or microspore mother cell.1 mm? The development of the microgametophyte (male gametophyte) in flowering plants involves three stages: • A diploid microsporocyte. 2. Do you estimate that the lily pollen is 0.05 mm. . or 0.

B.Plant Evolution 30/11/2009 • Each microspore divides by mitosis to form two haploid nuclei (the tube nucleus and the generative nucleus). • After landing on the stigma. the generative nucleus then divides by mitosis to form two haploid sperm nuclei. the pollen germinates forming a pollen tube. The Ovary . this is mature pollen in Lilium.

giving rise to four haploid megaspore nuclei (3 of which disintegrate).Plant Evolution 30/11/2009 The image below is an illustration of a cross section through Lilium ovary. How many ovules are in this ovary cross section? 2. the ovules. the ovules are attached in pairs to a central stalk. . note the small vascular strands in the central region. you may not always see all three pairs of ovules in one section. 3. A diploid megasporocyte (megaspore mother cell) undergoes meiosis. as might be seen using a scanning lens. In the Lilium ovary. and the protective ovary tissues around the outside. or 6? The development of the megagametophyte ("embryo sac") in most angiosperms involves three stages. In prepared slides. In the image to the right.

depending upon the species. the number of antipodals can vary from none to many.Plant Evolution 30/11/2009 The remaining megaspore nucleus undergoes three mitotic divisions to form eight haploid nuclei. Seed and Fruit Development A. Development of a Seed Capsella: a dicot . Nuclear migration and cytokinesis occur to form the mature megagametophyte While the above is "typical". III. The smallest megagametophyte only has an egg cell plus one polar nucleus.

This plant has numerous heart-shaped fruits ("purses") on a long stem.Plant Evolution 30/11/2009 Examine several slides of the cosmopolitan weed Capsella bursa-pastoris (Shepherd's purse). and fruit at more mature stages are at the bottom. Each of these fruits contains dozens of minute seeds.) The white flowers and developing fruits are at the very top of the plant. (See the illustration below. . Capsella seed development Review some of the basic parts of the developing seed using the illustrations below.

however. are exactly parallel to the embryonic axis. Use the following illustration to help understand these sections. you will note that many of the seeds have been cut at other angles. so that the stored food in the mature seed is in the cotyledons. and in the illustration below. (The same pattern is also true for the legumes. .) The sections from the previous illustrations. including the pea and bean seeds that you will study in the next section. As you examine the prepared slides.Plant Evolution 30/11/2009 Note that in this plant the endosperm is absorbed by the developing cotyledons.

Seed and Fruit Development • B. When you split open a pea pod. . with sepals and petals modified into several different shapes. Fruit Development Fruit development is a complex process. you can get a good impression of how the flower develops into the fruit. III. For example. Identify what the resulting section would look like if the section were made at the blue line. Three examples of the flower and then the fruit are found in the figures to the right.Plant Evolution 30/11/2009 The diagram above shows four of the infinite planes where the seed could be sectioned. The ovules are attached down a single row in the ovary so that the seeds develop in the same way. in peas the flower is "irregular" (that is. it has bilateral rather than radial symmetry). The angle of the cut is shown in blue. varying considerably among different plant groups.

Plant Evolution 30/11/2009 Complete your collection of life histories by adding two angiosperm sexual cycles. . the bush bean. The figure on the right is the sexual cycle of a dicot.

corn. .Plant Evolution 30/11/2009 Corn. Zea mays The images below show a monocot.

note that while the materials in the endosperm have been transported to the cotyledons during bean seed development (as in Capsella).Plant Evolution 30/11/2009 While completing the corn life cycle. C. the endosperm still exists in the mature corn kernel. Fruit Diversity .

These dispersal mechanisms range from structural modifications such as barbs that stick in fur. often because of the fruits. • Domestication: A variety of plants have been cultivated. who consume the whole fruit. on the right). . The original evolution of the fruit was for dispersal. For example. The Peach: an example of fruit dispersal mechanisms The outer layer ("skin") protects the next layer from drying out.Plant Evolution 30/11/2009 • Dispersal: Angiosperms have evolved a number of different methods for protection and dispersal of the next generation. The main central layer attracts animals. or buoyant structures that carry fruits and/or seeds through water. along with some "fertilizer". The pit is later deposited. to sweet fruits that are eaten by animals that disperse seeds (such as the peach. To continue. note the three layers of a peach on the image below. The inner layer ("pit") protects the seed during the process. click on the area of the image that was the ovule when this fruit was still a flower. after in has passed through the animal's gut.

(http://bio.rutgers.jpg) . Significance of grains Examine a corn "cob".edu/~gb101/lab8_angio_repro/cornkernelhigh.Plant Evolution 30/11/2009 3.

Plant Evolution 30/11/2009 .

. Examine the prepared slide of a corn kernel above.Plant Evolution 30/11/2009 The kernel is technically a fruit. not a seed. (The scientific name for corn is Zea mays). since the seed coat and fruit wall are fused.

The hierarchial nomenclature used in plant anatomy is often confusing. In this laboratory you will: . and is therefore outlined to the right. Note that both xylem and phloem tissues may also contain cell types that do not function in transport. Click here to get table. The cells responsible for long-range water transport have already been studied in Lab 7: the tracheids and vessel elements found in xylem. Introduction: The transport systems in plants are quite different than the circulatory system found in vertebrates. one to move water (and the minerals dissolved in it) from the roots to the leaves and the other to move organic compounds (mainly sucrose) from the "source" to the "sink". The cells responsible for long-range transport of organic compounds are found in the phloem. To investigate the structures associated with transport of water and of organic nutrients. To study water transport in plants. Vascular plants have two transport systems. 2.Biology Lab Practical Lab 9 Transport Systems in Plants Objectives: • • 1. such as the phloem fibers observed in Lab 7.

perform an experiment that suggests a mechanism for this movement 3. the vascular strands running longitudinally in the stem can be observed directly. congo red dye will be added to the water to trace the pathway of water up the stem.Angiosperm Reproduction 30/11/2009 1. thin-walled cells). Cross-sections of the stem demonstrate where in the stem the water is moving and obliquesections allow identification of the actual cells involved in water transport. Are the cells involved in water transport lignified? Lets start the experiment and see!!! Part I : Step 1: Impatients have translucent stems (due to the presence of large. Notice the strands in the picture to the right. Water Transport Pathway The purpose of this experiment is to examine the movement and pathway of water up an Impatiens stem. In the first part of the experiment. which cells. Click here to see how that was done. Cut off another centimeter of stem under water to ensure there are no air bubbles in the transport tissue that could disrupt water transfer. examine prepared slides to study the cellular structures in more detail II. and more specifically. confirm the path of water movement between the uptake by root hairs and the loss through stomata 2. . Therefore. Step 2: Cut the root system from the plant and immediately place the cut surface of the stem in water. are lignified. Click on the picture to label the strands with arrows. The second part of the experiment identifies which parts of the stem.

Angiosperm Reproduction 30/11/2009 Step 3: Quickly transfer the cut stem to a test tube containing a solution of Congo red and prepare your controls while you wait for water transport to occur up through the stem. Click here to observe the congo red setup. .

Angiosperm Reproduction 30/11/2009 .

it is first necessary to see if any of the cell walls of the plant are already stained red.Preparation of sections . This is control 1. (Congo red stains cellulose). however.Angiosperm Reproduction 30/11/2009 Step 4: Two controls are needed for this experiment. Add a drop of Congo red to a second • a. The second control is prepared in the same way. To do this mount a thinly cut section of stem from the piece cut off in step 2 on a microscope slide and view it under a dissecting microscope. this control is to check which parts of the stem Congo red will stain slide for control 2. Click on the following to observe each each part of step 4. Since the purpose of the experiment is to trace the movement of congo red dye to follow the pathway of water transport.

View Control 1 and 2 on the slide .Angiosperm Reproduction 30/11/2009 b.

View Control 1 under the dissecting scope .Angiosperm Reproduction 30/11/2009 c.

Angiosperm Reproduction 30/11/2009 d.View Control 2 under the dissecting scope .

cut cross-sections of the stem at various points along the stem. Also make longitudinal sections at the same points to identify the cell type. Click on the following to observe each part of step 5. Examine the location of the congo red.Angiosperm Reproduction 30/11/2009 Step 5: After the dye has moved to the stem apex. View slides of cross and oblique sections from each point . • a. Sections at various points along the stem b.

Angiosperm Reproduction 30/11/2009 .

. • Step1:Cut additional cross and longitudinal sections to stain lignified cell walls using phloroglucinol.Angiosperm Reproduction 30/11/2009 Part II: The purpose of the second part of the experiment is to determine whether or not the cells involved in water transport are lignified. Click on each step to observe the procedure.

.Angiosperm Reproduction 30/11/2009 • Step2:Soak sections in 2-3 drops of phloroglucinol for about 3 minutes.

Angiosperm Reproduction 30/11/2009 • Step3:Drain the stain off using a paper towel. .

(Lignin stains red) .Angiosperm Reproduction 30/11/2009 • Step4:Add 2-3 drops of concentrated HCl.

.Angiosperm Reproduction 30/11/2009 • Step5:Place a cover slip over the sections and examine it under the dissecting microscope.

Angiosperm Reproduction 30/11/2009 .

or negative pressure. If the above mechanism is correct. Begin Experiment: . This column is maintained by the hydrogen bonding between water molecules (cohesion) and the hydrogen bonding between water molecules and the molecules that make up the cell walls (adhesion). that pulls the water column up the plant. water evaporating through the stomata produces a tension. Water Transport Mechanism According to the transpiration-adhesion-cohesion-tension mechanism of water transport in xylem. The rate of movement can be determined by adding a dye to the water at the bottom of the pipette. then a twig that has been removed from the rest of the plant and attached to an artificial water column (as in a pipette) could pull water up the pipette.Angiosperm Reproduction 30/11/2009 II.

. Next. Manipulate the apparatus until it is completely filled with water . click on the yellow tubing to place it on the pipette.Angiosperm Reproduction 30/11/2009 1.NO air bubbles. It will fill with water. click on the pipette to place it in the dish pan. To do this. Place a transport apparatus (1 mL pipette with a piece of clear tubing on the larger end) under water using the large dish pan.

Add about 20 mL of red colored dye (Congo red) to the bottom of a small beaker. Insert the freshly cut end into the above tubing while under water. making certain that NO air bubbles are trapped within the assembly. For our purposes. wrap several strips of Parafilm around the stem before inserting it into the tubing. . Attach the twig firmly with wire using the pliers. Remember: NO air bubbles! Click on the test tube to add the Congo red. 3. This same set-up is repeated with a juniper stem without leaves to use as a control. Click on your set up to complete assembly. Click on the juniper twig to place it into the dish pan. Transfer the whole thing to the ring stand. assume it fits snugly. Next click on the pipette/twig assembly to place it into the test tube. transfer the assembly so that the tip is now into the dye.Angiosperm Reproduction 30/11/2009 2. Place the cut end of the juniper twig under water and cut off another centimeter to insure that there are no bubbles in the transport tissue that would disrupt water transport. Clamp it into place and turn on the light at the top of the assembly. 5. 4. If the twig does not fit tightly in the clear tubing. Click on the knife to cut off an additional cm under water. Next click on the twig again to insert it into the tubing. With your finger over the tip of the pipette. Click here to see the control's set-up.

At periodic intervals. look for a movement of dye up the pipette. . Click here to start the experiment.Angiosperm Reproduction 30/11/2009 6.

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NOTE: The movement of the dye observed takes approximately half an hour in real time. Click here to see the set-up and results of this experiment from a laboratory class. Click on either beaker in the experiment to get a close-up of the final results.

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A. The Root

1. Whole Roots Before examining the root ultrastructure, lets take a look at the parts of the whole root. We're going to examine the root system of a Cyperus(top right)

Angiosperm Reproduction

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and a Begonia (bottom right)

Angiosperm Reproduction

30/11/2009

. Click on the cyperus to examine its rooting structures under water.

Thinner secondary roots (thin black arrow) emerge from the older primary root. .Angiosperm Reproduction 30/11/2009 Next click on the roots to examine a single root shoot. Note that the Cyperus has a large primary root (thick black arrow).

The stem grew new root shoots. Now click on the Begonia roots. . Notice the numerous root hairs. This form of continuous development is characteristic of higher plant structures. The root hairs increase the surface area of the root.Angiosperm Reproduction 30/11/2009 Note how the secondary roots originate at a definite position behind the root apex and become progressively longer (older) as you go up the primary root. This plant had been clipped and the stem was placed in water.

label the parts of the root by clicking on the image to your right.Angiosperm Reproduction 30/11/2009 Below to your left. view the cross section of the root as if you were using a microscope.The xylem is in the center of the circular root. cortex. and endodermis at high magnification.sign. Identify the epidermis and the xylem at low magnification and identify the phloem cells. Xylem . Increase magnification by using the + sign and decrease magnification using the . Indentify: Epidermis at low magnification – The epidermis is a one-cell thick outer covering of the root. . Once you have examined the section. You can also move the stage of the microscope by clicking on the button on the far right and moving the hand.

Angiosperm Reproduction 30/11/2009 Identify: .

Angiosperm Reproduction 30/11/2009 • phloem cells – The phloem cells are the cluster of green-stained cells in the arms of the X-shaped xylem • Vessel Elements .The cortex is the intermediate portion of the bark. • endodermis at high magnification .The vessel elements are the large conducting cells in the xylem in the middle of the vascular region. . • cortex . between the epidermis and the vascular tissue.The endodermis is a single layer of cells that encircles the vascular cylinder.

sieve tube members. Click on the image to your right to identify the epidermis. cortex. You may then click on "high magnification" to label the parts of the vascular region. vessel elements and tracheids.The epidermis is a one-cell thick outer covering of the stem. and vascular bundles. Identify: Epidermis . the image to your left can be manipulated as if viewed through a microscope.The cortex is external from the vascular bundles and internal from the epidermis. It has a thick waxy cuticle and stomata.Angiosperm Reproduction 30/11/2009 Just as in the root exercise. cortex . Identify the phloem fibers. vascular bundles – .

Angiosperm Reproduction 30/11/2009 Identify: phloem fibers . The Leaf . – The tracheids are the smaller conducting cells located in the xylem. sieve tube members .The phloem fibers are clustered toward the outside of the vascular bundle. tracheids. C.The vessel elements are the larger. forming a cap that strengthens the stem. They are on the inner side of the phloem fiber cap.The seive tube members are located in the phloem. vessel elements . more efficient conducting cells in the xylem.

Whole Leafs Before we examine the ultrastructure of the leaf. lets take a look at a whole living leaf. Notice the two leafs on your right. The top leaf is from the plant zebrine and the bottom leaf is from the plant coleus. .Angiosperm Reproduction 30/11/2009 1.

It is also possible to visualize the stomata on the leaf's surface. each leaf was painted with clear nail polish on both sides. Click on the zebrina leaf to examine the leaf's surface. The film of nail polish was peeled off and mounted on a slide to examine under the microscope. Notice which surface the stomata are located and their distribution. an impression of the leaf cells on the surface will be left.Angiosperm Reproduction 30/11/2009 Notice the major and minor veins (vascular bundles) running throughout the leafs. . The major vein(s) are labeled with a large arrow(s) and the minor vein(s) are labeled with small arrow(s). If all goes well. To do so. Next click on the coleus leaf.

Angiosperm Reproduction 30/11/2009 .

Angiosperm Reproduction 30/11/2009 .

Identify: upper epidermis . Click on the image to your right to identify the upper epidermis.Angiosperm Reproduction 30/11/2009 Again.The upper epidermis is a one-cell thick outer covering of the top of the leaf. stoma. lower epidermis. guard cells. cuticle. the image to your left can be manipulated as if viewed through a microscope. palisade mesophyll (with chloroplasts). . and spongy mesophyll.

Guard cells are specialized lower epidermal cells that are responsible for opening and closing the stoma.Angiosperm Reproduction 30/11/2009 lower epidermis . spongy mesophyll .The cuticle is the waxy layer secreted by the upper epidermal cells to reduce water loss. guard cells .The palisade mesophyll is the layer of closely packed columnar cells near the upper epidermis. In this image only a small area is visible. Cuticle . Stoma . .The spongy mesophyll is the layer of loosely and irregularly arranged cells near the lower epidermis. palisade mesophyll (with chloroplasts) .The lower epidermis is a one-cell thick outer covering of the bottom of the leaf.The stoma is a minute air space opening at the base of the leaf.

Angiosperm Reproduction 30/11/2009 .

Chromosome Structure and Meiosis 30/11/2009 Objectives: The objectives of this lab are as follows: 1. To study the events associated with meiosis. To apply this knowledge to human genetics by analyzing a karyotype. 2. To review the structure of a chromosome. 3. Introduction: .

The full complement of 46 chromosomes is referred to as the diploid number (referring to the fact that each kind of chromosome is represented twice). While mitotic division may occur in almost any living cell of an organism. If the egg and sperm were both diploid (46 chromosomes each in the case of humans) then the resulting zygote would be tetraploid. Thus humans have 23 homologous pairs. They exist as homologous pairs (partners) that are similar in size and shape and carry the same kinds of genes. Fruit flies have 8. This is referred to as the haploid number. It resembles mitosis in many ways but the consequences of meiotic divisions are very different from those of mitotic divisions.Biology Lab Practical Lab 9 Meiosis is the second important kind of nuclear division. . This would be an intolerable situation. Each species has a characteristic number of chromosomes per somatic cell. meiosis occurs only in special cells. In higher organisms when an egg is fertilized the egg and sperm fuse to form a single cell called a zygote which develops into a new organism. meiosis is restricted to cells that form gametes (eggs and sperm). In animals. so a mechanism has evolved to insure that each gamete (egg or sperm) contains only one representative of each homologous pair (or half the diploid number). normal humans have 46.

Study the diagrammatic summary of cell division in Meiosis I and Meiosis II in your textbook before you begin. The lily flower has six anthers surrounding one carpel. However the DNA is only synthesized once (prior to Meiosis I). all the cells in the sac are in the same stage. and can potentially result in the production of four cells.Biology Lab Practical Lab 9 Haploid Egg + Haploid Sperm = Diploid Zygote The mechanism that makes this possible is meiosis. Part II: You are now ready to find the actual stages of meiosis. metaphase. Luckily. telophase) but as we shall see the events are somewhat different. REMEMBER: These slides are thin two-dimensional sections through three-dimensional reality. It is in these microsporangia that you will observe the stages of meiosis. Meiosis in the anther starts with the diploid microsporocyte. These cells are still attached in the microsporangium. The subdivisions of meiosis are named like the subdivisions of mitosis (prophase. Each nucleus has a diploid number of duplicated chromosomes. Therefore. . anaphase. Meiosis I and Meiosis II. Each anther has two pair of microsporangia ("pollen sacs"). Meiosis consists of two divisions. using the lily anther. Part I: Lets review the stages of Meiosis. 1. you will have to look at several different cells in each microsporangium to see exactly what is going on. in the early stages of meiosis.

Early prophase I . the chromosomes are long and slender. . The cross-section of the lily anther to the left is in the early stages of meiosis.In early prophase. and the bright red nucleoli are still present.Biology Lab Practical Lab 9 We will now begin to identify the different stages of meiosis. The nuclear region will be clear and the nuclear envelope is still present. Are the cells in: 1.

and condensed. thick.In late prophase I. The nucleolus is absent and the nuclear envelope has broken down.Biology Lab Practical Lab 9 2. . the chromosomes are short. Late prophase I .

During metaphase I the pairs of chromosomes are distinct and line up near the center of the cell.Biology Lab Practical Lab 9 3. . Metaphase I .

.Biology Lab Practical Lab 9 4.During anaphase I the chromosome pairs are separated and the two groups of chromosomes migrate towards opposite sides of the cell. Anaphase I .

By telophase the two groups of chromosomes have completely separated and are positioned at opposite sides of the cell. . Telophase I .Biology Lab Practical Lab 9 5.

The nuclear envelope begins to reform. Interkinesis . The cell plate becomes visible between the .Biology Lab Practical Lab 9 6.

images of single cells have been taken for identification of these stages. In Meiosis II. all the cells in a microsporangium are not in the same stage of division. .Biology Lab Practical Lab 9 duaghter cells after interkinesis. Therefore.

Prophase II . Metaphase II . . 2. the chromosomes are condensed and two new nuclear envelopes are present.During metaphase II the chromosomes are distinct and line up near the center of the cell.Biology Lab Practical Lab 9 Look for: 1.In prophase II.

Biology Lab Practical Lab 9 3. 4. Telophase II .During anaphase II the chromatid are separated and the migrate towards opposite sides of the cell. Anaphase II . .By telophase the two groups of chromosomes have completely separated and are positioned at opposite sides of the cell.

The cell plate becomes visible between the dividing cells.Biology Lab Practical Lab 9 5.In cytokinesis. . the nuclear envelope begins to reform. Cytokinesis II . Four cells are now visible.

. so severe that most afflicted infants die within a few weeks after birth. Autosomal trisomy that produces physical malformations... Each karyotype has been started for you. your patient list is growing. Your job is to construct the karyotypes of your patients in order to look for possible chromosomal abnormalities. You better get started. The first 12 chromosomes have been matched. and mental and developmental retardation.Biology Lab Practical Lab 9 Part III: You finally graduated top of your class and have taken a position as a cytogeneticist. Case 1: This is a female with Patau Syndrome. determine whether or not your patient has a chromosomal abnormality and if so.. and to diagnose the patient with the expected symptoms of their karyotype. Your job is to match the remaining 11 chromosomes. which one. .

Biology Lab Practical Lab 9 .

Biology Lab Practical Lab 9 Case 2: This is a normal male karyotype. Healthy male. .

Biology Lab Practical Lab 9 .

Healthy female.Biology Lab Practical Lab 9 Case 3: This is a normal female karyotype. .

Biology Lab Practical Lab 9 .

Biology Lab Practical Lab 9 Case 4: This is a male with the XYY karyotype. Trisomy of the sex chromosomes producing males with less severe abnormalities. though they often have poorly developed genetalia and subnormal intelligence. This genotype is significantly higher in individuals found in penal institutions compared to .

Biology Lab Practical Lab 9 the general public and has been suggested to predispose these men to aggressive behaviour .

Biology Lab Practical Lab 9 .

Biology Lab Practical Lab 9 Case 5: This is a male with Trisomy 22. Autosomal trisomy in which the fetus does not survive. .

Biology Lab Practical Lab 9 .

Biology Lab Practical Lab 9

Case 6: This is a male with Downs Syndrome. Autosomal trisomy associated with physical features including broad head, rounded face, perceptible epicanthic folds of the eyes, a flattened bridge of the nose, protruding tongue, small irregular teeth, and short stature. Mental retardations is also

Biology Lab Practical Lab 9

characteristic. It is also called mongolian idiocy.

Biology Lab Practical Lab 9

Biology Lab Practical Lab 9

Case 7: This is a male with Edwards Syndrome. Autosomal trisomy that produces physical malformations, and mental and developmental retardation, so severe that most afflicted infants die within a few weeks after birth.

Biology Lab Practical Lab 9

Biology Lab Practical Lab 9

Case 8: This is a female with Patau Syndrome. Autosomal trisomy that produces physical malformations, and mental and developmental retardation, so severe that most afflicted infants die within a few weeks after birth.

Biology Lab Practical Lab 9 .

Nondisjunction is a failure of chromosome or chromatid to separate to opposite poles during nuclear division. The red chromosomes represent maternal genes and blue chromosomes represent paternal genes. When nondisjunction occurs. The large chromosomes are homologous and the small chromosomes are homologous. Two homologous pairs of chromosomes are present within the cell depicted below. two chromosomes or chromatid go to one pole and none go to the other. Trisomy occurs during meiosis when nondisjunction occurs. it is essential to understand how they occur. .Biology Lab Practical Lab 9 Now that you have correctly determined each of the above karyotypes. The abnormalities viewed above are mostly cases of trisomies.

Biology Lab Practical Lab 9 .

Biology Lab Practical Lab 9 .

Turners syndrome). The gamete missing a chromosome could meet a normal gamete. as seen in the previous case studies (i. A normal gamete may also meet a normal gamete in which the patient would be a normal male or female.Biology Lab Practical Lab 9 Depending on which gamete meets which gamete. .e. The following illustrate these scenarios.e. The gamete with the extra chromosome could meet a normal chromosome. Down's syndrome). three scenarios are possible. In this case the patient would have a trisomy in their karyotype. The result of this match would be a patient with a monosomy (i.

Biology Lab Practical Lab 9 .

Biology Lab Practical Lab 9 .

.Biology Lab Practical Lab 9 You have now completed the meiosis laboratory.

30/11/2009 .

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