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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.

• 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.


• 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. In fact, you can buy a ruler mounted on a microscope slide that is especially designed for

this.

You will do this more simply by placing a piece of graph paper on a microscope slide and viewing it under

the microscope. Each box on the paper is a 1 mm square, but for microscopy, millimeters are too large of a unit of

measure. Microscopic objects are measured in micrometers.

1 mm = 1000 micrometers

Let's try this at our lowest magnification using the scanning lens (40 X total magnification).
Now estimate the field diameter in micrometers.
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, each time the magnification is increased, the block on the graph paper gets larger.
If we try to estimate field size using graph paper with the 40 X objective, the block will be so large that we

will not be able see the lines clearly and it will be difficult, if not impossible, to estimate the field size.

So, 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. Therefore, the field

diameter is inversely proportional to the magnification. We can use a mathematical formula to estimate the field

size at 40 X.
Let's do this using the numbers we just calculated for the 10 X objective. Remember, the field diameter

with the 10 X objective lens was 1700 micrometers. Now fill in the numbers in the equation.
Now solve the equation for the field diameter of the high magnification lens, you get the .

D. Image Formation

When a microscope magnifies an image it shifts the orientation of the object you are viewing.

For example, if you cut out the letter "e" from a newspaper and put it under the microscope what happens

to the orientation of the letter?


The "e" is now inverted and shifted from right to left. What happens if we move the specimen stage to the

right?

To simulate this, place your cursor on the letter "e". Hold down the left mouse button and slide the cursor

to the right.

Now move the cursor to the left.


What happened?

Now try this.

Place your cursor on the letter "e". Hold down the left mouse button and slide the cursor upward.

Now move the cursor downward.

What happened?

II. Cells

A. Plant Cells
1. Elodea: leaf cells
Elodea is a decorative aquatic plant often found in fish tanks. 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. If you examine the leaf

using the scanning lens you will find the midrib running down the center of the leaf. The midrib contains the main

vein for conducting materials to and from the leaf.

Now turn to higher magnification and examine a portion of the leaf away from the midrib. Can you identify

the chloroplasts in the upper photo and the cell walls in the lower photo?
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.
Look at the onion preparation with the condenser diaphragm wide open. 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.


In this example, 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.

B. An Animal Cell
You will use a cheek cell (simple squamous epithelium) as an example of an animal cell. 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. A

cover slip was then added. What do you see under the microscope?
The image has low contrast. You can increase the contrast by closing down the diaphragm. When the

diaphragm is in the closed position, you will have maximum contrast. When it is in the fully open position, you

have maximum resolution but the image has very low contrast. Most of the time, we compromise and close the

diaphragm about half way to maximize resolution and contrast as much as possible.

Now look at a higher magnification view of an epithelial cell.


What cell structures can you identify?

C. Euglena

a single-celled eukaryote
Euglena is a single-celled eukaryotic organism that is often called a "plant-like" organism since it is

photosynthetic.

Look at the swimming Euglena cells.

Now look at a single Euglena cell. What organelles can you see in the cell?
Flagellum –

Nucleus –

Eyespot –

Chloroplast –
D. Bacteria

single-celled prokaryotes
Bacteria are extremely small and often are not visible with the 10 X objective lens. The prepared slide of

bacteria has three types of bacteria on it.

The round bacteria are often clustered in groups of two, four or more cells.

Now find the rod-shaped bacteria. Notice that not all of the rods are of the same length.
Now find the spiral-shaped bacteria.

To estimate the sizes of these bacteria switch to a higher magnification image.

At this magnification the field diameter is about 45 micrometers.


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

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


3. Estimate the size of the round bacteria.
Cell Reproduction 30/11/2009

How do cells reproduce?

Introduction to the Lab:

The emphasis of this laboratory period will be on mitosis.

Mitosis is the sequence of events by which the nuclear material of one cell is distributed, by a process

involving chromosomes, into two equal parts.

At the right is a longitudinal section through an onion (Allium) root tip. The root tip is responsible for the

downward growth of the root and therefore, is one of the regions in the plant where cells are actively dividing and

elongating. 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, the chromosomes are fairly large and distinct, and this

species has a relatively small number of chromosomes.

Part 1.

a. Click on the root tip to magnify the image.

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 Structure 30/11/2009

a. Review the diagrammatic summary of cell division in your textbook before you begin.

b. View a video of mitosis in an animal cell

Part 3.

Microscopy Lab Now that you have seen how nuclei divide, you can begin the microscope exercises.

Identify the stage:

Interphase - At interphase the nuclear envelope is still intact, the nucleolus is present and

chromosomes are not distinct.

Prophase - During prophase the nuclear envelope disappears, the nucleolus is gone and

chromosomes are distinct and wound throughout the nucleus.

Metaphase – During metaphase the chromosomes are distinct and line up near the center of the

cell.
Cell Structure 30/11/2009

Anaphase - During anaphase the chromatids are separated and the two groups of chromosomes

migrate towards opposite sides of the cell.

Telophase/Cytokinesis - 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.

When you have identified at least one cell in each stage, proceed to the next section.

Part 4.

Mitosis in an Animal Cell.

Click to begin

Slides of whitefish blastulae will be used to show mitosis and cell division in animal cells.
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, there are some differences. See what differences you can detect.

Click on any of the slides at the right to magnify.

Use the scroll bars to move around the slide.

Place the cursor over a dividing cell and click once. Identify the stage of division.

Identify at least one cell in each stage.

Identify the stage:

Interphase - At interphase the nuclear envelope is still intact, the nucleolus is present and

chromosomes are not distinct.

Prophase - During prophase the nuclear envelope disappears, the nucleolus is gone and

chromosomes are distinct and wound throughout the nucleus.


Cell Structure 30/11/2009

Metaphase – During metaphase the chromosomes are distinct and line up near the center of the

cell.
Cell Structure 30/11/2009

Anaphase - During anaphase the chromatids are separated and the two groups of chromosomes

migrate towards opposite sides of the cell.

Telophase/Cytokinesis - 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.
Cell Structure 30/11/2009
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,

noting the transition from single-celled or multi-cellular organisms. (The animal-like

protists will be examined next semester.)

• To understand how the relative position of meiosis and fertilization varies in different life

cycles.

• To appreciate the tremendous reproductive capacity of organisms reproducing asexually.

This capacity may also be observed in some sexual cycles, as in the mushrooms.

Before you leave at the end of the laboratory period you should be able to tell a plant-like protist (alga)

from a fungus, and specify the kingdom of each. You should know how to recognize the haploid and diploid (and

dikaryotic) phases of each organism studied, and what cells were formed by mitosis and by meiosis.

Introduction:

Most plant and animal cells are diploid (2n), meaning that they contain two complete sets of their genetic

materials, located on two complete sets of chromosomes. Alternately, some plant and animal cells are haploid

(1n), containing only one complete set. [You may want to review the discussion of these terms in Lab 2.]
Cell Reproduction 30/11/2009

Dependent upon the organism, both diploid and haploid cells can divide by mitosis. In both cases, each

"daughter cell" has the same amount of genetic material (chromosome number) as the "mother cell".

During a second type of cell division called meiosis, 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.)

The diploid number is restored when two haploid cells fuse during fertilization.
Cell Reproduction 30/11/2009

The actual location of meiosis and fertilization in a sexual cycle depends upon the organism. For example,

the only haploid cells produced by humans (and other animals as well) are the sex cells (gametes). The gametes,

egg and sperm, are formed by meiosis; they immediately combine to restore the diploid condition by fertilization,

forming the zygote. The mature adult is then formed by a large number of mitotic divisions. Therefore, the relative

number of haploid cells is small, as indicated in the sexual cycle to the right.

On the other hand, many protists have a sexual cycle where the only diploid cell is the zygote. This

zygote immediately undergoes meiosis upon "germination"; all of the other cells are formed by mitosis - including

the cells that are capable of fusion again.

Compare these sexual cycles.

These two cycles can be abbreviated by simply showing meiosis and fertilization as gray arrows. On the

right, the only diploid cell is the zygote; on the left, the only haploid cells are the gametes. To help you keep track

of which cycle goes with each organism you will study in this lab, a simplified cycle will be found at the upper right

corner of each illustrated cycle.

In some instances the "many haploid cells" remain separate (e.g., Chlamydomonas), in other cases the

cells are attached to form a filament (e.g., Oedogoniium and Spyrogyra), and in still other cases the cells actually

form a multicellular organism (not seen in lab, but described in your textbook).
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, resulting in an "alternation of generations". We will study this cycle in the next two

laboratories.

Finally, the alternation of meiosis and fertilization, and the resultant recombination of genes, is associated

with the sexual cycle. 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, they are "clones"). This can be an

asexual reproduction of diploid organisms, as in the propagation of some commercial plants. It can be an asexual

reproduction of the haploid organisms, as the production of spores by many fungi. And some organisms, such as

yeast, can produce both diploid and haploid cells asexually.

Most biologists today subdivide all of life on earth into six kingdoms. The prokaryotic organisms, such as

the bacteria you observed in Laboratory 1, are in Kingdom Eubacteria and Kingdom Archaebacteria. Many if not all

of the simpler plant-like organisms ("algae") are placed with simpler animal-like organisms into a Kingdom

Protista; they are therefore called protists. The fungi that you will observe today are given their own kingdom.

I. Some Plant-Like Protists

Although the term "algae" is no longer used in classification, it is still a useful term to describe aquatic

photosynthetic organisms with little tissue differentiation. Chlamydomonas is a single-celled protist; Spirogyra and

Oedogonium are filamentous. Fucus is a macroscopic organism, placed in the plant kingdom in some textbooks.
Cell Reproduction 30/11/2009

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

1. Euglena

What characteristics do these cells have that are like plants?

Are like animals?

hints

Note that these cells divide by mitosis, and therefore asexually. 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.
Cell Reproduction 30/11/2009

Observe Paramecium under low and then high power.

What characteristics do these cells have that are like animals? Are like plants?

3. Chlamydomonas

This small organism requires that you use a high-power objective for viewing.

If you look carefully, you may observe in each tiny cell: a cup-shaped chloroplast with a round starch-like

product in the middle, two flagella, an eyespot (similar to Euglena), and you may even discern the centrally-located

nucleus.

Do you see any of these specialized structures in the cell below?

3. Chlamydomonas fusion

This small organism has at least two "mating types". Since you cannot tell one from the other microscopically,

they are called "plus" and "minus".

Click the thumbnail below to view a video of the first stage of mating, where plus and minus types "clump"

together.
Cell Reproduction 30/11/2009

Next, click the thumbnail below to view a video clip of "pairing", where two Chlamydomonas cells come

together to mate. Watch the two cells at the top of the window.

After pairing, fusion begins. On the right, each mating stage is shown in its correct temporal sequence.

Chlamydomonas cells have a sexual cycle, but may also reproduce clonally, by undergoing mitotic division, as

shown 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. What compensating process must occur in the diploid (2n)

cells?

Mitosis Meiosis

Chlamydomonas life cycle

In the figure below, identify which cells are haploid and which are diploid.

Click on the image at the right to begin.


Cell Reproduction 30/11/2009

B. Some filamentous algae

1. Oedogonium

Note the top image on the right, from a prepared slide of this alga. It shows the chain of cells that makes up

one filamentous Oedogonium organism.

In the middle image, note the clear, round structures within some of the cells; these are the eggs.

The lower image shows a fertilized egg, or zygote.

As in Chlamydomonas, 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, eventually becoming attached to

the lake bottom and differentiating into a new filament. This organism also reproduces asexually by simple

fragmentation or by the formation and liberation of single zoospores from vegetative cells.
Cell Reproduction 30/11/2009

Oedogonium is a filamentous organism. In this image, notice that

cells are connected end-to-end to form a long chain of cells. Also

note the small brown nucleus centrally located in each cell.

The bulb-like cells in the filament

are egg cells. Also notice the

smaller, sperm forming cells below the egg on the top

filament.

Zygotes are similar in size and shape

to unfertilized eggs. They are

distinguished by their thickened cell walls, increased starch content, and darker

color when stained.

2. Spirogyra
Cell Reproduction 30/11/2009

This is another filamentous alga, commonly found in ponds and puddles around New Jersey. From wet mounts,

and the figure below, you can see that it is also a long filament of cells.

Note the shape of the chloroplast(s) in the figure below. The protoplast of the cell is mostly transparent; therefore,

you can see both the front and back of the single spiral-shaped chloroplast that gives this genus its name.

Fusion in this genus begins as two filaments attach. In the image below, notice the small conjugation tube that

joins adjacent cells.


Cell Reproduction 30/11/2009

Conjugation is complete after the cytoplasm from the adjacent cells fuse to form zygotes.

The heavily stained ovoid structures in the figure below are zygotes. 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.

Spirogyra life cycle


Cell Reproduction 30/11/2009

C. Specialization in a large alga

The marine alga Fucus has adapted to life in the intertidal region: part of the day it is submerged, while other

times it is exposed to air.

a. Look at the images of Fucus below. Note the various specialized structures such as holdfast, air bladder,

and receptacles containing conceptacles.

Describe how these specializations are adaptations to an intertidal existence.


Cell Reproduction 30/11/2009

Fucus

b. Inside the receptacles are small pit-like areas that contain the male and female reproductive

structures. Examine images of these conceptacles below. 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?


Cell Reproduction 30/11/2009

The male conceptacle (above) can be identified by the many small, darkly-stained, antheridia. 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.

The female conceptacle is characterized by its large, bulb-shaped oogonia with egg or egg progenitor

cells inside. As in the male conceptacle, sterile hairs protect against dessication.

b. Male and female reproductive structures are located inside the conceptacles. Examine images of the

antheridia and oogonia below. 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

In these higher magnification images and diagrams, compare the small, slim antheridia (above) with the bigger,

rounder oogonium (below). Reproductive structures are stained red in the micrographs, sterile hairs appear blue.

Study the life cycle of Fucus below. Decide whether each structure is part of the haploid or diploid phase of the life

cycle.

II. Some Fungi


Cell Reproduction 30/11/2009

The sexual cycles of the fungi are basically the same as you have seen before, with the zygote being the only

diploid cell in the entire cycle. However, in fungi the fusion of the cytoplasm, or plasmogamy, is not immediately

followed by the fusion of the nuclei, or karyogamy. The stage between plasmogamy and karyogamy is called

dikaryotic, or (n + n), since each cell has two haploid nuclei: one from the plus strain and one from the

minus strain. Each mitotic division therefore results in two pair of nuclei that are dikaryotic, each having

one nucleus from the plus strain and one from the minus strain.

Fungal Life Cycle:

In the diagram above, each arrowhead represents an event in the life cycle of many fungal cells.

A. A Filamentous Fungus: Rhizopus (Black bread mold)


Cell Reproduction 30/11/2009

• On an amenable medium, Rhizopus can be seen with the naked eye, appearing as a fuzzy white mat

growing all over the surface. (And down into the medium. And even over the top of the petri dish

sometimes.) This is the mycelium, or "body" of a fungus. Each strand is called a hypha

(plural, hyphae).

• Through a microscope, one may note small, black balls growing at the ends of certain hyphae. These

are the sporangia. Sporangia can be seen in the image of Rhizopus at the right (top). Note that each

sporangium is composed of hundreds of smaller spores - the asexual reproductive cells of

the fungus.

• After the organisms in a culture initiate the sexual cycle, one may find zygotes (actually, zygospores

composed of numerous nuclei) in regions where the plus and minus join.

Rhizopus:

Rhizopus sporangium

as seen through 16x

objective. Rhizopus

zygosporangium as seen through 16x objective.


Cell Reproduction 30/11/2009

Rhizopus life cycle

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

B. Mushrooms

Examine the diagrams and images of the mushroom Coprinus below.

Basidiospores are formed by meiosis. Remember that the product of a meiotic division is four daughter

cells. Therefore, 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?)


Cell Reproduction 30/11/2009

Figure 1 above: Diagram of cross section through mushroom cap. Notice the gills radiating from the

central stalk. Figures 2 and 3: Higher magnification (as seen through 10x objective) through the basidiocarp.

Notice the basidiospores (stained red in Figure 2, above, and Figure 4, below) attached to gills at basidia.

Figure 4 above: Basidiocarp gills as seen through 40x objective. Notice the different stages of

development of basidiospores.

What is the ploidy of basidiospores?

Haploid or Diploid

Coprinus
Cell Reproduction 30/11/2009

In the life cycle below determine if each tissue is haploid, diploid, or dikaryotic (n + n). Remember that

plasmogamy refers to the fusion of the cytoplasm; karyogamy, of the nuclei.

Make a diagram like the one below, showing how one dikaryotic cell can divide by mitosis to form two

dikaryotic cells. (Click here to review mitosis).

C. A Cup Fungus
Cell Reproduction 30/11/2009

Examine the diagrams and images of Peziza (an ascomycete) that show longitudinal sections through the

fruiting structure, or "cup". In the lower image, look for the sac-like structures, or asci, holding the spores (each

ascus contains 8 ascospores, numbered in one ascus in the lower image). The eight ascospores are formed by

meiosis (to produce 4 cells) followed by a mitotic division (to double the number to 8).

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
Plant Evolution 30/11/2009

Plant Evolution

Objectives:

The objectives of this lab are as follows:

• 1. To look at cellular specialization in plants, with emphasis on lignified cells that function

in water transport and/or support.

• 2. To study the evolution of these vegetative structures in representative plants: a moss, a

fern, a gymnosperm, and both a woody and a nonwoody angiosperm.

• 3. To examine the evolution of sexual cycles in a moss, a typical fern, and a gymnosperm.

By the end of this exercise, you should be able to identify the various lignified cell types in cross sections

of stem. You should know how to tell a moss from a fern; a gametophyte from a sporophyte. You should

understand which cells are haploid and which diploid, and whether they are formed by mitosis, meiosis, or

fertilization. You should be able to apply this information to the reproductive cycle in gymnosperms such as pine.

Section 1:

The evolution of multicellular plants involved the modification of different cells to perform distinct

functions. 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.
Reproduction: Protists and Fungi 30/11/2009

• 1.Tracheids: first to evolve, combined the functions of support and water transport.

• 2.Vessel elements: subsequently evolved from tracheids, specialized for water transport.

• 3.Fibers:also evolved from tracheids, cells specialized for support only.


Reproduction: Protists and Fungi 30/11/2009

All three can be found in xylem, while fibers can be found in xylem, phloem, and other parts of the plant.

I. Evolution of Vegetative Structures

A. Moss

1. Observe the living green "leafy" gametophyte stage on the right. Remember that these organisms have

leaf-like, stem-like, and root-like structures. You can see more of the details by magnifying the image.
Reproduction: Protists and Fungi 30/11/2009

2. Now examine the cross section of a moss (Mnium) "stem". You can see the general structure of the "stem."

Magnify the image and identify the following structures.

• Find:

a. the outer epidermal cells - Epidermal cells are located on the outer wall of the moss stem.

b. the "leaves" attached to the epidermis - "Leaves" are an extension of the epidermis.

c. the multicellular cortex region - The cortex is composed of all the tissue from the epidermal

cells to the transport tissue.

d. the transport tissue - The transport tissue is located in the middle of the stem.
Reproduction: Protists and Fungi 30/11/2009

(Although some of the cell walls in the cortex are stained red, moss is a nonlignified plant.

The central portion of the "stem" is composed of much thinner cells. While the central cells are specialized

to transport water, they are not lignified so they are not called "vascular tissue." [And therefore, moss plants do

not have leaves, stems, and roots.]

B. Ferns
Reproduction: Protists and Fungi 30/11/2009

1. Examine the pictures of live ferns. How tall are these sporophytes? What structures are specialized for

photosynthesis?

2. Study the cross section of a fern rhizome (an underground stem) of the Cinnamon fern, Osmunda

cinnamomea. In the fern, the leaves extend in clusters from an upright position of the stem. Thus, you will find a

central vascular region surrounded by a ring of vascular bundles; each smaller bundle extends to a different leaf,

or frond. Magnify the image to identify the different structures.


Reproduction: Protists and Fungi 30/11/2009

• Find:

a. the central vascular region

b. the vascular bundles


Reproduction: Protists and Fungi 30/11/2009

Click here to enlarge the image yet again.

Within a vascular bundle find:

a. xylem tracheid cells


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As is the case with most ferns, the xylem in this fern is composed of tracheids only. (Remember, lignified

cell walls are stained red -- along with tannins and other materials with a net negative charge.)

C. Pine (a gymnosperm)

1. Examine the picture of a live pine tree. How tall is the sporophyte stage? Are there specialized

photosynthetic structures, and other portions of the plant to support them? Do these organisms have specific

structures that penetrate deep into the soil to obtain water?


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

• Under low magnification find:

a. the central pith

b. three layers (annual rings) of xylem

c. the bark

1. cork

2. cortex

3. phloem

Now click here to magnify the image even more. You should see more of the cellular detail.
Reproduction: Protists and Fungi 30/11/2009

You should notice that all of the cells with red cell walls look alike -- other than slight differences in

diameter in the early "spring" wood and the late "summer" wood found in each annual ring. Remember that in pine

wood, the only lignified cells are tracheids, cells that function both for water transport and support.

The cylindrical structures surrounded by green-staining cells are the resin canals. Under low

magnification you can see them in both the woody part of the twig as well as in the bark. This resin makes the cut

ends of twigs sticky, and are a defense mechanism in these plants.


Reproduction: Protists and Fungi 30/11/2009

• Under high magnification find:

a. the tracheids

b. the resin canals

c. the thin-walled phloem cells that transport "food" throughout the tree

d. 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, an alternative method can be used

called "maceration." The wood is treated with chemicals to make the cells come apart. Examine the macerated pine

wood on the right to see what the tracheids look like from the side. Note the bordered pits in the cell walls. Click on

the image to magnify it.

D. Angiosperms

I. A Woody Plant: Basswood (Tilia)

1. Examine the cross section through a basswood twig.


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Reproduction: Protists and Fungi 30/11/2009

• Find:

a. the central pith

b. three layers (annual rings) of xylem

c. the bark

1. phloem

2. cortex

3. cork

Notice that although the water conducting cells in pine wood were fairly uniform (Pinus stem), those in

angiosperm wood are composed of at least two cell types. Click here.
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Reproduction: Protists and Fungi 30/11/2009

The lignified cells with the largest diameters are the vessel elements. The lignified cells with smaller

diameters are either tracheids or fibers; the difference can only be determined from a side view (see diagram of

lignin cells in this lab's introductory page).

In the outer portion of the twig, the outermost layer is cork and the inner bark is phloem. But note that

there are lignified cells in the phloem region also! Click here.

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. Therefore, if you identify anything with red-stained cell walls as xylem, you may

be wrong! Lignified fiber cells can be found in the phloem as well.

It is easy to tell vessel elements in cross sections because of their size, but more difficult to distinguish a

tracheid from a fiber. Therefore, examine the prepared slide of macerated basswood . You can click on the image

to magnify it.
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Reproduction: Protists and Fungi 30/11/2009

• Identify:

a. a tracheid

b. a fiber

c. a vessel element

II. A nonwoody plant: Buttercup (Ranunculus)

Examine the Cross-Section of a Buttercup Ranunculus Stem.

• Under low magnification find:

1. the vascular bundles

2. the pith

3. the cortex

Using higher magnification find in the vascular bundle:


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1. vessel elements

2. fibers
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. The large diameters of the lignified cells on the inside are indicative

of vessel elements, and are therefore in the xylem. However, the cells on the outside have red-stained cell walls

that are very thick, and the hollow cavity is very narrow. Once again, you are seeing fibers. And, once again, since

this is the phloem on the outside of each bundle, these are phloem fibers.

Section 2

The evolution of plants also involved alterations in the sexual cycle. All plants have the sexual cycle that

alternates a diploid sporophyte generation and a haploid gametophyte generation. The sporophyte forms spores

by meiosis; the gametophyte forms gametes by mitosis.

In moss, the gametophyte is "dominant" (or predominant; you are more likely to encounter this phase of

the sexual cycle). In ferns and all other vascular plants, the sporophyte is predominant. In ferns (and other

"seedless vascular plants") the gametophyte is a free-living, autotrophic organism. While the fern sporophyte is

well adapted to land, the fern gametophyte is not. Therefore, 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. These changes will be observed in the coniferous gymnosperm, pine.

Changes in Sexual Cycles


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Reproduction: Protists and Fungi 30/11/2009

Remember that in the last lab you learned two types of sexual cycles.

• In one, fertilization is immediately followed by meiosis, the only diploid cell being the zygote (top).

The rest of the cells in this cycle are haploid.

• In the other, meiosis is immediately followed by fertilization, the only haploid cells being the

gametes (bottom). In this instance, the diploid phase is usually multicellular.

• A third cycle (center) occurs in all plants and some multicellular protists. In this cycle, haploid

spores are formed by meiosis. These spores divide by mitosis to produce a multicellular haploid phase

that produces gametes by mitosis. The zygote formed by the fusion of two gametes divides by mitosis

to produce a multicellular diploid phase.


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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.


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NOTE: Both the gametophyte and the sporophyte stage may be duplicated by asexual reproduction - the

resulting offspring being clones.

This basic alternation of generations has two variations. In the bryophytes such as moss, the

gametophyte generation is predominant ("dominant") and the sporophyte grows out of the gametophyte (right

bottom). In all vascular plants the sporophyte is predominant (left bottom); the gametophyte may be free-living,

as in ferns, or may be dependent upon the sporophyte, as in seed plants.

ALWAYS REMEMBER:

Mitosis: one diploid cell dividing to form two diploid cells - or - 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. Moss

1. Study the image of a moss antheridium. 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.


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Reproduction: Protists and Fungi 30/11/2009

2. Now study the image of a moss archegonium. If it is a good section, you will see a large cell (egg)

within a vase-like structure. 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, forming the zygote. Click on the image or the word "magnify"

to enlarge the image.


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Reproduction: Protists and Fungi 30/11/2009

Is this method of gamete transfer well adapted to a terrestrial habitat?

3. Examine the whole moss plant in the sporophyte stage. Do you think this brown structure can carry on

photosynthesis, or is it dependent upon "food" from the gametophyte?


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Label each stage as either:

a. sporophyte or..

b. gametophyte
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• Label:

a. antheridium

b. sperm

c. archegonium

d. egg

e. zygote

f. spore

g. protonema

NOTE:If you should discover living sporophytes outdoors, you can pass your hand across the top. The

results will be a cloud of fine particles. These are the spores formed by meiosis that will germinate to develop into

the new gametophyte generation.

B. Ferns
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1. Examine the live fern leaf ("frond") that has brown or black specks ("sori") on the underside. The small

brown or black cells are spores that, under the correct conditions, will germinate to form the gametophyte

generation. Some ferns, such as the cinnamon fern (previous section), have all of the sporangia on a separate

"fertile fond." The sporophyte cells are diploid; the spores (and gametophyte cells) are haploid. Were the spores

formed by mitosis or meiosis?


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2. Examine the images of the fern antheridium and archegonium. Click on the images to get a closer look

at the cells.
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Reproduction: Protists and Fungi 30/11/2009
Reproduction: Protists and Fungi 30/11/2009

Answer the following questions:

• A. Are the gametophyte plants haploid or diploid?

• B. 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?


Reproduction: Protists and Fungi 30/11/2009

Examine the drawing of the fern life cycle to the right.


Reproduction: Protists and Fungi 30/11/2009

• Label each stage as either...

a) sporophyte or..

b)gametophyte

Click here to reload the image. Now...

• Label:

a. antheridium

b. sperm

c. archegonium

d. egg

e. zygote

f. spore

C. Pine

In both of the plants you have observed so far, the spores formed by meiosis all look the same; the plants were

homosporous. A variation on this theme is where some spores are larger than others; these plants are

heterosporous. All gymnosperms and angiosperms are heterosporous.

Each scale in the smaller male "staminate" cones contains numerous diploid microsporocytes or

"microspore" mother cells that go through meiosis, each forming four haploid microspores. Each microspore

undergoes several mitotic divisions to form a microgametophyte, or pollen grain. Some of the nuclei, the sperm

nuclei, are capable of fusion with an egg nucleus.


Reproduction: Protists and Fungi 30/11/2009

The scale in the larger female ("ovulate") cone has two ovules, each containing a single megasporocyte

(megaspore) mother cell which goes through meiosis to form four haploid megaspores. In this case, three

megaspores degenerate while the fourth continues to divide mitotically, forming a multicellular

megagametophyte. This megagametophyte has two archegonia, each containing one egg.

• 1. Observe the image of the smaller male pine cone.


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, pollen is a MICROGAMETOPHYTE.)

You can see pollen being released from the male pine cones in the third image on the right.
Reproduction: Protists and Fungi 30/11/2009

The forth image on the right is a magnified picture of actual pollen grains.
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• 2. Now observe the image of the larger female (ovulate) pine cone, 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.


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. 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

• Label each stage as either..

a) sporophyte or..

b) gametophyte

Now click here to reload the image.

• Label:

a. megaspores

b. megagametophyte

c. archegonium

d. microspore

e. microgametophyte (= pollen)

f. pollen tube

g. embryo

h. seed coat

i. seedling

Pine is only one example of a gymnosperm. 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


Reproduction: Protists and Fungi 30/11/2009

This is a complicated lab, with lots of parts, so it is essential that you get an overview before try to sort

out the details.

• 1. Think about all of the figures that illustrate tracheids. 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. Think about all of the figures that illustrate vessel elements. 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. Think about all of the figures that illustrate fibers. 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. Compare the sexual cycles for moss and ferns. What do they have in common? How do they differ?

• 5. Compare the sexual cycles for ferns and pine. What do they have in common? How do they differ?

• 6. The "basic life cycle" illustrated in this lab is for a homosporous plant, such as most ferns. Change

this life cycle to represent the situation for a heterosporous plant, such as pine.
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Angiosperm Reproduction 30/11/2009

Angiosperm Reproduction

Objectives:

• To study the development of the different parts of the flower, and how these regions

further develop to form the fruit with its seeds.

• To practice forming three-dimensional images from two-dimensional observations.

• To observe some of the variation in different parts of sample fruits, and relate these

modifications to changes in function.

After completing the laboratory exercise you should know the parts of a flower and the functions of each.

You should be able to recall which part of the flower develops into each portion of a fruit. Again, you should be able

to recognize which cells are haploid and which diploid, and whether they are formed by mitosis or meiosis. Finally,

you should appreciate how plants have evolved different structures for pollen transfer and for fruit dispersal.

Introduction:

This lab is a continuation of the previous lab in which you studied some of the basic aspects of plant

evolution. Click here to briefly review the evolution of land plants. Next, review the generalized life cycle at the

beginning of Part II of lab 7.


Plant Evolution 30/11/2009

Finally, review the evolution of the gametophyte generation, starting as a free-living, multicellular,

photosynthetic organism in ferns. In gymnosperms like pine, the microgametophyte was reduced to a small, wind-

blown structure called "pollen". The megagametophyte was maintained within the sporophyte cone, and

surrounded by the sporophyte layer called the integument (later to become the seed coat). The megagametophyte

was still large enough to provide the nutrients for the young embryo in the mature seed. This trend of reducing the

gametophyte generation is continued in angiosperms. These and other trends in plant evolution are diagrammed

on the next page.

In this laboratory, you will observe several of the structures associated with the reproductive portion of

the plant life cycle. You will start by observing the development of the megagametophyte ("embryo sac") as well as

that of the microgametophyte ("pollen"). You will then study the subsequent development of the seed and fruit.

You will also examine examples of the different types of fruits to see how they have evolved different

structures for protection and dispersion.

I. The Flower

You have now come to a major evolutionary advancement of the plant kingdom. Review the main parts of

the flower below.

In the diagram, parts of the male reproductive structures, or stamen, are labeled in green. There are six

stamen in this flower.


Plant Evolution 30/11/2009

The female reproductive structures are labeled in blue. The stigma, style and ovary make up the single

pistil in this flower. Note that the ovary is not visible in the diagram, but is located in the center of the flower, at

the base of the style.

The diagram below illustrates the components associated with sexual reproduction.

On the left, label the parts of the stamen, the male reproductive structures.

On the right, label the external parts of the pistil (sometimes also called the "carpel"), the female

reproductive structure.

Sexual Structures of a Flower


Plant Evolution 30/11/2009

1.Anther

2. Filament

3. Stigma

4. Style

5. 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, you should know the function of

each:

• Sepal: To protect the developing flower bud.

• Petal: To attract pollinators with colors and/or odors; to provide a landing area for insect pollinators.

We can speculate that one of the functions of sepals is to hide developing flowers from pollinators. This

prevents colored petals from attracting insect visitors before pollen is ready for dispersal, 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, and so they are sometimes thicker, or fleshier, than petals. Are there other ways to

distinguish sepals from petals?


Plant Evolution 30/11/2009

The figure below depicts the internal features of the pistil after pollination. Label the parts and identify

each as belonging to either the sporophyte (diploid) or gametophyte (haploid) generation.

Flower Pistil After Pollination

1. Pollen grains

2. Stigma

3. Style

4. Pollen tube

5. Ovary

6. Ovule

7.

Gladeolus Flower Dissection

If the petals and sepals are removed from a gladeolus flower, the remaining

parts are the stamens and the pistil (sometimes called the carpel.)

When a longitudinal cut is made down the ovary, the line of ovules inside

becomes visible.

The following diagram shows the gladeolus flower dissected in this way.

A. The Anther
Plant Evolution 30/11/2009

Study the diagram below, which illustrates a cross section through flower sexual structures. Note the style

in the center of the circle of anthers. Each anther consists of two pollen sacs, one on each side of a vascular

bundle.

1. Stigma

2. Style

3. Ovary

4. Ovules

5. Anther

6. Filament

Cross Section through Flower Pistil and Anther


Plant Evolution 30/11/2009

1. Anther (pollen sacs are within the anthers, two pairs per anther.)

2. Style

Compare the diagram above with the prepared slide of the Lilium anther, to the right. This is a large

structure-- this image was captured with the 6x scanning lens! Scroll to find the style, anthers (and their pollen

sacs), filament, and vascular bundles.

A. The Anther, continued.

An anther with mature pollen is drawn in the diagram below.


Plant Evolution 30/11/2009

Two features of the pollen are worth noting: First, each pollen grain has two nuclei. (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.) Second, the cell wall has a definite structure. (Look for grains where the section was right across the upper

or lower surface of the pollen.)

Remember that pollen is a MICROGAMETOPHYTE formed from a MICROSPORE.

Examine a prepared anther using higher power by clicking on the image below to magnify.

Cross section through Lilium anther

In the high magnification image to the right

(http://bio.rutgers.edu/~gb101/lab8_angio_repro/lilyanther2.jpg), find pollen, pollen sacs, and vascular

bundles. Then estimate the size of the pollen grains.

If the field of view in the thumbnail image below is 1.2mm, how large is one pollen grain? Find a cluster of

grains that line up across the diameter of the field. Count the number of grains and divide this into the known field

of view.
Plant Evolution 30/11/2009

1.

2.

3.

You may review the procedures for estimating cell size using a known field of view in Laboratory 1.

Do you estimate that the lily pollen is 0.003 mm, 0.05 mm, or 0.1 mm?

Microgametophyte Development

The development of the microgametophyte (male gametophyte) in flowering plants involves three stages:

• A diploid microsporocyte, or microspore mother cell, divides to give rise to four haploid

microspores.
Plant Evolution 30/11/2009

• Each microspore divides by mitosis to form two haploid nuclei (the tube nucleus and the generative

nucleus); this is mature pollen in Lilium.

• After landing on the stigma, the pollen germinates forming a pollen tube; the generative nucleus

then divides by mitosis to form two haploid sperm nuclei.

B. The Ovary
Plant Evolution 30/11/2009

The image below is an illustration of a cross section through Lilium ovary, as might be seen using a

scanning lens.

In the Lilium ovary, the ovules are attached in pairs to a central stalk. In prepared slides, you may not

always see all three pairs of ovules in one section.

In the image to the right, note the small vascular strands in the central region, the ovules, and the

protective ovary tissues around the outside.

How many ovules are in this ovary cross section?

2, 3, or 6?

The development of the megagametophyte ("embryo sac") in most angiosperms involves three stages.

A diploid megasporocyte (megaspore mother cell) undergoes meiosis, giving rise to four haploid

megaspore nuclei (3 of which disintegrate).


Plant Evolution 30/11/2009

The remaining megaspore nucleus undergoes three mitotic divisions to form eight haploid nuclei.

Nuclear migration and cytokinesis occur to form the mature megagametophyte

While the above is "typical", the number of antipodals can vary from none to many, depending upon the

species. The smallest megagametophyte only has an egg cell plus one polar nucleus.

III. Seed and Fruit Development

A. Development of a Seed

Capsella: a dicot
Plant Evolution 30/11/2009

Examine several slides of the cosmopolitan weed Capsella bursa-pastoris (Shepherd's purse). This plant

has numerous heart-shaped fruits ("purses") on a long stem. Each of these fruits contains dozens of minute seeds.

(See the illustration below.) The white flowers and developing fruits are at the very top of the plant, and fruit at

more mature stages are at the bottom.

Capsella seed development

Review some of the basic parts of the developing seed using the illustrations below.
Plant Evolution 30/11/2009

Note that in this plant the endosperm is absorbed by the developing cotyledons, so that the stored food in

the mature seed is in the cotyledons. (The same pattern is also true for the legumes, including the pea and bean

seeds that you will study in the next section.)

The sections from the previous illustrations, and in the illustration below, are exactly parallel to the

embryonic axis. As you examine the prepared slides, however, you will note that many of the seeds have been cut

at other angles. Use the following illustration to help understand these sections.
Plant Evolution 30/11/2009

The diagram above shows four of the infinite planes where the seed could be sectioned. The angle of the

cut is shown in blue. Identify what the resulting section would look like if the section were made at the blue line.

III. Seed and Fruit Development

• B. Fruit Development

Fruit development is a complex process, varying considerably among different plant groups. Three

examples of the flower and then the fruit are found in the figures to the right.

For example, in peas the flower is "irregular" (that is, it has bilateral rather than radial symmetry), with

sepals and petals modified into several different shapes. The ovules are attached down a single row in the ovary so

that the seeds develop in the same way. When you split open a pea pod, you can get a good impression of how the

flower develops into the fruit.


Plant Evolution 30/11/2009

Complete your collection of life histories by adding two angiosperm sexual cycles.

The figure on the right is the sexual cycle of a dicot, the bush bean.
Plant Evolution 30/11/2009

Corn, Zea mays

The images below show a monocot, corn.


Plant Evolution 30/11/2009

While completing the corn life cycle, note that while the materials in the endosperm have been

transported to the cotyledons during bean seed development (as in Capsella), the endosperm still exists in the

mature corn kernel.

C. Fruit Diversity
Plant Evolution 30/11/2009

• Dispersal: Angiosperms have evolved a number of different methods for protection and dispersal of

the next generation. These dispersal mechanisms range from structural modifications such as barbs

that stick in fur, or buoyant structures that carry fruits and/or seeds through water; to sweet fruits

that are eaten by animals that disperse seeds (such as the peach, on the right).

• Domestication: A variety of plants have been cultivated, often because of the fruits. The original

evolution of the fruit was for dispersal. For example, note the three layers of a peach on the image

below.

The Peach:

an example of fruit dispersal mechanisms

The outer layer ("skin") protects the next layer from drying out. The main central layer attracts animals,

who consume the whole fruit. The inner layer ("pit") protects the seed during the process.

The pit is later deposited, along with some "fertilizer", after in has passed through the animal's gut.

To continue, click on the area of the image that was the ovule when this fruit was still a flower.
Plant Evolution 30/11/2009

3. Significance of grains

Examine a corn "cob". (http://bio.rutgers.edu/~gb101/lab8_angio_repro/cornkernelhigh.jpg)


Plant Evolution 30/11/2009
Plant Evolution 30/11/2009

The kernel is technically a fruit, not a seed, since the seed coat and fruit wall are fused.

Examine the prepared slide of a corn kernel above. (The scientific name for corn is Zea mays).
Biology Lab Practical Lab 9

Transport Systems in Plants

Objectives:

• 1. To study water transport in plants.

• 2. To investigate the structures associated with transport of water and of organic nutrients.

Introduction:

The transport systems in plants are quite different than the circulatory system found in vertebrates.

Vascular plants have two transport systems, 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 water transport have already been studied in Lab 7: the tracheids and vessel elements

found in xylem. The cells responsible for long-range transport of organic compounds are found in the phloem.

The hierarchial nomenclature used in plant anatomy is often confusing, and is therefore outlined to the

right. Click here to get table. Note that both xylem and phloem tissues may also contain cell types that do not

function in transport, such as the phloem fibers observed in Lab 7.

In this laboratory you will:


Angiosperm Reproduction 30/11/2009

1. confirm the path of water movement between the uptake by root hairs and the loss

through stomata

2. perform an experiment that suggests a mechanism for this movement

3. examine prepared slides to study the cellular structures in more detail

II. 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, congo red dye will be added to the water to trace the pathway of

water up the stem. Cross-sections of the stem demonstrate where in the stem the water is moving and oblique-

sections allow identification of the actual cells involved in water transport. The second part of the experiment

identifies which parts of the stem, and more specifically, which cells, are lignified. 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, thin-walled cells). Therefore, the

vascular strands running longitudinally in the stem can be observed directly. Notice the strands in the picture to

the right. Click on the picture to label the strands with arrows.

Step 2: Cut the root system from the plant and immediately place the cut surface of the stem in water.

Cut off another centimeter of stem under water to ensure there are no air bubbles in the transport tissue that could

disrupt water transfer. Click here to see how that was done.
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 set-

up.
Angiosperm Reproduction 30/11/2009
Angiosperm Reproduction 30/11/2009

Step 4: Two controls are needed for this experiment. Since the purpose of the experiment is to trace

the movement of congo red dye to follow the pathway of water transport, it is first necessary to see if any of the

cell walls of the plant are already stained red. 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 is control

1. The second control is prepared in the same way, however, this control is to check which parts of

the stem Congo red will stain (Congo red stains cellulose). Add a drop of Congo red to a second

slide for control

2. Click on the following to observe each each part of step 4.

• a.Preparation of sections
Angiosperm Reproduction 30/11/2009

b.View Control 1 and 2 on the slide


Angiosperm Reproduction 30/11/2009

c.View Control 1 under the dissecting scope


Angiosperm Reproduction 30/11/2009

d.View Control 2 under the dissecting scope


Angiosperm Reproduction 30/11/2009

Step 5: After the dye has moved to the stem apex, cut cross-sections of the stem at various points

along the stem. Examine the location of the congo red. Also make longitudinal sections at the same points to

identify the cell type. Click on the following to observe each part of step 5.

• a. Sections at various points along the stem

b. View slides of cross and oblique sections from each point


Angiosperm Reproduction 30/11/2009
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.

• Step1:Cut additional cross and longitudinal sections to stain lignified cell walls using

phloroglucinol.
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.


Angiosperm Reproduction 30/11/2009

• Step4:Add 2-3 drops of concentrated HCl. (Lignin stains red)


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
Angiosperm Reproduction 30/11/2009

II. Water Transport Mechanism

According to the transpiration-adhesion-cohesion-tension mechanism of water transport in xylem,

water evaporating through the stomata produces a tension, or negative pressure, that pulls the water

column up the plant. 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).

If the above mechanism is correct, 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. The rate of movement can

be determined by adding a dye to the water at the bottom of the pipette.

Begin Experiment:
Angiosperm Reproduction 30/11/2009

1. Place a transport apparatus (1 mL pipette with a piece of clear tubing on the larger end) under water

using the large dish pan. Manipulate the apparatus until it is completely filled with water - NO air bubbles.

To do this, click on the yellow tubing to place it on the pipette. Next, click on the pipette to

place it in the dish pan. It will fill with water.


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. Insert the freshly cut end into the above

tubing while under water, making certain that NO air bubbles are trapped within the assembly. If the twig does not

fit tightly in the clear tubing, wrap several strips of Parafilm around the stem before inserting it into the tubing.

Attach the twig firmly with wire using the pliers.

Click on the juniper twig to place it into the dish pan. Click on the knife to cut off an additional

cm under water. Next click on the twig again to insert it into the tubing. For our purposes, assume it

fits snugly.

3. Add about 20 mL of red colored dye (Congo red) to the bottom of a small beaker. With your finger over

the tip of the pipette, transfer the assembly so that the tip is now into the dye. Remember: NO air bubbles!

Click on the test tube to add the Congo red. Next click on the pipette/twig assembly to place it

into the test tube.

4. Transfer the whole thing to the ring stand. Clamp it into place and turn on the light at the top of the

assembly.

Click on your set up to complete assembly.

5. This same set-up is repeated with a juniper stem without leaves to use as a control.

Click here to see the control's set-up.


Angiosperm Reproduction 30/11/2009

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

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.


Angiosperm Reproduction 30/11/2009

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 30/11/2009

and a Begonia (bottom right)


Angiosperm Reproduction 30/11/2009

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


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). Thinner secondary roots (thin black arrow) emerge from the older primary 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 form of continuous development is characteristic of

higher plant structures.

Now click on the Begonia roots. This plant had been clipped and the stem was placed in water. The

stem grew new root shoots. Notice the numerous root hairs. The root hairs increase the surface area of the

root.
Angiosperm Reproduction 30/11/2009

Below to your left, view the cross section of the root as if you were using a microscope. Increase

magnification by using the + sign and decrease magnification using the - sign. You can also move the stage of the

microscope by clicking on the button on the far right and moving the hand. Once you have examined the section,

label the parts of the root by clicking on the image to your right. Identify the epidermis and the xylem at low

magnification and identify the phloem cells, cortex, and endodermis at high magnification.

Indentify:

Epidermis at low magnification – The epidermis is a one-cell thick outer covering of the root.

Xylem - The xylem is in the center of the circular root.


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 vessel elements are the large conducting cells in the xylem in the middle of

the vascular region.

• cortex - The cortex is the intermediate portion of the bark, between the epidermis and the vascular

tissue.

• endodermis at high magnification - The endodermis is a single layer of cells that encircles the

vascular cylinder.
Angiosperm Reproduction 30/11/2009

Just as in the root exercise, the image to your left can be manipulated as if viewed through a microscope.

Click on the image to your right to identify the epidermis, cortex, and vascular bundles. You may then click on

"high magnification" to label the parts of the vascular region. Identify the phloem fibers, sieve tube members,

vessel elements and tracheids.

Identify:

Epidermis - The epidermis is a one-cell thick outer covering of the stem. It has a thick waxy cuticle and

stomata.

cortex - The cortex is external from the vascular bundles and internal from the epidermis.

vascular bundles –
Angiosperm Reproduction 30/11/2009

Identify:

phloem fibers - The phloem fibers are clustered toward the outside of the vascular bundle, forming a cap

that strengthens the stem.

sieve tube members - The seive tube members are located in the phloem. They are on the inner side of

the phloem fiber cap.

vessel elements - The vessel elements are the larger, more efficient conducting cells in the xylem.

tracheids. – The tracheids are the smaller conducting cells located in the xylem.

C. The Leaf
Angiosperm Reproduction 30/11/2009

1. 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

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).

It is also possible to visualize the stomata on the leaf's surface. To do so, each leaf was painted with

clear nail polish on both sides. The film of nail polish was peeled off and mounted on a slide to examine under the

microscope. If all goes well, an impression of the leaf cells on the surface will be left. Click on the zebrina leaf to

examine the leaf's surface. Notice which surface the stomata are located and their distribution. Next click on the

coleus leaf.
Angiosperm Reproduction 30/11/2009
Angiosperm Reproduction 30/11/2009
Angiosperm Reproduction 30/11/2009

Again, the image to your left can be manipulated as if viewed through a microscope. Click on the image to

your right to identify the upper epidermis, lower epidermis, cuticle, guard cells, stoma, palisade

mesophyll (with chloroplasts), and spongy mesophyll.

Identify:

upper epidermis - The upper epidermis is a one-cell thick outer covering of the top of the leaf.
Angiosperm Reproduction 30/11/2009

lower epidermis - The lower epidermis is a one-cell thick outer covering of the bottom of the leaf.

Cuticle - The cuticle is the waxy layer secreted by the upper epidermal cells to reduce water loss. In this

image only a small area is visible.

guard cells - Guard cells are specialized lower epidermal cells that are responsible for opening and

closing the stoma.

Stoma - The stoma is a minute air space opening at the base of the leaf.

palisade mesophyll (with chloroplasts) - The palisade mesophyll is the layer of closely packed

columnar cells near the upper epidermis.

spongy mesophyll - The spongy mesophyll is the layer of loosely and irregularly arranged cells near the

lower epidermis.
Angiosperm Reproduction 30/11/2009
Chromosome Structure and Meiosis 30/11/2009

Objectives:

The objectives of this lab are as follows:

1. To review the structure of a chromosome.

2. To study the events associated with meiosis.

3. To apply this knowledge to human genetics by analyzing a karyotype.

Introduction:
Biology Lab Practical Lab 9

Meiosis is the second important kind of nuclear division. It resembles mitosis in many ways

but the consequences of meiotic divisions are very different from those of mitotic divisions. While

mitotic division may occur in almost any living cell of an organism, meiosis occurs only in special cells.

In animals, meiosis is restricted to cells that form gametes (eggs and sperm). Each species has a

characteristic number of

chromosomes per somatic cell.

Fruit flies have 8; normal humans

have 46. They exist as homologous

pairs (partners) that are similar in

size and shape and carry the same

kinds of genes. Thus humans have

23 homologous pairs. 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). 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. If the egg and sperm were both diploid (46

chromosomes each in the case of humans) then the resulting zygote would be tetraploid. This would

be an intolerable situation, 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). This is referred

to as the haploid number.


Biology Lab Practical Lab 9

Haploid Egg + Haploid Sperm = Diploid Zygote

The mechanism that makes this possible is meiosis. Meiosis consists of two divisions, Meiosis

I and Meiosis II, and can potentially result in the production of four cells. However the DNA is only

synthesized once (prior to Meiosis I). The subdivisions of meiosis are named like the subdivisions of

mitosis (prophase, metaphase, anaphase, telophase) but as we shall see the events are somewhat

different.

Part I:

Lets review the stages of Meiosis.

1. Study the diagrammatic summary of cell division in Meiosis I and Meiosis II in your

textbook before you begin.

Part II:

You are now ready to find the actual stages of meiosis, using the lily anther. The lily flower

has six anthers surrounding one carpel. Each anther has two pair of microsporangia ("pollen sacs"). It

is in these microsporangia that you will observe the stages of meiosis.

REMEMBER: These slides are thin two-dimensional sections through three-dimensional

reality. Therefore, you will have to look at several different cells in each microsporangium to see

exactly what is going on. Luckily, in the early stages of meiosis, all the cells in the sac are in the

same stage.

Meiosis in the anther starts with the diploid microsporocyte. Each nucleus has a diploid

number of duplicated chromosomes. These cells are still attached in the microsporangium.
Biology Lab Practical Lab 9

We will now begin to identify the different stages of meiosis. The cross-section of the lily

anther to the left is in the early stages of meiosis.

Are the cells in:

1. Early prophase I - In early prophase, the chromosomes are long and slender, and the bright red

nucleoli are still present. The nuclear region will be clear and the nuclear envelope is still present.
Biology Lab Practical Lab 9

2. Late prophase I - In late prophase I, the chromosomes are short, thick, and condensed. The

nucleolus is absent and the nuclear envelope has broken down.


Biology Lab Practical Lab 9

3. Metaphase I - During metaphase I the pairs of chromosomes are distinct and line up near the

center of the cell.


Biology Lab Practical Lab 9

4. Anaphase I - During anaphase I the chromosome pairs are separated and the two groups of

chromosomes migrate towards opposite sides of the cell.


Biology Lab Practical Lab 9

5. Telophase I - By telophase the two groups of chromosomes have completely separated and are

positioned at opposite sides of the cell.


Biology Lab Practical Lab 9

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

duaghter cells after interkinesis.

In Meiosis II, all the cells in a microsporangium are not in the same stage of division.

Therefore, images of single cells have been taken for identification of these stages.
Biology Lab Practical Lab 9

Look for:

1. Prophase II - In prophase II, the chromosomes are condensed and two new nuclear envelopes

are present.

2. Metaphase II - During metaphase II the chromosomes are distinct and line up near the center of

the cell.
Biology Lab Practical Lab 9

3. Anaphase II - During anaphase II the chromatid are separated and the migrate towards opposite

sides of the cell.

4. Telophase II - By telophase the two groups of chromosomes have completely separated and are

positioned at opposite sides of the cell.


Biology Lab Practical Lab 9

5. Cytokinesis II - In cytokinesis, the nuclear envelope begins to reform. The cell plate becomes

visible between the dividing cells. Four cells are now visible.
Biology Lab Practical Lab 9

Part III:

You finally graduated top of your class and have taken a position as a cytogeneticist. Your

job is to construct the karyotypes of your patients in order to look for possible chromosomal

abnormalities. Each karyotype has been started for you. The first 12 chromosomes have been

matched. Your job is to match the remaining 11 chromosomes, determine whether or not your patient

has a chromosomal abnormality and if so, which one, and to diagnose the patient with the expected

symptoms of their karyotype. You better get started, your patient list is growing.....

Case 1: 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
Biology Lab Practical Lab 9

Case 2: This is a normal male karyotype. Healthy male.


Biology Lab Practical Lab 9
Biology Lab Practical Lab 9

Case 3: This is a normal female karyotype. Healthy female.


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
Biology Lab Practical Lab 9

Now that you have correctly determined each of the above karyotypes, it is essential to

understand how they occur. The abnormalities viewed above are mostly cases of trisomies. Trisomy

occurs during meiosis when nondisjunction occurs. Nondisjunction is a failure of chromosome or

chromatid to separate to opposite poles during nuclear division. When nondisjunction occurs, two

chromosomes or chromatid go to one pole and none go to the other.

Two homologous pairs of chromosomes are present within the cell depicted below.

The large chromosomes are homologous and the small chromosomes are homologous. The

red chromosomes represent maternal genes and blue chromosomes represent paternal

genes.
Biology Lab Practical Lab 9
Biology Lab Practical Lab 9
Biology Lab Practical Lab 9

Depending on which gamete meets which gamete, three scenarios are possible. The gamete

with the extra chromosome could meet a normal chromosome. In this case the patient would

have a trisomy in their karyotype, as seen in the previous case studies (i.e. Down's syndrome). The

gamete missing a chromosome could meet a normal gamete. The result of this match would be

a patient with a monosomy (i.e. Turners syndrome). A normal gamete may also meet a normal

gamete in which the patient would be a normal male or female. The following illustrate these

scenarios.
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