Meiosis and Life Cycle Patterns Most organisms produce offspring by a process of sexual reproduction, in which a gamete from

one parent joins a gamete from the other parent to form a zygote (or fertilized egg). This process results in offspring that have a combination of parental chromosomes and provides a source of genetic variation for species. However, the chromosome number for a species is constant, so at some time prior to sexual reproduction, chromosome number must be reduced or the species chromosome number would double with each generation. Meiosis is the process that reduces the chromosome number by half. The process of meiosis accomplishes this reduction by first pairing, and then separating, homologous or matching pairs of chromosomes. Cells that contain pairs of homologous chromosomes are called diploid. When a cell has chromosome pairs, we refer to the diploid number of chromosomes. For humans, the diploid number of chromosomes is 46, comprised of 23 homologous pairs of chromosomes. Homologous chromosomes also form the basis of the variation we see in the genetic traits of living organisms. Meiosis, therefore, has a second function for living organisms: maintaining genetic variation. Meiotic products have half as many chromosomes as their parent cells, and are haploid. The nucleus of a haploid cell will have one set of chromosomes and no homologous pairs. A human gamete will have 23 chromosomes and no pairs. Meiosis involves two divisions. The first division, Meiosis I, involves pairing the homologous chromosomes and then separating the homologues into separate cells (which reduces the chromosome number by half). Because any cell division is preceded by DNA duplication, the second division of meiosis (Meiosis II) involves the separation of the individual duplicated chromosomes. In this laboratory you will observe slides of meiosis in the lily flower. You will use pop-bead chromosomes to simulate the process of meiosis, including simulating crossing over and recombination, and then compare, with diagrams, the process of oogenesis, which produces eggs, with spermatogenesis, which produces sperm in animals. You will also look at meiosis in the context of an organism's life cycle, comparing and contrasting the timing of meiosis in plants, animals, and haploid organisms. Exercise I Meiosis in the Lily anther Meiosis in plants occurs in structures called sporangia. The sporangia of flowering plants are located in the flower. Flowers have both male and female sporangia. The male sporangium is called the anther and the female sporangium is an ovule. For comparison, the sporangia of conifers are located in their cones. The brown fuzzy spots on the undersides of fern leaves are fern sporangia. Your instructor may show a series of slides showing meiosis in the anther (male sporangium) and ovule (female sporangium) of the lily, Lilium sp. before you try to find meiotic stages with the microscope slides. Since the lily is a plant, the meiotic products will not directly form gametes, but will develop by mitosis into structures (gametophytes) that will eventually contain sperm (the pollen grain) and egg (the embryo sac).

-1-

Materials Needed Slide projector and slides of meiosis in lily anthers and ovules Prepared slides of: Lilium anther, prophase I Lilium anther, first division (heterotypic division) Lilium anther, second division (homeotypic division) Lilium anther, pollen tetrad Procedure Observe the prepared slides and micrographs of the lily anther to locate the stages meiosis. Your instructor will provide you with an orientation to the structure of the anther so that you will know where to locate the meiotic figures. As you observe the different stages of meiosis be sure that you can recognize the events of both meiosis I and of meiosis II. The cells that do meiosis (the sporogenous tissue) in the anther are spherical. Each anther chamber has several cells, and each lily flower has six anthers. You may or may not find meiotic figures in all the anther sections on your slide. Although it would be ideal to have a slide with just one stage of meiosis, the sporogenous tissue of the anther does meiosis on its own time frame, not for the convenience of biology students trying to see the stages of meiosis in real cells. Meiosis I Recall that prior to meiosis, in premeiotic interphase, DNA duplication occurs so the all of the chromosomes in the cells about to do meiosis are duplicated. There are two slides for meiosis I: Lilium anther, prophase I and Lilium anther, first division (heterotypic division) Prophase I In prophase I, homologous chromosomes pair up in a process called synapsis. This uses proteins along the chromosomes to join the homologues together. The homologues literally join along their length. At several points, called chiasmata, the four chromatids of the paired homologous chromosomes intertwine. Chromatids of the homologous chromosomes can break at one or more chiasmata and exchange equivalent bits of DNA with their homologue. This exchange is called crossing over and is mediated by enzymes. If the exchanged pieces of the homologues were different forms of a gene, then recombination occurs. The sister chromatids now have some genetic variation; they are no longer precisely identical.

Early Prophase I

Synapsed Homologous Pairs

-2-

Metaphase I Homologous pairs of chromosomes, still synapsed, are moved to the equator by the spindle complex. The alignment is random; some "maternal" chromosomes will orient facing one pole along the equator; others face the opposite pole. Spindle microtubules just attach to homologous chromosomes as they find them from the respective poles of the cell. Anaphase I The homologous chromosomes are separated from each other and pulled toward opposite poles, officially reducing the chromosome number. Duplicated chromosomes are not affected during Anaphase I. The sister chromatids are still tightly bound to each other by their centromeres. No homologous chromosome pairs are present at the end of Anaphase I. Each cluster of chromosomes at the respective poles of the cell will have one of each type of homologous chromosome. The pairing and separation of homologues that you have just observed is the key to reducing chromosome number while maintaining all of the genetic information. Telophase I and Interkinesis Each nucleus that will form around the set of chromosomes at each pole will have half the number of chromosomes as the pre-meiotic cell, each with one set of the homologous chromosomes Cytokinesis typically will form two cells. (Each chromosome is still duplicated; this occurred in pre-meiotic interphase.) The cells are just preparing for the second division.

Metaphase I

Anaphase I

Telophase I

Meiosis II Meiosis II is just like a mitosis; duplicated chromosomes are distributed equally into new cells, each with the same number of chromosomes as the original cell The differences are that you are starting with two cells, and forming a total of four new cells, and the two cells contain no homologous chromosomes. Prophase II New spindle apparatus is formed in each of the two cells from telophase I The still-duplicated chromosomes stretch out and then recondense. Spindle microtubules from each pole attach to the kinetochores of each of the sister chromatids. Metaphase II The duplicated chromosomes are aligned along the equator by the spindle complex in each of the two cells.
-3-

Anaphase II Centromeres of sister chromatids are separated from each other and the now single chromosomes are pulled to the poles of the two cells. Telophase II and Cytokinesis Each new nucleus formed has half the number of the original chromosomes and each nucleus has one of each type of homologous chromosome. Cytokinesis results in a total of four new cells, the haploid products.

Prophase II

Metaphase II

Anaphase II

Telophase II

Exercise II Meiosis Simulation - Optional This exercise will give you the opportunity to review your understanding of meiosis by diagramming the process below. A clear understanding of the behavior of chromosomes, and in particular, homologous chromosomes and how they differ from sister chromatids, is essential to understanding the process of meiosis and its reduction of chromosome number. Recall that when the homologous chromosomes are synapsed during prophase I of meiosis, crossing-over, or recombination occurs. During crossing-over homologous chromosomes exchange small pieces of non-identical chromatids. The point where crossing-over occurs is called a chiasma, and several chiasmata occur in a single pair of homologous chromosomes. Recombination has genetic importance. All of the genes located on a single chromosome are inherited as one unit, so that the genes located on one chromosome are said to be "linked" together for purposes of inheritance. Crossing-over permits the separation of linked genes, which results in a greater diversity of gene combinations in the cells formed by meiosis. Because of crossing-over, no two meiotic products are exactly the same. Diagram the process of meiosis on the following page showing crossing-over by exchanging "bits" of your homologous chromosomes during prophase I.

-4-

Nucleus in Prophase I

-5-

Independent Assortment of Homologous Chromosomes during Meiosis A source of genetic variation in meiosis is the alignment of homologous chromosomes at the equator during metaphase I. Some maternal chromosomes will align towards one pole and some towards the other pole. Each pair of chromosomes assorts independently of any other pair, relative to maternal/paternal origin during each meiotic event. This is known as Independent Assortment. Diagram the possible ways that two pairs of homologous chromosomes can align at metaphase I and the resulting haploid products in telophase II below.
Premeiotic Interphase in G2

-6-

Exercise III: Meiosis and the Life Cycles of Organisms Just as each cell has a cell cycle, organisms have a life cycle. For most, the life cycle includes sexual reproduction. Meiosis is something that takes place at just one point in any sexually reproducing organism's life cycle. The timing of meiosis and whether the organism spends its assimilative life composed of diploid or haploid cells (or part of its life composed of haploid cells, and part with diploid) determines the life cycle pattern. Life cycles can be diploid, haploid or an alternation of generations. Diploid Life Cycle In animals, meiosis generally produces just haploid sex cells, or gametes, which at fertilization start the next generation. The only haploid cells of the animal are egg or sperm, and the respective maturation processes are called oogenesis and spermatogenesis. When gametes fuse, the zygote grows by mitosis producing the adult stage. All cells will be diploid and all cells are produced by mitosis. The animal life cycle is a diploid life cycle. Haploid Life Cycle In many protists, and some fungi, haploid gametes fuse to form a zygote, but the zygote immediately does meiosis forming single haploid cells. In protists, which remain single cell organisms, the nucleus is then haploid. At some time, a single cell will become a gamete and fuse with another to make a zygote, or haploid cells may do mitosis to make more individuals asexually. Haploid life cycles can be more complex. Fungi and some algae may make multicellular haploid organisms from the single-celled meiotic product by mitosis. At some time, special areas of the haploid body will become gamete-making structures (often called gametangia), and haploid gametes are then formed by mitosis. Alternation of Generations Most plants have both a multicellular haploid stage and a multicellular diploid stage in their life histories. This is called the alternation of generations. For most plants, either the diploid stage or the haploid generation predominates so most humans just haven't had the opportunity to become acquainted with these different forms of a plant's life. In plants, the structure in which meiosis occurs is called a sporangium. The multicellular diploid plants, or parts of plants that produce sporangia are called sporophytes (spore-making plant). Meiosis does not directly produce gametes, but produces haploid cells, called spores that in turn, grow, by mitosis, into multicellular haploid structures called gametophytes (gametemaking plant). Gametophytes eventually produce structures that make gametes by mitosis. Earlier you observed the process of meiosis in the male sporangium of the lily flower and saw the male spores that will develop into the male lily gametophytes. No matter how the life cycle varies, each gamete has a unique combination of chromosomes, each zygote will be unique, and genetic variation is both maintained and obtained within species. Compare three typical life cycle patterns shown and look at the timing of meiosis in the life cycle of each. Then compare the life cycle of the fern illustrated with the human life cycle to help you get a better idea of the alternation of generations life cycle typical of plants.

-7-

Diploid Life Cycle Human Life Cycle

Haploid Life Cycle

Alternation of Generations Fern Life Cycle

Fern Sporangia

Fern Gametophyte

-8-