EXPERIMENT 1: CELL DIVISION MITOSIS Introduction : Mitosis is the sequence of events by which the nuclear material of one cell

l is distributed, by a processing involving chromosomes, into two equal parts. At the right is a longitudinal section through an onion 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. Meiosis is more complex and involves two nuclear divisions. The two divisions are called Meiosis I and Meiosis II and they result in the production of four haploid gametes. This process allows increased genetic variation due to crossing over where genes can be exchanged. The process, like mitosis, depends on interphase to replicate the DNA. Meiosis begins with Prophase I. In this stage, homologous chromosomes move together to form a tetrad and synapsis begins. This is where crossing over occurs resulting in the recombination of genes. Metaphase I moves the tetrads to the metaphase plate in the middle of the cell, and Anaphase I reduces the tetrads to their original two stranded form and moves them to opposite poles. Telophase I then prepares the cell for its second division. Meiosis II generally resembles mitosis except that the daughter cells are haploid instead of diploid. DNA replication does not occur in Interphase II, and prophase, metaphase, anaphase, and telophase occur as usual. The only change is the number of chromosomes.

Objectives : 1. To learn how to used a light microscope in a proper ways. 2. To learn a technique of preparing a cells for the mitosis and meiosis experiment using a squash and colouring methods. 3. To understand the stages of the cells division in mitosis and meiosis.

Materials and apparatus : 1. The root tip of anions which was maintained in ethyl alcohol and glacial acetic 2. Aceto-orcien dye 3. 10 % HCl 4. 1 M HCl 5. 70 % alcohol 6. Light microscope 7. Alcohol lamp 8. Plate glass 9. Slide 10. Glass insert 11. Pin 12. matchsticks 13. filter paper Procedure : We were providing with roots tip of onion ( Allium cepa ) which was maintained in three part of ethyl alcohol and one part of glacial acetic acid about 4 hours. After that, the roots tip were kept in 70 % of alcohol at temperatures 40C. The roots tip must be hydrolysis in 1M of HCl at 600C about 6 minutes. 1. Aceto-osein dyeing a. 3 or 4 of onion roots tip were placed in glass insert and and a few drops of acetoorcien acid. b. the roots tip was heat up with on alcohol lamp within 2-3 minutes. c. The glass insert was sealed and let it about 10 minutes. If the dyeing unsatisfactory, the steps a-c were repeated. 2. Squash method a. The roots tip was placed on the clean slide. The tapered root tip was cut about 1-2mm and the excess part was removed.

b. The end of the squash was squashed until fines with the pin. A few drop of asetoorsien dye was placed on slide. The excess of the dye was cleaned by using the filter paper. c. The slide was closed by glass insert, then the glass insert was knocked slowly using a pin to wean the meristem tissues. d. The slide was heated for a few times e. The slide was placed in the fold of filter paper, the the slide is given a little push using a thumbnail to complete the isolation of cells f. The slide was studied carefully under the light microscope and the stages of the cell division that can be seen under the light microscope were draw. g. From the observation, the numbers of the chromosomes of somatid cell that can be seen was noted. h. For the meiosis, the sample of slides given was studied undrf the light microscope and the stages of the cell division that can be seen under the light microscope were draw.

Discussion : A. Mitosis Mitosis is nuclear division plus cytokinesis, and produces two identical daughter cells during prophase, , metaphase, anaphase, and telophase. Interphase is often included in discussions of mitosis, but interphase is technically not part of mitosis, but rather encompasses stages G1, S, and G2 of the cell cycle. During the interphase The cell is engaged in metabolic activity and performing its prepare for mitosis (the next four phases that lead up to and include nuclear division). Chromosomes are not clearly discerned in the nucleus, although a dark spot called the nucleolus may be visible. The cell may contain a pair of centrioles (or microtubule organizing centres in plants) both of which are organizational sites for microtubules. In the prophase stage, Chromatin in the nucleus begins to condense and becomes visible in the light microscope as chromosomes. The nucleolus disappears. Centrioles begin moving to opposite ends of the cell and fibers extend from the centromeres. Some fibers cross the cell to form the mitotic spindle. At the metaphase stage, Spindle fibres align the chromosomes along the middle of the cell nucleus. This line is referred to as the metaphase plate. This organization helps to ensure that in the next phase, when the chromosomes are separated, each new nucleus will receive one copy of each chromosome. During the anaphase, the paired chromosomes separate at the kinetochores and move to opposite sides of the cell. Motion results from a combination of kinetochore movement along the spindle microtubules and through the physical interaction of polar microtubules. Telophase stage starts when Chromatids arrive at opposite poles of cell, and new membranes form around the daughter nuclei. The chromosomes disperse and are no longer visible under the light microscope. The spindle fibers disperse, and cytokinesis or the partitioning of the cell may also begin during this stage.

After complete the mitotic phase, the cell will go through the cytokenesis.In animal cells, cytokinesis results when a fiber ring composed of a protein called actin around the center of the cell contracts pinching the cell into two daughter cells, each with one nucleus. In plant cells, the rigid wall requires that a cell plate be synthesized between the two daughter cells. B. Meiosis In prophase I, the DNA coils tightly and individual chromosomes become visible under the light microscope. Homologus chromosomes become closely associated in synapsis and they exchange segments by crossing over. By Metaphase I, the nuclear membrane has disappeared and the centrioles form spindle fibres attach to only side of each chromosomes and the two homologous chromosomes attach to microtubules orienteering from opposite poles. Each pair of homologous then lines up on the metaphase plate. Either maternal or parental may orient toward a given pole. In anaphase I, the microtubules of the spindle fibre shorten and pull the chromosomes toward the poles, taking both sister chromatids with them. Each pole ends up with a complete haploid set of chromosomes consisting one of member of the homologous pair. During the telophase I, the nuclear membrane reform around the daughter nuclei. Each daughter nucleus contain two sister chromatids for each chromosomes, attached to a common centromere. Because of the crossing over, the two sister chromatid are not identical. During the prophase II, the nuclear envelope breaks down and a new spindle form. In Metaphase II, spindle fibres bind to both sides of the centromeres. During the anaphase II, the spindle fibres contract and the sister chromatids move toward opposite poles. In telophase II, nuclear envelopes reform around the sets of daughter chromosomes.

C. Differences between mitosis and meiosis

The differences between mitosis and meiosis are as follows:

No Mitosis 1. 2. 3. Mitosis takes place within somatic cells (cells that make up the body). One single division of the mother cell results in two daughter cells. A mitotic mother cell can either be haploid or diploid. The number of chromosomes per 4. nucleus remains the same after division. It is preceded by a S-phase in 5. which the amount of DNA is duplicated. 6. In mitosis, there is no pairing of homologous chromosomes. There is no exchange of DNA 7. (crossing-over) between chromosomes. 8. The centromeres split during anaphase. The genotype of the daughter cells 9. is identical to that of the mother cells. 10. After mitosis, each daughter cell has exactly same DNA strands.

Meiosis Meiosis takes place within gamete cells (sex cells). Two divisions of the mother cell result in four meiotic products or haploid gametes. A meiotic mother cell is always diploid. The meiotic products contain a haploid (n) number of chromosomes in contrast to the (2n) number of chromosomes in mother cell. In meiosis, only meiosis I is preceded by a S-phase. During prophase I, complete pairing of all homologous chromosomes takes place. There is at least one crossing-over or DNA exchange per homologous pair of chromosomes. The centromeres do separate during anaphase II, but not during anaphase I. Meiotic products differ in their genotype from the mother cell. After meiosis, each daughter cell has only half of the DNA strands

Conclusion : 1. The number of chromosomes of onion is16. 2. Mitosis has a four stage which is prophase, metaphase, anaphase and telophase.

3. Meiosis has a eight stage which is prophase I, metaphase I, anaphase I, telophase I, prophase II, methaphase II, anaphase II and telophase II.