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Lab Three: Mitosis and Meiosis
Introduction
There are two types of nuclear division, mitosis and meiosis. Mitosis is usually used for the growth and replacementof somatic cells, while meiosis produces the gametes or spores used in an organism’s reproduction.Mitosis is the first of these studied in this lab. It is easily observed in cells that are growing at a rapid pace such aswhitefish blastula or onion root tips, which are used in this lab. The root tips contain an area called the apicalmeristem that has the highest percentage of cells undergoing mitosis. The whitefish blastula is formed directly after the egg is fertilized. This is a period of rapid growth and numerous cellular divisions where mitosis can be observed.Just before mitosis the cell is in interphase. In this part of the cell cycle the cell will have a distinct nucleus andnucleoli where the thin threads of chromatin are duplicated. After duplication the cell is ready to begin mitosis andits starts with a step called prophase. In prophase, the chromatin thicken into distinct chromosomes and the nuclear envelope breaks open releasing them into the cytoplasm. The first signs of the spindle begin to appear. Next the cell begins metaphase, where the spindle attaches to the centromere of each chromosome and moves them to the samelevel in the middle of the cell. This level position is called the metaphase plate. Anaphase begins when thechromatids are separated and pulled to opposite poles. Then, the final stage is telophase. The nuclear envelope isreformed and the chromosomes gradually uncoil. Cytokinesis may occur, in which case, a cleavage furrow will formand the two daughter cells will separate.Meiosis is more complex and involves two nuclear divisions. The two divisions are called Meiosis I and Meiosis IIand they result in the production of four haploid gametes. This process allows increased genetic variation due tocrossing 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 andsynapsis begins. This is where crossing over occurs resulting in the recombination of genes. Metaphase I moves thetetrads to the metaphase plate in the middle of the cell, and Anaphase I reduces the tetrads to their original twostranded form and moves them to opposite poles. Telophase I then prepares the cell for its second division. MeiosisII generally resembles mitosis except that the daughter cells are haploid instead of diploid. DNA replication does notoccur in Interphase II, and prophase, metaphase, anaphase, and telophase occur as usual. The only change is thenumber of chromosomes.The process of crossing over can be easily studied in
Sordaria fimicola
, an ascomycete fungus.
Sordaria
form a setof eight ascospores called an ascus. They are contained in a perithecium until they are mature and ready for release.Crossing over can be observed in the arrangement and color of these asci. If an ascus has four tan ascospores in arow and four black ascospores in a row (4:4 arrangement), then no crossing over had taken place. However, if theasci has black and tan ascospores in sets of two (2:2:2:2 arrangement) or two pairs of black ascospores and four tanascospores in the middle (2:4:2 arrangement), then crossing over had taken place.
Hypothesis
Mitosis occurs in whitefish blastula and onion root tip, and it is easily observable. Meiosis and crossing over occursin the production of gametes and spores.
Materials
This lab requires prepared slides of whitefish blastula, onion root tips, and Sordaria, pencil, paper, a lightmicroscope, and a chromosome simulation kit.
Methods
 Exercise 3A.1: Observing Mitosis
Prepared slides of whitefish blastula and onion root tips were observed under the 10X and 40X objectives. A cell ineach stage of mitosis were identified, and then sketched.
 
 Exercise 3A.2: Time for Cell Replication
Using a high power objective, every cell in a field of view was observed. Each cell was counted as being in one of the stages of mitosis and recorded. At least 200 cells and 3 fields of vision were counted and recorded. Next, the percentage of cells in each stage was recorded and the amount of time spent in each phase was calculated.
 Exercise 3B.1: Simulation of Meiosis
In this part of the lab, a chromosome simulation kit was used to demonstrate meiosis. Two strands of the same color were connected to simulate DNA replication in both of the homologous pairs. Next, the chromosomes wereentwined to represent synapsis. Sections of beads were switched between the pairs as in crossing over and werealigned at the equator. Next, anaphase was simulated as the homologous pairs were separated and then telophase wassimulated by pushing the chromosomes into two separate cells (circles).Meiosis II was simulated as well. The DNA is not replicated in Interphase II. The chromosomes again move to theequator and in Anaphase II the two chromatids were separated and moved to opposite poles. Telophase II separatesthem into four different cells.
 Exercise 3B.2: Crossing Over during Meiosis in Sordaria
Prepared slides of 
Sordaria fimicola
were observed under a light microscope. Over 100 asci were identified as either 4:4 or asci showing crossover and recorded. The percentage of each and the map units were calculated.
Results
Whitefish Blastula
 
Onion Root TipTable 3.1: Time for Cell ReplicationNumber of Cells
Field1Field2Field3TotalPercentageTime
Interphase
42364712561.27%14 hours 42 minutes
Prophase
1013184120.10%4 hours 49 minutes
Metaphase
654157.35%1 hour 46 minutes
Anaphase
23273.43%49 minutes
Telophase
754167.84%1 hour 59 minutes204
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