Analyzing observed inherited phenotype proportions of sepia and wild type Drosophila of the F 2 generation and comparing observed

and expected phenotypic proportions

Vishal Oza 12/01/10 Green Hope High School AP Biology 4th period, Mr. Jordan

Abstract

The main idea of this experiment is to test and gather sufficient evidence to conclude that the monohybrid cross between sepia and wild type drosophila will yield proportions of 0.25% and 0.75% respectively. To test this phenomenon of inheritance when crossing heterozygous drosophila, we crossed the male wild (red eyes) type with the female sepia type (brown eyes) which produced the F1 (se+se) generation, approximately 10 days later the F1 drosophila had mated and produced the F2 (se+se x se+se) . The flies were then recorded for their phenotypes fly by fly, the results the experiment yielded was approximately 18.75% on average were sepia type, and approximately 81.25% on average were wild type. This is close to the expected value for the proportions of the phenotypes for this cross, which is wild to sepia, a 3 to 1 ratio, this also proves that in fact sepia is the recessive phenotype and wild eyes the dominant phenotype.

Introduction

The central problem surrounding this experiment is whether or not the phenotypic ratios of wild and sepia type drosophila fall within a close range to the 3 to 1 ratio respectively, therefore the percentage of flies should fall within 25% for sepia eyes and 75% for the wild type eyes. The idea of inheritance is the interest in this lab; according to the crosses the inherited phenotype of sepia should have an expressed recessive outcome on the offspring, and the inherited phenotype of wild type eyes should have an expressed dominant outcome on the offspring. The mode of inheritance in the F2 sepia wild cross is a monohybrid, meaning that there is a single pair of contrasting characteristics involved. The phenotypes sepia and wild type eyes are not sex-linked because the number of female flies was equal to the number of male flies; therefore the characteristic cannot be inherited through the Xchromosome. The main hypothesis of the experiment is how these traits are inherited. By looking at patterns in traits between the F1 and F2 generation, and by determining these patterns of inheritance; we can then hypothesize about how the traits are inherited. We figured out previously that the trait was not sex-linked, and we also hypothesized that for the offspring the sepia will be a recessive trait while the wild type will be a dominant trait. The overall motivation of the experiment is to identify which trait is dominant and which is recessive, and then hypothesize about the method of inheritance. The chi-square test is used to test our null hypothesis and to see if our data has sufficient evidence to conclude that our data is correct and can be accepted , to do this we calculate our chi square value and compare that to the critical value, which is derived from the df (degrees of freedom) and the probability, the df =n-1 which is 2-1 in this case, due to the fact that there are only 2 choices for phenotypes and the probability is 0.5 so according to the chart the critical value is 3.84, if the chi-square value is less than 3.84 we fail to reject the null hypothesis and if it is greater than or equal to 3.84 we reject the null hypothesis. The drosophila fly that was used in this experiment is appropriate due to its small life cycle, very limited food and space requirements, it also produces many offspring and can be easily immobilized (College Board, 2001 revised). The drosophila also have only 4 pairs of chromosomes 3 pairs of autosomes and 1 pair of sex chromosomes (Campbell-Reece, 2002).The life cycle of a drosophila fly is also only 12 days long, with 2 days being a egg, 2 days I the first instar larva and the same amount of time for the second and third instar larva stage , and later 2 more days for the stage of a pupa, and finally a fully metamorphosed adult (college board, 2001 revised). Thomas Hunt Morgan an embryologist at Columbia University first associated genes with chromosomes in the early 1900s (Campbell-Reece,

2002) this later showed that genes also had a specific location on the chromosome based on how they were inherited. Independent assortment and crossing over also show how recombination occurs and is also applicable to the flies, for explaining inheritance of certain traits. The law of segregation that Mendel theorized stated that the genes were segregated and became part of separate sex cells and after fertilization at the developmental stages the dominant trait hides the recessive trait and the dominant trait is what was expressed as the phenotype (Henig, Robin Marantz, 2009). Genetic maps created by Sturtevant to find the loci of genes on a chromosome, and their distances (frequencies) this can be determined from recombination data; he then found that the distance between the two genes is where crossing over takes place and therefore the relative locations on a chromosome. The linkage maps also mapped the recombination frequencies, which helped in determining the purpose and effect of the gene. The distances between the genes were expressed in map units (Gelbart WM, 1993) which is also a method to measure distances on a drosophila chromosome.

Materials and methods Paint brush Flies sepia se(3) and wild type + 17-2010 sep 30 Medium (flakes, water, yeast) flakes: Instant drosophila medium formula 4-24 Fly Nap PAT NO. 4,224,898 Funnel Isopropyl Alcohol Petri dish Vials (for housing the medium) Spongy covers Fume hood Dissecting microscope Carolina Biological Supply Company (source of chemicals and materials) Morgue (flask with alcohol for disposal of flies)

Obtain a container of sepia x wild drosophila and a Fly Nap anesthetic and apply to a paint brush, then put inside container where the flies are being contained and keep container under fume hood, leave brush in container until the flies are no longer mobile. Then prepare the medium for the flies, take 1 scoop of flakes, 1 scoop of water, and 1 pinch yeast and mix together. Then place the immobilized flies in a Petri dish, and place under a dissecting microscope, and then begin recording sex, presence of wings, and eye color fly by fly. Afterwards place 10 females and 10 males of the F1 generation into the newly prepared medium. Later generate a hypothesis about what a prediction could be for the proportion of the F2 generation phenotypes. Approximately 10 days after the transfer record the number for the flies that are alive, and then dispose of the flies by putting them into the morgue. After about 2 days observe and record the phenotypes of the flies of the F 2 generation and then dispose by putting them into the morgue. Repeat the process of recording and disposing of the flies in two, four, eight, and 11 days from the current date. After gathering all the data for the F2 total up all the flies for each of the four dates and calculate the

proportion of sepia type and wild type eyes out of the total number of flies. Then perform the chi square test statistic analysis: ( )

by plugging in the expected values which are derived from the 3 to 1 ratio, and the observed values. Once calculated compare that to the critical value based on degrees of freedom (df) = n-1 and the probability on the chart. If the chi square value is less than the critical value then we fail to reject the null hypothesis, but if the chi square value is greater than or equal to the critical value than we reject the null hypothesis (College Board, 2001 revised)

Results F1 generation 10/22/10 male 0 10 10 female 0 9 9 Total 0 19 19

Sepia type Wild type total

F1 generation 11/01/10

Sepia type Wild type total

male 0 5 5

female 0 5 5

Total 0 10 10

F2 generation 11/03/10

Sepia type Wild type total

male 3 6 9

female 2 17 19

Total 5 23 28

F2 generation 11/05/10

Sepia type Wild type total

male 5 18 23

female 2 21 23

Total 7 39 46

F2 generation 11/09/10

Sepia type Wild type total

male 2 12 14

female 6 25 31

Total 8 37 45

F2 generation 11/12/10

Sepia type Wild type total

male 2 12 14

female 5 10 15

Total 7 22 29

Proportion of phenotype chart:

Date 11/3/10 11/5/10 11/9/10 11/12/10

Sepia type proportion 5/28 7/46 8/45 7/29

(

Wild type proportion 23/28 39/46 37/45 22/29

)

Proportion of phenotype=

Chi-square calculations table: phenotype observed expected sepia 27 37 wild 121 111 df=1 and probability is 0.5 so critical value = 3.84 3.6<3.84(on chart) Conclusion: Since the chi square value is less than the critical value, we fail to reject the null hypothesis. Some errors could be that the company sold us the wrong flies and possibly the 11/12 data containing F 3 flies. We could have improved the experiment by preventing the medium from sticking to the sides of the vials so the flies do not get stuck to it. o-e -10 10 (o-e)2 100 100 (o-e)2/e 2.7 0.9 X2=3.6

Proportion of the phenotypes of F2 Drosophila at various dates 0.90 proportion of phenotype to total 0.80 0.70 0.60 0.50 0.40 0.30 0.20 0.10 0.00 proportion of sepia type Drosophila proportion of wild type Drosophila

Sources AP biology lab manual (college board, 2001) revised Campbell-Reece, 2002 Henig, Robin Marantz (2009). The Monk in the Garden : The Lost and Found Genius of Gregor Mendel, the Father of Genetics. Houghton Mifflin. ISBN 0-395-97765-7. "The article, written by a monk named Gregor Mendel..."