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Gregor Mendel proposed the law of independent assortment, which states that alleles of different genes assort independently of one another during gamete formation. Mendel’s experiments of mixing one trait, a monohybrid cross resulted in a 3:1 ratio between dominant and recessive phenotypes, in the inheritance of a single trait. While his experiment of mixing two traits, a dihybrid cross in which two characteristics were observed, the F1 generation only expressed the dominant phenotype, and in the F2 generation, both dominant and recessive traits were expressed and four phenotypic categories with a ratio of 9:3:3:1 were noticed. This is true for genes that are not linked to each other. Thus the purpose of this experiment was to determine if the recessive genes of vestigial wings and ebony body are not linked in Drosophilia melanogaster. Our experiment aims to prove that genes are not linked, so the null hypothesis would imply that there is to be no significant difference between what is observed and what is expected, resulting in a 9:3:31 ratio. Thereby illustrating Medelian genetics on independent assortment.
Drosophilia melanogaster, known as fruit flies (fly singular) are eukaryotic organisms that are found near unripe or rotted fruit. Drosophilia are small insects, and thus use minimal space when compared to other species. Fruit flies have a small number of chromosomes (four pairs), which are easily located in the large salivary gland cells. The diet of fruit flies is simple and does not require extraneous foods or materials. Additionally, the life cycle of fruit flies is quite short, lasting approximately 37 days. From an egg to an adult, it takes approximate-
Page 2 of 7 ly 10 days at room temperature (25°C). The short generation time of fruit flies makes them an ideal study for this experiment, as multiple generations and variations can be studied and examined in a short period of time. The life cycle of a fruit fly starts when an impregnated female fruit fly lays an egg. Fruit flies lay only one egg at a time. The egg hatches in 22 hours, and the larvae eats and grows for four days. The larvae goes through three larva stages called instars before entering the pupa stage. In the pupa stage, the pupal case forms, darkens and hardens for 4-6 days. Finally, the pupa ecloses into the adult stage. Males fruit flies have a smaller body with a rounded black tip at the end of their body. Females, on the other hand, have a pointed abdomen and are often lighter than males DISCUSSION
The results of the parental cross (WWbb & wwBB), demonstrate that the wild wings / wild body is the dominant allele as no vestigial wings / ebony body were seen in the F1 generation. Calculations from our F2 data suggest that the ratio of the wild wing / wild body, to vestigial wings / wild body, to wild wings / ebony body, to vestigial wings / ebony body is the same as the expected, which is 9:3:3:1. The chi square test does not prove that the hypothesis is correct, but it evaluates whether or not the data and the hypothesis have a good fit. So we determined the degrees of freedom (df ). The (df ) is a measure of the number of categories that are independent of each other. Based on the result that was calculated, our df was = 3, and the Chi-square value was 4.1 in which p = 0.5-0.1. Showing that the calculated result we obtained, was greater than 0.5. Since our result was greater than 0.5, we accepted the null hypothesis. This suggests that the experimental data did not differ significantly from the expected ratio. Thus any other difference
Page 3 of 7 between them is due to random chance, or the abundance of food, protected shelter, lack of prey and the reproduction of more offsprings. Based on this we can say, the result of this experiment confirms Mendel’s law of independent assortment.
While the overall class’s data did not yield the expected phenotypic ratio of 9:3:3:1. Using the Chi-square test value which was = 17.4, and the degree of freedom was = 3 we determined the value of (p). p = <.005, thus it did not fit the theoretical expectation so we had to reject the null hypothesis. According to the given result, these numbers indicate that there might be a reason to doubt the hypothesis because the observed data differed significantly from the expected. It implies that the difference is not only due to random chance sampling, and might be due to lower statistics than the critical value causing the null hypothesis to be rejected. Other reasons for this outcome range from numerous potential errors. It is possible we did not count, and or observe all the flies in the vials. Flies often “escape” despite best efforts to contain them. It is very likely they flew away. We may have had flies with vestigial wings / ebony body misrepresented for wild wings / ebony body and vice versa for wild wings / wild body and vestigial wings / wild body due to human error. It is possible that we may have done our count too early, because some flies were still in their pupae stage and thus yet to mature to an adult. If the count was done some days later we might have had different ratios, which would have supported the null hypotheses. Other errors could have been the result of difference in food amount or the reproduction of less offspring than others.
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RESULTS The Dosophilia melanogaster parents of our F1 generation were homozygous wild wings / ebony body (WWbb) and vestigial wings / wild body (wwBB). Resulting in F1 generations being heterozygous wild wings / wild body (WwBb) as seen on the punette square as shown in Figure 1a.
Fig. 1a PARENTS wB Punette Square (Phenotypes) of Parents Wb WwBb Wb WwBb
Figure 1b shows the characteristics of the F1 generation
FIG. 1b Recorded Fly characteristics of F1 Generation Cross PHENOTYPE WILD WINGS/ WILDBODY
GENOTYPE WwBb MALE 50% = 50 FEMALE 50% = 50
To test the Mendel’s law of segregation we crossed the F1 progeny and collected the data of the resulting F2 Flies, as shown in Figure 2a. Fig. 2a Observed Data of Phenotypic Characters of F2 Generation
Page 5 of 7 PHENOTYPES MY GROUP’S OBSERVED DATA
57 28 23 10
OVERALL CLASS OBSERVED TO TAL
750 205 215 51
WILD WINGS / WILD BODY
VESTIGIAL WINGS / WILD BODY
WILD WINGS / EBONY BODY
VESTIGIAL WING / EBONY BODY
Chi-Square value of the X(pt sub) Distribution
p 0.995 df 1 2 3 4 5 6 7 8 9 10 0.000 0.010 0.072 0.207 0.412 0.676 0.989 1.344 1.735 2.156 0.000 0.051 0.216 0.484 0.831 1.237 1.690 2.180 2.700 3.247 0.016 0.211 0.584 1.064 1.610 2.204 2.833 3.490 4.168 4.865 0.455 1.386 2.366 3.357 4.351 5.348 6.346 7.344 8.343 9.342 2706 4.605 6.251 7.779 9.236 10.645 12.017 13.362 14.684 15.987 3.841 5.991 7.815 9.488 11.070 12.592 14.067 15.507 16.919 18.307 5.024 7.378 9.348 11.143 12.832 14.449 16.013 17.535 19.023 20.483 6.635 9.210 11.345 13.277 15.086 16.812 18.475 20.090 21.666 23.209 7.879 10.597 12.838 14.860 16.750 18.548 20.278 21.955 23.589 25.188 1 2 3 4 5 6 7 8 9 10 0.975 0.9 0.5 0.1 0.05 0.025 0.01 0.005 df
To determine the statistical relevance of the data, we performed the Chi square test on our F2 data as shown in Figure 2b, using the Chi-square table above
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Fig. 2b Chi-Square of Dihybrid Cross: My Groups Data OBSERVED EXPECTED (O-E) 57 28 23 10 66 22 22 7 -9 6 1 3 (O-E)^2 81 36 1 9 (O-E)^2/E 1.2 1.6 0.045 1.3
WILD / WILD VESTIGIAL / WILD
WILD / EBONY
Chi- square (X^2) = 4.1 Degrees of freedom = 3 Using the Chi- square chart, we found (p) p = 0.5-0.1 Since our (p) value was greater than 0.5, we concluded that it is not possible to reject the null hypothesis on the basis of this experiment.
To determine the statistical relevance of the data, we performed the Chi square test on the overall class’s F2 data as shown in Figure 3.
Fig. 3 Chi-Square of Dihybrid Cross; Overall Class’s Data
OBSERVED EXPECTED (O-E) (O-E)^2 (O-E)^2/E
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OBSERVED 750 215 205 51 EXPECTED 687 229 229 76 (O-E) 63 -14 -24 -25 (O-E)^2 3969 196 576 625 X^2 =17.365 (O-E)^2/E
LD / WILD
STIGIAL / WILD
STIGIAL / EBONY
Chi- square (X^2) = 17.4 Degrees of freedom = 3 Using the Chi- square chart, we found (p) p = <.005 Since our (p) value was less than 0.5, we concluded that it is not possible to accept the null hypothesis on the basis of this experiment so we rejected it.
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