AP Biology The Science of Drosophila Genetics Lab Report Title Analysis of Traits and Comparison of Observed Data with

Expected Data in Understanding the Life Cycle of Fruit Flies

David Wong

Abstract Drosophila melanogaster is the name of a fruit fly, a two-winged insect which lives on fruit (Arizona). Fruit flies play a major role in genetics where biologists can study transposable elements, heat shock responses, and oncogenes (EVDOTEK). The main objective is to understand the life cycle of the fruit flies by studying them across several generations. In the results, there are Punnett squares showing P, F1, and F2 generational crosses. With recorded data, the Chi square analysis tests the null hypothesis using observed and expected data. The data results show that the observed ratios match with the class expected ratios. In the conclusion section, the null hypothesis for the red and sepia crosses are accepted, but the hypothesis for the red-vestigial and sepia-normal dihybrid crosses were rejected.

Introduction Because of its short 14-day life cycle, the fruit fly was among the first chosen for study and analysis. Studying fruit flies helps increase understanding of biological functions in cells. Based on early studies and research, scientists were able to further understand gene linkage, crossing over, and mutational events, which not only apply to the fruit fly but to various other animals as well. Currently, scientists are studying transposable elements, heat shock responses, and oncogenes in which they hope to understand human diseases such as cancer (EDVOTEK). The fruit fly has four basic stages in its life cycle and goes through a complete metamorphosis (Woodrow). Stages in this life cycle are egg, larva, pupa, and adult. The length of the entire life cycle mainly depends on the temperature but typically lasts for 15 days at 20°C and 10 days at 25°C (EDVOTEK). Fruit flies lay their eggs in rotten fruit. Next after the egg is fertilized, the larva hatches within a few days. The larva has a total of two molting periods and three instars (periods of growth before and after molting, each lasting a couple days) until it reaches the pupa stage (Woodrow). The pupa stage is when the major adult organs more fully develop. Finally, the adult fly may push itself out of the pupa casing and live for up to several weeks (EDVOTEK). The main objective is to understand the life cycle of the fruit flies by studying them across several generations. This introduces basic concepts such as understanding genetics and the inheritance of traits (Arizona). It involves the separation of flies by

Document1

Page 1 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

gender and then followed by observations under the microscope to identify any genderspecific differences (Arizona). The lab also introduces the observation of heredity in fruit flies and, based on data, Punnett square constructions of wild-type and mutated flies. Included in the report is the Chi square analysis, which finally tests the null hypothesis using data and declares whether to accept or reject the hypothesis.

Null Hypothesis If the observed data was representative of the expected data, a Chi square analysis would show no significant differences between the two.

Materials & Methods Materials: 1) Live fruit flies as specimen (EDVO-Kit #287) 2) Anesthetic for flies 3) Vials for containment and separation 4) Microscope for observation 5) Blank paper as microscope slide 6) Pointer (Pencil / Wooden Stick) for moving specimen Methods / Procedure: 1) Separate fruit flies from original vial with eggs and medium into new vial with adult flies only 2) Sedate the fruit flies with anesthetic 3) Put fruit flies onto blank paper 4) Put blank paper under microscope 5) Observe fruit flies for gender-specific differences such as sex combs in males and larger body shape in females 6) Separate fruit flies into corresponding male vial or female vial The time requirement gives EDVOTEK three weeks for the shipment of the live flies. After receiving the live flies, allow several hours for setting up the initial parental crosses. It is expected to perform one cross per class period; at this pace, it will take several weeks to complete (EDVOTEK).

Document1

Page 2 of 16

AP Biology The Science of Drosophila Genetics Lab Report Results Red Male × Sepia Female R = Dominant red-eye trait (wild type) r = Recessive sepia-eye trait (mutant) P Generation: [RR] × [rr] Offspring:

David Wong

F1 Generation R R
r

F2 Generation R r
R

Rr

Rr

RR

Rr

r

Rr

Rr

r

Rr

rr

Data

F1 1 Red Male : 1 Red Female

F2 3 Red : 1 Sepia 1 Male : 1 Female 37 Red Male 34 Red Female 10 Sepia Male 16 Sepia Female

Expected

Actual

42 Red Male 23 Red Female

Document1

Page 3 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

Red Female × Sepia Male R = Dominant red-eye trait (wild type) r = Recessive sepia-eye trait (mutant) P Generation: [rr] × [RR] Offspring:

F1 Generation r r
R

F2 Generation R r
R

Rr

Rr

RR

Rr

R

Rr

Rr

r

Rr

rr

Data

F1 1 Red Male : 1 Red Female

F2 3 Red : 1 Sepia 1 Male : 1 Female 38 Red Male 35 Red Female 11 Sepia Male 15 Sepia Female

Expected

Actual

49 Red Male 51 Red Female

Document1

Page 4 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

Red Male × White Female W+ = Dominant Red-eyed trait (wild type) W = Recessive white-eyed (mutant) Xw Xw = White-eyed female (mutant) Xw+ Xw = Red-eyed heterozygote female (wild type) Xw Y = White-eyed male (mutant) Xw+ Y = Red-eyed male (wild type) P Generation: [Xw+ Y] × [Xw Xw] Offspring:

F1 Generation Xw+ Y
Xw
Xw Xw+ Xw Xw+ Xw Xw Y Xw Y

F2 Generation Xw Y
Xw+ Xw+ Xw
Xw Xw Xw Xw+ Y Xw Y

Data

F1 1 White Male : 1 Red Female 42 Red Male 23 Red Female

F2 1 Red : 1 White 1 Male : 1 Female N/A

Expected Actual

Document1

Page 5 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

White Male × Red Female W+ = Dominant Red-eyed trait (wild type) W = Recessive white-eyed (mutant) Xw Xw = White-eyed female (mutant) Xw+ Xw = Red-eyed heterozygote female (wild type) Xw Y = White-eyed male (mutant) Xw+ Y = Red-eyed male (wild type) P Generation: [Xw Y] × [Xw+ Xw+] Offspring:

F1 Generation Xw Y
Xw+ Xw+ Xw
Xw+ Xw+ Xw Xw+ Y Xw+ Y

F2 Generation Xw+ Y
Xw+ Xw+ Xw+
Xw Xw+ Xw Xw+ Y Xw Y

Data

F1

F2 1 Red Male : 1 White Male 4 Red Female : 0 White Female 1 Male : 1 Female 63 Red Male 28 Red Female 47 White Male 21 White Female

Expected

1 Red Male : 1 Red Female

Actual

91 Red Male 91 Red Female

Document1

Page 6 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

Red Vestigial Female x Sepia Normal Male R = Dominant red-eye trait (wild type) r = Recessive sepia-eye trait (mutant) vg+ = Dominant normal wings (wild type) vg = Recessive vestigial wings (mutant) P Generation: [r r vg+ vg+] × [R R vg vg] Offspring: [R r vg+ vg] × [R r vg+ vg] (P Generation offspring all [R r vg+ vg])

F1 Generation
r vg+ R vg
R r vg+ vg

F2 Generation
r vg+
R r vg+ vg

r vg+
R r vg+ vg

r vg+
R r vg+ vg

R vg+ R vg+
R R vg+ vg+

R vg
R R vg+ vg

r vg+
R r vg+ vg+

r vg
R r vg+ vg

R vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

R vg

R R vg+ vg

R R vg vg

R r vg+ vg

R r vg vg

R vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

r vg+

R r vg+ vg+

R r vg+ vg

r r vg+ vg+

r r vg+ vg

R vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

r vg

R r vg+ vg

R r vg vg

r r vg+ vg

r r vg vg

Data

F1

F2 9 Red Normal : 3 Red Vestigial : 3 Sepia Normal : 1 Sepia Vestigial 1 Male : 1 Female 79 Red Normal Male 67 Red Normal Female 49 Red Vestigial Male 48 Red Vestigial Female

Expected

1 Red Normal Male : 1 Red Normal Female

Actual

31 Red Normal Male 19 Red Normal Female 20 Red Vestigial Male 18 Red Vestigial Female

Document1

Page 7 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

Red Vestigial Male × Sepia Normal Female R = Dominant red-eye trait (wild type) r = Recessive sepia-eye trait (mutant) vg+ = Dominant normal wings (wild type) vg = Recessive vestigial wings (mutant) P Generation: [R R vg vg] × [r r vg+ vg+] Offspring: [R r vg+ vg] × [R r vg+ vg] (P Generation offspring all [R r vg+ vg])

F1 Generation
R vg r vg+
R r vg+ vg

F2 Generation
R vg
R r vg+ vg

R vg
R r vg+ vg

R vg
R r vg+ vg

R vg+ R vg+
R R vg+ vg+

R vg
R R vg+ vg

r vg+
R r vg+ vg+

r vg
R r vg+ vg

r vg+

R r vg+ vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

R vg

R R vg+ vg

R R vg vg

R r vg+ vg

R r vg vg

r vg+

R r vg+ vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

r vg+

R r vg+ vg+

R r vg+ vg

r r vg+ vg+

r r vg+ vg

r vg+

R r vg+ vg

R r vg+ vg

R r vg+ vg

R r vg+ vg

r vg

R r vg+ vg

R r vg vg

r r vg+ vg

r r vg vg

Data

F1

F2 9 Red Normal : 3 Red Vestigial : 3 Sepia Normal : 1 Sepia Vestigial 1 Male : 1 Female 12 Red Normal Male 16 Red Normal Female 4 Red Vestigial Male 0 Red Vestigial Female

Expected

1 Red Normal Male : 1 Red Normal Female

Actual

25 Red Normal Male 26 Red Normal Female 3 Red Vestigial Male 0 Red Vestigial Female

Document1

Page 8 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

Chi Square Analysis of Red Male × Sepia Female ∑ [( ) ]

O = the observed frequencies C = the calculated frequencies Degrees of Freedom = (# of Phenotypic Classes) – 1 = 1 Table 1: Phenotype Red Eyes Sepia Eyes Totals Table 2: P = 0.99 P = 0.95 Observed 71 26 97 Calculated 72.75 24.25 97 -1.75 1.75 — ( ) 3.0625 3.0625 — [( ) ]

0.04209622 0.12628866

— P = 0.90 P = 0.80 P = 0.70 P = 0.50 P = 0.30 P = 0.20 P = 0.05 P = 0.01 Degrees of — — — — → ← — — — — Freedom 0.000157 0.000393 0.0158 1 0.0642 0.148 0.455 1.074 1.642 3.841 6.635 2 3 4 5 6 7 8 9 10 20 30 50 100 0.020 0.115 0.297 0.554 0.872 1.239 1.65 2.09 2.56 8.26 14.95 29.71 70.07 0.103 0.352 0.711 1.145 1.635 2.167 2.733 3.325 3.940 10.851 18.493 34.764 77.930 0.211 0.584 1.004 1.610 2.204 2.833 3.490 4.168 4.865 12.443 20.599 37.689 82.945 0.446 1.005 1.649 2.343 3.070 3.822 4.594 5.380 6.179 14.578 23.364 41.449 87.945 0.713 1.424 2.195 3.000 3.828 4.671 5.527 6.393 7.267 16.266 25.508 44.313 92.129 1.386 2.366 3.357 4.351 5.348 6.346 7.344 8.343 9.342 19.337 29.336 49.335 99.334 2.408 3.665 4.878 6.064 7.231 8.383 9.524 10.656 11.781 22.775 33.530 54.723 3.219 4.642 5.989 7.289 8.558 9.803 11.030 12.242 13.442 25.083 36.250 58.164 5.991 7.815 9.488 11.070 12.592 14.067 15.507 16.919 18.907 31.400 43.773 67.505 9.210 11.345 13.277 15.086 16.812 18.475 20.090 21.666 23.209 37.566 50.892 76.154

103.906 111.677 124.342 135.806

Document1

Page 9 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

Chi Square Analysis of Red Female × Sepia Male ∑ [( ) ]

O = the observed frequencies C = the calculated frequencies Degrees of Freedom = (# of Phenotypic Classes) – 1 = 1 Table 1: Phenotype Red Eyes Sepia Eyes Totals Table 2: P = 0.99 P = 0.95 Observed 73 26 99 Calculated 74.25 24.75 99 -1.25 1.25 — ( ) 1.5625 1.5625 — [( ) ]

0.021043771

— P = 0.90 P = 0.80 P = 0.70 P = 0.50 P = 0.30 P = 0.20 P = 0.05 P = 0.01 Degrees of — — — → ← — — — — — Freedom 0.000157 0.000393 0.0158 1 0.0642 0.148 0.455 1.074 1.642 3.841 6.635 2 3 4 5 6 7 8 9 10 20 30 50 100 0.020 0.115 0.297 0.554 0.872 1.239 1.65 2.09 2.56 8.26 14.95 29.71 70.07 0.103 0.352 0.711 1.145 1.635 2.167 2.733 3.325 3.940 10.851 18.493 34.764 77.930 0.211 0.584 1.004 1.610 2.204 2.833 3.490 4.168 4.865 12.443 20.599 37.689 82.945 0.446 1.005 1.649 2.343 3.070 3.822 4.594 5.380 6.179 14.578 23.364 41.449 87.945 0.713 1.424 2.195 3.000 3.828 4.671 5.527 6.393 7.267 16.266 25.508 44.313 92.129 1.386 2.366 3.357 4.351 5.348 6.346 7.344 8.343 9.342 19.337 29.336 49.335 99.334 2.408 3.665 4.878 6.064 7.231 8.383 9.524 10.656 11.781 22.775 33.530 54.723 3.219 4.642 5.989 7.289 8.558 9.803 11.030 12.242 13.442 25.083 36.250 58.164 5.991 7.815 9.488 11.070 12.592 14.067 15.507 16.919 18.907 31.400 43.773 67.505 9.210 11.345 13.277 15.086 16.812 18.475 20.090 21.666 23.209 37.566 50.892 76.154

103.906 111.677 124.342 135.806

Document1

Page 10 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

Chi Square Analysis of Red Vestigial Female x Sepia Normal Male ∑ Table 1: Phenotype Red Normal Red Vestigial Sepia Normal Sepia Vestigial Totals Table 2: P = 0.99 P = 0.95 Observed 148 97 0 0 245 Calculated 137.8125 45.9375 45.9375 15.3125 245 10.1875 51.0625 -45.9375 -15.3125 — ( ) [( ) ] [( ) ]

103.7851563 2607.378906 2110.253906 234.4726563 —

0.753089569 56.75926871 45.9375 15.3125

— P = 0.90 P = 0.80 P = 0.70 P = 0.50 P = 0.30 P = 0.20 P = 0.05 P = 0.01 Degrees of — — — — — — — — — → Freedom 0.000157 0.000393 0.0158 1 0.0642 0.148 0.455 1.074 1.642 3.841 6.635 2 3 4 5 6 7 8 9 10 20 30 50 100 0.020 0.115 0.297 0.554 0.872 1.239 1.65 2.09 2.56 8.26 14.95 29.71 70.07 0.103 0.352 0.711 1.145 1.635 2.167 2.733 3.325 3.940 10.851 18.493 34.764 77.930 0.211 0.584 1.004 1.610 2.204 2.833 3.490 4.168 4.865 12.443 20.599 37.689 82.945 0.446 1.005 1.649 2.343 3.070 3.822 4.594 5.380 6.179 14.578 23.364 41.449 87.945 0.713 1.424 2.195 3.000 3.828 4.671 5.527 6.393 7.267 16.266 25.508 44.313 92.129 1.386 2.366 3.357 4.351 5.348 6.346 7.344 8.343 9.342 19.337 29.336 49.335 99.334 2.408 3.665 4.878 6.064 7.231 8.383 9.524 10.656 11.781 22.775 33.530 54.723 3.219 4.642 5.989 7.289 8.558 9.803 11.030 12.242 13.442 25.083 36.250 58.164 5.991 7.815 9.488 11.070 12.592 14.067 15.507 16.919 18.907 31.400 43.773 67.505 9.210 11.345 13.277 15.086 16.812 18.475 20.090 21.666 23.209 37.566 50.892 76.154

103.906 111.677 124.342 135.806

Document1

Page 11 of 16

AP Biology The Science of Drosophila Genetics Lab Report Chi Square Analysis of Red Vestigial Male × Sepia Normal Female ∑ Table 1: Phenotype Red Normal Red Vestigial Sepia Normal Sepia Vestigial Totals Table 2: P = 0.99 P = 0.95 Observed 28 4 0 0 32 Calculated 18 6 6 2 32 10 -2 -6 -2 — ( 100 4 36 4 — ) [( ) ]

David Wong

[(

) ]

6 2

— P = 0.90 P = 0.80 P = 0.70 P = 0.50 P = 0.30 P = 0.20 P = 0.05 P = 0.01 Degrees of — — — — — — — — — → Freedom 0.000157 0.000393 0.0158 1 0.0642 0.148 0.455 1.074 1.642 3.841 6.635 2 3 4 5 6 7 8 9 10 20 30 50 100 0.020 0.115 0.297 0.554 0.872 1.239 1.65 2.09 2.56 8.26 14.95 29.71 70.07 0.103 0.352 0.711 1.145 1.635 2.167 2.733 3.325 3.940 10.851 18.493 34.764 77.930 0.211 0.584 1.004 1.610 2.204 2.833 3.490 4.168 4.865 12.443 20.599 37.689 82.945 0.446 1.005 1.649 2.343 3.070 3.822 4.594 5.380 6.179 14.578 23.364 41.449 87.945 0.713 1.424 2.195 3.000 3.828 4.671 5.527 6.393 7.267 16.266 25.508 44.313 92.129 1.386 2.366 3.357 4.351 5.348 6.346 7.344 8.343 9.342 19.337 29.336 49.335 99.334 2.408 3.665 4.878 6.064 7.231 8.383 9.524 10.656 11.781 22.775 33.530 54.723 3.219 4.642 5.989 7.289 8.558 9.803 11.030 12.242 13.442 25.083 36.250 58.164 5.991 7.815 9.488 11.070 12.592 14.067 15.507 16.919 18.907 31.400 43.773 67.505 9.210 11.345 13.277 15.086 16.812 18.475 20.090 21.666 23.209 37.566 50.892 76.154

103.906 111.677 124.342 135.806

Document1

Page 12 of 16

AP Biology The Science of Drosophila Genetics Lab Report Conclusion

David Wong

The hypotheses for the red and sepia crosses are valid. Both crosses had one degree of freedom. The Chi square value of the red male × sepia female cross was 0.168 or between 0.148 and 0.455, so it was below the 3.841 value in the P column of 0.05. The Chi square value of the red female × sepia male cross was 0.084 or between 0.0642 and 0.148, so it was below the 3.841 value in the P column of 0.05. Based on the Chi square analysis of the observed data and the expected data showing that the Chi square values did not exceed the 3.841 values in the P columns, the null hypothesis of those crosses should be accepted. However, the hypotheses for the red-vestigial and sepia-normal dihybrid crosses were invalid. Both the crosses had four categories, so the degree of freedom is three. In the first cross of red-vestigial female x sepia-normal male, the Chi square value was 118.8, which overly exceeded the 7.815 value under the P column for three degrees of freedom. In the red-vestigial male × sepia-normal female cross, the Chi square value was . This value also exceeds the 7.815 value under the P column for three degrees of freedom. This means that the observed data for both crosses were highly significant, and thus the null hypotheses should be rejected. The hypotheses for the red-vestigial and sepia-normal dihybrid crosses were rejected because their observed data had unusually high numbers of red-eyed flies but it had zero sepia-eyed flies. According to the ratios, it was expected that there would be some sepia-eyed flies. This means that there were significant differences between the observed and expected data, which can be as a result of error or chance. Data can be affected by sources of error. For example, a source of error would be a misidentification or miscategorization of a fly’s gender or trait, thus affecting the number in each original category. Study Questions: 1) If the genes analyzed by the dihybrid cross were linked on the same chromosome, how would that affect your results? If the locations of those genes are near enough so that recombination is more unlikely, then the offspring may instead have a pattern of alleles matching their parents’. 2) Why is it important that the parental females be virgins? The parental females must be virgins because they will usually mate only once in their lifetime. 3) Why are the adult flies removed? They must be removed so that they can be separated by gender and further examined under the microscope to note any distinctive features.

Document1

Page 13 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

4) Why are Drosophila favorite experimental models for geneticists? Fruit flies are easy to study because they are small, easily handled, short-lived, and different depending on gender, making it is easy to separate by male or female. 5) Why is it necessary to analyze a large number of offspring before making conclusions about genotypic and phenotypic ratios? A large number of offspring will be more likely to resemble the expected data and less likely to be affected by chance. 6) Why did you collect data for both males and females? Sex-linked crosses involve data specifically for males and females. 7) If you were a molecular biologist and had obtained two gene clones for eye color, one isolated from pure-bred, wild-type, red-eyed flies and the other from the mutant, sepia-eyed flies, how would you analyze these genes to identify the type of gene mutation? To identify the type of gene, the biologist can mate the flies together and look at the generational data from F1 to F2. Generally, the mutant alleles are recessive to the wild-type alleles. Then by analyzing the higher and lower numbers of traits, it is possible to figure out whether it is a wild-type or mutant gene. 8) What possible genotypes and phenotypes would you observe if the F2 flies from the dihybrid cross were allowed to randomly mate? Genotypes: Phenotypes: 1 × [R R vg+ vg+] 9 Red Normal 2 × [R R vg+ vg] : 1 × [R R vg vg] 3 Red Vestigial 2 × [R r vg+ vg+] : 4 × [R r vg+ vg] 3 Sepia Normal 2 × [R r vg vg] : 1 × [r r vg+ vg+] 1 Sepia Vestigial 2 × [r r vg+ vg] 1 × [r r vg vg]

Document1

Page 14 of 16

AP Biology The Science of Drosophila Genetics Lab Report

David Wong

9) Assume that during a hypothetical monohybrid cross, the double recessive gene was lethal and expressed itself early in the fly development. Would that skew the results in a large population? Would the recessive eventually disappear from the population? A lethal gene may skew the results more than not because it would be much rarer to find a living specimen with the lethal double recessive gene. However, this does not mean that the recessive genes will disappear from the population because the recessive genes may still be present in heterozygous flies for which their genes are unexpressed and nonlethal. Thus, the heterozygous flies may mate and give offspring to continue passing on the recessive gene. 10) Based on the example in the background section, set up a Punnett square to predict the genotypic and phenotypic ratios obtained from a dihybrid cross in the F1 generation and F2 generation. (HINT: Red eyes + vestigial wings = RRww. Sepia eyes + normal wings = rrWW. The genotypes for all F1 flies are RrWw. Therefore, each F1 fly can produce 4 types of gametes. Determine these and set up a 4×4 Punnett square.) See Results section. 11) Use the χ2 statistic to determine the values for the monohybrid, dihybrid, and sex-linked crosses for your individual data and for the class data. Are the hypotheses valid? See Results section. The hypotheses for the red and sepia crosses are valid while the hypotheses for the red-vestigial and sepia-normal dihybrid crosses were invalid. However, Chi square analysis could not be done for the sex-linked crosses because data for the F2 generation were not available.

Works Cited “An Introduction to Drosophila Melanogaster”. University of Arizona. <http://biology.arizona.edu/sciconn/lessons2/geiger/intro.htm>. EDVO-Kit #287: The Science of Drosophila Genetics. Bethesda: EDVOTEK, Inc., 19911998. <http://www.edvotek.com/287.html>. “Life Cycle of the Fruit Fly”. Woodrow Wilson Biology Institute. <http://www.woodrow.org/teachers/bi/1994/life_cycle.html>.

Document1

Page 15 of 16

AP Biology The Science of Drosophila Genetics Lab Report Comments N/A

David Wong

Document1

Page 16 of 16

Sign up to vote on this title
UsefulNot useful