Drosophila Genetics

Adam Brantley Vaun S, Megan F, Taylor J. Period #2, 9th Grade Biology

Introduction:
The Common Fruit Fly: Drosophila melanogaster, or the common fruit fly, is the main subject of this lab. Drosophila melanogaster literally means dark-bellied dew lover in Greek [1]. Drosophila can be found all over the world, being more common in tropical areas. Their average lifespan is approximately 39 days, and each fly completes four different life-stages throughout the course of its life. These include the egg, larva, pupa, and adult fly stages, respectively [2]. The first stage of its life is the egg stage (visual 1). Pregnant females may lay up to 400 eggs into rotting fruit, and these eggs hatch within a period of only 12-15 hours. The next stage of their life, the larva stage, lasts 4 days, and during this time the young Drosophila molt twice into their 2nd instar and 3rd instar stages (visual 2). During its larvae stage, Drosophila feeds on fruit-decomposing microorganisms, as well as fruit itself [2]. After it completes its larval stage, Drosophila then begins its 4 day period as a pupa, during the course of which it will encapsulate itself in a cocoon of sorts and undergo metamorphosis (visual 3). Afterwards, it will emerge from the cocoon as an adult fly (visual 4). Drosophila melanogaster is ideally suited for the study of Genetics because of their short life cycle. An entirely new generation of adult drosophila can emerge in just 10 days following the pregnant females laying of fresh eggs. Because of this, scientists can study the traits passed on to the drosophilas offspring without having to wait unreasonable amounts of time for those offspring to mature, as would be the case with most organisms.

(visual 1) [4]

Drosophila melanogaster egg [3]

(visual 2)

Drosophila melanogaster larvae

(visual 3)

Drosophila melanogaster pupae [5]

(visual 4)

Drosophila melanogaster adult [6]

Key Terms: Gregor Mendel: Father of genetics. Between 1856 and 1863, Gregor Mendel studied the inheritance of certain traits in pea plants. His experiments led to the observation that the inheritance of physical characteristics in organisms follows certain laws. His research shaped the modern-day study of genetics [7]. Gene: A specific sequence of DNA, controlling the transmission and manifestation of physical traits in organisms [8]. Allele: Any of the forms of a gene that an organism may carry. Different alleles produce different phenotypes [4]. For example, a single gene, depending on what allele is expressed can code for either red eyes or sepia eyes. Dominant Allele: An allele expressed even when heterozygous with a recessive allele [9]. For example, both the genotypes AA and Aa will code for the A alleles resulting phenotype (dominant alleles are conveyed in writing by capital letters). Recessive Allele: An allele that is not expressed when heterozygous with a dominant allele. For example, only the genotype aa will code for the a alleles resulting phenotype (recessive alleles are conveyed in writing by lower-case letters). Homozygous: A genotype consisting of a set of identical alleles. For example, AA is a homozygous dominant genotype, and aa is a homozygous recessive genotype. Heterozygous: A genotype consisting of a set of different alleles. For example, Aa is a heterozygous genotype. Phenotype: The physical expression of a genotype. For example, blue eyes, brown eyes and green eyes are all different phenotypes.

 Genotype: The specific set of alleles for a certain phenotype that an organism possesses. For example, AA, Aa and aa are all different genotypes. Punnett Square: A diagram predicting the possible genotypes and/or phenotypes inherited across generations. For a Punnett square to function correctly, the genotypes of the male and female organisms being crossed must be known. For example: A A AAAAa a Aa Aa Dihybrid Cross: A Punnett square that predicts the possible genotypic and phenotypic expressions in an organism. For example, a Dihybrid cross may calculate the possible genotypes and/or phenotypes of the offspring of two organisms with the genotypes Aa BB and Aa Bb. For example: AB aB AB aB

A BAA BBAa BBAA BBAa BB A b AA Bb Aa Bb AA Bb Aa Bb a B Aa BB aa BB Aa BB aa BB a b Aa Bb aa Bb Aa Bb aa Bb

Traits To Be Examined: In this experiment, we will be examining the eye color and wing shape traits of the common fruit fly. Our 1st generation consists of a group of both male and female flies, all of them having the genotype Rr Ww, which codes for red eyes and the wings (visual 5) . The recessive alleles in these genotypes, when homozygous with another recessive allele, code for sepia (brown) eyes and apterous (wingless) flies (visuals 6, 7, 8). So, the offspring of these heterozygous flies could express any one of 4 different phenotypes: They could have either red eyed and wings, sepia eyed and wings, red eyed and wingless or sepia eyed and wingless traits. According to the Punnett square in the predictions segment of this report, when these 1st generation flies reproduce sexually their offspring of the 2nd generation could express any of the following genotypes:

Red Eyes, Wings (visual 5) ~~~~~ RR WW Rr WW RR Ww Rr Ww Red Eyes, Wingless (visual 6) ~~~~~ RR ww Rr ww

Sepia Eyes, Wings (visual 7) ~~~~~ rr WW rr Ww Sepia Eyes, Wingless (visual 8) ~~~~~ rr ww

(visual 5)

Red-eyed, winged [10]

(visual 6)

Red-eyed, apterous [10]

(visual 7)

Sepia-eyed, winged [10]

(visual 8)

Sepia-eyed, apterous [10]

Research Question:
Will the 2nd generation offspring of 2 parent Drosophila melanogaster, both heterozygous for eye color and wing shape, correspond to the expected 9:3:3:1 ratio as predicted by a dihybrid Punnett square?

Hypothesis:
I hypothesize that the offspring of the two heterozygous parent flies will be approximately 9/16 red-eyed and winged, 3/16 red-eyed and apterous, 3/16 sepia-eyed and winged, and 1/16 sepia-eyed and apterous. This is because, according to Gregor Mendels laws of genetics, traits are passed from generation to generation in a very specific way. If each parent

is heterozygous for both traits, and they can pass on only one of their two different alleles for each trait, then the phenotypes of the of the flys progeny should conform to the predicted 9:3:3:1 ratio.

Predictions:
If Gregor Mendels proposed laws of genetics are correct, then the offspring of the two parent flies, heterozygous for both eye color and wing shape, should conform, if not exactly, to the following predictions: RW Rw rW rw

R WRR WWRR WwRr WWRr Ww R w RR Ww RR ww Rr Ww Rr ww r W Rr WW Rr Ww rr WW rr Ww r w Rr Ww Rr ww rr Ww rr ww

Genotypes:

RR WW = 1/16 ~~~ RR Ww = 1/8 ~~~ Rr WW = 1/8 ~~~ Rr Ww = 1/4

RR ww = 1/16 ~~~ rr WW = 1/16 ~~~ Rr ww = 1/8 ~~~ rr Ww = 1/8 ~~~ rr ww = 1/16 Phenotypes: Red-eyed & winged = 1/4 ~~~~~ Red-eyed & apterous = 1/8

Sepia-eyed & winged = 1/8 ~~~~~ Sepia-eyed & apterous = 1/16

Materials:
Vial & cotton stopper ; Microscope (magnifying glass) ; Microscope slides ; FlyNap (fly anesthetic, 50% Triethylamine, 25% Ethanol, 25% Fragrances) [11] ;

2 live Drosophila melanogaster, 1 males, 1 females. ALL FLIES MUST BE HETEROZYGOUS FOR EYE COLOR AND WING SHAPE!!! ; Instant Drosophila medium ; Fly brushes (small brushes used to brush drugged flies onto microscope slides) .

Procedure:
1. Practice anesthetizing with inexpensive drosophila to perfect dosage amount (to assure that you do not overdose and kill the expensive heterozygous flies). The anesthetizing process includes dipping a wand into the FlyNap solution, shaking it

off, gently pulling back the cotton stopper and inserting it into the vial for approximately four minutes, or until the flies have all fallen under the influence of the drug. Experiment until you have perfected the technique. 2. Prepare multiple culture vials containing Drosophila medium.

3. Anesthetize heterozygous drosophila, sex them, and place 6-14 drosophila (preferably with equal amounts of both males and females) in one of the set up vials. Make sure to record exactly how many flies you isolate (for this experiment, my group isolated 6 female and 7 male Drosophila). 4. After one week, remove the adult Drosophila from the original vial and place them in a separate vial to live out their days. 5. 4 days after pulling the 1st generation flies out of the vial, we anesthetized the young adult flies and sorted them into four groups, depending on their phenotypes (see page 4 of this report). 6. 9 days after pulling the 1st generation flies out of the vial, we counted the phenotypes of the flies again, recording how many of each of the four types we found (ideally, one would count the phenotypes of the Drosophila every other day for ten days after pulling out the 1st generation flies). 7. Compare the actual phenotypic results to the expected results (see page 5 of this report).

Data:
GROUP DATA: Phenotype Red Eyes, Wings Red Eyes, Apterous Sepia Eyes, Wings Sepia Eyes, Apterous # Observed Ratio % 82 18 17 8 82\12565% 18/12515% 17/12514% 8/125 6%

CLASS DATA: Phenotype # Observed Ratio %

Red Eyes, Wings Red Eyes, Apterous Sepia Eyes, Wings Sepia Eyes, Apterous

549 99 81 30

549/75972% 99/759 13% 81/759 11% 30/759 4%

Analysis:

GROUP DATA:

CLASS DATA:

Conclusion:
If Gregor Mendel's laws of genetics are correct, then the 2nd generation drosophila phenotypes should conform to the predicted 9:3:3:1 ratio. In percentages, this would translate to 56% : 19% : 19% : 6 %. The actual results of my lab group yielded percentages of 65%, 15%, 14 % and 6%. Though not in complete correspondence to the predicted ratio, the group data is quite similar to it. Normally, the larger the sample of Drosophila observed, the closer the results should be to the expected phenotypic ratios. It is similar to flipping a coin: if a coin is flipped three times, the result may be three heads or three tails. But the more times a coin is flipped, the more likely the ratio will be 1:1. However, the flies that the entire class counted (759) had phenotypic percentages of 72%, 13%, 11%, and 4%. So, even though there were 634 more Drosophila examined in the class set than in the group set, the class results were actually not as close to the predicted results as were those of the smaller sample. This may be due to major miscounting on the part of other groups.

Possible Sources of Error:

A few possible sources of error in this experiment include the following. There may have been some miscounting during the recording of the Drosophila's phenotypic data. Though my group had a very specific way of examining the Drosophila's traits, I suspect that other groups were less consistent. They may have mistaken sepia eyes for red eyes, apterous flies for being winged (some apterous flies

have small nubs that people may have mistaken for wings), etc. This would explain why the class results were so radically different from the predicted results. We did not finish sort all the 2nd generation flies, which may have caused the data to become slightly different than what it what have been had we sorted them all. Selective sorting may be a possible source of error in this lab; instead of pulling flies out and counting them at random, some of us may have been specifically searching for all of the apterous flies, all of the sepia-eyed flies, all the red-eyed and winged flies, etc, which may have altered the results.

Research Implications:
These type of experiments are important to our understanding of genetics in that, from their results, we can learn how to calculate the possible genotypic and phenotypic outcomes of reproduction between any two organisms, not just Drosophila melanogaster. Because the property of heredity, or the passing on of genetic traits from one generation to another is shared by all living organisms, we can carry out genetic experiments with species other than humans and still apply the results to our own race. This experiment also relates to Charles Darwin's theory of Evolution by Natural Selection. For example, because wings are a useful part of Drosophila's everyday life, the species evolved so that almost all of its kind has wings (Drosophila's unicellular ancestors obviously did not begin with wings). By conducting experiments in the field of genetics, we can not only predict possible physical outcomes of 2nd generation offspring, we can also attempt to predict some of the likely courses that evolution may take in the future.

Literature Cited:
1. 2. 3. 4. http://www.greek-language.gr/greekLang/index.html Jarod Raithel. Meet Drosophila melanogaster: The Common Fruit Fly. PowerPoint presentation. http://upload.wikimedia.org/wikipedia/commons/c/cc/Drosophila_egg.png http://132.236.112.18/fruitfly/shaker/development/img/larvae_all3_0.8x_web_sm.jpg

5. http://www.exploratorium.edu/imaging_station/gal_media/drosophila/drosophila_pupae/droso phila_pupae_cat.jpg 6. 7. 8. 9. http://student.biology.arizona.edu/honors2006/group09/Drosophila_melanogaster.jpg http://en.wikipedia.org/wiki/Gregor_mendel http://www.merriam-webster.com/dictionary/ http://www.biochem.northwestern.edu/holmgren/Glossary/Definitions/

10. http://www.exploratorium.edu/exhibits/mutant_flies/mutant_flies.html

11. http://www.carolina.com/product/flynap+anesthetic+kit.do