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GENETIC LABWORK

MONOHYBRID CROSSES In Drosophila melanogaster

BY:

MISRAI FARAUK 100210103057

BIOLOGY EDUCATION STUDY PROGRAM DEPARTMENT OF MATHEMATIC AND SCIENCE EDUCATION FACULTY OF TEACHER TRAINING AND EDUCATION 2011

I.

TITLE: Monohybrid crosses in Drosophila melanogaster

II.

PURPOSE

1. Know the way to breed Drosophila melanogaster.

2. Exercise to monohybrid crosses.


3. Calculate the phenotype ratio of F1 and F2 offspring.

III.

BASIC THEORY

Monohybrid Cross is a method of determining the inheritance pattern of a trait between two single organisms. Monohybrid inheritance is the inheritance of a single characteristic. The different forms of the characteristic are usually controlled by different alleles of the same gene. For example, a monohybrid cross between two purebreeding plants (homozygous for their respective traits), one with yellow seeds (the dominant trait) and one with green seeds (the recessive trait), would be expected to produce an F1 (first) generation with only yellow seeds because the allele for yellow seeds is dominant to that of green. A monohybrid crosses compares only one trait. (Wikipedia, 2011). Monohybrid cross is a cross between two species of the same with a different nature. Monohybrid cross is highly related to Mendel's laws I or the so-called law segregation. This law reads, "In the formation of gametes for genes that are couples would be segregated into two chicks". First learned of Mendel's monohybrid nature at the time of crossing experiments on peas (Pisum sativum). So far in the monohybrid crosses always apply the law of Mendel I. I Mendel's laws apply in F1 x F1 gametogenesis has genotif heterozygous. The gene is located in the same locus on the chromosome, at the time of gametogenesis will one allel separate genes, each of which goes to one of gametes (Orphan, 1986). The person who first cross-breeding experiment is George Mendel (1822-1884). Previously, Mendel called genes as determinants. Then the chromosome (loss of chromatin that will appear during mitosis and serves as a carrier of the gene) was discovered by Wilhelm Roux and strengthened by experimental T. Boveri and W.S. Sutton (1902) who proved that genes are part of the chromosome. This theory is known as the chromosome theory. Genes inherited from parents to offspring through the gametes (Suryo, 2008).

When Mendel crossed tall pea plants with a trunked dwarf, all descendants of the first (F1) have high stem. A sign that the nature of high-defeating nature of the dwarf. High trait is the dominant trait. Plinkagerty that was defeated called recessive plinkagerties. When the first offspring of plants pollinated left alone, obtained the second offspring of plants with high stem ratio : stem dwarf (Suryo, 2008). First Mendel's laws apply in F1 x F1 gametogenesis has genotype heterozygous. The gene is located in the same locus on the chromosome, at the time of gametogenesis will one allel separate genes, each of which goes to one of gametes (Yatim,1986). Monohybrid In Plants, character height rod to rod dominant force on the generally low plants, including corn. In the presence of corn is also known characters such as sugarcane stem growth. In the bread mold Neurospora mycelium is also known that the red color of characters against a white dominant. Monohybrid In Animals. In guinea pigs, as well as in other animals, dominant gene causing abnormal pigmentation and its allel causing an albino. Normal pigmented guinea pig who is a furry black. Black guinea pig mated with an albino guinea pig. Her children are all black. If the child was married each other it will produce a black: white 3: 1. Monohybrid In Humans, such synthetic chemicals called PTC, there is a class of people who can taste it will feel bitter and a class of people who can not taste it will feel the amber. Bitter taste due to a dominant gene. In addition to tasting many other properties that follow the nature of monohybrid crosses, namely: polydactyly, phenylketonuria, Huntington's chorea brown teeth, crostic fibrosis. (Anonymous, 2011 ). Some important things about monohybrid crosses: All individual F1 is uniform. If it seems outright dominance, then individual F1 has a dominant In the F1 heterozygote forms gametes, there was segregation of alleles, If dominance is fully visible, then the crosses monohybrid produce

phenotype like its parent. so that gametes have only one allele only. offspring with a ratio of 3:1 (Change, 2008). To demonstrate Mendel, we often use Drosophilla melanogaster as experimental animals. Before we conducted experiments using Drosophilla melanogaster, we first have to know the characteristic of this insect. Drosophila melanogaster is a species of fruit flies that can be found in rotting fruit. Drosophila has

been

used

for

years

in

the

study

of

genetics

and

animal

behavior.

Here is the classification of Drosophila melanogaster (Borror, 1992): Kingdom Phyllum Class Order Family Genus : Animalia : Arthropods : Insecta : Diptera : Drosophilidae : Drosophila

Species: Drosophila melanogaster

In addition, Drosophila also classified into sub-orders Cyclophorpha (grouping of flies that contained pupa third instar skin, has a jaw hooks) and included in the series Acaliptrata imago is hatched to get out of the anterior part of the pupa. There are 12 reasons why Drosophila used for genetic experiments. The advantages of these flies, among others: 1. Easily obtained. Drosophilla melanogaster is insects that are cosmopholite. In Indonesia there are over 600 species of fruit flies of the family Drosophilidae. Thus it is not all that fruit flies found that the type Drosophilla melanogaster, but this species can be developed alone. 2. Easily maintained. Drosophilla melanogaster are usually kept in a jam jar containing medium banana tape. Besides it does not require sterile conditions as in microbiological experiments. However, should all the tools sterilized materials in advance to avoid fungal contamination. Mold growth can be prevented by administering NA-benzoate and tape to the process of fermentation. 3. Low maintenance costs. Because the medium of food was simple then treatment is not too expensive. But we must diligently replace the medium every two weeks. 4. Not much need a place. Mutant stock is usually kept in jars of jam. One bottle can contain about 300500 adult flies. Bottles can be stored on one shelf.
5. Visible size.

Although only the size of 2-5 mm only, morphology and properties of mutants can be observed with the naked eye.
6. Not dangerous.

Fruit flies do not transmit diseases or intermediate hosts.


7. Have many descendants.

A female fly can produce 500 eggs during her life.

8. Have a short life cycle.

Life cycle ranges from 10-15 days.

9. Variances.

These flies have a phenotype that is easy to distinguish on eye color, hair form, body color and wing shape.

10. Simple chromosome.

Only had 4 pairs of chromosomes alone, making it easy to count. In addition to the saliva gland chromosomes are giant-sized politen and stratiated array that is useful to identify where the gene.

11. Has many mutants.

The history of biological research suggests that Drosophilla sp. Organisms are the richest mutants. It is therefore appropriate once used as experimental animals in the genetics.

12. Male flies no crossovers.

Drosophilla sp. Males do not have crossovers. Similarly, the displacement cross can be calculated more precisely. On the other animals or in plants, a parental crossovers (Genetic Team, 2011). The other common trait of Drosophila melanogaster are: 1. Body color is brownish-yellow with black rings on the back of the body. 2. The size is small, between 3-5 mm. 3. The edge of the wing veins (costal vein) has two parts interuptus close to his body. 4. Tentacle (Arista) typically in the form of feathers, has 7-12 branching. 5. Cross vein posterior generally straight, not curved. 6. Compound eyes slightly ellipse-shaped and colored red. 7. There ocelli eye on the top of the head with a smaller size than the compound eyes. Elliptical head. 8. Thorax hairy-fur with a white base color, while the five-segmented abdomen and black stripes. 9. The wing length, colored transparent, and the position stems from the thorax. (Ardiawan, 1990). Differences male and female Drosophila melanogaster: 1. size of the female fly flies are usually larger than males. 2. Caudal parts of the body shape / abdominal tip rounded and blunt male flies, the flies were females slightly elongated and tapering. Abdomen male flies relative narrow and cylindrical, while the female fly abdomen enlarged and form a spherical or oval. 3. Body colour black pigment in the fly male is more dominant than the female fly. In male flies the abdomen there are five bands that extends up to go to the ventral body, whereas in female flies, there are seven black bands present only in the dorsal section. 4. Sex comb or comb sex only male flies have sex comb of stiff black hair 10 of the last tarsus forefoot (Genetic Team, 2011).

male ()

Female ()

Source: (McClean, 1997).

Fly Drosophila life cycle is relatively short, consisting of the stages as follows:

Source: (McClean, 1997). a. Individual adult females lay eggs two days after leaving the pupa. This spawning period lasts more or less for 1 week, with the number of eggs 50 to 75 grains / day. Eggs laid on the surface of the food. The shape is oval, has a structure like a hook that serves as a float to prevent it from sinking into the liquid food. Its diameter shaped 0.5 mm so that it can be seen with the naked eye. Eggs stage lasted for approximately 24 hours (Geiger, 2002) Drosophila egg is coated by two layers, namely a thin vitellin membrane surrounding the cytoplasm and a thin but strong membrane (chorion) on the outside and there were two in stalk anterior. thin chorion has a tough outer skin of the egg (Borror, 1992). At the anterior end there mikrophyle, where spermatozoa into the egg. Although many sperm into the mikrophyle but only

one that can fertilized with pronuleus female and the other immediately wiil absorb in the development of embryonic tissue (Borror, 1992). b. The development begins soon after fertilization, which consists of two periods. First, the embryonic period in the egg at the time of fertilization until the young larvae hatch from the eggs and this occurs within approximately 24 hours. And at times like this, larvae did not stop, stop to eat (Silvia, 2003). The second period is the period after hatching from the egg and is called the development postembrionic divided into three stages: larva, pupa, and imago (sexually phase with the development of the wing). Other formations in sexual development occurs during adulthood (Silvia, 2003). c. Drosophila larvae are white, segmented with a length of about 4.5 mm, shaped like worms, and probing with a black mouth near the head. For respiratory problems in the trachea, there is a pair of spiracles which are both located on the anterior and posterior ends (Silvia, 2003). The larvae live in the food and feeding activity is very high. In the larval stage occurs two times the change of skin, and the period between the turn of the skin called instar stage. Thus, the known three instar stages, i.e. before the first change of skin, between the period change of skin, and after the turn of the second. At the end of the third instar stage, larvae out of the media toward the food to a more dry to develop into pupae. Overall larval stage takes approximately one week (Geiger, 2002). When the cuticle is no longer soft, young larvae periodically sheds its skin to reach adult size. Discarded the old cuticle and integument expanded with new high speed feed. During the period change of skin, called instar larvae. The first instar larvae after hatching until the first change of skin. And indications instar larvae are the size and number of teeth in the mouth black. After the second change of skin, the larvae (third instar) eating until ready to form pupae. In the last stage, the third instar larvae to the top surface of the medium crawl food to a dry place and stop moving. And if it can be summarized, in Drosophila, destruction of larval cells occurs in prose turn of the skin (molting) which took place four times with three stadia instar: from larval instar 1 to instar II, of the larval instar to instar II, III, of instars III to pupa and from pupa to imago (Ashburner, 1985). During the meal, the larvae create channels in the medium, and if there are many channels of the growth in culture can be said to be going well. Mature larvae usually crawl up on the wall of the bottle or on a tissue paper in a bottle. And here the larvae will attach themselves to a dry place with liquid glue like produced by the salivary glands and then form a pupa (cocoon).

d. Pupa has a hard cuticle and dark. Length about 3 mm. This stage lasts about 5 days. When Drosophila larvae form pupae shell, his body shortens, becomes hard and pigmented cuticle, without the head and wings are called larval instar 4. Pupa formation characterized by the formation of the head, wing pads, and legs. Puparium (outer form of the pupa) using a cuticle on the third instar. At this stage pupae, larvae in an inactive state, and in these circumstances, the larvae change into adult flies (Ashburner, 1985). The structure of adults was evident during the period of pupa dormancy in small part the same network as in the embryonic stage. Tissue restrictions pre adult (before adulthood) is called anlagen. The main function of the pupa is for development outside of the anlagen to the adult form (Silvia, 2003). e. Adult in Drosophila melanogaster in a cycle life of about 9 days old. Once out of the pupa, the fruit fly color is still pale, translucent color body and wings do not inflate. This situation will change in a few hours. Meanwhile, the female flies reached age sex mature within 12-18 hours and can survive for about 26 days (Geiger, 2002) Series of Sex in Drosophila melanogaster Sex linkage initially discovered by TH Morgan in experiments on Drosophila melanogaster, He get white-eyed flies. These flies are mutant (gene change) because normal red-eyed flies. When white-eyed male fly flies mated with normal females (redeyed), then all red-eyed offspring. And if the fly is mated F1, the F1 offspring 3 shows the ratio of red-eyed: one-eyed white. From this comparison, obtained evidence that red is dominant over white, in addition, all red-eyed F2 flies all, while half of male flies with red eyes and some white-eyed. From here drawn the conclusion that the gene resesive only show its effect on male flies only. Morgan argues therefore that the gene that determines eye color is found on chromosome-X (Suryo, 2008). If a dominant gene W determine color red eyes, and its allel recessive w for white eyes, all red-eyed female fly offspring, whereas half of the number of male flies with red eyes and white-eyed half again. Because the male flies have only one X chromosome, whereas in the Y-chromosome gene then there is no male flies are homozigotic (Suryo, 2008). Because the gene that determines eye color is located on chromosome-X, of course, can occur with the white-eyed female fly genotype ww. This occurs when flies a red-eyed males mated with red-eyed flies with the genotype Ww. So obtained half of the number of children and half female fly from the number of children male flies have white eyes (Suryo, 2008).

Factors affecting growth in the life cycle of Drosophila melanogaster including the following: Temperature Environment Drosophila melanogaster during the 8-11 day cycles in ideal conditions. Ideal conditions in question is the temperature around 25-28 C. At this temperature the flies will have one round in an optimal cycle. While at low temperatures or around 180C, the time required to complete its life cycle is relatively long, slow, which is about 1820 days. At a temperature of 30 C, adult flies that grow will be sterile. The availability of Food Media The number of eggs of Drosophila melanogaster issued will be reduced if the food shortage. Adult fruit flies that lack of food will produce small-sized larvae. These larvae are capable of forming a pupa is small, but often fail to develop into mature individuals. Some may become adults who can only produce a few eggs. Viability of eggs was also influenced by the type and amount of food eaten by the larvae of females. Level Density Bottle Maintenance Medium bottles should be filled with a medium fruit enough and not too dense. In addition, fruit flies which were bred in the bottle should not be too much, quite a few pairs only. In Drosophila melanogaster with the ideal conditions where there is enough space (not too dense) adult individuals can live to about 40 days. However, if conditions are too dense medium bottle will cause the decline in egg production and the increasing number of deaths in adult individuals. Intensity of Light

Drosophila melanogaster prefers dim light and will experience slower growth during his stay in a dark place. A chi square (X2) statistic is used to investigate whether two distributions of categorical variables differ from each other. In statistical testing we always use a null hypothesis that there is no difference between the distributions. (note: Chi square tests can only be used on actual numbers and not on percentages, proportions, means, etc.) In our case, we can use the actual observed number of flies of each type as our observed values. We can find the expected number of flies of each type for a 3:1 ratio by using the same number of flies (100 flies) and dividing by 4 to give us expected values of 75:25 for a 3:1 ratio of 100 flies (McClean, 1997).

Calculate the chi square statistic x2 by completing the following steps:


1. For each observed number in the table subtract the corresponding expected

number (O E).
2. Square the difference [ (O E)2 ]. 3. Divide the squares obtained for each cell in the table by the expected number

for that cell [ (O - E)2 / E ].


4. Sum all the values for (O - E)2 / E. This is the chi square statistic.

For our example, the calculation would be: Observed Expected (O E) (O E)2 (O E)2/ E Wild type 70 Mutant type totals 100 100 x2 = 1.31 Having now obtained our chi square statistic x2 = 1.31, we look up in a table of the Chi Square X2 distribution the probability attached to it. Before we can do this, however, we need to know the degrees of freedom. When a comparison is made between one sample and another, a simple rule is that the degrees of freedom equal (number of columns minus one) x (number of rows minus one) not counting the totals for rows or columns. For our data this gives (2-1) x (2-1) = 1. Entering the Chi square distribution table with 1 degree of freedom and reading along the row we find our value of x2 (1.31) lies between 0.455 and 2.706. The corresponding probability is 0.5<P<0.1. This is well below the conventionally accepted significance level of 0.05 or 5%, so the null hypothesis that the two distributions are the same is verified. In other words, when the computed x2 statistic exceeds the critical value in the table for a 0.05 probability level, then we can reject the null hypothesis of equal distributions. Since our x2 statistic (1.31) did not exceed the critical value for 0.05 probability level (3.841) we can accept the null hypothesis that a ratio of 70:30 is the same as a 75:25 ratio (within 5% error). And here is Chi Square distribution table: Df 0.5 1 2 0.10 0.05 0.02 5.412 7.824 0.01 6.635 9.210 0.001 10.827 13.815 30 75 25 5 5 25 25 0.31 1.00

0.455 2.706 3.841 1.386 4.605 5.991

3 4 5

2.366 6.251 7.815 3.357 7.779 9.488

9.837

11.345 16.268

11.668 13.277 18.465

4.351 9.236 11.070 13.388 15.086 20.517 Source: (McClean, 1997).

To put this into context, it means that we do have a 3:1 ratio of wild type to mutant offspring and that these offspring could have come from a cross between two heterozygotes. To make the chi square calculations a bit easier, plug your observed and expected values into the following applet (McClean, 1997).

IV.

OBSERVATION METHOD

IV.1

Materials And Equipments

Binocular microscope Culture bottles with sponge stoppers HVS paper labels Petri dish that has a layer of cotton as a place of re-anesthesis or

re-etherizer

Ether solution Pipette Bottle containing the alcohol or detergent to kill the flies that are

no longer needed

Small brush no 1 Stock of Drosophila melanogaster strains of normal with

phenotip pure, white eye mutant flies and mutant flies purple or (turn fur)

Medium of banana-cassava fermentation Preventive substances fungal growth (Na-Benzoat)

IV.2

Work procedures

1. Establishment of a medium banana cassava fermentation.

Take 5 pieces of king banana and tape with comparisons: tape = 3:1

Pulverize both ingredients in a glass beaker

Give Na-Benzoate to prevent mold growth

Enter a medium banana 2 tablespoons tape into each culture bottle.

Make paper pupasi into culture bottles that had contained food medium.

Close the culture bottles so as not to be contaminated with wild flies.

2. . Etherization

Examine the culture bottle, if it flies at the mouth of the bottle, knock slowly so fly falls into the bottom of the bottle.

Put 3-5 drops of ether on a piece of cotton.

Enter the cotton with ether, and let stand for 20 seconds or flies do not move

Put the flies that had been drugged on a sheet of white paper so that the flies are easily observed.

Separate living flies with flies that die (if any).

Re-etherisasi for a few If the flies, has regained consciousness before the trial is completed

Flies that are no longer used must be killed in a bottle containing alcohol or detergent

3. Creating a cross Requirement to make a cross between two varieties should be virgin female flies. For the first do isolation female process:

After anesthetized, remove all imago of the cooking culture bottle / that already contains the pupa, then write down the date.

After 8 days, remove any newly hatched flies from the pupa, then select females.

Obtained virgin flies. To get virgin female fly pupa is by doing isolation.

crosses 1
Take two bottles of cultured normal type flies (wild), purple type (purple eye)

Anesthesis fly on the two bottle

Separate according to gender

Take 3 normal type female flies and 3 types of purple eye (Pm) and its opposite

Close the culture with sponge cover

Give information about the kinds of crosses, date and the name

Store culture bottles in place that has the specified

On day 7 move all parental flies. Record when flies first appear

On day 10 anesthesis flies and count the number of flies emerging. Distinguish between sex and count flies

Make a list of data

Parental crosses (P)

Make a cross between a fly flies F1

After anesthetized, take 5 tail male and female fly flies

Enter on the bottles containing the culture medium was new and had been given a paper pupasi

Store in a place that has been determined and write data on table

On day 7 remove parental

On day 10 will get the flies in F2

Anesthesised , separate sexes, count them, and record fenotipnya (normal or purple)

Crosses II

Cross the eyes of five flies a white male with normal female fly.

Perform the same experiment as in crossing

Calculate the F1 offspring

Perform a cross between F1 flies, flies that is crosses 5 male and 5 female flies

Calculate the F2 offspring should get a descent of at least 100 flies

Make a list

V.

OBSERVATION RESULT

Table of life cycle in monohybrid crosses Date 1 28 Oktober 2011 2 29 Oktober 2011 3 30 Oktober 2011 Abdomen female mutant purple become big (pregnant) Explanations 3 Female mutant purple and 3 male normal enter to bottle There is no changes Picture -

4 31 Oktober 2011

Female mutant lay eggs

Eggs colour is white transparant

The size is 0,5 mm

5 1 November 2011

There are larvae that crawl on wall and paper of bottle. Shape like worm, white transparant. Active, approximately 2 mm called larvae instar I

6 2 November 2011

The amount of larvae more increase. Its length approximately 5 mm In antherior there are stalk like anthena Passive and called larvae instar II

7 3 November 2011

Larvae become big and long

There stalk like anthena in antherior Calle larvae instar III

8 4 November 2011

Larvae stagnant and attached at wall or paper. The colour is white Called prepupa

9 5 November 2011

Prepupa become pupa The colour become brown Parental is realised

10 6 November 2011 11 7 November 2011

The colour of pupa become black in part of body The imago out from pupa The colour is pale Wings is folded And become adult imago

Table of F1 from bottle 2

Sex of fruit fly Total

Male normal 4

Male mutant

Female normal 52

Female mutant 9

Total

71

VI.

ANALYSIS

Monohybrid cross is a cross between two species of the same with a different nature. Monohybrid cross is highly related to Mendel's laws I or the so-called law segregation. This law reads, "In the formation of gametes for genes that are couples would be segregated into two chicks". A monohybrid crosses compares only one trait.

In this labwork activity about monohybrid crosses we use mutant fruit fly and normal fruit fly. The type of mutant fruit fly that we use is purple (Pm). We breed the fruit fly in 2 culture bottles. First bottle contain 3 male mutant purple (Pm) and 3 female normal, and the other bottle contain 3 female mutant purple (Pm) and 3 male normal. The fruit fly that we use is virgin fruit fly in female, because the female fruit fly can store sperm in spermatheca in long enough time and in order to fruit fly can crosses with fruit fly that have determined. So fruit fly not in pregnant condition when fruit fly will crosses.

Drosophila melanogaster for stock maintenance can be used various kinds of mediums. Medium initially used was a mixture of green banana and cassava tape with a ratio of 6: 1. Medium was used for over 15 years. In 1984 started to use some medium that can be attempted to maintain other types Drosiphila and recent years have used a new recipe. This is caused by the quality of the tape and bananas are not uniform, so the gain medium need for felt more solid and reliable. New recipes that will be used is a modification of an existing recipe and adapted to the condition of Indonesia.

Usually the fruit fly (Drosophila melanogaster) bred in bottles of medium, the medium may consist of: molasses, Banana with a mixture of fermented cassava with a 6:1 ratio. Type of medium the most widely used is the medium that consists of a mixture of bananas with cassava. This type of medium is also typically used for maintenance.

The materials used to make the culture medium of Drosophila melanogaster in this experiment is ripe plantains as a food ingredient like by Drosophila melanogaster, antifungal to control mold growth, fermipan to convert complex sugars into simple sugars and to grow the mushrooms as Drosophila melanogaster, palm sugar as source

of sugar or carbohydrates, in order to solidify the medium, sorbic acid benzoate to prevent contaminants from the outside and distilled water as a solvent. The cycle of life in monohybrid crosses between mutant and normal fruit fly is longer than last observation that have we done early. It because in monohybrid crosses we use virgin fruit fly in order to know that the offspring of fruit fly derive from mutant and normal. Not like fruit fly that we use in experiment of life cycle, because maybe the offspring do not derive from fruit that culture in bottle but it can cause by that female store sperm in spermatecha at that time. And here is life cycle of monohybrid crosses between mutant and normal fruit fly. In first day 3 Female mutant purple and 3 male normal enter to bottle, then there is no changes in second day because the fruit fly is adaptation to new environment. In third day abdomen female mutant purple become big (pregnant), then female mutant lay eggs, eggs colour is white transparent, the size is 0,5 mm that occur in 4th day. There are larvae that crawl on wall and paper of bottle in 5th, the larvae shape is like worm, the color is white transparent, active move to anywhere, and approximately 2 mm called larvae instar I. Then in 6th day the amount of larvae more increase, its length approximately 5 mm and in antherior there are stalk like anthena that have function to help pupa not sink in medium when in pupa stadium, larvae is passive and called larvae instar II occur in 7th. Larvae become big and long, there is stalk like anthena in antherior and also have spiracle antherior, and the size of larvae is the biggest than larvae instar I and larvae instar II and called larvae instar III that occur in 8th day. Larvae stagnant and attached at wall or paper, the color is white, and called prepupa occur in 9th day. In 10th day prepupa become pupa, the color become brown then parental is realized in order to not mix with the offspring. In last day the color of pupa become black in part of body then in 11th day, the imago out from pupa he color is pale, wings is folded at first time. Then young imago become adult imago and the amount is increased. In crosses process that have we done, the result show that in first bottle (3 male mutant purple (Pm) and 3 female normal) nothing changes that occur in fruit fly. And in second bottle we obtain 71 fruit flies, that consist of male normal (4), male mutant (6), female normal (52), and female mutant (9).

In first bottle there is no changes that occur in fruit fly that we crosses. We guess that the male mutant purple is steril and the female normal is fertil. Its because we can see in second bottle fruit fly normal male which come from one parents that

crosses by mutant purple can produce offspring. While in mutant steril may be when scientist make mutant Drosophila ,chemical substance influence reproductive organ of Drosophila .

And it can be influenced by condition bottle, such as food in medium that contaminated by fungi can make fruit fly dont want to crosses or marry. And also the condition of temperature, because recently is wet season so the temperature increase than normal, it can reach 28-30 C. According to literature, if temperature reach 30 or more it can cause fruit fly be steril. And our group think that we do not make mistake when determine sex of fruit fly in bottle one, because between mutant male and normal female is distinctly appears.

In second bottle that contain 71 fruit flies, consist of male normal (4), male mutant (6), female normal (52), and female mutant (9). In this case, we can make possibility of the genotype in fruit fly. Usually gene in mutant is dominant and normal is recessive. So we can make crosses:

P:

Mutant purple female () >< (XPrXPr)

Normal male () (XprY) Xpr, Y

G: F1:

XPr XPr Xpr, XPrY

Mutant purple female () : 50 % Mutant purple male() : 50 %

But in this case all F1 is not mutant but also normal. So the possibility of phenotype in mutant female is heterozygote. P: Mutant purple female () (XPrXpr) G: F1: XPr, Xpr XPrXpr, XPrY, XprXpr, XprY >< Normal male () (XprY) Xpr, Y

Mutant purple female () : 25 %

Mutant purple male() Normal female () Normal male ()

: 25 % : 25% : 25%

From result of observation we can do Chi square test with temporary hypothesis:

H0 : fruit fly that produce in male fruit fly are 50% male normal, 50% female normal, 50% mutant male, and 50% mutant female. H1 : fruit fly that produce in F1 those are 50% male normal, 50% female normal, 50% mutant male, and 50% mutant female.

H0 can accepted if the value is more than 0,05 and Ho is rejected if the value less than 0,05. And below is Chi square table of second bottle.

Count table of chi square. Normal


Observed value (A) Expected value (H)

Mutant 15 17,75 2,75 2,25 5,06 0,28

Total 71 71

56 53,25 2,75 2,25 5,06 0,09

|A-H| |A-H| - 0,5 {|A-H| - 0,5}2 {|A-H| - 0,5}2 H

0,37

Used Yates correction, deviation value is mines by 0,5.

Number of class: 2 Degree of freedom: n 1 = 2 1 = 1

Chi square value: 0,37 According to table of chi square, 0,37 located between probability 70% and 50% and more near with 50% probability, with chi square value between 0,15 and 0,46. So from the result chi square value 0,37 more than critical point 0,05 (0,37 > 0,05), so in our observation about monohybrid crosses accepted the hypothesis.

But in our experiment maybe there unknown factor that we do not concern about it, for example mistake to determined sex of male and female fruit fly, although we have try to determined exactly. Than the condition of medium in bottle that contaminated by fungi make food supply in medium less than enough to preqursor fruit fly do mating process. Because usually the female fruit fly do not want lay egg if the condition of merdium in bad. And other factor like conditon of environment such as temperature, light of sun, also influence the mating process.

VII.

CONCLUSION

Monohybrid cross is a cross between two species of the same with a different

nature. And monohybrid cross compares only one trait.


A

monohybrid

crosses

in

the

fly

Drosophila

melanogaster

done by crossing different properties from each other. The result of monohybrid crosses that we observed in bottle 1 nothing changes that occur in fruit fly, it may cause by many factor like steril, chemical subtances in mutant etc.
The result of monohybrid crosses that we observed in bottle 2 produce 71 fruit

flies, that consist of male normal (4), male mutant (6), female normal (52), and female mutant (9).
The results of monohybrid crosses fly Drosophila melanogaster can

evidenced by chi-square test.


The value of chi-square test in second bottle is 0,37 that more than 0.05 (0,37 >

0,05) so the value is accepted hypothesis.

VIII. REFERENCES

Ardiawan.

2009.

Drosophila

melanogaster.

[online]

http://

ardiawan1990.multiply.com/journal/item/21 Filipina. Accessed at Jember on November 6, 2011. Ashburner, Michael. 1989. Drosophila, A Laboratory Handbook. USA: Coldspring Harbor Laboratory Press. Borror.J.D,Triplehorn. 1992. Pengenalan Pengajaran Serangga.Yogyakarta: Gadjah Mada University Press Campbell,Reece,Mitchell. 2004. BIOLOGI JILID I Edisi kelima. Jakarta : Penerbit Erlangga Change. 2008. Persilangan Monohibrid. [online] http://erikarianto.wordpress.com. Accessed at Jember on November 6, 2011. Geiger, Pete. 2002. An introduction to Drosophila melanogaster. [online] http://biology.arizona.edu/sciconn/lessons2/Geiger/intro.htm. Jember on November 6, 2011. Genetic Team. 2011. Genetic Labwork Module. Jember: Jember University. Orphans, Wildfire. 1986. Genetics. New York: Publishers Tarsito. McClean, Philip. 1997. Simple Mendelian Genetics in Drosophil.[online] Accessed at

http://www.ndsu.edu/pubweb/~mcclean/plsc431/linkage/linkage1.htm. Accessed at Jember on November 6, 2011. Silvia, Triana. 2003. Pengaruh Pemberian Berbagai Konsenterasi Formaldehida Terhadap Perkembangan Larva Drosophila. Bandung : Jurusan Biologi Universitas Padjdjaran.

Strickberger, Monroe, W. 1962. Experiments in Genetics with Drosophila.London: John Wiley and Sons, inc. Suryo, H.2004. Genetika manusia. Yogyakarta: Gadjah Mada University Press. Wikipedia. 2011. Monohybrid Crosses. [online]

http://en.wikipedia.org/wiki/Monohybrid cross . Accessed at Jember on November 4, 2011. Yatim, wildan. 1986. Genetika. Bandung: Penerbit Tarsito.
IX.

ADDITION
Question

1. Bar eye in Drosophila is caused by a dominant gene B. While the normal eye by its recessive allele b. Normal eyed male flies in the cross with a bar-eyed females, its F1 offspring consist of: 100% bar-eyed females Bar-eyed males 50% 50% of normal-eyed males Determine parental genotype generation! Answer: The crosses from case above: P: Bar eye female () (XBXb) G: F1: XB, Xb XBXB, XBYb, XBY, XbY >< Bar eye male () (XBY) XB, Y

XBXB: Bar eye female () 100 % XBYb: Bar eye female () XBY : Bar eye male () 50 % XbY : normal male () 50 %

2. In humans muselar pseudo hypertrophic muscular dystrophy (PHMD) is a condition in which muscles become weak and unable to function progressively and gradually lead to death in the teen years. This disease is caused by a recessive gene and only suffered by boys. Give your opinion about genetic diseases on top! Answer: The major possibility that influence is Y chromosome that less countan genes, because Y chromosome is shorter than X chromosome , chromosome Y only have in male , so, trait from parental will inheritatance to their male offspring that contain Y chromosome. if occur marriage between male that suffer muselar pseudo hypertrophic muscular dystrophy with normal female, so the trait of father will inheritance to male children. The crosses from case above: P: normal female () (X X) G: F1: X X X , XhY >< PHMD male () (XhY) Xh, Y

X X : normal female () XhY: PHMD male ()