Genetics Lab, February 2012

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Modo de Herencia in Drosophila Through Vestigial Wings and Yellow Body Mutations
Wesemann, Charles A. Abstract:
Throughout the process of cross linking genes we can observe the way genes are hereditable and can be observed from P1 (parental genes) and observed in F2 (second generation genes). An example of such study is best carried out in Drosophila (fruit flies). Through high reliability of no crossing over of Drosophila genes, we will observe a line of heritage of two mutations: Crippled Wings (CW) and Yellow Body (YB). Using a direct hereditable group versus reciprocate hereditable group, we expect to view differences in the way the F2 will come out and we will try to predict the quantity and proportions of gene expression. Finally the obtained information will be quantified and analyzed through X2 (Chi-squared) equation.

Introduction:
The Drosophila is a very useful and interesting model group to study. It is known for its high productivity rate in reproduction, its consistency with high numbers of offspring and short gestation periods. It is also vey easy to keep in labs and very easy to handle. They require basic food medium for nutrition. Drosophila has been used as the model of genetic studies for a very long time. For these main reasons is that Drosophila Melanogaster is the ideal species to use as a model to study. (Skillcorn) The used model of study during this report will be the use of vestigial wings (Fig.1) and yellow body (Fig2) (Exploratorium). These mutations in Drosophila are recessive and can be sex linked but do not necessarily mean they are exclusively sex linked.

Fig. 1 Female with vestigial wings

Figure 2 Female (left) and male (right) Yellow body with white-eye The means of inheritance that are going to be studied is direct inheritance and reciprocate inheritance. These have unique ways of how the genes are transferred from one generation to another. A direct 1

Genetics Lab, February 2012 inheritance is viewed such in Figure 3. Reciprocate inheritance is viewed in such a manner as presented in Figure 4. (Hunt Morgan)

Issue 1 to observe and handle the Drosophila; ether to help sleep the Drosophila in order to handle them; a fine brush, to move he Drosophila without damaging the specimen; anesthetizing chamber, a small chamber where the Drosophila can be anesthetized easily. The P1 Drosophila was given to us with the knowledge that they were homozygote with the genes presented. At first we place ether into the anesthetizer. Later on we take the Drosophila and transfer them into the anesthetizer appropriately. After being anesthetized we separated them through sex differentiation and mutation under the microscope, after observing and annotating the observation and classification of each mutation and sex. Those who do not express mutations are known as wild type (WT). This process is repeated for F1 and F2. Figure 5 presents a visual summary of the observable and desired experiment. The figure presents the example of vestigial wings but does not express the study of 2 different genetic mutations.

Fig. 3 Direct inheritance using whiteeye

Fig. 4 Reciprocal inheritance example using white eye.

Materials and Methods:

During the experiment we used the most easy and accessible methods available to us. The items we used were: a telescopic microscope, in order Fig. 5 Example inheritance. of cross-linking

Sex is differentiation was based 2

Genetics Lab, February 2012 mostly on certain sex markers. Males are smaller in bodies than females. Males also have a dark end thorax while females are more stripped. There is also the observation of the genitalia area of Drosophila is a key component. (Fig. 6) (University of Dundee) Mutation 1 (CW) WT Vestigial Mutation 2 (YB) Yellow WT

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Sex R

Total 10 3 13

Fig. 6 Female (left) and Male (right) After all the data collection the information will be used to determine the importance of the mode of inheritance through X2 (Chi-squared).

Results:
The data collected from the parental Drosophila is compiled to show that the direct inheritance shows that males had the vestigial wing mutation and females had the yellow body mutation. This would mean that the genotypic cross here is wwBB x WWbb. Where W is wild type wings and w is vestigial wings, B is wild type body and b is yellow body. Table 1. Collected data of P1 Direct Mutation Mutation Sex Total 1 (CW) 2 (YB) Vestigial WT 5 WT Yellow 3 D 8 The data obtained from the reciprocate inheritance showed that males had the yellow mutation and females had the vestigial mutation. So the genotypic cross would look something like this WWbb x wwBB. Table 2. Collected Reciprocate data of P1

When we observe and collected the data for the F1 progeny we can then determine the type of inheritance we are observing. First of all we can determine if the gene is sex linked, autosomal and maybe even if it is co dominant or recessive. Most mutations in Drosophila tend to be recessive. Since then we observed that the F1 showed distinct inheritance of genes. The calculated view of the F1 progeny in direct inheritance shows that all progeny in F1 should be WT for wings and only males would be yellow body. Therefor the expression yellow body, based on the observed is sex linked, expressed in the X chromosome of Drosophila. This assumption can be obtained through Punnet square analysis (Apendix) using the observed data of F1 and P1. Correcting then the genotypic expression of P1 as XBYww x XbXbWW. As shown the data collected of F1 progeny in direct inheritance (Table 3). The genotypic cross of F1 direct is XbYWw x XBXbWw. Table 3. Collected data F1 Direct Mutation Mutation Sex Total 1 (CW) 2 (YB) WT Yellow 10 WT WT 12 FD 22 In the reciprocal inheritance the same methods of observation and Punnet analysis was made. The observed inheritance of F1 in Drosophila with reciprocal inheritance showed that the entire population would be heterozygous and showed only WT characteristics (Table 4). The parental is also corrected to being XbYWW x 3

Genetics Lab, February 2012 XBXBww, that way the genotypic cross of F1 reciprocate is then expressed as XBYWw x XBXbWw. Table 4. Collected data F1 Reciprocate Mutation Mutation Sex Total 1 (CW) 2 (YB) WT WT 6 WT WT 7 FR 13 Knowing the genotypes from the F1 progeny helped allot in the process of sorting out the final expression of the F2 predicted. Through the observations of P1 and F1 we can easily see the final expression of the genes that were going to be expressed. The expected ratio of the F2 direct inheritance are male: 3WT: 1Vestigial: 3Yellow: 1Vestigial and Yellow; females: 3WT: 1Vestigial: 3Yellow: 1Vestigial and Yellow. In a complete distribution we could say that the Drosophila total would be a 6:2:6:2 distribution. The data observed and collected from the direct inheritance is shown in Table 5. These numbers will be used in X2 analysis. Table 5. Collected data F2 Direct Mutation Mutation Sex Total 1 (CW) 2 (YB) WT WT 78 Vestigial WT 29 WT Yellow 56 Vestigial Yellow 14 WT WT 66 Vestigial WT 20 WT Yellow 63 Vestigial Yellow 27 The F2 of the reciprocate inheritance is repeated as the F2 for direct inheritance. The method of using the Punnet square analysis we can obtain the expected ratio of distribution of genetic mutations. The ratio that we expected to be observed is: males,

Issue 1 3WT: 3 Yellow: 1 Vestigial: 1 Yellow and Vestigial; females, 6WT: 2Vestigial. In total the distribution of the observed would be 9:3:3:1. This type of distribution is identical to the Mendel Distribution. The data obtained from the observation is placed on Table 6. Table 6. Collected data F2 Reciprocate Mutation Mutation Sex Total 1 (CW) 2 (YB) WT WT 53 Vestigial WT 23 WT Yellow 65 Vestigial Yellow 10 WT WT 105 Vestigial WT 30 The last of the collected data is the X (chi-squared) analysis. This analysis will be calculated with the data obtained of the F2 progenies. First of we use the Chi-squared equation:
2

The next data we will obtain will be the expected values of the F2 progenies. This will be done through the multiplication of the total number of progeny with the fraction of the distribution of expected mutations. After this, the data will be summed up and the total will give us the value of Chi squared that will tell us if the value of the reciprocate inheritance and direct inheritance is significant or not. This is compared and determined by a value table that already postulates this. This information is also dependent in degrees of freedom.

Genetics Lab, February 2012

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X P-value

Direct 3.14 0.4

Reciprocate 5.88 0.35

df 3

Literatures:
Works Cited
Exploratorium. Microscope Image Station. 2012. 5 March 2012 <http://www.exploratoriu m.edu/imaging_station/g allery.php?Asset=%3CI %3EDrosophila %20melanogaster%3C/I %3E%20yellow%20white %20mutants&Group=&C ategory=%20Fruit %20flies&Section=Model %20Organisms>. Hunt Morgan, Thomas. A CRITIQUE OF THE THEORY OF EVOLUTION. 24 February 1916. 5 March 2012 <http://www.gutenberg.o rg/files/30701/30701h/30701-h.htm>. Reece, JB. biology. Campbell, 2005. Skillcorn, John. The Fruit Fly. 21 September 2002. 5 March 2012 <http://dendroworld.co.u k/BDGarchive/general.ht ml>. University of Dundee. Sex Determination. 2007. 5 March 2012 <http://learning.covcolleg e.ac.uk/content/Jorum/GM B_Autosomal-and-sexlinkage_LM1.2_20Jan08/page75.htm >.

Discussion:
The observed data presented suggests that the Chi-squared value gave us that the P-values of both direct and reciprocate inheritance are within the parameters of that H0 (null hypothesis) cannot be rejected cause the values shown are within the acceptable parameters. This means that the obtain values and calculated values both show a strong correlation and give a relationship that strongly shows that the expected and observed values are in the acceptable range of normality. Given the observed data we can see that the proportions of the direct inheritance shows a 6:2:6:2 ratio and in the reciprocate inheritance we observe a 9:3:3:1, classic Mendel like distribution. The interesting thing that is learned during the Punnet square distribution is the fact of Vestigial wings being a recessive autosomal gene and Yellow Body is a sex linked gene.

Conclusion:
In conclusion we can observe and say firmly and surely that the data presented is useful and perfect in determining the mode of inheritance. The techniques and the procedures taken in consideration for this experiment express the most quantified and visual data that could be expected from this experiment. After the experiment one learns to observe and better understand how Drosophila Melanogaster is the ideal model species in genetics. Its easy use and fast pace study is incredible and ideal for the

Genetics Lab, February 2012

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Appendix:
Direct inheritance: P1: XBYww x XbXbww XbW XbW XbW XbW XBw XBXbWw XBXbWw XBXbWw XBXbWw XBw XBXbWw XBXbWw XBXbWw XBXbWw F1: XbYWw x XBXbWw XBW XBw XbW Xbw XbW XBXbWW XBXbWw XbXbWW XbXbWw Xbw XBXbWw XBXbww XbXbWw XbXbww YW XBYWW XBYWw XbYWW XbYWw Yw XBYWw XBYww XbYWw XbYww Yw XbYWw XbYWw XbYWw XbYWw Yw XbYWw XbYWw XbYWw XbYWw

Reciprocate inheritance: P1: XbYWW x XBXBww XBw XBw XBw XBw XbW XBXbWw XBXbWw XBXbWw XBXbWw XbW XBXbWw XBXbWw XBXbWw XBXbWw F1: XBYWw x XBXbWw XBW XBw XbW Xbw XBW XBXBWW XBXBWw XBXbWW XBXbWw XBw XBXBWw XBXBww XBXbW XBXbww YW XBYWW XBYWw XbYWW XbYWw Yw XBYWw XBYww XbYWw XbYww YW XBYWw XBYWw XBYWw XBYWw YW XBYWw XBYWw XBYWw XBYWw