1X. MECHANISMS OF EVOLUTION Evolution can be described as changes in the frequency of alleles in a population. In this lab you will examine the mechanisms that can lead to these frequency changes. There are five major factors that contribute to the process of evolution. They are: 1) Natural Selection - Differe survival and reproduction of genotypes. 2) Migration - The arrival and incorporation of new individuals from another population. 3) Genetic Drift - Random fluctuations in allelic frequencies due to chance (expected to have significant impact only when the population is small). 4) Nonrandom mating ~ Often referred to as sexual selection, this is when individuals with a certain inherited characteristic are more likely to secure a mate than other individuals. 5) Mutation - The creation of an allele due to a random change in DNA structure (mutation rates are usually very low fora given gene, though can increase upon exposure to radiation and certain molecules). You will examine the ways that natural selection, migration and genetic drift can affect allele frequencies, but first it is necessary to build an understanding of how these changes are measured by looking at a population that is not changing. These populations are said to be in equilibrium. The English mathematician G.H. Hardy and the German physician W. Weinberg simultaneously developed a way to measure the changes in the frequency of alles in a population. This shows that heredity itself does not cause changes in the population. Outside forces such as those listed above are what lead to the changes in allele frequency. ‘The Hardy-Weinberg theorem uses p to denote the frequency of the dominant trait and q to denote the frequency of the recessive trait. With traits that only have two alleles it then follow that p + q must be equal to 1. ptq=i This also means that if we know the frequency of one allele then we can determine the frequency of the other. So, ifq = .3 then we can calculate p by subtracting q from 1. This would mean that 1 -.3 would be equal to p. Or, 10.3 0.7. Meaning that p is 0.7. ‘A population is in Hardy-Weinberg equilibrium when gene frequencies remain constant generation after generation (i.e. no evolution) and equilibrium is expected to occur if there are no evolutionary processes acting on the populations. In order to compare observed genotypic numbers to expected genotypic numbers, we used the following equations to calculate expected phenotypic numbers. ‘Squaring each side of the equation p + q = 1 gives the following equation and allows the calculation of expected genotypic frequency under conditions for Hardy-Weinberg equilibrium: ptt 2pq+q=1.0To determine the possible allelic combinations we can then calculate the frequency of homozygous dominant, p?, homozygous recessive, q?, and heterozygous, 2pq, individuals. From the example the frequency of the homozygous dominant would be p* or, (0.7) = 0.49. Part C: Testing HW and Natural Sele During this lab exercise, you will investigate factors that influence genetic variation within species. You will examine traits known to be determined by genes. The general hypothesis to be investigated is: Environmental conditions cause changes in gene frequencies. You will analyze a laboratory experiment that tests the effect of a specific environmental change on a single set of traits (fruit fly wing shape). The experiment has already been performed and you will tabulate flies based on wing shape, and then analyze these results. METHODS AND MATERIALS To test the hypothesis, the fruit fly, Drosophila melanogaster, was used. The test populations have two types of wings ~ normal and vestigial. Vestigial wings are tiny and do not allow flight. These phenotypic variations are determined genetically from both alleles of one autosomal gene, To change the independent variable in this test of the hypothesis, sticky paper was introduced as the environmental factor which could give one phenotype an advantage over the other. The two cases of the independent variable are: population cage A without sticky paper and population cage B with sticky paper. ‘To measure the dependent variable, phenotypic frequencies were determined by examining samples of flies from each population cage four generations after the population cages wereestablished. The number of normal wing flies and the number of vestigial wing flies were tabulated for each cage. Between 100 and 125 flies were tabulated in each cage. Both cages were started with parents that were heterozygotes for normal wings and vestigial wings. Normal wings are dominant to vestigial wings. Let "p" represent the frequency of the dominant allele (in this case, normal wings) and let "q represent the frequency of the recessive allele (in this case, vestigial wings). The genotype frequency of the heterozygous parents for these populations was "2pq’. Fifty heterozygous flies were added to each cage at the beginning of the experiment. A chi-square test was used to compare observed and expected number of flies. To compare the expected frequencies to the observed number of flies tabulated from the cages, expected frequencies were converted to expected number of flies. This was done for each phenotype in each cage by using the following equation: expected number of flies of |= the expected frequency of __X the total number of flies a given genotype given genotype tabulated from that cage So if the expected frequency of a genotype is .25 and 100 flies are counted, 25 of the 100 flies should have the given genotype if the population is in HW equilibrium. By comparing the observed frequencies after four generations to those expected under Hardy- Weinberg equilibrium in Cage B, we test whether sticky paper is causing genetic change within the population. By comparing the observed frequencies after four generation to those expected under Hardy-Weinberg equilibrium in Cage A, we test whether variables other than the sticky paper are influencing genetic change within the population. Other variables were controlled in investigation by keeping the amount of food, amounts of egg-laying space provided, temperature, and light the same in each cage. Also, cages were designed to prevent flies from entering or leaving the cage. Phenotype vs. Genotype Flies with genotype p*have normal wings, but flies with pq also have normal wings. There is no difference between p?flies and pq flies. Flies with genotype q” have vestigial wings. This means that it was necessary to calculate the expected phenotype frequency rather than the genotype frequency. The phenotype frequency for normal winged flies can be calculated as p*+2pqAll of the original flies of the parental generation were all heterozygous (pq), meaning the allele frequencies of the parental generation were p=0.5, and q= 0.5. This should produce offspring in the following frequencies: p (0.5) (0.5) p (0.5) p? (.25) pq (.25) 4 (0.5) ba (.25) a (.25) Offspring Genotype Frequencies: pq = (.25) +(.25) = 2pq=.5 @=.25 Offspring Phenotype Frequencies: p?+ 2pq=.75 (normal wings) q?=.25 (vestigial wings) Chi-square Calculation: Use the data provided in the tables on the following pages to calculate the Chi-square value for each cage. Chi-Square = (Jad — bel - 0.5TY (T) (atb)(a+c)(b+d)(c+d) Df= (# phenotypes-1)(# populations -1)D: RESULTS Title: Population A Calculated Expected | Calculated Expected Actual - Frequency based on H-W | Number based on H-W | Observed Wing Phenotype ee 7 75 77 Normal wing p2#2pq 23 Vestigial wing oe |q2_ 1.0 100 Total cai Title: Population B Calculated Expected Calculated Expected Actual 7 Frequency based on H-W | Number based on H-W Observed Wing Phenotype comnts .75 Normal wing a p2+2pq -25 Vestigial wing ou 2 1.0 100 100 Total if there was a difference between the expected allele You will use Chi square calculation to determi frequencies to be and the observed frequencies.Title: Cage A Wing Type Observed Expected Total (T) Normal (a) 7 (b) (atbtctd) Vestigal ( ee (a) Cage A Chi-square p-value Title: Cage B Wing Type Observed Expected Total (T) Normal fa) 43 (b) (atb+cea) Vestigal () 57 (a) ‘Cage B Chi-square p-valu ‘Answer the questions on the following page in a word document and upload to turnitin.comAnswer the following questions for both Cage A and Cage B: 1. What is the Chi-Square value calculated for each cage? 2. Whats the critical value for each cage? 3. Whats the p-value or range of p-values associated with your Chi-square value for each cage? 4. What is the null hypothesis for each cage? Was the null hypothesis rejected or not? Why or why not? Answer the following general questions: 5. Was the hypothesis supported? Why or why not? 6. What can you conclude about HW equilibrium based on the results collected and the statistical test performed?