LABORATORY 8 - Population Genetics and Evolution

LABORATORY 8: POPULATION GENETICS AND EVOLUTION

OVERVIEW
In this activity you will learn about the Hardy-Weinberg law of genetic equilibrium and study the relationship between evolution and changes in allele frequency by using your class as a sample population. pp. 448-449 6th ed. Campbell, Reece

OBJECTIVES
Before doing this laboratory you should understand: - how natural selection can alter allelic frequencies in a population - the Hardy-Weinberg equation and its use in determining the frequency of alleles in a population - the effects on the allelic frequencies of selection against the homozygous recessive or other genotypes After doing this laboratory you should be able to: - calculate the frequencies of alleles and genotypes in the gene pool of a population using the HardyWeinberg formula - discuss natural selection and other causes of microevolution as deviations from the conditions required to maintain Hardy-Weinberg equilibrium

INTRODUCTION
In 1908, G.H. Hardy and W. Weinberg independently suggested a scheme whereby evolution could be viewed as changes in the frequency of alleles in a population of organisms. They reasoned that if A and a are alleles for a particular gene locus and each diploid individual has two such loci, then p can be designated as the frequency of the A allele and q as the frequency of the a allele. Thus, in a population of 100 individuals (each with two loci) in which 40% of the loci are A, p would be 0.40. The rest of the loci (60%) would be a, and q would equal 0.60 (i.e., p + q = 1.0). These are referred to as allele frequencies. If certain conditions are met, the frequency of the possible diploid combinations of these alleles (AA, Aa, aa) should equal p2 + 2 pq + q2 = 1.0. Hardy and Weinberg also argued that if five conditions are met, the population's allele and genotype frequencies will remain constant from generation to generation. These conditions are as follows: 1. 2. 3. 4. 5. Large population size. The effects of chance on changes in allele frequencies is greatly reduced. Random mating. Individuals show no mating preference for a particular phenotype. There is no mutation of the alleles. No differential migration occurs (no immigration or emigration). There is no selection. All genotypes have an equal chance of surviving and reproducing.

Basically, the Hardy-Weinberg equation describes the status quo. If the five conditions are met, then no change will occur in either allele or genotype frequencies in the population. Of what value is such a rule? It provides a yardstick by which changes in allele frequency, and therefore evolution, can be measured. One can look at a population and ask: Is evolution occurring with respect to a particular gene locus? Since evolution is difficult if not impossible to observe in a natural population, we will simulate the evolutionary process using the class as a reproducing population. The purpose of this simulation is to provide an opportunity for students to question experience, and test some of the basic tenets of population genetics and evolutionary biology.

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LABORATORY 8 - Population Genetics and Evolution

EXERCISE 8A: ESTIMATING ALLELE FREQUENCIES FOR A SPECIFIC TRAIT WITHIN A SAMPLE POPULATION Using the class as a sample population, the allele frequency of a gene controlling the ability to taste the chemical PTC (phenylthiocarbamide) could be estimated. A bitter-taste reaction to PTC is evidence of the presence of a dominant allele. The inability to taste the chemical depends on the presence of homozygous recessive alleles (aa). (Instead of PTC tasting, other traits, such as tongue-rolling, may be used.) To estimate the frequency of the PTCtasting allele in the population, one must find p. To find p, one must first determine q (the frequency of the nontasting PTC allele), because only the genotype of the homozygous recessive individuals is known for sure (Why?). PROCEDURE 1. Press the control paper to the tip of your tongue. 2. Next, press the PTC taste-test paper to the tip of your tongue. In comparison to the control, is the PTC bitter? PTC tasters will sense a bitter taste. For the purposes of this exercise these individuals are considered to be tasters. Record your values in Table 8.1. 3. Calculate below the values of p and q based on the class data. Remember the definitions of p and q and the order in which they must be determined. (Hint: Use the Hardy-Weinberg equation...what does p + q equal?) You should then record these numbers in Table 8.1. Calculations:

Table 8.1: Phenotypic Proportions of Tasters and Nontasters and Frequencies of the Determining Alleles Raw Data Phenotypes Allele Frequency Based on the H-W Equation p q # Tasters # Nontasters % Tasters % Nontasters Class Population North American Population 55 45

TOPICS FOR DISCUSSION (don't forget to show calculations) 1. What is the percentage of heterozygous tasters in your class? ______________

2. What percentage of the North American population is heterozygous for the taster trait? ________

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LABORATORY 8 - Population Genetics and Evolution

EXERCISE 8B: CASE STUDIES CASE I (A Test of an Ideal Hardy-Weinberg Population) You as a class will become a population of randomly mating heterozygous individuals with an initial gene frequency of 0.5 for the dominant allele A and the recessive allele a. NOTE: Be sure to READ the directions before beginning. Look at Figure 8.1. It is important for everyone to be mating within the same generation (i.e., do not mate your F2 genotype with someone's F1 genotype.) Why is this important? 1. Each of you will assume the initial genotype 'Aa.' Pick up four cards, two of which have the letter A printed on them and the other two the letter a. These four cards represent the products of meiosis. 2. Choose any student in the class. The sex of your mate does not matter, nor does the genotype. 3. Place your four allele cards behind your back and randomly shuffle them. Both you and your partner should then present one card. This is the genotype of your first offspring. One of you should record on your data page. Place all four cards behind your backs again, shuffle, and present a card. This is the genotype of your second offspring. The other partner should record this on their data page. Figure 8.1

Student 1 ( Aa ) Products of Meiosis

Student 2 ( Aa ) Products of Meiosis

Contributes A

Contributes a

First offspring is Aa. (Student 1 assumes this genotype)

RESHUFFLE FOUR CARDS

Student 1 ( Aa ) Products of Meiosis

Student 2 ( Aa ) Products of Meiosis

Contributes a

Contributes a

Second offspring is aa. (Student 2 assumes this genotype)

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LABORATORY 8 - Population Genetics and Evolution

4. To maintain a constant population size, the parent genotype dies. You assume the genotype of one of your two offspring, and your partner then assumes the other offspring's genotype. In the example in Figure 8.1, student 1 becomes genotype Aa and obtains cards A, A, a, a; student 2 becomes aa and obtains cards a, a, a, a. To do this, you may have to go to the central allele supply and pick up a new cards. 5. Proceed to a new individual. Follow the exact same mating procedures, being sure to record the new genotype after each generation. After five generations, return to your seats. USE THE FOLLOWING METHOD TO CALCULATE THE ALLELE FREQUENCY FOR DATA SHEET! Allele Frequency: The allele frequencies, p and q, should be calculated for the population after five generations of random mating. Number of A alleles present at the fifth generation Number of offspring with genotype AA _________ X 2 = __________ A alleles Number of offspring with genotype Aa _________ X 1 = __________ A alleles a) Total = __________A alleles

p=

TOTAL number of A alleles TOTAL number of alleles in the population

Number of a alleles present at the fifth generation Number of offspring with genotype aa _________ X 2 = __________ a alleles Number of offspring with genotype Aa _________ X 1 = __________ a alleles b) Total = __________a alleles

q=

TOTAL number of a alleles TOTAL number of alleles in the population

** TOTAL number of alleles in the population is = a) Total # of A alleles + b) Total # of a alleles

ANALYSIS OF RESULTS
1. What does the Hardy-Weinberg equation predict for the new p and q (with respect to the old p and q)?

2. Do the results you obtained in this simulation agree? _____ If not, give possible reasons for the discrepancy.

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LABORATORY 8 - Population Genetics and Evolution

3. What major assumption(s) were not strictly followed in this simulation?

4. What is genetic drift?

CASE II (Selection) Now that you have the facts of life well in hand, we can begin to modify our simulation, making it more realistic to enable us to investigate some basic questions about selection and gene frequencies. In humans, there are several genetic conditions that have been thoroughly investigated. One good example is sickle-cell anemia. This is a single allele trait; the homozygous recessive genotype is lethal, and the afflicted individual usually dies prior to reaching reproductive maturity. Both the homozygous dominant and the heterozygote survive. At this point, we will consider both the homozygous dominant and the heterozygote phenotypically identical. In other words, we are selecting against the homozygote recessive 100% of the time. The procedure is similar to Case I. Start again with your initial genotype (Aa), and choose the genotype of your offspring as in Case I. This time, however, there is one important difference. Every time your offspring is aa, it dies. Since we want to maintain a constant population size, the same two parents must try again until they produce a surviving offspring. It is important to remember that an aa individual cannot mate. Proceed through five generations, selecting against the homozygous recessive offspring. Return to your seat. Now, add up the genotype frequencies that exist in the population and calculate the new p and q. (AGAIN, USE THE ALLELE FREQUENCY METHOD ON PAGE 4) 1. How do the new p and q compare to the values in Case I? How has the population changed? Explain using your class data.

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LABORATORY 8 - Population Genetics and Evolution

2. What would happen to p and q if we went another five generations?

3. In a large population, what are the chances of completely eliminating a deleterious recessive allele?

CASE III (Heterozygote Advantage) From Case II, it is easy to see that the lethal recessive allele rapidly decreases in the population. However, data from many human populations show an unexpectedly high frequency of the sickle-cell allele present in some populations. In other words, our simulation does not accurately reflect the real situation. In actuality, the heterozygote is slightly more resistant to a deadly form of malaria than the homozygous dominant individual. In other words, there is a slight selection against the homozygous dominant individual as compared to the heterozygote. This fact is easily incorporated into our simulation. In this round, keep everything the same as in Case II, except that if your offspring if AA, flip a coin. Heads he lives, tails he dies of malaria, and the same parents must mate again until they get a viable offspring. Go through five generations, starting again with your initial genotype. The genotype aa dies 100% of the time. After five generations, return to your seat. Total the class genotypes and calculate p and q. Now, starting with the genotype you last had, go through five more generations, and again total the genotypes and calculate p and q. (AGAIN, USE THE ALLELE FREQUENCY METHOD ON PAGE 4) If time permits, another five generations is extremely informative. 1. How do p and q in Case II compare with Case I and Case III? Explain.

2. Do you think the recessive allele will be completely eliminated in either Case II or Case III?

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LABORATORY 8 - Population Genetics and Evolution

3. What is the importance of heterozygous advantage in maintaining genetic variation in populations?

CASE IV (Genetic Drift, optional) In Case I, the role of small populations in genetic drift was suggested. It is possible to use our simulation to look at this phenomenon in more detail. Assume your F5 genotype. Randomly divide the lab into several smaller populations (for example, a class of 30 could be divided into 3 populations of 10 each). When the populations are reproductively isolated by mountain ranges, oceans, or imaginary walls, total the genotypic and allelic frequencies in each population. Now go through five generations as in Case I. Record the new genotypic frequencies, and calculate the new p and q for each population. (AGAIN, USE THE ALLELE FREQUENCY METHOD ON PAGE 4) 1. How did the initial genotypic frequencies of the populations compare?

2. What do your results indicate about the importance of population size as an evolutionary force?

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LABORATORY 8 - Population Genetics and Evolution

HARDY-WEINBERG PROBLEMS - Use the Hardy Weinberg equation to calculate the following. 1. Dark eyes are dominant to light eyes. In a population of 1000 individuals, 360 show the recessive phenotype. How many individuals would you expect to be homozygous dominant and heterozygous for this trait?

2. The ability to roll one's tongue is dominant to a lack of this ability. In a population of 500 individuals 75% show the dominant phenotype. How many individuals would you expect to be homozygous dominant and heterozygous for this trait?

3. Having a widow's peak is dominant to lacking a widow's peak. In a population of 1000 individuals, 510 show the dominant phenotype. How many individuals would you expect of each of the possible three genotypes for this trait?

4. In the U.S. about 16% of the population if Rh-. Rh- is recessive to Rh+. If the student population of a high school is 2000, how many students would you expect for each of the three possible genotypes?

5. In Nigeria, 4% of the newborn babies have sickle cell anemia. Out of a random population of 1000 newborn babies, how many would you expect for each of the three possible genotypes?

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LABORATORY 8 - Population Genetics and Evolution

DATA SHEET
show one sample calculation on another sheet CASE I - Hardy-Weinberg Equilibrium Initial: p = 0.5, q = 0.5 CASE III - Heterozygote Advantage Initial: p = 0.5, q = 0.5

My initial genotype: _Aa_ F1_____ F2_____ F3_____ F4_____ F5_____ note - only F5s genotype need to be shown Final: AA ____ Aa _____ p _____ q _____

My initial genotype: _Aa_ F1_____ F6_____ F2_____ F7_____ F3_____ F8_____ F4_____ F9_____ *F5_____ *F10_____ note - only F5 and F10s genotype need to be shown Final (after 5 generations): AA ____ Aa _____ aa _____ p _____ q _____ Final (after 10 generations): AA ____ Aa _____ aa _____ p _____ q _____

aa _____

CASE II - Selection Initial: p = 0.5, q = 0.5

My initial genotype: _Aa_ F1_____ F2_____ F3_____ F4_____ F5_____ Final: AA ____ Aa _____ p _____ q _____

aa _____

CASE IV - Genetic Drift initial aa final aa

AA group 1 group 2 group 3 group 4 group 5

Aa

AA

Aa

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LABORATORY 8 - Population Genetics and Evolution

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