Hardy Weinberg Equilibrium Gregor Mendel (1822-1884) Wilhem Weinberg G. H. Hardy (1877 - 1947) (1862 – 1937) Recall from Previous Lectures Darwin’s Observation Evolution acts through changes in allele frequency at each generation Leads to average change in characteristic of the population Lectures 4-11: Mechanisms of Evolution (Microevolution) • • • • • • Hardy Weinberg Principle (Mendelian Inheritance) Genetic Drift Mutation Recombination Epigenetic Inheritance Natural Selection These are mechanisms acting WITHIN populations, hence called “population genetics”—EXCEPT for epigenetic modifications, which act on individuals in a Lamarckian manner 4 Major Evolutionary Mechanisms acting at the population level, changing allele frequencies: • • • • Genetic Drift Natural Selection Mutation Migration Testing for Hardy-Weinberg equilibrium can be used to assess whether a population is evolving The Hardy-Weinberg Principle • A population that is not evolving shows allele and genotypic frequencies that are in Hardy Weinberg equilibrium • If a population is not in Hardy-Weinberg equilibrium, it can be concluded that the population is evolving Fig. 23-5a MAP AREA •What is a “population?” A group of individuals within a species that is capable of interbreeding and producing fertile offspring Beaufort Sea Porcupine herd range (definition for sexual species) Fortymile herd range Fig. 23-5 Porcupine herd MAP AREA Beaufort Sea Porcupine herd range Fortymile herd range Fortymile herd Hardy-Weinberg Principle • Mathematical description of Mendelian inheritance Hardy-Weinberg Equilibrium • According to the Hardy-Weinberg principle, frequencies of alleles and genotypes in a population remain constant from generation to generation • In a given population where gametes contribute to the next generation randomly, allele frequencies will not change • Allelic and genotypic frequencies follow the transmission rules of Mendelian inheritance, which maintains constant proportions in a population across generations In the absence of Evolution… Patterns of inheritance should always be in “Hardy Weinberg Equilibrium” Following the transmission rules of Mendel Requirements of HW Violation Evolution Large population size Genetic drift Random Mating Inbreeding & other No Mutations Mutations No Natural Selection Natural Selection No Migration Migration An evolving population is one that violates Hardy-Weinberg Assumptions “Null Model” • No Evolution: Null Model to test if no evolution is happening should simply be a population in Hardy-Weinberg Equilibrium • No Selection: Null Model to test whether Natural Selection is occurring should have no selection, but should include Genetic Drift – This is because Genetic Drift is operating even when there is no Natural Selection Hardy-Weinberg Theorem In a non-evolving population, frequency of alleles and genotypes remain constant over generations important concepts • gene: A region of genome sequence (DNA or RNA), that is the unit of inheritance , the product of which contributes to phenotype • locus: Location in a genome (used interchangeably with “gene,” if the location is at a gene… but, locus can be anywhere, so meaning is broader than gene) • loci: Plural of locus • allele: Variant forms of a gene (e.g. alleles for different eye colors, BRCA1 breast cancer allele, etc.) • genotype: The combination of alleles at a locus (gene) • phenotype: The expression of a trait, as a result of the genotype and regulation of genes (green eyes, brown hair, body size, finger length, cystic fibrosis, etc.) important concepts • allele: Variant forms of a gene (e.g. alleles for different eye colors, BRCA1 breast cancer allele, etc.) • We are diploid (2 chromosomes), so we have 2 alleles at a locus (any location in the genome) • However, there can be many alleles at a locus in a population. – For example, you might have inherited a blue eye allele from your mom and a brown eye allele from your dad… you can’t have more alleles than that (only 2 chromosomes, one from each parent) – BUT, there could be many alleles at this locus in the population, blue, green, grey, brown, etc. • Alleles in a population of diploid organisms A2 Eggs A1 A3 A1 A2 A1 A4 A2 Sperm A1 A3 A4 A1 A1 Random Mating (Sex) Zygotes A1A1 A1A3 • Genotypes A1A1 A1A1 A2A4 A3A1 A2 Eggs A1 A3 A1 A2 A1 A4 So then can we predict the % of alleles and genotypes in the population at each generation? A2 Sperm A1 A3 A4 A1 Zygotes A1 A1A1 A1A3 A1A1 A1A1 A2A4 A3A1 Hardy-Weinberg Theorem In a non-evolving population, frequency of alleles and genotypes remain constant over generations Fig. 23-6 Alleles in the population Frequencies of alleles p = frequency of CR allele = 0.8 q = frequency of CW allele = 0.2 Gametes produced Each egg: 80% chance 20% chance Each sperm: 80% chance Hardy-Weinberg proportions indicate the expected allele and genotype frequencies, given the starting frequencies 20% chance • By convention, if there are 2 alleles at a locus, p and q are used to represent their frequencies • The frequency of all alleles in a population will add up to 1 – For example, p + q = 1 If p and q represent the relative frequencies of the only two possible alleles in a population at a particular locus, then for a diploid organism (2 chromosomes), (p + q) 2 = 1 = p2 + 2pq + q2 = 1 – where p2 and q2 represent the frequencies of the homozygous genotypes and 2pq represents the frequency of the heterozygous genotype What about for a triploid organism? What about for a triploid organism? • (p + q)3 = 1 = p3 + 3p2q + 3pq2 + q3 = 1 Potential offspring: ppp, ppq, pqp, qpp, qqp, pqq, qpq, qqq How about tetraploid? You work it out. Hardy Weinberg Theorem ALLELES Probability of A = p Probability of a = q p+q=1 GENOTYPES AA: p x p = p2 Aa: p x q + q x p = 2pq aa: q x q = q2 p2 + 2pq + q2 = 1 More General HW Equations • One locus three alleles: (p + q + r)2 = p2 + q2 + r2 + 2pq +2pr + 2qr • One locus n # alleles: (p1 + p2 + p3 + p4 … …+ pn)2 = p12 + p22 + p32 + p42… …+ pn2 + 2p1p2 + 2p1p3 + 2p2p3 + 2p1p4 + 2p1p5 + … … + 2pn-1pn • For a polyploid (more than two chromosomes): (p + q)c, where c = number of chromosomes • If multiple loci (genes) code for a trait, each locus follows the HW principle independently, and then the alleles at each loci interact to influence the trait ALLELE Frequencies Frequency of A = p = 0.8 Frequency of a = q = 0.2 p+q=1 Expected GENOTYPE Frequencies AA: p x p = p2 = 0.8 x 0.8 = 0.64 Aa: p x q + q x p = 2pq = 2 x (0.8 x 0.2) = 0.32 aa: q x q = q2 = 0.2 x 0.2 = 0.04 Allele frequencies remain the same at next generation p2 + 2pq + q2 = 0.64 + 0.32 + 0.04 = 1 Expected Allele Frequencies at 2nd Generation p = AA + Aa/2 = 0.64 + (0.32/2) = 0.8 q = aa + Aa/2 = 0.04 + (0.32/2) = 0.2 Hardy Weinberg Theorem ALLELE Frequency Frequency of A = p = 0.8 Frequency of a = q = 0.2 p+q=1 Expected GENOTYPE Frequency AA: Aa: aa : pxp= p2 = 0.8 x 0.8 = 0.64 p x q + q x p = 2pq = 2 x (0.8 x 0.2) = 0.32 qxq= q2 = 0.2 x 0.2 = 0.04 p2 + 2pq + q2 = 0.64 + 0.32 + 0.04 = 1 Expected Allele Frequency at 2nd Generation p = AA + Aa/2 = 0.64 + (0.32/2) = 0.8 q = aa + Aa/2 = 0.04 + (0.32/2) = 0.2 Similar example, But with different starting allele frequencies p q p2 2pq q2 Fig. 23-7-4 80% CR ( p = 0.8) 20% CW (q = 0.2) Sperm CR (80%) 64% ( p2) CR CR 16% (qp) CR CW CW (20%) 16% ( pq) CR CW 4% (q2) CW CW 64% CR CR, 32% CR CW, and 4% CW CW Gametes of this generation: 64% CR + 16% CR = 80% CR = 0.8 = p 4% CW + 16% CW = 20% CW = 0.2 = q Genotypes in the next generation: 64% CR CR, 32% CR CW, and 4% CW CW plants Perform the same calculations using percentages Fig. 23-7-1 80% CR (p = 0.8) 20% CW (q = 0.2) Sperm CR (80%) CW (20%) 64% (p2) CRCR 16% (pq) CRCW 16% (qp) CRCW 4% (q2) CW CW Fig. 23-7-2 64% CRCR, 32% CRCW, and 4% CWCW Gametes of this generation: 64% CR + 16% CR = 80% CR = 0.8 = p 4% CW + 16% CW = 20% CW = 0.2 = q Fig. 23-7-3 64% CRCR, 32% CRCW, and 4% CWCW Gametes of this generation: 64% CR + 16% CR = 80% CR = 0.8 = p 4% CW + 16% CW = 20% CW = 0.2 = q Genotypes in the next generation: 64% CRCR, 32% CRCW, and 4% CWCW plants Calculating Allele Frequencies from # of Individuals • The frequency of an allele in a population can be calculated from # of individuals: – For diploid organisms, the total number of alleles at a locus is the total number of individuals x 2 – The total number of dominant alleles at a locus is 2 alleles for each homozygous dominant individual – plus 1 allele for each heterozygous individual; the same logic applies for recessive alleles Calculating Allele and Genotype Frequencies from # of Individuals AA 120 Aa 60 aa 35 (# of individuals) #A = (2 x AA) + Aa = 240 + 60 = 300 #a = (2 x aa) + Aa = 70 + 60 = 130 Proportion A = 300/total = 300/430 = 0.70 Proportion a = 130/total = 130/430 = 0.30 A + a = 0.70 + 0.30 = 1 Proportion AA = 120/215 = 0.56 Proportion Aa = 60/215 = 0.28 Proportion aa = 35/215 = 0.16 AA + Aa + aa = 0.56 + 0.28 +0.16 = 1 Applying the Hardy-Weinberg Principle • Example: estimate frequency of a disease allele in a population • Phenylketonuria (PKU) is a metabolic disorder that results from homozygosity for a recessive allele • Individuals that are homozygous for the deleterious recessive allele cannot break down phenylalanine, results in build up mental retardation • The occurrence of PKU is 1 per 10,000 births • How many carriers of this disease in the population? – Rare deleterious recessives often remain in a population because they are hidden in the heterozygous state (the “carriers”) – Natural selection can only act on the homozygous individuals where the phenotype is exposed (individuals who show symptoms of PKU) – We can assume HW equilibrium if: • There is no migration from a population with different allele frequency • Random mating • No genetic drift • Etc So, let’s calculate HW frequencies • The occurrence of PKU is 1 per 10,000 births (frequency of the disease allele): q2 = 0.0001 q = sqrt(q2 ) = sqrt(0.0001) = 0.01 • The frequency of normal alleles is: p = 1 – q = 1 – 0.01 = 0.99 • The frequency of carriers (heterozygotes) of the deleterious allele is: 2pq = 2 x 0.99 x 0.01 = 0.0198 or approximately 2% of the U.S. population Conditions for Hardy-Weinberg Equilibrium • The Hardy-Weinberg theorem describes a hypothetical population • The five conditions for nonevolving populations are rarely met in nature: – – – – – No mutations Random mating No natural selection Extremely large population size No gene flow • So, in real populations, allele and genotype frequencies do change over time DEVIATION from Hardy-Weinberg Equilibrium Indicates that EVOLUTION Is happening 4 Major Evolutionary Mechanisms: • • • • Genetic Drift Natural Selection Mutation Migration Hardy-Weinberg across a Genome • In natural populations, some loci might be out of HW equilibrium, while being in Hardy-Weinberg equilibrium at other loci • For example, some loci might be undergoing natural selection and become out of HW equilibrium, while the rest of the genome remains in HW equilibrium Allele A1 Demo Examples of Deviation from Hardy-Weinberg Equilibrium What would Genetic Drift look like? • Most populations are experiencing some level of genetic drift, unless they are incredibly large Examples of Deviation from Hardy-Weinberg Equilibrium Generation 1 Generation 2 Generation 3 Generation 4 AA 0.64 0.63 0.64 0.65 Aa 0.32 0.33 0.315 0.31 aa 0.04 0.04 0.045 0.04 Is this population in HW equilibrium? If not, how does it deviate? What could be the reason? Examples of Deviation from Hardy-Weinberg Equilibrium Generation 1 Generation 2 Generation 3 Generation 4 AA 0.64 0.63 0.64 0.65 Aa 0.32 0.33 0.315 0.31 aa 0.04 0.04 0.045 0.04 This is a case of Genetic Drift, where allele frequencies are fluctuating randomly across generations Examples of Deviation from Hardy-Weinberg Equilibrium AA 0.64 Aa 0.32 aa 0 Is this population in HW equilibrium? If not, how does it deviate? What could be the reason? Examples of Deviation from Hardy-Weinberg Equilibrium AA 0.64 Aa 0.32 aa 0 Here this appears to be Directional Selection favoring AA Or… Negative Selection disfavoring aa Examples of Deviation from Hardy-Weinberg Equilibrium AA 0.25 Aa 0.70 aa 0.05 Is this population in HW equilibrium? If not, how does it deviate? What could be the reason? Examples of Deviation from Hardy-Weinberg Equilibrium AA 0.25 Aa 0.70 aa 0.05 This appears to be a case of Heterozygote Advantage (or Overdominance) Examples of Deviation from Hardy-Weinberg Equilibrium AA 0.10 Aa 0.10 aa 0.80 Is this population in HW equilibrium? If not, how does it deviate? What could be the reason? Examples of Deviation from Hardy-Weinberg Equilibrium AA 0.10 Aa 0.10 aa 0.80 Selection appears to be favoring aa How can you tell whether a population is out of HW Equilibrium? Example: Does this population remain in Hardy Weinberg Equilibrium across Generations? Generation 1 Generation 2 Generation 3 AA 0.25 0.20 0.10 Aa 0.50 0.60 0.80 aa 0.25 0.20 0.10 Generation 1 Generation 2 Generation 3 AA 0.25 0.20 0.10 Aa 0.50 0.60 0.80 aa 0.25 0.20 0.10 In this case, allele frequencies (of A and a) did not change. ***However, the population did go out of HW equilibrium because you can no longer predict genotypic frequencies from allele frequencies For example, p = 0.5, p2 = 0.25, but in Generation 3, the observe p2 = 0.10 How can you tell whether a population is out of HW Equilibrium? 1. When allele frequencies are changing across generations 2. When you cannot predict genotype frequencies from allele frequencies (means there is an excess or deficit of genotypes than what would be expected given the allele frequencies) • One generation of Random Mating could put a population back into Hardy Weinberg Equilibrium Summary (1) A nonevolving population is in HW Equilibrium (2) Evolution occurs when the requirements for HW Equilibrium are not met (3) HW Equilibrium is violated when there is Genetic Drift, Migration, Mutations, Natural Selection, and Nonrandom Mating Hardy Weinberg Equilibrium Gregor Mendel (1822-1884) Wilhem Weinberg G. H. Hardy (1877 - 1947) (1862 – 1937) 1. Nabila is a Saudi Princess who is arranged to marry her first cousin. Many in her family have died of a rare blood disease, which sometimes skips generations, and thus appears to be recessive. Nabila thinks that she is a carrier of this disease. If her fiancé is also a carrier, what is the probability that her offspring will have (be afflicted with) the disease? (A) 1/4 (B) 1/3 (C) 1/2 (D) 3/4 (E) zero The following are numbers of pink and white flowers in a population. Generation 1: Generation 2: Generation 3: Pink 901 1204 1510 White 302 403 504 2. Which of the following is most likely to be TRUE? (A) The heterozygotes are probably pink (B) The recessive allele here (probably white) is clearly deleterious (C) Evolution is occurring, as allele frequencies are changing greatly over time (D) Clearly there is a heterozygote advantage (E) The frequencies above violate Hardy-Weinberg expectations The following are numbers of purple and white peas in a population. Generation 1: Generation 2: Generation 3: (A1A1) Purple 360 100 0 (A1A2) Purple 480 200 100 (A2A2) White 160 200 300 3. What are the genotype frequencies at each generation? (A) Generation 1: 0.30, 0.50, 0.20 Generation 2: 0.20, 0.40, 0.40 Generation 3: 0, 0.333, 0.666 (B) Generation 1: 0.36, 0.48, 0.16 Generation 2: 0.10, 0.20, 0.20 Generation 3: 0, 0.10, 0.30 (C) Generation 1: 0.36, 0.48, 0.16 Generation 2: 0.20, 0.40, 0.40 Generation 3: 0, 0.25, 0.75 (D) Generation 1: 0.36, 0.48, 0.16 Generation 2: 0.36, 0.48, 0.16 Generation 3: 0.36, 0.48, 0.16 4. From the example on the previous slide, what are the frequencies of alleles at each generation? (A) Generation1: Dominant allele (A1) = 0.6, Recessive allele (A2) = 0.4 Generation2: Dominant allele = 0.4, Recessive allele = 0.6 Generation3: Dominant allele = 0.125, Recessive allele = 0.875 (B) Generation1: Dominant allele = 0.6, Recessive allele = 0.4 Generation2: Dominant allele = 0.6, Recessive allele = 0.4 Generation3: Dominant allele = 0.6, Recessive allele = 0.4 (C) Generation1: Dominant allele = 0.6, Recessive allele = 0.4 Generation2: Dominant allele = 0.5, Recessive allele = 0.5 Generation3: Dominant allele = 0.25, Recessive allele = 0.75 (D) Generation1: Dominant allele = 0.4, Recessive allele = 0.6 Generation2: Dominant allele = 0.5, Recessive allele = 0.5 Generation3: Dominant allele = 0.25, Recessive allele = 0.75 5. From the example two slides ago, which evolutionary mechanism might be operating across generations? (A) Mutation (B) Selection favoring A1 (C) Heterozygote advantage (D) Selection favoring A2 (E) Inbreeding Answers: 1. Parents: Aa x Aa = Offspring: AA (25%), Aa (50%), aa (25%) Answer = A 2. A 3. C 4. A 5. D