#### Transcript GLYPHOSATE RESISTANCE Background / Problem

Lecture 14: Population structure and Population Assignment October 12, 2012 Lab 7 Update u Corrected instructions for lab 7 will be posted today u Problem 1: consider relative levels of F-statistics as well as significance from bootstrapping u Up to 3 points extra credit if problem 2 is done correctly u See lab open hours schedule on lab web page u Caveat: exams and class usage of lab u Other computers are available: see Hari or me Population structure from worldwide human population Population = subpopulation. Group = Regions Eurasia East Asia Oceania America Africa Lab 7 Revised Problem 1 Problem 1. File human_struc.xls contains data for 10 microsatellite loci used to genotype 41 human populations from a worldwide sample. a.) Convert the file into Arlequin format and perform AMOVA based on this grouping of populations within regions using distance. How do you interpret these results? Report values of the phi-statistics and their statistical significance for each AMOVA you run. b.) Do you think that any of these regions can justifiably be divided into subregions? Pick a region, form a hypothesis for what would be a reasonable grouping of populations into subregions, then run AMOVA only for the region you selected using distance measures. Was your hypothesis supported by the data? c.) GRADUATE STUDENTS: Which of the 5 initially defined regions has the highest diversity in terms of effective number of alleles? What is your biological explanation for this? Lab 7 Original Problem 2 (worth 8 points if you answer this). Use Structure to further test the hypotheses you developed in Problem 1. a.) Calculate the posterior probabilities to test whether: i. All populations form a single genetically homogeneous group. ii. There are two genetically distinct groups within your selected region iii. There are three genetically distinct groups within your selected region. b.) Use the ΔK method to determine the most likely number of groups. How does this compare to the method based on posterior probabilities? c.) How do the groupings of subpopulations compare to your expectations from Problem 1? d.) Is there evidence of admixture among the groups? If so, include a table or figure showing the proportion of each subpopulation assigned to each group. e.) GRADUATE STUDENTS: Provide a brief, literature-based explanation for the groupings you observe. Last Time u Sample calculation of FST u Defining populations on genetic criteria: introduction to Structure Today Interpretation of F-statistics More on the Structure program Principal Components Analysis Population assignment FST: What does it tell us? Degree of differentiation of subpopulations Rules of thumb: 0.05 to 0.15 is weak to moderate 0.15 to 0.25 is strong differentiation >0.25 is very strong differentiation Related to the historical level of gene exchange between populations May not represent current conditions FST is related to life history Seed Dispersal Gravity Explosive/capsule Winged/Plumose 0.446 0.262 0.079 Successional Stage Early 0.411 Middle 0.184 Late 0.105 Life Cycle Annual Short-lived Long-lived 0.430 0.262 0.077 (Loveless and Hamrick, 1984) Structure Program One of the most widely-used programs in population genetics (original paper cited >8,000 times since 2000) Very flexible model can determine: The most likely number of uniform groups (populations, K) The genomic composition of each individual (admixture coefficients) Possible population of origin A simple model of population structure Individuals in our sample represent a mixture of K (unknown) ancestral populations. Each population is characterized by (unknown) allele frequencies at each locus. Within populations, markers are in Hardy-Weinberg and linkage equilibrium. Roughly speaking, the model sorts individuals into K clusters so as to minimize departures from HWE and Linkage Equilibrium. Slide adapted from Jonathan Pritchard, 2007 presentation to Conservation Genetics meeting More on the model... Let A1, A2, …, AK represent the (unknown) allele frequencies in each subpopulation Let Z1, Z2, … , Zm represent the (unknown) subpopulation of origin of the sampled individuals Assuming Hardy-Weinberg and linkage equilibrium within subpopulations, the likelihood of an individual’s genotype in subpopulation k is given by the product of the relevant allele frequencies: Pr(Gi | Zi= k, Ak) = loci PPl Where Pl is probability of observing genotype l at a particular locus in subpopulation k Slide adapted from Jonathan Pritchard, 2007 presentation to Conservation Genetics meeting Probability of observing a genotype in a subpopulation Probability of observing a genotype at locus l by chance in population is a function of allele frequencies: Pl = p 2 i Homozygote Pl = 2 pi p j Heterozygote m P = Õ Pl for m loci l=1 Assumes unlinked (independent loci) and HardyWeinberg equilibrium If we knew the population allele frequencies in advance, then it would be easy to assign individuals. If we knew the individual assignments, it would be easy to estimate frequencies. In practice, we don’t know either of these, but the following MCMC algorithm converges to sensible joint estimates of both. Slide adapted from Jonathan Pritchard, 2007 presentation to Conservation Genetics meeting MCMC algorithm (for fixed K) Start with random assignment of individuals to populations Step 1: Gene frequencies in each population are estimated based on the individuals that are assigned to it. Step 2: Individuals are assigned to populations based on gene frequencies in each population. And this is repeated... …Estimation of K performed separately. Slide adapted from Jonathan Pritchard, 2007 presentation to Conservation Genetics meeting Admixed individuals are mosaics of ancestry from the original populations Ancestral Populations Slide adapted from Jonathan Pritchard, 2007 presentation to Conservation Genetics meeting The two basic ancestry models used by structure. No Admixture: each individual is derived completely from a single subpopulation Admixture: individuals may have mixed ancestry: some fraction qk of the genome of individual i is derived from subpopulation k. The admixture model allows for hybrids, but it is more flexible and often provides a better fit for complicated structure. This is what we used in lab. Slide adapted from Jonathan Pritchard, 2007 presentation to Conservation Genetics meeting Notes on Estimating the Number of Subpopulations (k) Likelihood-based method is the simplest, but likelihood often increases continuously with k More variability at values of k beyond “natural” value Evanno et al. (2005) method measures change in likelihood and discounts for variation Use biological reasoning at arriving at final value Priors based on population locations, other information Often need to do hierarchical analyses: break into subregions and run Structure separately for each Inferred human population structure Africans Europeans MidEast Cent/S Asia Asia Oceania America Each individual is a thin vertical line that is partitioned into K colored segments according to its membership coefficients in K clusters. Rosenberg et al. 2002 Science 298: 2381-2385 Structure is Hierarchical: Groups reveal more substructure when examined separately Rosenberg et al. 2002 Science 298: 2381-2385 Alternative clustering method: Principal Components Analysis Structure is very computationally intensive Often no clear best-supported K-value Alternative is to use traditional multivariate statistics to find uniform groups Principal Components Analysis is most commonly used algorithm EIGENSOFT (PCA, Patterson et al., 2006; PloS Genetics 2:e190). Eckert, Population Structure, 5-Aug-2008 49 Principal Components Analysis Efficient way to summarize multivariate data like genotypes Each axis passes through maximum variation in data, explains a component of the variation http://www.mech.uq.edu.au/courses/m ech4710/pca/s1.htm How do we identify population of origin? Human Population Assignment with SNP Assayed 500,000 SNP genotypes for 3,192 Europeans Used Principal Components Analysis to ordinate samples in space High correspondence betweeen sample ordination and geographic origin of samples Novembre et al. 2008 Nature 456:98 Individuals assigned to populations of origin with high accuracy Likelihood Approaches Allow evaluation of alternative hypotheses by comparing their relative likelihoods given the evidence L( H1 , H 2 | E ) P ( E | H1 ) P( E | H 2 ) In a population assignment or forensic context, definition of the competing hypothesis is the most essential component Population Assignment: Likelihood Assume you find skin cells and blood under fingernails of a murder victim Victim had major debts with the Sicilian mafia as well as the Chinese mafia Can population assignment help to focus investigation? P(G | H1 ) L( H1 , H 2 | G ) LR , P(G | H 2 ) What is H1 and what is H2? Population Assignment: Likelihood "Assignment Tests" based on allele frequencies in source populations and genetic composition of individuals Likelihood-Based Approaches Calculate likelihood that individual genotype originated in particular population Assume Hardy-Weinberg and linkage equilibria Genotype frequencies corrected for presence of sampled individual Usually reported as log10 likelihood for origin in given population relative to other population Implemented in ‘GENECLASS’ program (http://www.montpellier.inra.fr/URLB/gene class/geneclass.html) Pk l p 2 il for homozygote AiAi in population l at locus k Pkl 2 pil p jl for heterozygote AiAj in population l at locus k m P Pk k 1 for m loci Power of Population Assignment using Likelihood Assignment success depends on: Number of markers used Polymorphism of markers Number of possible source populations Differentiation of populations Accuracy of allele frequency estimations Rules of Thumb (Cornuet et al. 1999) for 100% assignment success, for 10 reference populations need: 30 to 50 reference individuals per population 10 microsatellite loci HE > 0.6 FST > 0.1 Knowing what you know about human population genetics, is it worth the effort to assign our skin sample to Asian or Sicilian populations? Rules of Thumb (Cornuet et al. 1999) for 100% assignment success, for 10 reference populations need: 30 to 50 reference individuals per population 10 microsatellite loci HE > 0.6 FST > 0.1 Population Assignment Example: Wolf Populations in Northwest Territories Wolf populations sampled on island and mainland populations in Canadian Northwest Territories Immigrants detected on mainland (black circles) from Banks Island (white circles) Carmichael et al. 2001 Mol Ecol 10:2787 Population Assignment Example:Fish Stories Fishing competition on Lake Saimaa in Southeast Finland Contestant allegedly caught a 5.5 kg salmon, much larger than usual for the lake Compared fish from the lake to fish from local markets (originating from Norway and Baltic sea) 7 microsatellites Lake Saimaa Market Based on likelihood analysis, fish was purchased rather than caught in lake -