Presentation at SMBEBA

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demography and linked selection in maize and teosinte Jeffrey Ross-Ibarra @jrossibarra UC Davis photo by Jason Wallace

Transcript of Presentation at SMBEBA

demography and linked selection in maize and teosinte

Jeffrey Ross-Ibarra @jrossibarra

UC Davis

photo by Jason Wallace

Photo: https://www.flickr.com/photos/croptrust/

Beadle 1979 Field Museum of Natural History Bulletin

Meyerowitz 1994 Current Biology Duvick et al. 1999 US 6639132 B1

maize origins

Tripsacum extinct maizeF1F1

teosinte (Z. mays ssp. parviglumis)

maize (Z. mays ssp. mays)

George Beadle

van Heerwaarden 2011 PNAS Matsuoka et al 2002 PNAS

Zea mays ssp. parvgilumis

Zea mays ssp. mexicana

Zea mays ssp. mays

1250m

Title Text Title TextPhotos: Matt Hufford, Brandon Gaut, John Doebley

M. D. PURUGGANAN AND D. Q. FULLER

Figure 4. Comparison of evolutionary rate estimates. Box plots of the rates of evolution in (A) log (darwins) and (B) log (haldanes) fordomestication (DOM) as well as plants (PLAN) (from Bones and Farres 2001) and anthropogenic (AN) and natural (NAT) conditions for wildanimal species (Hendry et al. 2008). The asterisk indicates domestication rates under the assumption of the shortened 2000-year periodfor legume species. The vertical lines give the estimate ranges, whereas the boxes span the minimum and maximum quartile range. Thehorizontal line within the box gives the median rate.

while for grain/seed increase is 0.68 Β± 0.15 Γ— 10βˆ’3 haldanes.Using the log (rate) estimates, we find that the evolutionary ratesare not significantly different (t = 0.63, df = 18, P < 0.54). Incontrast, however, making the assumption of an ∼2000-year do-mestication period for legumes leads to a nearly twofold higherestimate for grain/seed size increase across all species (mean =1.15 Γ— 10βˆ’3 Β± 0.26 haldanes), but this rate difference betweennonshattering and seed size increase is still not significant (t =0.34, df = 18, P < 0.74). This analysis does not reveal a sig-nificantly higher rate for nonshattering compared to grain sizeevolution.

EVOLUTIONARY RATES DURING DOMESTICATION

ARE SIMILAR TO THOSE EXPERIENCED BY WILD

SPECIES

The real interest in calculating evolutionary rates for these dif-ferent domestication traits lies in the comparison with estimatesobserved in wild species that are subject to natural selection. Therehave been extensive compilations of phenotypic rates of evolu-tion from contemporary microevolutionary (Hendry and Kinnison1999; Bone and Farres 2001; Kinnison and Hendry 2001; Hendryet al. 2008) and paleontological data (Gingerich 2001), and wecan use these to compare domesticated versus wild species.

The rate estimates for evolution during domestication fallwithin the range observed in these microevolutionary studies, buton the lower side of the range, either those for plants (Bone andFarres 2001) or under natural and anthropogenic conditions inanimals (Hendry et al. 2008,) see Figure 4. Comparison with theplant data (Bone and Farres 2001), for example, reveals that themean rates of evolution based on log-transformed data are muchhigher than those observed for the rate of evolution during domes-tication (see Fig. 4 and Table 4). Indeed, the mean evolutionaryrates we observe during domestication are significantly lower thanmean rates of phenotypic evolution of plant species in the wild(t = 7.48, df = 91, P < 0.0001).

Most of the studies that document rapid evolution in wildplant species, however, represent cases of very strong selection(e.g., growth in serpentine soils, herbicide resistance, Bone andFarres 2001). To examine evolution of other wild species, we alsocompare our results to data from wild animal species (Hendry et al.2008). Moreover, the large size of the dataset (Hendry et al. 2008)allows us to focus on allochronic data (e.g., change across a timeseries, which is comparable to the our archaeological data), and topartition the data for wild species into those that have experiencednatural versus anthropogenic conditions. This comparison is basedon the assumption that, in contrast to the compiled data from plantsunder strong selection pressures (Bone and Farres 2001), the ratesof evolution under less-stringent selective conditions are at leastbroadly comparable between plants and animals.

Not surprisingly, the mean rates of evolution for wild speciesunder anthropogenic conditions are higher than those under nat-ural conditions (see Fig. 4 and Table 4). Nevertheless, the meanrate of evolution under domestication is significantly lower than

Table 4. Mean rates of evolution.

Mean Rates Rate RateΓ—103

(darwins) (haldanes)

DomesticationOverall 144.56Β±33.55 0.73Β±0.14Overall1 200.82Β±44.04 1.17Β±0.26Nonshattering 835.44Β±196.13 0.98Β±0.16Grain/seed size 107.92Β±20.7 0.69Β±0.16Grain/seed size1 158.35Β±32.15 1.21Β±0.29

NaturalPlant 8893.8Β±2604.8 33.4Β±10.5Anthropogenic animal 5906.7Β±781 71.3Β±18.4Natural animal 626.2Β±86.4 29.5Β±5.4

1Rates calculated under the assumption that legume domestication occurred

over a 2000-year period.

1 7 8 EVOLUTION JANUARY 2011

log

Dar

win

s (e

/106

yrs

)

Purugganan and Fuller 2010 Evolution Benz 2009 Hist. Maize

F1

F2

Beadle:

1/500

4-5 genes

Briggs et al. 2007 Genetics Wang et al. 2005 Nature

1 2 3 4 5

6 7 8 9 10

tb1gt1

Figure 1.Phenotypes. a. Maize ear showing the cob (cb) exposed at top. b. Teosinte ear with the rachisinternode (in) and glume (gl) labeled. c. Teosinte ear from a plant with a maize allele of tga1introgressed into it. d. Close-up of a single teosinte fruitcase. e. Close-up of a fruitcase fromteosinte plant with a maize allele of tga1 introgressed into it. f. Ear of maize inbred W22(Tga1-maize allele) with the cob exposed showing the small white glumes at the base. g. Earof maize inbred W22:tga1 which carries the teosinte allele, showing enlarged (white) glumes.h. Ear of maize inbred W22 carrying the tga1-ems1 allele, showing enlarged glumes. For highermagnification copies of f–h see Supplementary Information.

Wang et al. Page 10

Nature. Author manuscript; available in PMC 2006 May 23.

NIH

-PA Author Manuscript

NIH

-PA Author Manuscript

NIH

-PA Author Manuscript

tga1Figure 2 Map of parviglumis Populations and Hopscotch allele frequency. Map showing the frequencyof the Hopscotch allele in populations of parviglumis where we sampled more than 6 individuals. Size ofcircles reflects number of individuals sampled. The Balsas River is shown, as the Balsas River Basin isbelieved to be the center of domestication of maize.

as our independent trait for phenotyping analyses. SAS code used for analysis is available athttp://dx.doi.org/10.6084/m9.figshare.1166630.

RESULTSGenotyping for the Hopscotch insertionThe genotype at the Hopscotch insertion was confirmed with two PCRs for 837 individualsof the 1,100 screened (Table S1 and Table S2). Among the 247 maize landrace accessionsgenotyped, all but eight were homozygous for the presence of the insertion Withinour parviglumis and mexicana samples we found the Hopscotch insertion segregatingin 37 (n = 86) and four (n = 17) populations, respectively, and at highest frequencywithin populations in the states of Jalisco, Colima, and Michoacan in central-westernMexico (Fig. 2). Using our Hopscotch genotyping, we calculated diVerentiation betweenpopulations (FST) and subspecies (FCT) for populations in which we sampled sixteenor more chromosomes. We found that FCT = 0, and levels of FST among populationswithin each subspecies (0.22) and among all populations (0.23) (Table 1) are similar togenome-wide estimates from previous studies Pyhajarvi, HuVord & Ross-Ibarra, 2013.Although we found large variation in Hopscotch allele frequency among our populations,BayEnv analysis did not indicate a correlation between the Hopscotch insertion andenvironmental variables (all Bayes Factors < 1).

Vann et al. (2015), PeerJ, DOI 10.7717/peerj.900 8/21

Studer et al. 2011 Nat. Gen.; Vann et al. PeerJ 2015 Wills et al. 2013 PLoS Gen

T/TM/TM/M

T/TM/TM/M

A B

T/TM/TM/M

T/TM/TM/M

A B

3’ UTR

5’ control region

n=23 n=15

landraces teosinte

inbred lines, 3-5X coverage

Chia et al. 2012 Nat. Gen

Hufford et al. 2012 Nat. Gen.

strong selection (s~0.015) on ~1,100 genes 11M maize-teosinte shared SNPs

3,000 fixed SNPs

Tim Beissinger

teos

inte

nuc

leot

ide

dive

rsity

mai

ze n

ucle

otid

e di

vers

ity

distance to nearest substitution (cM)

Tim Beissinger Wallace et al. 2014 PLoS Gen.

Hard sweeps?

Interpretation

Hard sweeps were not widespread during maize domestication

Hernandez et al., 2011

Sattath et al., 2011

Other species

9distance to substitution (cM)mai

ze n

ucle

otid

e di

vers

ity

Sattah et al. 2011 PLoS Gen.

nucl

eotid

e di

vers

ity

distance to nearest substitution (cM)

distance to nearest substitution (kb)

scal

ed d

iver

sity

Ne ~ 150,000 Ne ~ 10,000

Ne ~ 2,000,000 Ne ~ 600,000

Sattah et al. 2011 PLoS Gen. Williamson et al. 2014 PLoS Gen Hernandez et al. 2011 Science

maize

Hufford et al. 2012 Nat. Gen.

teosinte

Tajima’s D

Tim Beissinger

rela

tive

nucl

eotid

e di

vers

ity

distance to gene (cM)

teosinte Ne~150,000maize Ne~50,000

200-2000bp

Nordborg et al. 1996 Genetical Res.Tim Beissinger

het. selection (hs)deleterious mut. rate ΞΌ

recombination rB ⇑ e

οΏ½P

iΒ΅iti

(ti+ri(1οΏ½ti))2

B =

rela

tive

dive

rsity

distance to gene (M)

observedbest fit

maizeteosinte

teosinte 2Ns=1maize 2Ns=1

s (h=0.1)

Tim Beissinger

Fitness effects of new mutations

maize

3.6E-7

3.6E-9

teosinte

Β΅

t = hs 2.3E-7

6E-9

maize

Hufford et al. 2012 Nat. Gen.

teosinte

25% SNPs unique36% SNPs unique

Tajima’s D

Wright et al. 2005 Science Tenaillon et al. 2004 MBE

Eyre-Walker et al. 1996 PNAS

Β΅, r

teos

inte

maize

DATAte

osin

te

maize

MODEL

Tim Beissinger

0.05Na

Na

Na 3Na

150,000 450,000

Nm~1.5

Kate Crosby Hearne et al. 2014 http://hdl.handle.net/11529/10034

4K samples ~1M SNPs

Ne ~106

time (years BP)Li Wang

multiple sequential markovian coalescent

Ne~108

CIMMYT 1984

49M ha of open-pollinated maize 22,000 plants/ha. Nc ~1012

min. 10 plants/ha to reseed Ne ~5 x 108

Tim Beissinger

0.05Na

Na

Na 3Na

nucl

eotid

e di

vers

ity

distance to gene (cM)

sing

leto

n di

vers

ity

distance to gene (cM)

Tim Beissinger

Mexico highland6,000 BP

S. America lowland

6,000 BP

S. America Highland

4,000 BP

Mexico lowland

9,000 BP

Matsuoka et al. 2002; Piperno 2006 Perry et al. 2006; Piperno et al. 2009

Li Wang Hearne et al. 2014 http://hdl.handle.net/11529/10034

●

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0.039 βˆ’ 8.8eβˆ’07 β‹… x, r2 = 0.0529y = 0.039 βˆ’ 8.8eβˆ’07 β‹… x, r2 = 0.0529y = 0.039 βˆ’ 8.8eβˆ’07 β‹… x, r2 = 0.0529y = 0.039 βˆ’ 8.8eβˆ’07 β‹… x, r2 = 0.0529y = 0.039 βˆ’ 8.8eβˆ’07 β‹… x, r2 = 0.0529

0.00

0.02

0.04

0.06

0 2000 4000 6000 8000geographical distance to Balsas Valley (km)

perc

enta

ge o

f pol

ymor

phic

sites

elevation100020003000

country_name●

●

●

Central AmericaMEXICOSouth America

Lowland Highland

NA

NB

NCN1 N2

N2P

tDtE

tF

NA

NB

NCN1 N2

N2P

tDtE

tF

tmex

Nmex

NA

NB

NCN1 N2

tDtE

tF

N3 N4

NC = Ξ±NA

N1 = Ξ²NC

N2 = (1-Ξ²)NC

N2P = Ξ³N2

NC = Ξ±NA

N1 = Ξ²NC

N2 = (1-Ξ²)NC

N2P = Ξ³N2

NC = Ξ±NA

N1 = Ξ²1NC

N2 = (1-Ξ²1)NC

N3 = Ξ²2N2

N4 = (1-Ξ²2)N2

N4P = Ξ³N4

tG

N4PLowland Highland mexicana

Meso-americaLowland Lowland Highland

Model IA Model IB Model II

Mesoamerica South America

Figure 2 Models of historical population size for lowlandand highland populations. Parameters in bold were esti-mated in this study. See text for details.

(Figure 2).We calculated the observed joint frequency distributions

(JFDs) using only the GBS data set due to its lower level ofascertainment bias. A subset of synonymous and noncodingSNPs were utilized that had οΏ½ 15 individuals without missingdata in both lowland and highland populations and did not vi-olate HWE. A HWE cut-off of P < 0.005 was used for eachsubpopulation due to our under-calling of heterozygotes.

We obtained similar results under more or less stringentthresholds for significance (P < 0.05 β‡  0.0005; data notshown), though the number of SNPs was very small at P <0.05.

Parameters were inferred with the software οΏ½aοΏ½i (Gutenkunstet al. 2009), which uses a diffusion method to calculate an ex-pected JFD and evaluates the likelihood of the data assumingmultinomial sampling. We did not use the β€œfull” model thatincorporates all four populations because parameter estimationunder this model is computationally infeasible.

Model IA This model is applied separately to both theMesoamerican and the S. American populations. We assumethe ancestral diploid population representing parviglumis fol-lows a standard Wright-Fisher model with constant size. Thesize of the ancestral population is denoted by NA. At tD gen-erations ago, the bottleneck event begins at domestication, andat tE generations ago, the bottleneck ends. The populationsize and duration of the bottleneck are denoted by NB andtB = tD οΏ½ tE , respectively. The population size recovers toNC = ↡NA in the lowlands. Then, the highland populationis differentiated from the lowland population at tF generationsago. The size of the lowland and highland populations at timetF is determined by a parameter οΏ½ such that the populationis divided by οΏ½NC and (1 οΏ½ οΏ½)NC ; our conclusions hold if

we force lowland population size to remain at NC (data notshown).

We assume that the population size in the lowlands is con-stant but that the highland population experiences exponentialexpansion after divergence: its current population size is οΏ½times larger than that at tF .

Model IB We expand Model IA for the Mesoamericanpopulations by incorporating admixture from the teosintemexicana to the highland Mesoamerican maize population.The time of differentiation between parviglumis and mexicanaoccurs at tmex generations ago. The mexicana population sizeis assumed to be constant at Nmex. At tF generations ago, theMesoamerican highland population is derived from admixturebetween the Mesoamerican lowland population and a portionPmex from the teosinte mexicana.

Model II The final model includes the Mesoamericanlowland, S. American lowland and highland populations. Thismodel was used for simulating SNPs with ascertainment bias(see below). At time tF , the Mesoamerican and S. Americanlowland populations are differentiated, and the sizes of popu-lations after splitting are determined by οΏ½1. At time tG, the S.American lowland and highland populations are differentiated,and the sizes of populations at this time are determined byοΏ½2. As in Model IA, the S. American highland population isassumed to experience population growth with the parameter οΏ½.

Estimates of a number of our model parameters were avail-able from previous work. NA was set to 150,000 using esti-mates of the composite parameter 4NAΒ΅ β‡  0.018 from parvig-lumis (Eyre-Walker et al. 1998; Tenaillon et al. 2001, 2004;Wright et al. 2005; Ross-Ibarra et al. 2009) and an estimateof the mutation rate Β΅ β‡  3 β‡₯ 10

οΏ½8 (Clark et al. 2005) persite per generation. The severity of the domestication bottle-neck is represented by k = NB/tB (Eyre-Walker et al. 1998;Wright et al. 2005), and following Wright et al. (2005) we as-sumed k = 2.45 and tB = 1, 000 generations. Taking into ac-count archaeological evidence (Piperno et al. 2009), we assumetD = 9, 000 and tE = 8, 000. We further assumed tF = 6, 000for Mesoamerican populations in Models IA and IB (Piperno2006), tF = 4, 000 for S. American populations in Model IA(Perry et al. 2006; Grobman et al. 2012), and tmex = 60, 000,Nmex = 160, 000 (Ross-Ibarra et al. 2009), and Pmex = 0.2(van Heerwaarden et al. 2011) for Model IB. For both Mod-els IA and IB, we inferred three parameters (↡, οΏ½ and οΏ½), and,for Model II, we fixed tF = 6, 000 and tG = 4, 000 (Piperno2006; Perry et al. 2006; Grobman et al. 2012) and estimatedthe remaining four parameters (↡, οΏ½1, οΏ½2 and οΏ½).

4

Lowland Highland

NA

NB

NCN1 N2

N2P

tDtE

tF

NA

NB

NCN1 N2

N2P

tDtE

tF

tmex

Nmex

NA

NB

NCN1 N2

tDtE

tF

N3 N4

NC = Ξ±NA

N1 = Ξ²NC

N2 = (1-Ξ²)NC

N2P = Ξ³N2

NC = Ξ±NA

N1 = Ξ²NC

N2 = (1-Ξ²)NC

N2P = Ξ³N2

NC = Ξ±NA

N1 = Ξ²1NC

N2 = (1-Ξ²1)NC

N3 = Ξ²2N2

N4 = (1-Ξ²2)N2

N4P = Ξ³N4

tG

N4PLowland Highland mexicana

Meso-americaLowland Lowland Highland

Model IA Model IB Model II

Mesoamerica South America

Figure 2 Models of historical population size for lowlandand highland populations. Parameters in bold were esti-mated in this study. See text for details.

(Figure 2).We calculated the observed joint frequency distributions

(JFDs) using only the GBS data set due to its lower level ofascertainment bias. A subset of synonymous and noncodingSNPs were utilized that had οΏ½ 15 individuals without missingdata in both lowland and highland populations and did not vi-olate HWE. A HWE cut-off of P < 0.005 was used for eachsubpopulation due to our under-calling of heterozygotes.

We obtained similar results under more or less stringentthresholds for significance (P < 0.05 β‡  0.0005; data notshown), though the number of SNPs was very small at P <0.05.

Parameters were inferred with the software οΏ½aοΏ½i (Gutenkunstet al. 2009), which uses a diffusion method to calculate an ex-pected JFD and evaluates the likelihood of the data assumingmultinomial sampling. We did not use the β€œfull” model thatincorporates all four populations because parameter estimationunder this model is computationally infeasible.

Model IA This model is applied separately to both theMesoamerican and the S. American populations. We assumethe ancestral diploid population representing parviglumis fol-lows a standard Wright-Fisher model with constant size. Thesize of the ancestral population is denoted by NA. At tD gen-erations ago, the bottleneck event begins at domestication, andat tE generations ago, the bottleneck ends. The populationsize and duration of the bottleneck are denoted by NB andtB = tD οΏ½ tE , respectively. The population size recovers toNC = ↡NA in the lowlands. Then, the highland populationis differentiated from the lowland population at tF generationsago. The size of the lowland and highland populations at timetF is determined by a parameter οΏ½ such that the populationis divided by οΏ½NC and (1 οΏ½ οΏ½)NC ; our conclusions hold if

we force lowland population size to remain at NC (data notshown).

We assume that the population size in the lowlands is con-stant but that the highland population experiences exponentialexpansion after divergence: its current population size is οΏ½times larger than that at tF .

Model IB We expand Model IA for the Mesoamericanpopulations by incorporating admixture from the teosintemexicana to the highland Mesoamerican maize population.The time of differentiation between parviglumis and mexicanaoccurs at tmex generations ago. The mexicana population sizeis assumed to be constant at Nmex. At tF generations ago, theMesoamerican highland population is derived from admixturebetween the Mesoamerican lowland population and a portionPmex from the teosinte mexicana.

Model II The final model includes the Mesoamericanlowland, S. American lowland and highland populations. Thismodel was used for simulating SNPs with ascertainment bias(see below). At time tF , the Mesoamerican and S. Americanlowland populations are differentiated, and the sizes of popu-lations after splitting are determined by οΏ½1. At time tG, the S.American lowland and highland populations are differentiated,and the sizes of populations at this time are determined byοΏ½2. As in Model IA, the S. American highland population isassumed to experience population growth with the parameter οΏ½.

Estimates of a number of our model parameters were avail-able from previous work. NA was set to 150,000 using esti-mates of the composite parameter 4NAΒ΅ β‡  0.018 from parvig-lumis (Eyre-Walker et al. 1998; Tenaillon et al. 2001, 2004;Wright et al. 2005; Ross-Ibarra et al. 2009) and an estimateof the mutation rate Β΅ β‡  3 β‡₯ 10

οΏ½8 (Clark et al. 2005) persite per generation. The severity of the domestication bottle-neck is represented by k = NB/tB (Eyre-Walker et al. 1998;Wright et al. 2005), and following Wright et al. (2005) we as-sumed k = 2.45 and tB = 1, 000 generations. Taking into ac-count archaeological evidence (Piperno et al. 2009), we assumetD = 9, 000 and tE = 8, 000. We further assumed tF = 6, 000for Mesoamerican populations in Models IA and IB (Piperno2006), tF = 4, 000 for S. American populations in Model IA(Perry et al. 2006; Grobman et al. 2012), and tmex = 60, 000,Nmex = 160, 000 (Ross-Ibarra et al. 2009), and Pmex = 0.2(van Heerwaarden et al. 2011) for Model IB. For both Mod-els IA and IB, we inferred three parameters (↡, οΏ½ and οΏ½), and,for Model II, we fixed tF = 6, 000 and tG = 4, 000 (Piperno2006; Perry et al. 2006; Grobman et al. 2012) and estimatedthe remaining four parameters (↡, οΏ½1, οΏ½2 and οΏ½).

4

Table 1 FST of synonymous and noncoding GBS SNPs

Mesoamerica S. America

Lowlands Highlands Lowlands Highlands

Mesoamerica Lowlands –

Highlands 0.0244 –

S. America Lowlands 0.0227 0.0343 –

Highlands 0.0466 0.0534 0.0442 –

be that the adaptive mutation occurs in a lowland area, sub-sequently migrating into the highlands. The calculation abovedoes not account for these points, but the approximation is quitegood, as verified by exact numerical calculation of the chanceof fixation of a mutation as a function of the location whereit first appears (see Figure A1); for theoretical treatment seeBarton (1987).

Migration It is harder to intuit a corresponding expressionfor the chance that an allele established by selection in onehighland population moves to the other.

For maize in the Andean highlands to have inherited ahighland-adapted allele from the Mesoamerican highlands,those Andean plants must be directly descended from highlandMesoamerican plants that lived more recently than the appear-ance of the adaptive allele. In other words, the ancestral lin-eages along which the modern Andean plants have inheritedat that locus must trace back to the Mesoamerican highlands.If the allele is neutral in the lowlands, we can treat the move-ment of these lineages as a neutral process, using the frame-work of coalescent theory (Wakeley 2005). To do this, we needto follow all of the N ⇑ 2.5β‡₯ 10

6 lineages backwards. Thesequickly coalesce to fewer lineages; but this turns out to not af-fect the calculation much. Assuming demographic stationarity,the motion of each lineage can be modeled as a random walk,whose displacement after m generations has variance mοΏ½2, andfor large m is approximately Gaussian. If we assume that lin-eages move independently, and Zn is the distance to the furthestof n lineages, then Zn

pmοΏ½2

(

p2 log n +

p2/ log n) with

very high probability (Berman 1964).Since this depends only on the logarithm of n, the number

of lineages, the practical upshot of this is that the most distantlineage is very unlikely to be more than about 6 times moredistant than the typical lineage, even among 10

7 lineages. Lin-eages are not independent, but this only makes this calculationconservative.

ResultsSamples and data

We sampled 94 maize landraces from four distinct regions inthe Americas (Table S1; Figure 1): the lowlands of Mesoamer-ica (Mexico/Guatemala; n = 24) and northern S. America

Table 2 Estimated parameters of population size model

Mesoamerica Model IA Model IB

Likelihood οΏ½5592.80 Likelihood οΏ½4654.79

NC 138,000 NC 225,000

N1 52,440 N1 171,000

N2 85,560 N2 54,000

N2P 85,560 N2P 54,000

S. America Model IA Model II

Likelihood οΏ½3855.28 Likelihood οΏ½8044.71

NC 78,000 NC 150,000

N1 75,660 N1 96,000

N2 2,340 N2 54,000

N2P 205,920 N3 51,300

N4 2,700

N4P 145,800

(n = 23) and the highlands of Mesoamerica (n = 24) andthe Andes (n = 23). Samples were genotyped using theMaizeSNP50 Beadchip platform (β€œMaizeSNP50”; n = 94) andgenotyping-by-sequencing (β€œGBS”; n = 87). After filteringfor Hardy-Weinberg genotype frequencies and minimum sam-ple size at least 10 in each of the four populations (see Materi-als and Methods) 91,779 SNPs remained, including 67,828 and23,951 SNPs from GBS and MaizeSNP50 respectively.

Population structure

We performed a STRUCTURE analysis (Pritchard et al. 2000;Falush et al. 2003) of our landrace samples, varying the numberof groups from K = 2 to 6 (Figure 1B, Figure S3). Most lan-draces were assigned to groups consistent with a priori popu-lation definitions, but admixture between highland and lowlandpopulations was evident at intermediate elevations (β‡  1700m).Consistent with previously described scenarios for maize dif-fusion (Piperno 2006), we find evidence of shared ancestry be-tween lowland Mesoamerican maize and both Mesoamericanhighland and S. American lowland populations. Pairwise FST

among populations reveals low overall differentiation (Table 1),and the higher FST values observed in S. America are consis-tent with the decreased admixture seen in STRUCTURE. Ar-chaeological evidence supports a more recent colonization ofthe highlands in S. America (Piperno 2006; Perry et al. 2006;Grobman et al. 2012), suggesting that the observed differentia-tion may be the result of a stronger bottleneck during coloniza-tion of the S. American highlands.

Population differentiation

To provide a null expectation for allele frequency differentia-tion, we used the joint site frequency distribution (JFD) of low-

7

Table 1 FST of synonymous and noncoding GBS SNPs

Mesoamerica S. America

Lowlands Highlands Lowlands Highlands

Mesoamerica Lowlands –

Highlands 0.0244 –

S. America Lowlands 0.0227 0.0343 –

Highlands 0.0466 0.0534 0.0442 –

be that the adaptive mutation occurs in a lowland area, sub-sequently migrating into the highlands. The calculation abovedoes not account for these points, but the approximation is quitegood, as verified by exact numerical calculation of the chanceof fixation of a mutation as a function of the location whereit first appears (see Figure A1); for theoretical treatment seeBarton (1987).

Migration It is harder to intuit a corresponding expressionfor the chance that an allele established by selection in onehighland population moves to the other.

For maize in the Andean highlands to have inherited ahighland-adapted allele from the Mesoamerican highlands,those Andean plants must be directly descended from highlandMesoamerican plants that lived more recently than the appear-ance of the adaptive allele. In other words, the ancestral lin-eages along which the modern Andean plants have inheritedat that locus must trace back to the Mesoamerican highlands.If the allele is neutral in the lowlands, we can treat the move-ment of these lineages as a neutral process, using the frame-work of coalescent theory (Wakeley 2005). To do this, we needto follow all of the N ⇑ 2.5β‡₯ 10

6 lineages backwards. Thesequickly coalesce to fewer lineages; but this turns out to not af-fect the calculation much. Assuming demographic stationarity,the motion of each lineage can be modeled as a random walk,whose displacement after m generations has variance mοΏ½2, andfor large m is approximately Gaussian. If we assume that lin-eages move independently, and Zn is the distance to the furthestof n lineages, then Zn

pmοΏ½2

(

p2 log n +

p2/ log n) with

very high probability (Berman 1964).Since this depends only on the logarithm of n, the number

of lineages, the practical upshot of this is that the most distantlineage is very unlikely to be more than about 6 times moredistant than the typical lineage, even among 10

7 lineages. Lin-eages are not independent, but this only makes this calculationconservative.

ResultsSamples and data

We sampled 94 maize landraces from four distinct regions inthe Americas (Table S1; Figure 1): the lowlands of Mesoamer-ica (Mexico/Guatemala; n = 24) and northern S. America

Table 2 Estimated parameters of population size model

Mesoamerica Model IA Model IB

Likelihood οΏ½5592.80 Likelihood οΏ½4654.79

NC 138,000 NC 225,000

N1 52,440 N1 171,000

N2 85,560 N2 54,000

N2P 85,560 N2P 54,000

S. America Model IA Model II

Likelihood οΏ½3855.28 Likelihood οΏ½8044.71

NC 78,000 NC 150,000

N1 75,660 N1 96,000

N2 2,340 N2 54,000

N2P 205,920 N3 51,300

N4 2,700

N4P 145,800

(n = 23) and the highlands of Mesoamerica (n = 24) andthe Andes (n = 23). Samples were genotyped using theMaizeSNP50 Beadchip platform (β€œMaizeSNP50”; n = 94) andgenotyping-by-sequencing (β€œGBS”; n = 87). After filteringfor Hardy-Weinberg genotype frequencies and minimum sam-ple size at least 10 in each of the four populations (see Materi-als and Methods) 91,779 SNPs remained, including 67,828 and23,951 SNPs from GBS and MaizeSNP50 respectively.

Population structure

We performed a STRUCTURE analysis (Pritchard et al. 2000;Falush et al. 2003) of our landrace samples, varying the numberof groups from K = 2 to 6 (Figure 1B, Figure S3). Most lan-draces were assigned to groups consistent with a priori popu-lation definitions, but admixture between highland and lowlandpopulations was evident at intermediate elevations (β‡  1700m).Consistent with previously described scenarios for maize dif-fusion (Piperno 2006), we find evidence of shared ancestry be-tween lowland Mesoamerican maize and both Mesoamericanhighland and S. American lowland populations. Pairwise FST

among populations reveals low overall differentiation (Table 1),and the higher FST values observed in S. America are consis-tent with the decreased admixture seen in STRUCTURE. Ar-chaeological evidence supports a more recent colonization ofthe highlands in S. America (Piperno 2006; Perry et al. 2006;Grobman et al. 2012), suggesting that the observed differentia-tion may be the result of a stronger bottleneck during coloniza-tion of the S. American highlands.

Population differentiation

To provide a null expectation for allele frequency differentia-tion, we used the joint site frequency distribution (JFD) of low-

7

Hufford et al. 2013 PLoS Genetics van Heerwaarden et al. 2011 PNAS Takuno et al. 2015 bioRxiv http://dx.doi.org/10.1101/013607

Mexico highlands Andean highlands

Li Wang

Highland (>1800m)

Lowland (<1700m)

30 genomes ~25X

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β—βˆ’1

0

1

Andean SAlow MexHigh MexLow GuaHigh swUS

Tajim

aD

factor(pop)AndeanGuaHighMexHighMexLowSA_LowSW_US

Tajim

a’s D

SAm Low

Gua High

SAm High

Mex High

Mex Low

SW High

Rodgers-Melnick et al. 2015 PNAS

26

Figure S30. The mean GERP rates within annotated codons across the genome. Bars give the mean rates at all first and second codon positions or all third codon positions (red and orange bars) or the mean rates at those positions with known segregating polymorphisms (blue and green bars).

Figure S31. Percent difference between the proportion of polymorphisms at sites with GERP above a threshold in hotspots ( ΞΈ 1) vs. the rest of the genome (ΞΈ 2).

A B

N = 744,505 N = 506,898

r = -0.8

Mea

n G

ERP

Mea

n G

ERP

MAFchromosome

Jinliang Yang

Li Wang

●

●

●

●

●

1500000

2000000

2500000

3000000

Andes SA_Low MexHigh MexLow SW_US GuaHigh

load

popAndesGua_HighlandMex_HighlandMex_LowlandSA_LowlandSW_US

●

●

●

2500000

3000000

3500000

4000000

4500000

Andes SA_Low MexHigh MexLow SW_US GuaHigh

num

ber o

f var

iabl

e si

tes

with

GER

P >0

popAndesSA_LowlandMex_HighlandMex_LowlandSW_USGua_Highland

Variable(Sites(with(Posi0ve(GERP(

all deleterious (GERP>0) SNPsNon7reference(homozygous(sites(with(Posi0ve(GERP(●

●

●

●

190000

210000

230000

Andes SA_Low MexHigh MexLow SW_US GuaHigh

load

popAndesGua_HighlandMex_HighlandMex_LowlandSA_LowlandSW_US

●

●

●

200000

220000

240000

Andes SA_Low MexHigh MexLow SW_US GuaHigh

num

ber o

f non

Ref

hom

o si

tes

with

GER

P >

0

popAndesGua_HighlandMex_HighlandMex_LowlandSA_LowlandSW_US

pop Andes SA_Lowland Mex_HighlandMex_LowlandSW_US Gua_HighlandAndesSA_Lowland 0.0022Mex_Highland 0.0411 0.1201Mex_Lowland 0.004 0.2684 0.3961SW_US 0.0022 0.197 0.2424 0.5346Gua_Highland 0.0179 0.2738 0.131 0.1964 0.131

homozygous deleterious SNPs

SAm Low

Gua High

SAm High

Mex High

Mex Low

SW High

SAm Low

Gua High

SAm High

Mex High

Mex Low

SW High

Fu et al. 2014 AJHG

Paul Bilinski

2.50

2.75

3.00

3.25

3.50

3.75

MH ML SAH SAL mexicana parviglumis1C

Gen

ome

Size

(Gb)

Altitudehighland

lowland

β€’ limited impact of (hard) sweeps on linked diversity

β€’ strong role for background selection

β€’ efficacy of selection in maize changed with Ne

β€’ recent demography affects genetic load among populations

conclusions

Acknowledgements

Hufford Lab (Iowa State)Matt Hufford

Li Wang

Ross-Ibarra Lab (UC Davis)Tim Beissinger (USDA U. Missouri)

Kate CrosbyArun Durvasula

Graham Coop (UC Davis)