Population genetics of marmosets in Asian primate research centers and loci associated with epileptic risk revealed by whole-genome sequencing

The common marmoset(Callithrix jacchus)has emerged as a valuable nonhuman primate model in biomedical research with the recent release of high-quality reference genome assemblies.Epileptic marmosets have been independently reported in two Asian primate research centers.Nevertheless,the population genetics within these primate centers and the specific genetic variants associated with epilepsy in marmosets have not yet been elucidated.Here,we characterized the genetic relationships and risk variants for epilepsy in 41 samples from two epileptic marmoset pedigrees using whole-genome sequencing.We identified 14558184 single nucleotide polymorphisms(SNPs)from the 41 samples and found higher chimerism levels in blood samples than in fingernail samples.Genetic analysis showed fourth-degree of relatedness among marmosets at the primate centers.In addition,SNP and copy number variation(CNV)analyses suggested that the WW domain-containing oxidoreductase(WWOX)and Tyrosine-protein phosphatase nonreceptor type 21(PTPN21)genes may be associated with epilepsy in marmosets.Notably,KCTD18-like gene deletion was more common in epileptic marmosets than control marmosets.This study provides valuable population genomic resources for marmosets in two Asian primate centers.Genetic analyses identified a reasonable breeding strategy for genetic diversity maintenance in the two centers,while the case-control study revealed potential risk genes/variants associated with epilepsy in marmosets.

Molecular phylogeography and population genetic structure of the planktonic copepod Calanus sinicus Brodsky in the coastal waters of China

Planktonic copepod Calanus sinicus is the dominant meso-zooplankton in the Northwest Pacific Ocean. To better understand its population dynamics and phylogeographic patterns, 243 C. sinicus individuals were collected from seven locations across the shelf waters of China and its population genetics was studied by mitochondrial DNA cytochrome oxidase I（mtCOI） sequences analyses. Thirty-nine different sequences, or haplotypes, were detected with moderate haplotype diversity（h=0.749） and low nucleotide diversity（π=0.003） for all populations. The evolutionary divergence between geographic populations varied from 0.24% to 0.37%, indicative of very limited genetic differentiation. Visualized minimum spanning network（MSN） and phylogenetic analysis of all the detected haplotypes did not reveal any clear phylogeographic pattern. Furthermore, AMOVA data showed no significant spatial population differentiation existed among the individuals collected across China shelf waters. Pairwise FST values showed that population collected from northwest of the East China Sea（ECS） displayed a low difference to other populations. Mismatch distribution analyses and neutrality tests indicated that C. sinicus might undergo a demographic/population expansion. No significant population genetic structuring was detected, indicating an extensive gene flow among the C. sinicus populations. Our results provide molecular evidence for the hypothesis that C. sinicus in the northwestern South China Sea in winter is transported from the East China Sea and the Yellow Sea by the China Coastal Current during the northeast monsoon period.

Population genetic studies in the genomic sequencing era

Recent advances in high-throughput sequencing technologies have revolutionized the field of population genetics. Data now routinely contain genomic level polymorphism information, and the low cost of DNA sequencing enables researchers to investigate tens of thousands of subjects at a time. This provides an unprecedented opportunity to address fundamental evolutionary questions, while posing challenges on traditional population genetic theories and methods. This review provides an overview of the recent methodological developments in the field of population genetics, specifically methods used to infer ancient population history and investigate natural selection using large-sample, large-scale genetic data. Several open questions are also discussed at the end of the review.

Population Genetic Analysis of Sillago nigrofasciata (Perciformes:Sillaginidae) Along the Coast of China by Sequencing Mitochondrial DNA Control Region

Sillago nigrofasciata, a small to moderate size nearshore species, is newly found along the eastern and southern coasts of China. The present study is carried out in order to analyze the population genetics of the S. nigrofasciata. The control region sequence of mitochondrial DNA revealing 73 haplotypes were obtained from 162 individuals collected at 8 locations along the coast of China. The whole S. nigrofasciata population along the coast of China showed a low nucleotide diversity(0.012) and a high population diversity(haplotype diversity)(0.943), and all the 8 local populations showed low nucleotide diversities(0.014 – 0.001), suggesting the protective measures are effective. The haplotypes of the 8 local populations were widely distributed in haplotype network diagram and neighbor-joining phylogenetic tree, while no branch associating with sampling locations was detected. Recent gene flow analysis showed asymmetric gene exchanges among local populations. The pairwise FST values and unweighted pair-group method with arithmetic mean(UPGMA) tree revealed a certain amount of genetic difference among local populations. Moreover, analysis of molecular variance(AMOVA) reflected genetic differences between hypothetical subdivision groups. Neutral test and mismatch distribution of pairwise nucleotide suggested S. nigrofasciata may have experienced recent population expansion events. The historical geographic events associating with ice age may be the main explanation to the heterogeneity among local populations with short geographic distances, and the homogeneity among local populations with long geographic distances.

Genetic diversity and population genetic structure of Python bivittatus in China

The Burmese python （Python bivittatus） has recently suffered large population declines in the wild in China due to illegal capture, overexploitation, deforestation and the loss of its natural habitat. Greater knowledge of the genetic diversity and structure of wild P. bivittatus populations is needed to help ensure its effective management. In this study, we investigated the genetic diversity and population genetic structure of wild P. bivittatus in China in detail. 109 P. bivittatus individuals from five distribution areas in Guangdong （GD）, Guangxi （GX）, Hainan （HN）, Fujian （FJ） and Yunnan （YN） province of China were collected, and their genetic structure and diversity were analyzed. Eight highly polymorphic microsatellite loci were utilized to reveal high levels of genetic diversity in the P. bivittatus population. Genetic diversity was highest in GX, and lowest in GD. All geographic populations demonstrated a bottleneck effect indicating recent population decline. Fst and AMOVA analyses revealed that there was moderate genetic differentiation among the five populations, and that only 10.59 % of total genetic diversity occurred among populations. Fst values between pop- ulations were positively correlated with their geographical distances. Genetic structure analyses revealed that the HN, GX and GD populations, which were geographically closest, were assigned to a genetic cluster, while the YN and FJ populations constituted a single cluster, respectively.

Reveal the Population Genetic Characteristics of Bombay Duck (Harpadon nehereus) in Coastal Waters of China with Genotyping-by-Sequencing Technique

Bombay duck(Harpadon nehereus)is an economically important species in the estuarine and coastal offshore waters of the Indo-West Pacific.This study aims to reveal the genome-wide genetic characteristics of five populations of H.nehereus from the coastal areas of China by using the genotyping-by-sequencing technique.After strict filtering,32088 high-quality single-nucleotide polymorphism markers were detected and analyzed.The average observed heterozygosity and expected heterozygosity ranged from 0.41651 to 0.56725 and from 0.30998 to 0.45531,respectively,indicating that heterozygosity excess occurred in H.nehereus populations.The nucleotide diversity ranged from 0.63664 to 0.74868,which was larger than those from other marine fishes.No obvious genetic structure was detected among H.nehereus populations,and the genetic variation originated within individuals.Extensive gene exchange caused by longshore currents in the reproductive season might be the primary reason for the weak genetic differentiation.Among various environmental factors,water temperature might be the key element affecting the genetic structure of H.nehereus.Due to the destruction and overfishing of spawning grounds,the fishery resources declined in recent years.This study could serve as a reference for the resource protection and rational utilization of H.nehereus.

Microsatellite marker development and population genetic analysis revealed high connectivity between populations of a periwinkle Littoraria sinensis(Philippi,1847)

Periwinkle Littoraria sinensis is a widely distributed gastropod in rocky intertidal zone of the northwestern Pacifi c.To examine genetic diversity and genetic connectivity among coastal populations of L.sinensis in China,1636 pairs of primers were successfully designed using whole-genome shotgun sequencing,de novo assembly,and a bioinformatics pipeline QDD.Twelve highly variable polymorphic markers were selected to genotype 351 individuals from 15 populations.Data of nine microsatellite loci were retained for population genetic analysis,and weak genetic differentiation among populations were detected,suggesting high gene flow among populations.The long planktonic larval duration of L.sinensis might have played an important role in the high gene flow among populations.A tendency of genetic differentiation between north and south populations of L.sinensis was detected,which might be resulted from isolation due to lowered sea level in the last glacial maximum.Furthermore,the newly founded populations along the coast of Jiangsu Province were closely related to populations to the south of the Changjiang(Yangtze)River estuary,suggesting that the main source of the newly founded populations is from natural rocky populations south of the estuary.

Molecular phylogenetics and population demographic history of Amphioctopus fangsiao,inferred from mitochondrial and microsatellite DNA markers

Amphioctopus fangsiao(Cephalopoda:Octopodidae)is an important commercial species in the coastal waters of China.In recent years,however,the resource of A.fangsiao have declined because of habitat destruction and overfishing.To analyze the genetic variations of A.fangsiao caused by the fluctuation of resources,the population genetic structure of nine sampling locations collected from the Bohai Sea to the South China Sea were investigated,using mtDNA COI fragments and microsatellite DNA.The results of F-statistics,AMOVA,STRUCTURE and PCA analyses showed three phylogeographic clades(Clades A,B and C),revealing limited genetic exchange between north and south populations.These clades diverged in 2.23(Clades A and B)and 3.67(Clades A,B and C)million years ago,during the dramatic environmental fluctuations,such as sea level and temperature changes,have exerted great influence on the survival distribution pattern of global organisms.Our results for low genetic connectivity among A.fangsiao populations provide insights into the development of management strategies,that is,to manage this species as separate management unit.

Haplotype and Genetic Analysis of 41 Y-STR Loci in the Wuwei Han Population from Gansu Province,China

Objective Y-Chromosomal short tandem repeat polymorphism(Y-STR)analysis plays an indispensable role in the identification of male individuals,population genetics,and biogeographic research.While profiles of many populations based on Y-STR markers in human genomes are ample,haplotype data for the Wuwei Han are still scarce.Methods In this study,2180 unrelated Wuwei Han male individuals residing in Gansu Province,China were collected and genotyped using the novel Microreader™40Y Plus ID system.Phylogenetic relationship reconstructions,multidimensional scaling(MDS),and heatmap analysis were performed based on the genetic distance(Rst)values between our studied population and other populations of the Ymax module in the Y-STR Haplotype Reference Database(YHRD).Results A total of 2129 unique haplotypes were obtained,and the haplotype diversity(HD)and discrimination capacity(DC)for the Wuwei Han were 0.9999 and 0.9931,respectively.Conclusion Our results demonstrate that the Wuwei Han population had intimate genetic relationships with East Asians,especially the geographically close Han populations.Overall,this Y-Chromosomal assay gives valuable information about paternal lineages in male individual tracking and genealogical database construction.

Genetic variation of Laminaria japonica (Phaeophyta) populations in China as revealed by RAPD markers

For the population genetics analysis of the naturally grown brown seaweed Laminaria japonica （Laminariales,Phaeophyta） sampled from Dalian,Yantai,Weihai,Rongcheng and Qingdao in China,ten primers were employed to produce 88 bands as revealed by randomly amplified polymorphic DNA （RAPD） markers,and all these bands were polymorphic.According to these band patterns,there were 94 distinct phenotypes occurred in 100 samples indicating the high heterozygosity of this kelp.Dalian population samples showed the highest percentage of polymorphism （71.67%）,and also the higher diversity estimated on the basis of the Shannon’s index （8.498）,suggesting that this population could be chosen as the best resource for genetic breeding.The highest diversity of Yantai population possibly resulted from the introduction of L.longissima used for interspecific cross breeding with L.japonica cultivated in China.From Dalian southwards to Qingdao,the genetic variation of the five populations became less with a decrease in latitude,possibly due to the natural selection especially of high temperature.The genetic distance （Φ ST values） of the five populations was a little significantly correlated with the geographical distance （r=0.496） at P =0.05 by Mantel’s test.Weihai,Rongcheng and Yantai populations were closely grouped genetically together by Neighbor-joining cluster analysis probably in that the dispersal of the kelp by propagules more easily occurring in the range of relatively short distance.The analysis of molecular variance （AMOVA） also demonstrated that the relatively higher variation occurred among populations （71.49%） at an extremely significant level （P 0.000 1）.All these evidence showed that there was a relatively distinct genetic differentiation among the sampled kelp populations,and L.japonica grown in China was also rather heterozygous in heredity.

Molecular Population Genetics of Rice Domestication

Domestication is a selection process that genetically modifies species to meet human needs. A most intriguing feature of domestication is the extreme phenotypic diversification among breeds. What could be the ultimate source of such genetic variations？ Another notable outcome of artificial selection is the reduction In the fitness of domesticated species when they live In the wild without human assistance. The complete sequences of the two subspecies of rice cultlvars provide an opportunity to address these questions. Between the two subspecies, we found much higher rates of non-synonymous （N） than synonymous （S） substitutions and the N/S ratios are higher between cultivars than between wild species. Most interestingly, substitutions of highly dissimilar amino acids that are deleterious and uncommon between natural species are disproportionately common between the two subspecies of rice. We suggest strong selection in the absence of effective recombination may be the driving force, which we called the domestication-associated Hill-Robertson effect. These hitchhiking mutations may contribute to some fitness reduction in cultivars. Comparisons of the two genomes also reveal the existence of highly divergent regions in the genomes. Haplotypes in these regions often form highly polymorphic linkage blocks that are much older than speciation between wild species. Genes from such regions could contribute to the differences between indica and japonica and are likely to be involved in the diversifying selection under domestication. Their existence suggests that the amount of genetic variation within the single progenitor species Oryza rufipogon may be insufficient to account for the variation among rice cultivars, which may come from a more inclusive gene pool comprising most of the A-genome wild species. Genes from the highly polymorphic regions also provide strong support for the Independent domestication of the two subspecies. The genomic variation in rice has revealing implications for studying the genetic basis of indica-japonica differentiation under rice domestication and subsequent improvement.

Population genetic structure of the biological control agent Macrolophus pygmaeus in Mediterranean agroecosystems

Biological control of agricultural pests relies on knowledge of agroecosystem functionality, particularly when affected by the use of mass-produced biological agents. Incorporating pre- and/or post-release information such as genetic diversity and struc- ture on these agents using molecular-based approaches could advance our knowledge of how they perform in agroecosystems. We evaluated the population genetics of Macrolo- phus pygmaeus, the most widely used predatory mirid against many arthropod pests of greenhouse crops in the Mediterranean region, using the mitochondrial Cytb sequence and microsatellite data, and population genetics and phylogeny approaches. We investigated commercially mass-produced insects （i.e., commercial insects either mass-reared in the laboratory for many generations, or purchased by farmers and released in the greenhouses） and ＂wild＂ insects （i.e., that occur naturally outside or are collected in nature for release in the greenhouses）. The mirids were mainly collected in agroecosystems in which solana- ceous plants are grown in northern Spain, southern France and Greece. Both molecular markers and approaches distinguished 2 genetically differentiated populations. The less genetically diverse population, hereafter named the ＂commercial＂ strain included all indi- viduals from laboratory mass-rearings and most releases of commercially bred individuals. The most genetically diverse population mainly comprised individuals originating from noncultivated environments, or from releases of＂wild＂ individuals. Rare examples of hy- bridization between M. pygmaeus from the 2 populations were observed and asymmetric gene flow was revealed. These findings provide new insights into what happens to M. pyg- maeus released in the agroecosystems we studied, and show that it is possible to monitor some commercial strains.

Genetic Differentiation of Japanese Sardinella(Sardinella zunasi) Populations in the Northwest Pacific Revealed by ISSR Analysis

Knowledge of population genetic structure plays an important role in fisheries management.In this research,In-ter-Simple-Sequence-Repeat(ISSR)markers were employed to evaluate the genetic structure of Japanese sardinella(Sardinella zun-asi)populations in the Northwest Pacific.Eighty seven individuals from 5 locations were screened using 4 highly polymorphic prim-ers.A total of 173 polymorphic loci were detected out of 191 loci amplified.Small but significant genetic differentiation was de-tected between the Chinese and Japanese populations by both AMOVA and pairwise FST analyses,which was further supported by cluster analysis.We consider that climate change during glaciations should be responsible for the genetic differentiation.Isolation by geographic distance among populations was observed,indicating that the distance might also lead to the genetic differentiation.However,no genetic structure was found within the populations off both the Chinese and Japanese coasts,indicating a high-level along-coast gene flow,which might result from ocean current transport and common ground for over-wintering.

An interpretation of the genetic polymorphism and population genetic background of Ankang Han population via a novel InDel panel

In this research,genotyping data of 43 InDel loci in 311 Han individuals in Ankang City,Shaanxi Province,China were detected using a self-developed five-dye multiplex amplification panel.The allelic frequencies and forensic parameters of all InDel loci were calculated.The combined power of discrimination and probability of exclusion values were 0.99999999999999999882739 and 0.999887424,respectively,which demonstrated that this 43-InDel panel was powerful for individual identifications in Ankang Han population.Moreover,genetic distances,pairwise F_(ST)values,principal component analyses,phylogenetic trees and STRUCTURE analyses were performed to investigate the genetic affinities between Ankang Han and reference groups.Population genetic investigations indicated that Ankang Han population had a close genetic relationship with Southern Han population compared with other reference groups.

Population genetics and forensic efficiency of 30 InDel markers in four Chinese ethnic groups residing in Sichuan

Sichuan Province is located at the transitional junction regions of the Qinghai-Tibet Plateau and the low-altitude plains.It also serves as the corridor of Sino Tibetan-speaking population migration and expansion since neolithic expansion of Proto-Tibeto-Burman populations from Middle/Upper Yellow River during Majiayao period(3300-2000 BC).However,the population structure and the corresponding genetic diversity of forensic-related markers in this region remain unclear.Thus,we genotyped 30 insertion-deletion(InDel)markers in 444 samples from four ethnic groups(Han,Tibetan,Hui and Yi)from Sichuan Province using the Investigator■ DIPplex kit to explore the characteristics of population genetics and forensic genetic focuses.All the loci were found to be in Hardy-Weinberg Equlibrium(HWE)after applying a Bonferroni correction and no pairwise loci showed prominent linkage disequilibrium.The combined matching probability(CMP)and the combined power of discrimination(CPD)are larger than 1.8089×10^(11)and 0.9999999995,respectively.Principal component analysis,multi-dimen-sional scaling plots and Neighbour-Joining tree among 65 worldwide populations indicated that Sichuan Hui and Han are genetically close to Hmong-Mien and Tai-Kadai-speaking popula-tions,and Sichuan Tibetan and Yi bear a strong genetic affinity with Tibeto-Burman-speaking populations.The model-based genetic structure further supports the genetic affinity between the studied populations and linguistically close populations.

The Hardy-Weinberg principle and its applications in modern population genetics

The discovery of the Hardy-Weinberg principle marked the beginning of the field of population genetics.Over the past hundred years,it has provided a starting point for many population genetic investigations.In this review,the Hardy-Weinberg principle,its statistical testing,and several of its applications in various modern population genetic research areas,including allelic variability and selection in the human leukocyte antigen region,microsatellite genotyping error detection,and accuracy of haplotype estimation,are discussed.

DISTRIBUTION OF ALPHA-1-ANTITRYPSIN VARIANT ETOKYO AND ITS IMPLICATION IN HUMAN POPULATION GENETICS

Ⅰ. INTRODUCTION Alpha-1-antitrypsin (A1AT; locus symbol, PI) is one of the main protease inhibitors in serum. Up to now there are at least 50 A1AT variants that have been found in human populations (Cox, D. W., private communication). The distribution of A1AT variants appears highly racial specificity and geographical variability. This

Genetic diversity analysis of spawner and recaptured populations of Chinese shrimp(Fenneropenaeus chinensis)during stock enhancement in the Bohai Bay based on an SSR marker

Eight microsatellite markers were used to analyze genetic diversity, level of inbreeding, and effective population size of spawner and recaptured populations of Chinese shrimp（Fenneropenaeus chinensis） during stock enhancement in the Bohai Bay in 2013. A total of 254 and 238 alleles were identified in the spawner and recaptured populations, respectively, and the numbers of alleles（N_a） were 8–63 and 6–60, respectively. The numbers of effective alleles（N_e） were 2.52–21.60 and 2.67–20.72, respectively. The polymorphism information content ranged from 0.529 to 0.952. The observed heterozygosity（H_o） values（0.638–0.910 and 0.712–0.927） were lower than the expected heterozygosity（H_e） values（0.603–0.954 and 0.625–0.952）, which indicated that the two populations possessed a rich genetic diversity. In 16 tests（2 populations×8 loci）, 13 tests deviated from the HardyWeinberg equilibrium. F_（is） values were positive at seven loci and the inbreeding coefficients（F） of the two populations estimated by trio ML were 13.234% and 11.603%, suggesting that there was a relatively high degree of inbreeding. A certain level of inbreeding depression had occurred in the Chinese shrimp population. F_（st） values ranged from 0 to 0.059, with a mean of 0.028, displaying a low level of genetic differentiation in the two populations. Effective population sizes（3 060.2 and 3 842.8） were higher than the minimum number suggested for retaining the evolutionary potential to adapt to new environmental conditions. For enhancement activity in 2014,the ideal number of captured shrimp spawners should have ranged from 7 686 to 19 214 to maintain genetic diversity and effective population size. Further strategies to adjust the balance of economic cost, fishing effort and ideal number of shrimp spawners to maintain a satisfactory effective population size for ensuring the sustainability of Chinese shrimp are proposed.

Genetic variation and population structure of the mushroom Pleurotus ferulae in China inferred from nuclear DNA analysis

To investigate the genetic diversity of an edible fungus Pleurotus ferulae, a total of 89 wild samples collected from six geographical locations in the Xinjiang Uygur Autonomous Region of China and two geographical locations in Italy, were analyzed using three DNA fragments including the translation elongation factor（EF1α）, the second largest subunit of t he RNA polymerase II（RPB2） and the largest subunit of the RNA polymerase II（RPB1）. The results indicated relatively abundant genetic variability in the wild resources of P. ferulae. The analysis of molecular variance（AMOVA） showed that the vast majority of the genetic variation was found within geographical populations. Both the Chinese populations and the Italian populations of P. ferulae displayed a limited genetic differentiation. The degree of differentiation between the Chinese populations and the Italian populations was obviously higher than that between the populations from the same region, and moreover the genetic differentiation among all the tested populations was correlated to the geographical distance. T he phylogeny analyses confirmed that samples from China and Italy belonged to another genetic group separated from Pleurotus eryngii. They were closely related to each other but were clustered according to their geographical origins, which implied the Chinese populations were highly differentiated from the Italian populations because of distance isolation, and the two populations from different regions might be still in the process of allopatric divergence.

Genetic Diversity and Population Structure in Landraces and Improved Rice Varieties from India

A set of 50 rice genotypes comprising landraces, local selections, and improved varieties were characterized using simple sequence repeat（SSR） and inter simple sequence repeat（ISSR） markers to study genetic diversity and population structure. Following unweighted pair group method with arithmetic mean based clustering using binary data of polymorphic markers, the genotypes were grouped into 5 clusters and 11 sub-clusters, whereas population structure analysis separated 50 rice genotypes into 5 sub-populations. Grouping of rice genotypes showed better resemblance with the pedigree information of the genotypes. Both genetic diversity and population structure analysis separated majority of the improved varieties from landraces and local selections. Some of the SSR markers amplified unique alleles which were specific to a particular genotype and could distinguish them from the rest. The results indicate that these rice genotypes exhibit a higher genetic diversity and can be very useful in rice improvement program.