Comparison of SSR and SNP Markers in Estimation of Genetic Diversity and Population Structure of Indian Rice Varieties

Simple sequence repeat (SSR) and Single Nucleotide Polymorphic (SNP), the two most robust markers for identifying rice varieties were compared for assessment of genetic diversity and population structure. Total 375 varieties of rice from various regions of India archived at the Indian National GeneBank, NBPGR, New Delhi, were analyzed using thirty six genetic markers, each of hypervariable SSR (HvSSR) and SNP which were distributed across 12 rice chromosomes. A total of 80 alleles were amplified with the SSR markers with an average of 2.22 alleles per locus whereas, 72 alleles were amplified with SNP markers. Polymorphic information content (PIC) values for HvSSR ranged from 0.04 to 0.5 with an average of 0.25. In the case of SNP markers, PIC values ranged from 0.03 to 0.37 with an average of 0.23. Genetic relatedness among the varieties was studied; utilizing an unrooted tree all the genotypes were grouped into three major clusters with both SSR and SNP markers. Analysis of molecular variance (AMOVA) indicated that maximum diversity was partitioned between and within individual level but not between populations. Principal coordinate analysis (PCoA) with SSR markers showed that genotypes were uniformly distributed across the two axes with 13.33% of cumulative variation whereas, in case of SNP markers varieties were grouped into three broad groups across two axes with 45.20% of cumulative variation. Population structure were tested using K values from 1 to 20, but there was no clear population structure, therefore Ln(PD) derived Δk was plotted against the K to determine the number of populations. In case of SSR maximum Δk was at K=5 whereas, in case of SNP maximum Δk was found at K=15, suggesting that resolution of population was higher with SNP markers, but SSR were more efficient for diversity analysis.     

30个粳稻品种SSR标记遗传多样性分析

选用分布于水稻12条染色体上的64对SSR引物,对江苏省育成以及日本引进的粳稻品种共30份材料进行遗传多样性分析。结果表明,有50对SSR引物在30个品种间表现为多态性。共检测到140个等位基因,每对引物的等位基因数变幅为2～5个,平均为2．8个。有效等位基因为94．336个,平均为1．887。每个SSR位点的多态性信息量（PIC）变化范围为0．064～0．752,平均为0．410。30个品种间的遗传相似系数变幅为0．386～0．956之间,平均值为0．719,且81．4％的供试品种其遗传相似系数在0．600～0．800之间,亲缘关系较近;以遗传相似系数为原始数据,按UPGMA方法将30个品种划分为3大类群,结合系谱分析结果表明,江苏省育成的水稻品种遗传多样性不够丰富,多数品种间的亲缘关系较近,欲进一步提高江苏省水稻产量还需拓宽亲本选择范围,扩大遗传背景。     

303份甘薯地方种SSR遗传多样性与群体结构分析

利用SSR分子标记,对我国303份甘薯地方种进行了遗传多样性和群体结构分析。进一步明确了甘薯地方种间的遗传多样性和亲缘关系,为优异资源挖掘和品种改良提供了参考。利用SSR建立研究材料的0~1数据库,通过NTSYS-pc2.10软件计算Nei72遗传距离矩阵,将遗传距离矩阵导入MEGA 6.06,计算平均遗传距离和聚类分析;并利用STRUCTURE2.3.4对303份地方种进行群体结构分析。结果表明:30对SSR引物共检测出203条多态性位点,每对引物检测到1~14条多态性条带,平均每对引物获得6.77条。303份材料的平均遗传距离为0.564,聚类分析在遗传距离为0.477处可以把303份材料分成11个类群,其中第Ⅺ类群在遗传距离为0.452处可分为3个亚群。群体结构分析将303份材料划分成了5个稳定的群体,群体结构划分与聚类有相似的结果,其中70份材料Q值小于0.6,属于混合亚群。     

黑龙江省近年审定水稻品种基于SSR标记的遗传多样性分析

为评估黑龙江省水稻品种的遗传基础,利用24个用于水稻DNA指纹图谱构建的SSR标记以及其他均匀分布于水稻12条染色体的38个SSR标记,对黑龙江省近年审定的73个水稻常规稻品种进行遗传多样性分析。结果表明,在62个SSR标记位点中,共检测到142个等位基因,平均每个标记2.3个,多态性比率平均为71.0%,多态性频率变幅为0~0.775,平均值为0.246。供试品种间两两遗传相似系数的平均值为0.759,变幅为0.622~0.966,且96.4%的品种间遗传相似系数在0.66~0.86之间,表明供试的73个品种亲缘关系较近。通过SSR标记基因型聚类分析将这些品种划分为6个类群,与系谱分析趋势一致,类群间的差异主要表现在生育期和米质方面。综上所述,黑龙江省近年审定的水稻品种遗传基础狭窄,在育种中需要导入新的种质资源,加强种质资源创新,以期丰富水稻品种的遗传多样性,进一步提高水稻产量和抗性。     

宁夏89份粳稻种质遗传多样性的SSR分析

选用分布于水稻12条染色体上的47对引物对宁夏89份粳稻种质材料进行SSR分析,以探讨宁夏粳稻品种的遗传多样性水平.结果表明,(1)47个位点上共检测到204个具有多态性的等位片段,每对引物检测出2～9个多态性片段,平均为4.16个;(2)聚类分析显示,89份材料的相似系数为0.63～0.91,平均为0.79.在相似系数0.718处聚为8个类群.研究表明,宁夏粳稻种质之间的相似性较高,遗传差异较小,遗传背景比较单一.     

Genetic Diversity and Population Structure in Aromatic and Quality Rice (Oryza sativa L.) Landraces from North-Eastern India

The North-eastern (NE) India, comprising of Arunachal Pradesh, Assam, Manipur, Meghalaya, Mizoram, Nagaland, Sikkim and Tripura, possess diverse array of locally adapted non-Basmati aromatic germplasm. The germplasm collections from this region could serve as valuable resources in breeding for abiotic stress tolerance, grain yield and cooking/eating quality. To utilize such collections, however, breeders need information about the extent and distribution of genetic diversity present within collections. In this study, we report the result of population genetic analysis of 107 aromatic and quality rice accessions collected from different parts of NE India, as well as classified these accessions in the context of a set of structured global rice cultivars. A total of 322 alleles were amplified by 40 simple sequence repeat (SSR) markers with an average of 8.03 alleles per locus. Average gene diversity was 0.67. Population structure analysis revealed that NE Indian aromatic rice can be subdivided into three genetically distinct population clusters: P1, joha rice accessions from Assam, tai rices from Mizoram and those from Sikkim; P2, chakhao rice germplasm from Manipur; and P3, aromatic rice accessions from Nagaland. Pair-wise F_ST between three groups varied from 0.223 (P1 vs P2) to 0.453 (P2 vs P3). With reference to the global classification of rice cultivars, two major groups (Indica and Japonica) were identified in NE Indian germplasm. The aromatic accessions from Assam, Manipur and Sikkim were assigned to the Indica group, while the accessions from Nagaland exhibited close association with Japonica. The tai accessions of Mizoram along with few chakhao accessions collected from the hill districts of Manipur were identified as admixed. The results highlight the importance of regional genetic studies for understanding diversification of aromatic rice in India. The data also suggest that there is scope for exploiting the genetic diversity of aromatic and quality rice germplasm of NE India for rice improvement.     

设施用厚皮甜瓜品种SSR标记遗传多样性分析

使用分布于甜瓜12条染色体上的72对SSR引物,对我国中东部设施内栽培的30个厚皮甜瓜品种进行分析;56对SSR引物在30个品种间表现为多态性。共检测到138个等位变异,每对引物的等位变异数变幅为2～6个,平均为2.6个。有效等位变异为86.16个,平均为2.25。每个SSR位点的多态性信息量(PIC)变化范围为0.045～0.725,平均为0.390。30个品种间遗传相似系数变幅为0.274～0.974之间,平均值为0.665,且90.4%的供试品种其遗传相似系数在0.474～0.824之间,亲缘关系较近;以遗传相似系数为原始数据,按UPGMA方法将30个品种划分为3大类群,结合系谱分析结果表明,我国中东部设施适宜种植的甜瓜品种遗传多样性不够丰富,多数品种间的亲缘关系较近,欲进一步提高中东部地区设施甜瓜产量和品质还需要拓宽亲本选择范围,扩大遗传背景。     

Analysis of Genetic Diversity and Population Structure of Rice Germplasm from North-Eastern Region of India and Development of a Core Germplasm Set

The North-Eastern region (NER) of India, comprising of Arunachal Pradesh, Assam, Manipur, Meghalaya, Mizoram, Nagaland and Tripura, is a hot spot for genetic diversity and the most probable origin of rice. North-east rice collections are known to possess various agronomically important traits like biotic and abiotic stress tolerance, unique grain and cooking quality. The genetic diversity and associated population structure of 6,984 rice accessions, originating from NER, were assessed using 36 genome wide unlinked single nucleotide polymorphism (SNP) markers distributed across the 12 rice chromosomes. All of the 36 SNP loci were polymorphic and bi-allelic, contained five types of base substitutions and together produced nine types of alleles. The polymorphic information content (PIC) ranged from 0.004 for Tripura to 0.375 for Manipur and major allele frequency ranged from 0.50 for Assam to 0.99 for Tripura. Heterozygosity ranged from 0.002 in Nagaland to 0.42 in Mizoram and gene diversity ranged from 0.006 in Arunachal Pradesh to 0.50 in Manipur. The genetic relatedness among the rice accessions was evaluated using an unrooted phylogenetic tree analysis, which grouped all accessions into three major clusters. For determining population structure, populations K = 1 to K = 20 were tested and population K = 3 was present in all the states, with the exception of Meghalaya and Manipur where, K = 5 and K = 4 populations were present, respectively. Principal Coordinate Analysis (PCoA) showed that accessions were distributed according to their population structure. AMOVA analysis showed that, maximum diversity was partitioned at the individual accession level (73% for Nagaland, 58% for Arunachal Pradesh and 57% for Tripura). Using POWERCORE software, a core set of 701 accessions was obtained, which accounted for approximately 10% of the total NE India collections, representing 99.9% of the allelic diversity. The rice core set developed will be a valuable resource for future genomic studies and crop improvement strategies.     

Genetic Diversity and Population Structure Analysis in Indian Mustard Germplasm Using Phenotypic Traits and SSR Markers

Plant Molecular Biology Reporter - Indian mustard is an economically important oilseed crop in India; therefore, exploring the genetic diversity of various germplasm collections is quite relevant...     

基于SSR标记的荔枝种质遗传多样性分析

从53对本课题组自主开发设计的SSR特异引物对中筛选出22对多态性引物,对46份荔枝材料的基因组DNA进行扩增,共得到54个等位基因,其中每对引物的平均等位基因数为2.4,共获得23个特异性标记,各标记的平均观察杂合度值、平均期望杂合度和PIC值分别为0.451、0.355和0.507。其中L it6位点各数值最高,是最理想的选择标记。利用22对多态性引物对46份荔枝种质进行聚类分析,构建树状图,结果表明:大多数种质相似系数在0.63～0.95之间,说明它们之间的亲缘关系较近,并发现淮枝(广东)与淮枝(海南)两者可能为同名异物品种。     

东南亚国家引进水稻种质的遗传多样性和遗传结构分析

用72对SSR引物对316份东南亚不同地理来源的水稻种质资源进行遗传多样性和遗传结构分析。结果表明,共检测到387个等位基因,平均每对引物检测到5.375个,变幅在2~12之间;其中频率5%的稀有等位基因有293个,占全部等位基因数的75.7%。Nei基因多样性指数(He)变化幅度在0.055~0.855之间,平均为0.623。He以菲律宾的种质资源最为丰富(He=0.619),其余国家的He大小依次是越南(He=0.515)老挝(He=0.467)柬埔寨(He=0.455);聚类分析显示分为籼粳稻两大群体,地理分组不是特别清晰。AMOVA分析表明,遗传变异主要来源于不同地理类群间,且遗传分化极显著。遗传结构分析结果显示K=2时,有相对明显的遗传结构。其次是K=5时,显示菲律宾群体分为了3个比较明显的小类群,存在较为明显的遗传分化结构。东南亚引进水稻种质资源丰富的遗传多样性和遗传结构为后期水稻育种的亲本选择提供依据。     

Assessment of Genetic Diversity and Population Genetic Structure of Corylus mandshurica in China Using SSR Markers

Corylus mandshurica, also known as pilose hazelnut, is an economically and ecologically important species in China. In this study, ten polymorphic simple sequence repeat (SSR) markers were applied to evaluate the genetic diversity and population structure of 348 C. mandshurica individuals among 12 populations in China. The SSR markers expressed a relatively high level of genetic diversity (Na = 15.3, Ne = 5.6604, I = 1.8853, Ho = 0.6668, and He = 0.7777). According to the coefficient of genetic differentiation (F _st = 0.1215), genetic variation within the populations (87.85%) were remarkably higher than among populations (12.15%). The average gene flow (Nm = 1.8080) significantly impacts the genetic structure of C. mandshurica populations. The relatively high gene flow (Nm = 1.8080) among wild C. mandshurica may be caused by wind-pollinated flowers, highly nutritious seeds and self-incompatible mating system. The UPGMA (unweighted pair group method of arithmetic averages) dendrogram was divided into two main clusters. Moreover, the results of STRUCTURE analysis suggested that C. mandshurica populations fell into two main clusters. Comparison of the UPGMA dendrogram and the Bayesian STRUCTURE analysis showed general agreement between the population subdivisions and the genetic relationships among populations of C. mandshurica. Group I accessions were located in Northeast China, while Group II accessions were in North China. It is worth noting that a number of genetically similar populations were located in the same geographic region. The results further showed that there was obvious genetic differentiation among populations from Northeast China to North China. Results from the Mantel test showed a weak but still significant positive correlation between Nei’s genetic distance and geographic distance (km) among populations (r = 0.419, P = 0.005), suggesting that genetic differentiation in the 12 C. mandshurica populations might be related to geographic distance. These data provide comprehensive information for the development of conservation strategies of these valuable hazelnut resources.     

Evaluation of Genetic Diversity and Population Structure Analysis among Germplasm of Agaricus bisporus by SSR Markers

Agaricus bisporus is a popular edible mushroom that is cultivated worldwide. Due to its secondary homothallic nature, cultivated A. bisporus strains have low genetic diversity, and breeding novel strains is challenging. The aim of this study was to investigate the genetic diversity and population structure of globally collected A. bisporus strains using simple sequence repeat (SSR) markers. Agaricus bisporus strains were divided based on genetic distance-based groups and model-based subpopulations. The major allele frequency (MAF), number of genotypes (NG), number of alleles (NA), observed heterozygosity (HO), expected heterozygosity (HE), and polymorphic information content (PIC) were calculated, and genetic distance, population structure, genetic differentiation, and Hardy–Weinberg equilibrium (HWE) were assessed. Strains were divided into two groups by distance-based analysis and into three subpopulations by model-based analysis. Strains in subpopulations POP A and POP B were included in Group I, and strains in subpopulation POP C were included in Group II. Genetic differentiation between strains was 99%. Marker AB-gSSR-1057 in Group II and subpopulation POP C was confirmed to be in HWE. These results will enhance A. bisporus breeding programs and support the protection of genetic resources.     

308个糯玉米审定品种SSR标记遗传多样性分析

随着中国糯玉米产业的蓬勃发展,每年通过审定的糯玉米品种逐渐增多,为了解其育成品种的遗传多样性和亲缘关系,本研究以308个糯玉米审定品种为材料,利用SSR标记从审定年份和适宜种植区2个角度分析近年来糯玉米审定品种遗传多样性特点和发展趋势.结果 表明,308份糯玉米审定品种在40个SSR标记上共检测出529个等位基因、14...     

基于SSR标记的新疆野杏群体遗传多样性及遗传结构

利用27对SSR分子标记对新疆4个野杏群体遗传多样性和遗传结构进行分析,评价新疆野杏遗传多样性水平和分化程度,为新疆野杏合理保护与利用提供科学依据。结果显示：（1）27对SSR引物共检测到431个等位基因（N_a）,各位点平均等位基因数（N_a）和多态性信息含量（PIC）分别为15.96和0.84;物种水平上Shannons信息指数（I）和期望杂合度（H_e）分别为2.21和0.78。（2）群体水平上等位基因数（N_a）、有效等位基因（N_e）、Shannons信息指数（I）、期望杂合度（H_e）和观察杂合度（H_o）分别为10.98、5.85、1.92、0.79和0.55;其中新源县野杏群体遗传多样性最丰富,巩留县群体遗传多样性最低。（3）基于F统计量分析的遗传分化系数（F_st）为0.05,基因流（N_m）为5.26;分子方差分析显示新疆野杏群体大部分遗传变异来自群体内（95.4%）,群体间的遗传变异仅占4.6%。（4）新疆野杏群体遗传距离为0.06~0.49,平均为0.24;遗传相似度为0.61~0.94,平均为0.80;遗传相似度的聚类分析和遗传距离的主坐标分析结果一致,均将供试4个群体划分为两组;Mantel检测显示,新疆野杏群体遗传距离与地理距离无显著相关（r=0.332,P＝0.16）。研究表明,新疆野杏资源具有丰富的遗传多样性,群体遗传分化程度较低,群体间遗传距离较小,这与新疆野杏群体的大小和悠久的演化历史以及群体间频繁的基因交流相关。     

Genetic Diversity and Population Differentiation of Liaoning Weedy Rice Detected by RAPD and SSR Markers

Weedy rice refers to populations of usually annual Oryza species that diminish farmer income through reduction of grain yield and lowered commodity value at harvest. The genetic diversity and population genetic structure of weedy rice in Liaoning Province were studied by RAPD and SSR markers. The results indicate that the level of genetic diversity of Liaoning weedy rice is very low, with polymorphic loci being only 3.70% (RAPDs) and 47.62% (SSRs). On the other hand, high genetic differentiation was found among populations, in particular between two regions (Shenyang and Dandong), with Fst values of 0.746 (RAPDs) and 0.656 (SSRs), suggesting that more than two thirds of the genetic variation resides among regions. Combined with our investigations of cultural traditions, the low level of genetic diversity in Liaoning Province is attributed to its narrow genetic background enhanced by exchanges of cultivar seeds, whereas the high genetic differentiation between the two regions is most likely the result of different founding parents and gene flow from local rice varieties to weedy rice. The rice cultivars in the two regions are all local varieties and are different genetically. A comparison of the two marker systems demonstrates that SSR is more informative and powerful in terms of the assessment of genetic variability, although both RAPD and SSR provide useful genetic information on weedy rice.     

Genetic Diversity of Global Aromatic Rice Varieties

Genetic diversity of 434 rice accessions collected from 16 countries, including 345 fragrance rice varieties and 89 non fragrance rice varieties, have been analyzed. A total of 573 alleles were detected by using 77 simple sequence repeats (SSR) primer pairs covering all rice 12 chromosomes. The value of allelic polymorphism information content (PIC) ranged from 0090 to 0845, with an average of 0516 per locus; Gene diversity (GD) varied from 0091 to 0859, with an average of 0573; The mean value of major allele frequencies (MAF) was 0540, covering from 0251 to 0953. In addition, 434 rice accessions are divided into three sub populations by cluster and population structure analysis, and FST between sub populations showed a mean of -0116, ranging from -0623 to 0494; The score of genetic distance calculated by Nei′s method appeared from 0207 to 0355. Major allele frequencies within each sub population distributed from 0408 to 0746, and gene diversity level from 0354 to 0699, while PIC from 0320 to 0658. Sequencing 6 mitochondrion genes in 18 rice varieties exhibited no different in 5 genes, whereas Mit4 contains a 3 SNPs in the gene body, which acts as an important marker to understanding the relationship between Indica/Japonica differentiation and the evolution of fragrant gene. Finally, genetic diversity and mitochondrion gene sequencing would help to know about the origin of fragrant resource and benefit rice breeding.     

Analysis of Genetic Diversity and Population Structure in Sophora japonica Linn. in China With Newly Developed SSR Markers

Sophora japonica is a medium-sized deciduous native tree to China. Its flower buds and fruits have special medicinal value. By using 26 highly variable nuclear microsatellites, newly reported here, we assessed the genetic diversity and population structure of 10 representative populations (180 individuals in total) within the species. The results revealed a relatively high genetic diversity in S. japonica (N_a?=?8.00, H_e?=?0.74, PIC?=?0.80, I?=?1.67), and reduced genetic variation and heterozygote excess were detected in landrace populations (N_a?=?5.59, H_e?=?0.71, A_r?=?4.11, F_IS?=???0.043) when compared with those semi-wild population (N_a?=?9.7, H_e?=?0.74, A_r?=?8.80, F_IS?=?0.082). A significant bottleneck was detected in two Landraces populations. Moderate differentiation and frequent gene flow were detected among all populations (F_ST?=?0.079, P?<?0.05, N_m?=?2.893). Mantel test detected a marginal significant pattern of isolation by distance (r?=?0.009, P?=?0.50), and strong differentiation was observed between most southern and northern populations both by the STRUCTURE clustering and principal coordinate analysis (PCoA), indicating that geographic isolation played a key role in the genetic differentiation of the species. The results from this study will benefit the breeding and conservation of S. japonica, other congener species, and tree species with similar life history traits.

     

中国水稻地方品种与选育品种遗传多样性的比较分析

利用98对SSR标记对202份中国水稻地方品种和选育品种的遗传多样性进行比较分析.结果显示供试品种具有较丰富的遗传多样性,共检测到等位基因1350个,每个位点的等位基因数(Na)变化范围为3～ 39,平均14个;Nei基因多样性指数变化范围(He)为0.125 ～0.955,平均0.733;多态信息量(PIC)变化范围为0.122 ～0.953,平均0.680;稀有等位基因数(Nr)913个;等位基因丰度(Rs)8.33.栽培稻地方品种和选育品种遗传多样性差异明显,地方品种等位基因数、Nei基因多样性指数、多态信息量、稀有等位基因数和等位基因丰度(Na=1219,He=0.747,PIC=0.710,Nr=756,Rs =8.50)均高于选育品种(Na =919,He =0.704,PIC =0.650,Nr=529,Rs =7.01).各染色体组水平的遗传多样性分析表明,选育品种仅在1号染色体上的遗传多样性高于地方品种,进一步分析显示选育品种的遗传改良在基因组水平上具有区间特异性.