DNA profiling and genetic diversity of Korean soybean (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill) landraces by SSR markers

Approximately 7,000 accessions of Korean soybean (Glycine max (L.) Merrill) landraces, largely composed of three collections, the Korea Atomic Energy Research Institute’s soybean (KAS), the Korean Crop Experiment Station’s soybean (KLS) and the Korean Agricultural Development and Technology Center’s soybean (KADTC) collections, have been conserved at the Rural Development Administration (RDA) genebank in Korea. The accessions within collections were classified based on their traditional uses such as sauce soybean (SA), sprouted soybean (SP), soybean for cooking with rice (SCR), and OTHERS. A total of 2,758 accessions of Korean soybean landraces were used to profile and to evaluate genetic structure using six SSR loci. A total of 110 alleles were revealed by at the six SSR loci. The number of alleles per SSR locus ranged from 9 to 39 in Satt187 and Satt_074, respectively. The number of alleles ranged from 87 in the KADTC collection to 96 in the KLS collection, and from 63 in the SCR group to 95 in the SP group. Nei’s average genetic diversity ranged from 0.68 to 0.70 across three collections, and 0.64 to 0.69 across the usage groups. The average between-group differentiation (G _st) was 0.9 among collections, and 4.1 among the usage groups. The similar average diversity among three collections implies that the genetic background of the three collections was quite similar or that there were a large number of duplicate accessions in three collections. The selection from the four groups classified based upon usage may be a useful way to select accessions for developing a Korean soybean landrace core collection at the RDA genebank. DNA profile information of accessions will provide indications of redundancies or omissions and aid in managing the soybean collection held at the RDA genebank. The information on diversity analysis could help to enlarge the genetic diversity of materials in breeding programs and could be used to develop a core collection.     

Molecular diversity and genome-wide linkage disequilibrium patterns in a worldwide collection of <Emphasis Type="Italic">Oryza sativa</Emphasis> and its wild relatives

Genetic diversity analysis within a species is vital for understanding evolutionary processes at the population and genomic levels. We report a detailed study of molecular diversity, polymorphism and linkage disequilibrium in three groups of rice (Oryza) germplasm accessions based on 176 SSR markers. The first group included 65 rice (O. sativa L.) accessions introduced from seven countries, including five regions of China. The second group included 58 US rice varieties released in the past 25 years. The third group consisted of 54 accessions of rice wild relatives represented by ten different species. The number of alleles per SSR marker ranged from 4 to 32 with a mean of 16 alleles and the polymorphism information content values ranged from 0.43 to 0.91 with a mean of 0.70. The variation in SSR alleles was a significant contribution to the genetic discrimination of the 177 accessions within the three Oryza groups. Analysis of molecular variance identified deviation from Hardy–Weinberg equilibrium. Principal coordinates analysis clearly separated the accessions into their respective three groups. Neighbor-joining phylogenetic cluster reflects the ordination of each accession. Linkage disequilibrium (D′) averaged 0.75 in wild Oryza spp., and about 0.5 in both US and international O. sativa accessions. Our results showed that LD among adjacent loci in both O. sativa and Oryza spp. accessions is strong enough to be detecting marker-trait association via genome-wide scans.     

用于中国小豆种质资源遗传多样性分析SSR分子标记筛选及应用

以375份小豆核心种质为试验材料, 利用从小豆及其近缘种SSR引物中筛选出的13对引物进行遗传多样性分析。检测结果显示, 小豆种质资源具有丰富的遗传多样性, 共检测到133个等位变异, 每对SSR引物检测到等位变异4~19个, 平均10.23个, 国内各省多态信息含量(PIC)平均为0.561, 多态位点比例(P)平均为93.523%。聚类结果表明, 小豆资源遗传关系与生态分区间有明显的联系, 且东北地区资源与中南部资源遗传关系较近。湖北、安徽、陕西3省资源的PIC较高, 且基本位于主坐标三维图的中心区域, 推断湖北、安徽、陕西是中国栽培小豆的起源地或多样性中心。该结果有助于更好地对小豆种质资源进行收集、保护和利用。     

陕西省野生大豆种质资源的SSR遗传多样性研究

为了研究陕西地区野生大豆的遗传多样性特点,利用SSR分子标记分析了陕西省6个野生大豆(Glycine soja)天然种群和1个栽培大豆(Glycine max)种群的遗传结构与遗传多样性。结果显示：13个位点共检测出113个等位基因,平均每个位点的等位基因数(A)为8.69个,等位基因数目范围为4~13个,有效等位基因数(N_e)范围为2.135(Satt590)~9.385 (Satt487),平均有效等因基因数为5.623;观察杂合度(H_o)变化范围为0.033~0.121,平均为0.080;预期杂合度(H_e)的变化范围为0.312~0.658,平均为0.482;种群平均Shannon遗传多样性指数(I)为0.657;野生大豆种群基因多样度比率(F_ST)为0.465。该研究显示,陕西省野生大豆具有较高水平的遗传多样性,野生大豆的遗传多样性普遍高于栽培大豆;随着海拔的不断升高,野生大豆遗传多样性变低;陕西中部、南部的野生大豆种质资源丰富、种群具有较高的遗传多样性,推测该区域为陕西省野生大豆的遗传多样性中心。     

Analysis of population structure and genetic diversity in balloon flower (Platycodon grandiflorum (Jacq.) A. DC.) germplasm using Simple Sequence Repeat (SSR) markers

Balloon flower (Platycodon grandiflorum A. DC) is widely distributed in South Korea and there are some local landraces that are cultivated as a vegetable crop or medicinal plant. Making use of the gene resources of wild-type and landraces is a way to increase the genetic diversity of the cultivars. However, few tools or information are available on an efficient identification system for maintaining and management of these landraces. To improve the genetic resources for balloon flower, 22 simple sequence repeat (SSR) markers, also known as microsatellite markers, were evaluated in a collection of 42 balloon flower landraces, 34 of which were from Korea and eight from China. All microsatellite markers produced the 107 alleles ranging from 2 to 10 with a mean of 4.864 alleles per each locus (N_A). The values of observed heterozygosity (H_O) and expected heterozygosity (H_E) ranged from 0.00 to 0.667 (mean of 0.285) and from 0.024 to 0.741 (mean of 0.416), respectively. An average value of polymorphic information contents (PIC) were 0.382 with a range of 0.023 to 0.703. Results of population structure and phylogenetic and principal coordinate analysis (PCoA) indicated that P. grandiflorum germplasm formed two largely distinct clusters according to their origins and the genetic differentiation. There was a high level of genotypic diversity at broad geographic regions between Korea and China, but the low genetic differentiation was found within the collections from Korea. The results of the genetic diversity will be useful for the selection of the parents for developing balloon flower breeding and the multi-locus SSR markers developed herein will be a valuable resource for germplasm assessments, evaluation of genetic diversity, and population genetic studies of balloon flower.     

绿豆SSR标记的开发及遗传多样性分析

SSR标记以其数量丰富、多态性好、共显性遗传等优点在基础研究和育种工作中发挥了重要作用,但目前绿豆基因组中的SSR标记依然较少。本研究将磁珠富集法和测序技术相结合高通量检测绿豆基因组SSR位点,鉴定出3,275,355个SSR位点,开发了2742个SSR标记。选取其中157个SSR进行PCR验证,发现有90个(57.33%)标记在10份材料中表现出多态性。挑选40个条带清晰、多态性高、染色体上均匀分布的标记对90份绿豆资源进行遗传多样性分析,单个位点检测到的等位变异数为2~8个,平均为3.0个,有效等位基因数为1.31~4.21个,平均为2.16。Nei’s基因多样性指数在0.23~0.76之间,平均为0.51。多态性信息含量为0.22~0.72,平均为0.43。聚类分析将90份材料分为2个类群,包含4个组。第I组主要由北方资源组成,第Ⅱ组种质来源较为分散,第Ⅲ组主要由山东的资源构成,第Ⅳ组包含多数河北的种质资源。本研究开发的多态性SSR标记不仅可以用于绿豆种质资源的遗传多样性分析,也将在高密度遗传图谱构建、基因定位和分子标记辅助育种中发挥重要作用。     

Genetic diversity and differentiation of Chinese wild soybean germplasm (G. soja Sieb. & Zucc.) in geographical scale revealed by SSR markers

Wild soybean (Glycine soja), as the progenitor of soybeans (G. max), is widely distributed in China and has been collected as a supplementary germplasm pool of soybeans. In this study, 375 wild soybean accessions from a set of genebank core collection were analysed for genetic diversity by using 42 simple sequence repeat primer pairs. The mean allele number per locus was 19.62. Ten‐percent unique alleles involving 35 or 83.33% loci differentiated among the geographical regions. The mean gene diversity (h) per locus was 0.89. A very low mean coefficient of gene differentiation (G_ST = 0.08) for geographical regions and a high mean within‐region gene diversity (H_S = 0.81) were observed, indicating that most genetic diversity existed within the regions. There was an obvious relationship between genetic distance and geographical distance. The results showed multiple centers of genetic diversity for Chinese wild soybean in North China, the Huanghe River Valley, and Central China as well as the Changjiang River Valley, implicating multiple site origins of soybeans within China.     

Density enhancement of a faba bean genetic linkage map (Vicia faba) based on simple sequence repeats markers

Genetic mapping for faba bean lags far behind other major crops. Density enhancement of the faba bean genetic linkage map was carried out by screening 5,325 genomic SSR primers and 2033 expressed sequence tag (EST)‐SSR primers on the parental cultivars '91825' and 'K1563'. Two hundred and fifteen genomic SSR and 133 EST‐SSR primer pairs that detected polymorphisms in the parents were used to screen 129 F₂ individuals. This study added 337 more SSR markers and extended the previous linkage map by 2928.45 cM to a total of 4516.75 cM. The number of SSR markers in the linkage groups varied from 12 to 136 while the length of each linkage group ranged from 129.35 to 1180.21 cM. The average distance between adjacent loci in the enhanced genetic linkage map was 9.71 cM, which is 2.79 cM shorter than the first linkage map of faba bean. The density‐enhanced genetic map of faba bean will be useful for marker‐assisted selection and breeding in this important legume crop.     

Microsatellite (SSR) variation in a collection of Malus (apple) species and hybrids

A collection of 142 accessions of 23 Malus species, derived hybrids and cultivar accessions from the USDA-ARS Plant Genetic Resources Unit's core collection, which represents an extensive range of Malus species, was screened with a set of previously described SSR (simple sequence repeat) markers. The markers were used to determine genetic identities, estimate genetic diversity, identify genetic relationships among the accessions, and determine the utility of SSR primers developed from Malus ×domestica for making genetic assessments across the whole Malus genus. All eight primer pairs amplified multiple fragments when used in polymerase chain reactions with DNA from these accessions. High levels of variation were detected with a mean of 26.4 alleles per locus and a mean direct count heterozygosity across all eight loci equal to 0.623. The eight primer pairs used in this study unambiguously differentiated all but five pairs of accessions in this collection of 142 accessions of 23 Malus species, derived hybrids and cultivars. These SSR data were not useful in identifying genetic relationships among this diverse collection of accessions, with the majority of the accessions not clustering in ways concordant with taxonomic information and/or geographic origin. The resulting phenogram resolved only two meaningful clusters, for the taxonomically isolated Section Chloromeles and for M. fusca accessions, reflecting genetic relationships arising from geographic origin. The detection of identical accessions in the collection, which were previously considered to be unique, highlights the critical need to further bolster collections of certain Malus species. This revised version was published online in July 2006 with corrections to the Cover Date.     

小豆遗传差异、群体结构和连锁不平衡水平的SSR分析

利用57对小豆SSR标记和31对绿豆SSR标记,用5份日本材料作对照,对249份中国小豆种质进行遗传差异、群体结构和连锁不平衡(LD)分析。结果表明,共检测到630个等位变异,SSR位点等位变异数在2~17之间,遗传多样性指数范围为0.024~0.898,平均为0.574。15个不同地理来源群体间表现出显著的遗传多样性差异,其中中国云南最高,河北、天津最低。聚类分析将254份材料划分为3个类群,在一定程度上和地理生态环境相关。LD分析显示和其他作物相比,小豆LD衰减距离较短,最大衰减距离为5.8 cM (R²>0.1),基因组LD平均衰减距离小于1 cM (R²>0.1,P<0.001)。     

Molecular diversity and population structure of the Ethiopian lentil (<Emphasis Type="Italic">Lens Culinaris</Emphasis> Medikus) genotype assessment using SSR markers

Knowledge of genetic diversity in germplasm is essential for formulating effective germplasm collection, conservation, utilization strategies in and crop improvement programs. It also provides an opportunity to take corrective steps infusing new genes to avoid risks associated with a narrow genetic bases. Genetic diversity analysis of 119 lentil genotypes of including 83 germplasm and 36 exotic genotypes from International Center for Agricultural Research in the Dry Areas was studied using 27 primers of simple sequence repeat (SSR) marker. Molecular analysis of variance showed variations of 82% within and 18% of the among population variance was explained. Degree of polymorphism observed among the populations was 100%. A total 122 alleles were detected, with 2 to 7 alleles per locus, with a mean of 4.52 alleles per locus. The estimated gene diversity value for 27 loci was 0.64. The average Shannon’s information index value of 1.19 was obtained showed the existence of high genetic variation within the genotypes. The genetic similarity indices ranged from 0.21 to 1.00. The SSR markers showed an average polymorphic information content (PIC) value of 0.58. Cluster analysis grouped the genotypes into five major clusters as distinct genetic populations. Diversity analyses revealed the existence of a high level of genetic variation among genotypes. This molecular diversity information provides a basis for future germplasm collection, utilization, and conservation strategies in gene banks and introducing exotic germplasm to widen the genetic base of the current lentil breeding population.     

Genetic linkage map of Chinese native variety faba bean (Vicia faba L.) based on simple sequence repeat markers

Simple sequence repeat (SSR) marker is a powerful tool for construction of genetic linkage map which can be applied for quantitative trait loci (QTL) and marker‐assisted selection (MAS). In this study, a genetic map of faba bean was constructed with SSR markers using a 129 F₂ individuals population derived from the cross of Chinese native variety 91825 (large seed) and K1563 (small seed). By screening 11 551 SSR primers between two parents, 149 primer pairs were detected polymorphic and used for F₂ population analysis. This SSR‐based genetic linkage map consisted of 15 linkage groups with 128 SSR. The map encompassed 1587 cM with an average genetic distance of 12.4 cM. The genetic map generated in this study will be beneficial for genetic studies of faba bean for identification of marker‐locus‐trait associations as well as comparative mapping among faba bean, pea and grasspea.     

Cross‐genera amplification of informative microsatellite markers from common bean and scarlet runner bean for assessment of genetic diversity in mungbean (Vigna radiata)

Mungbean (V. radiata) is an important Asiatic legume supplying inexpensive protein to a vast majority of vegetarian masses. To increase markers repertoire in mungbean, a study was conducted to analyse 384 microsatellite markers derived from common bean, scarlet runner bean and adzuki bean for their transferability and polymorphism. The results showed that 87 (24.71%) primer pairs could amplify DNA loci of 20 mungbean genotypes including one accession of V. trilobata, while 52 showed reliable banding and polymorphism. These showed different degrees of variability at each locus producing 250 alleles with the number of alleles varying from 2 to 9. The major allele frequency varied from 0.17 to 0.95, while the polymorphic information content of SSRs ranged between 0.09 and 0.86 with an average of 0.60 ± 0.16. UPGMA revealed three major clusters accommodating ~95% of the accessions while one accession of V. trilobata (‘NSB‐007’) did not group with any other genotype describing the discriminating power of informative microsatellites. This study identified a set of useful microsatellite markers to accelerate the genetic studies and breeding programme of mungbean.     

国内外蚕豆核心种质SSR遗传多样性对比及微核心种质构建

运用24对SSR引物, 对国内外1075份初级地理蚕豆核心种质的遗传多样性分析显示, 等位变异数、有效等位变异数及Shannon’s信息指数分别为8.54、2.26和1.02;对全部参试资源进行聚类分析, 没有发现明显的群体结构, 表明初级地理核心种质的代表性较好, 遗传背景较广泛。之后采用每个类内随机抽样的方法构建含有129份国内资源和63份国外资源的蚕豆微核心种质, 等位基因变异数、有效等位变异数和Shannon’s信息指数的保留比例分别为87.32%, 101.26%, 101.82%;经t检验得出微核心种质与全部参试资源群体间遗传多样性差异不显著, 表明构建的微核心种质的遗传多样性可以代表初级地理蚕豆核心种质。     

Simple sequence repeats (SSRs) in faba bean: new loci from Orobanche-resistant cultivar 'Giza 402'

Genomic DNA from an inbred line from the Orobanche crenata resistant cultivar 'Giza 402' was restricted and enriched for AG-containing simple sequence repeats (SSRs). The overall SSR enrichment rate was 54.6%. Of 73 selected SSR markers that successfully amplified in 'Giza 402' 54 were polymorphic among a group of 10 Egyptian and Spanish faba bean genotypes extensively used in Orobanche crenata resistance/susceptibility studies and linkage map development. The polymorphism information content ranged from 0.16 to 0.72 and Unweighted Pair-Group Method using Arithmetic averages cluster analysis based on these loci placed all resistant genotypes descended from the parental line F 402 in the same group. Successful amplification from 28 primer pairs was also observed in five European faba bean genotypes. The SSR loci described here will be extremely useful in studies focusing on Orobanche crenata resistance, genetic diversity studies and will provide additional co-dominant loci to complement existing faba bean linkage maps based primarily on dominant markers.     

普通菜豆根系相关性状的关联分析

幼苗期根系发育对作物的生长发育具有重要作用。利用生长袋纸培系统对324份普通菜豆种质的主根长、根干重、根体积、根表面积等9个根系相关性状进行表型鉴定,并结合覆盖全基因组、有多态性的116对SSR标记,利用MLM（Q+K）模型进行表型和标记的关联分析。表型分析表明,324份材料的9个根系相关性状表型变异丰富,平均变异系数的变动范围是10.09%~37.03%;基因型分析表明,116个多态性SSR标记共检测到919个等位变异位点,每个标记的平均基因多样性指数为0.59,多态性信息含量（PIC）平均值为0.54,显示这些标记具有较高的基因多样性;群体结构分析表明,供试材料分为两个亚群,与普通菜豆起源于两个基因库对应;关联分析结果显示,以P<0.01作为显著条件,共检测到48个显著标记位点,其中有10个位点同时与2个以上性状相关联,有5个位点与前人研究结果一致。研究结果为进一步理解普通菜豆根系的遗传机理提供了理论参考,也为分子标记辅助选择改良普通菜豆根系奠定了基础。     

Genetic effects of introgression genomic components from Sea Island cotton (<Emphasis Type="Italic">Gossypium barbadense</Emphasis> L.) on fiber related traits in upland cotton (<Emphasis Type="Italic">G</Emphasis>. <Emphasis Type="Italic">hirsutum</Emphasis> L.)

The germplasm with exotic genomic components especially from Sea Island cotton (Gossypium barbadense L. Gb) is the dominant genetic resources to enhance fiber quality of upland cotton (G. hirsutum L., Gh). Due to low efficiency of phenotypic evaluation and selection on fiber quality, genetic dissection of favorable alleles using molecular markers is essential. Genetic dissection on putative Gb introgressions related to fiber traits were conducted by SSR markers with mapping populations derived from a cross between Luyuan343 (LY343), a superior fiber quality introgression line (IL) with genomic components from Gb, and an elite Upland cotton cv. Lumianyan#22 (LMY22). Among 82 polymorphic loci screened out from 4050 SSRs, 42 were identified as putative introgression alleles. A total of 29 fiber-related QTLs (23 for fiber quality and six for lint percentage) were detected and most of which clustered on the putative Gb introgression chromosomal segments of Chr.2, Chr.16, Chr.23 and Chr.25. As expected, a majority of favorable alleles of fiber quality QTLs (12/17, not considering the QTLs for fiber fineness) came from the IL parent and most of which (11/12) were conferred by the introgression genomic components while three of the six (3/6) favorable alleles for lint percentage came from the Gh parent. Validation of these QTLs using an F₈ breeding population from the same cross made previously indicated that 13 out of 29 QTLs showed considerable stability. The results suggest that fiber quality improvement using the introgression components could be facilitated by marker-assisted selection in cotton breeding program.     

Transferability of rice SSR markers to Miscanthus sinensis, a potential biofuel crop

Miscanthus sinensis (M. sinensis) is a perennial C₄ photosynthesis grass, with high yield, high efficiency of water usage, low fertilizer requirement, tolerance to extreme environments, and is one of the plant species with good biofuel potential. Simple sequence repeat (SSR) markers are highly informative and widely used in plant genetic studies. In this study, 88 SSR primer pairs derived from the rice genome, including 47 EST-SSRs (eSSRs) and 41 genomic SSRs (gSSRs), were evaluated for cross-species transferability to M. sinensis. Forty-one SSR primer pairs in total could successfully amplify DNA fragments in M. sinensis, showing an overall transferability rate of 46.6 % between rice and M. sinensis. The transferability rate of eSSR (51.1 %) was higher than that of gSSR (41.5 %). A total of 140 SSR loci and 340 alleles in the set of rice and M. sinensis germplasm collections were detected. Nei’s gene diversity varied from 0 to 0.72 and averaged 0.35. Shannon’s information index varied from 0 to 1.49 and averaged 0.56. The observed heterozygosity ranged from 0 to 0.95 and averaged 0.08. Thirty-nine loci (27.86 %) were shown heterozygosity out of 140 SSR loci. A dendrogram based on genetic distance showed a significant geographic differentiation. Our results indicated that 46.6% of the rice SSR markers are transferable to M. sinensis, and are useful for germplasm evaluation and genetic analysis in M. sinensis.     

Genetic diversity of natural populations of Atriplex halimus L. in Morocco: An isoenzyme-based overview

Based on polyacrylamide gel electrophoresis, nine natural populations of Atriplex halimus L., a perennial shrub, collected in different regions of Morocco, were studied for their genetic variation using isoenzyme polymorphism of the highly active enzyme systems: esterases (EST), acid phosphatases (ACP) and glutamate oxaloacetate transaminase (GOT). Different allozyme frequencies from 7 different loci were obtained for all populations of this halophyte species. High levels of genetic diversity were revealed. The mean number of alleles per locus (A = 1.9–2.0), the percentage of polymorphic loci (p = 71.4–85.7) and the mean expected heterozygosity (H_e = 0.339–0.385) showed an important variability in all populations. Gene diversity was essentially explained by the within population component. The between populations differentiation accounted for 8% of the whole diversity (F_ST, averaged over all loci, is 0.08). The relationships among the 9 populations were inferred from the Nei’s genetic distances. Four major groups were formed. The northern population ‘Tanger’, forming a unique group, was highly divergent from the other groups. It appeared that the genetic distance between all groups was related to the geographic distance that separates them. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characterization of novel sugarcane expressed sequence tag microsatellites and their comparison with genomic SSRs

Microsatellites or simple sequence repeats (SSRs) are one of the most suitable markers for genome analysis as they have great potential to aid breeders to develop new improved sugarcane varieties. The development of SSR derived from expressed sequence tags (EST) opens new opportunities for genetic investigations at a functional level. In the present work, the polymorphism obtained with a subset of 51 EST–SSRs derived from sucest was compared with those generated by 50 genomic SSRs (gSSR) in terms of number of alleles, polymorphism information content, discrimination power and their ability to establish genetic relationships among 18 sugarcane clones including three Saccharum species (S. officinarum, S. barberi, S. sinense). The majority of EST–SSRs loci had four to six alleles in contrast to the seven to nine observed for the gSSRs loci. Approximately, 35% of the gSSRs had PIC values around 0.90 in contrast to 15% of the EST–SSRs. However, the mean discrimination power of the two types of SSR did not differ significantly as much as the average genetic similarity (GS) based on Dice coefficient. The correlation between GS of the two types of SSRs was high (r = 0.71/P = 0.99) and significant. Although differences were observed between dendrograms obtained with each SSR type, both were in good agreement with pedigree information. The S. officinarum clone IJ76‐314 was grouped apart from the other clones evaluated. The results here demonstrate that EST–SSRs can be successfully used for genetic relationship analysis, extending the knowledge of genetic diversity of sugarcane to a functional level.