“十五”大豆创新种质和1968—1995年间育成品种的SSR遗传结构及遗传多样性分析

对22份“十五”攻关培育的创新种质和22份大豆育成品种进行了24个SSR标记的分析比较,目的是在分子水平上阐明创新种质的遗传结构特点,为拓宽我国大豆育成品种遗传基础及亲本选择提供理论依据。本研究在24个SSR位点共发现231个等位变异,其中15.8%（36个等位变异）为创新种质所特有,特别是在与大豆胞囊线虫紧密连锁的Satt309位点上验证了一个我国独有的等位变异。结合UPGMA和Model-based聚类结果,将创新种质和育成品种分为4组,第Ⅰ组由13份来自东北和山西的创新种质组成;第Ⅱ组由8份来自东北的育成品种组成;第Ⅲ组由8份来自黄淮海和南方的大豆种质组成,其中创新种质和育成品种各为4份;第Ⅳ组由4份育成品种组成,分别来自吉林、黑龙江、河南和山西。遗传多样性分析结果表明,利用国外种质和野生大豆创造的创新种质丰富了东北地区育成品种的遗传多样性。因此,应加强利用国外种质、我国栽培大豆地方品种和野生大豆等优异资源,在创造优异大豆新种质的同时,拓宽我国大豆的遗传基础。     

国外种质对中国大豆育成品种遗传贡献的分子证据

用SSR标记对32份中国大豆品种与40份国外引进大豆育成品种祖先亲本的遗传多样性进行分析,以明确引进国外大豆种质对中国大豆育种的遗传贡献。结果表明,在22个SSR位点共检测到170个等位变异,中国大豆和引进国外大豆平均等位变异数分别为6.0和6.9个,遗传多样性指数都为0.71,国外品种中检测到48个特有等位变异,而中国大豆中仅检测到22个,且共有等位变异在中外大豆中的分布频率差异较大。聚类分析也发现中国育成品种与国外引进大豆存在较大差异。遗传组成分析发现,Amsoy和十胜长叶2个国外种质的引入使5个中国大豆育成品种增加了23个国外种质特有等位变异;其在育成品种中的保留比例为29.13%,但不同遗传背景中保留的等位变异不同,说明国外种质在中国大豆育种中起着重要作用,而且仍有很多特有等位变异没有被利用,可以继续作为亲本在中国大豆改良中发挥作用。     

"十五"大豆创新种质和1963-1995年间育成品种的SSR遗传结构及遗传多样性分析

对22份"十五"攻关培育的创新种质和22份大豆育成品种进行了24个SSR标记的分析比较,目的是在分子水平上阐明创新种质的遗传结构特点,为拓宽我国大豆育成品种遗传基础及亲本选择提供理论依据。本研究在24个SSR位点共检测出231个等位变异,其中15.8%(36个等位变异)为创新种质所特有,特别是在与大豆胞囊线虫紧密连锁的Satt309位点上验证了一个我国独有的等位变异。结合UPGMA和Model-based聚类结果,将创新种质和育成品种分为4组,第Ⅰ组由13份来自东北和山西的创新种质组成;第Ⅱ组由8份来自东北的育成品种组成;第Ⅲ组由8份来自黄淮海和南方的大豆种质组成,其中创新种质和育成品种各为4份;第Ⅳ组由4份育成品种组成,分别来自吉林、黑龙江、河南和山西。遗传多样性分析结果表明,利用国外种质和野生大豆创造的创新种质丰富了东北地区育成品种的遗传多样性。因此,应加强利用国外种质、我国栽培大豆地方品种和野生大豆等优异资源,在创造优异大豆新种质的同时,拓宽我国大豆的遗传基础。     

分析中国栽培大豆遗传多样性所需SSR引物的数目

我国拥有极其丰富的大豆资源。传统的方法是根据农艺性状来分析其遗传变异，但农艺性状受自然环境和人为因素影响明显。随着大豆育成品种的增加，有限的表型变异已难以详细阐明我国2万余份大豆品种的遗传变异情况，需要从DNA分子水平深入研究我国大豆资源遗传变异分布规律。本研究以190份为大豆为初选核心种质的一个无偏样本。用60对SSR引物扩增，获得606个等位变异，平均每个位点有10个等位变异。位点多态信息量范围从0．55到0．99，平均为0．83。对190份大豆相似系数矩阵的标准误分析表明。SSR引物数增加到50左右时。再增加引物，标准误变化很小。共表型矩阵之间的相关性测验显示，当等位变异数达到570以上，相互之间相关性极显著。从实验材料中选取东北春大豆类型作为一个小样本进行共表型矩阵相关性分析也有类似结果。用SSR方法分析中国栽培大豆(G．max)遗传变异关系时，只有等位变异数达到一定的范围时，才能真实地反映出品种之间的遗传变异关系。当群体的遗传变异范围变得相对较小时。分析个体之间的遗传变异关系所需的等位变异数目也相应降低。结合SSR位点在大豆基因组中的分布和基因多样性水平。能够找到分析栽培大豆遗传多样性的核心SSR引物。只有获得等位变异数在570以上。才能客观地反映出中国栽培大豆遗传变异关系。     

发掘大豆资源中灰斑病1号生理小种抗性优异等位变异

通过发掘大豆资源中抗灰斑病1号生理小种的优异等位变异和载体材料,为开展抗灰斑病品种分子设计育种提供理论基础。以205份大豆资源构建的自然群体为试验材料,对其进行灰斑病1号生理小种的抗性鉴定;利用117对SSR标记进行全基因组扫描,分析群体的遗传多样性和群体结构,应用GLM和MLM程序对标记与大豆灰斑病抗性开展关联分析。结果表明：205份大豆资源对灰斑病1号生理小种抗性遗传变异系数为20.90%;2个模型共检测到与灰斑病1号生理小种抗性关联的位点7个,表型变异解释率在7.58%~16.06%;发掘到增效等位变异36个,其中效应值较大的等位变异为Satt244-230（26.16）、Satt142-154（21.94）和Satt244-186（20.19）,携带上述3个等位变异的载体材料均为野生资源,3个典型材料分别为12C8646、12C8670和12C6175;育成品种中具有最大增效值的等位变异为Satt142-189（8.94）,有7个品种携带该等位片段,均为黑龙江品种,典型载体材料为东农43。 上述信息可用于分子标记辅助选择育种和抗源筛选。     

贵州省部分大豆种质资源SSR遗传多样性分析

利用微卫星分子标记(SSR)对来自贵州省32个县(市)的115份大豆地方种质资源的遗传多样性进行了研究.结果表明,参试的115份大豆种质在8个位点共检测到56个等位变异,等位变异数在5～9个之间,平均每个位点的等位变异数为7个.聚类分析表明,115份大豆种质资源间的遗传距离变异范围为0.07～0.58,可将其分为11个类型,并发现其中5个材料与其它种质有明显差异.通过本研究为进一步开展贵州省大豆种质资源的遗传多样性研究提供了参考.     

基于分子和表型性状的大豆骨干品种遗传多样性分析

绥农14是集优质、高产、抗病、广适应性于一身的大豆优良品种,对绥农14系谱亲本进行分子和表型的遗传基础解析,为有目的地选择杂交亲本拓宽遗传基础提供理论指导.利用包含有生长性状、产量性状、品质性状、抗逆性状、固氮性状在内的50个表型性状和550个微卫星位点对绥农14系谱亲本进行分析.550个SSR位点共检测出等位变异1 494个,平均每个SSR位点的等位变异为2.716 4,平均PIC值为0.445 0,其中30个多态性高的位点可作为评价大豆种质资源遗传多样性的重要位点;连锁群CI的多态性位点比例最高为0.961 5,连锁群A2的保守片断最多为11个,构建绥农14系谱亲本的指纹图谱最少需2个位点.50个表型性状共检测到等位变异255个,每个位点的平均等位变异为5.1个,平均PIC值为0.683 2,主成分分析结果表明,6个主成分的累积贡献率在80.1％以上,分析每个主成分的组成发现,产量性状、生长性状,品质性状、抗逆性状、固氮性状在分析大豆种质资源遗传多样性时均具有重要的作用,进行每一类性状的主成分分析,选出重要性状作为大豆综合性状考察的主要指标.基于SSR的UPGMA聚类结果与基于农艺数据的UPGMA聚类结果的相关系数仅为0.393 0,2种聚类方法都只能在一定程度上揭示品种间的亲缘关系,因此,在进行种质资源遗传多样性研究时应将分子数据分析与表型性状解析相结合.     

黑龙江省主栽大豆品种遗传多样性和群体结构分析

利用187对SSR标记对近25年（1992-2017）在黑龙江省栽培的202个大豆品种进行遗传多样性和群体结构分析。结果表明,从试验材料的基因组DNA中扩增出多态性位点808个,平均每对引物扩增出多态性位点4.42个;多态性位点最多的引物是satt703和satt311,均为10个;等位变异频率最高的引物是satt417和satt575,等位变异频率均为99.5%。供试品种间的遗传相似系数为0.283~0.930,平均值为0.519。同一个育种单位育成的部分品种具有较高的遗传相似性。群体结构、主坐标分析和NJ聚类将202个品种划分的结果是一致的,均为3个类群。类群中的部分材料血缘不是独立的,而是相互渗透的。     

油菜自育与其他主栽品种的遗传多样性和遗传关系分析

利用SSR标记研究甘蓝型油菜自育杂交种(陕西省杂交油菜研究中心培育的品种及杂交组合)与其他主栽品种的遗传多样性和遗传关系,为种质创新、材料选择和育种设计提供参考。47对引物在62份样品中检测到50个多态性位点,23份自育杂交种及39份其他主栽品种的遗传多样性为中度丰富,自育杂交种等位变异数、基因型数、观察杂合度、期望杂合度及多态性信息含量等值相对较低;69%的杂交品种杂合率大于40%;发现其他主栽品种对自育杂交种有补充等位变异的位点10个、含重要高频率等位变异的位点14个。遗传相似系数(GS)分析表明自育杂交种内部总体为中高度相似性(0.7GS0.8),其他主栽品种内部及两组之间为中度相似性(0.6GS0.7)。主坐标分析和遗传分化系数显示两组间有明显遗传分化。系统聚类在GS值0.66处将全部样品主要分为两类,自育杂交种、黄淮区及长江中游区品种总数96%在第一类,第二类主要为长江上游区和下游区品种。自育杂交种的遗传多样性相对较低,其与黄淮区和长江中游区品种遗传关系较近,与大部分长江上游区和下游区品种遗传关系较远;可利用遗传关系远的品种创制材料,用前述补充等位变异进行分子标记辅助选择,以高效拓宽现有育种资源的遗传基础。     

中国春大豆品种聚类分析及主成分分析

本文对８９个中国春大豆品种的１８个数量性状进行了聚类,逐步判别和主成分分析,聚类分析用“遗传距离”定量测定品种间的遗传差异,并根据“遗传距离”将春大豆品种分为六类：东北早熟春大豆,东北中熟春大豆,黄淮春（夏）大豆,长江春大豆,西北春大豆,黄淮晚熟春（夏）大豆及南方春大豆。在聚类分析基础上用逐步判别分析选出分枝数,节数,荚数,百粒重,单株粒重,粒形指数,生育后期,全生育期,蛋白质含量等１１个对品种分     

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.     

Evaluation of soybean germplasm conserved in NIAS genebank and development of mini core collections

Genetic variation and population structure among 1603 soybean accessions, consisted of 832 Japanese landraces, 109 old and 57 recent Japanese varieties, 341 landrace from 16 Asian countries and 264 wild soybean accessions, were characterized using 191 SNP markers. Although gene diversity of Japanese soybean germplasm was slight lower than that of exotic soybean germplasm, population differentiation and clustering analyses indicated clear genetic differentiation among Japanese cultivated soybeans, exotic cultivated soybeans and wild soybeans. Nine hundred ninety eight Japanese accessions were separated to a certain extent into groups corresponding to their agro-morphologic characteristics such as photosensitivity and seed characteristics rather than their geographical origin. Based on the assessment of the SNP markers and several agro-morphologic traits, accessions that retain gene diversity of the whole collection were selected to develop several soybean sets of different sizes using an heuristic approach; a minimum of 12 accessions can represent the observed gene diversity; a mini-core collection of 96 accession can represent a major proportion of both geographic origin and agro-morphologic trait variation. These selected sets of germplasm will provide an effective platform for enhancing soybean diversity studies and assist in finding novel traits for crop improvement.     

Genetic diversity of Chinese Spring Soybean Germplasm Revealed by SSR Markers

To use, maintain and increase crop germplasm collections efficiently, it is important to assess the diversity of these collections. In this study, 1383 accessions of Chinese spring sowing soybean ( Glycine max ) were used for SSR analysis. In total, 1111 alleles were detected among these collections with an average number of alleles (NA) of 18.52 per locus. The genetic diversity index (PIC value) varied from 0.456 to 0.928 with an average of 0.815. Intensive breeding of cultivars have led to a decrease of genetic diversity. Random-repeated sampling within landraces of different geographical regions suggested that the ranking of both average NA and PIC values among different geographical regions were North spring soybean (Nsp) > South spring soybean (Ssp) > Northeast spring soybean (NEsp), but because of the uneven distribution of SSR variation patterns, the differences between them did not reach a significant level. There was a relationship between genetic distances and geographical distances among soybean populations from different regions, indicating a certain degree of geographical differentiation among Chinese soybean germplasm collections.     

豫南地区野生大豆种质资源的SSR遗传多样性分析

为了研究豫南地区的野生大豆种质资源分布和种群间的亲缘关系,利用SSR分子标记采用20对引物对豫南地区12份野生大豆进行鉴定和遗传多样性分析。结果表明,20对引物对12份野生大豆共扩增63个等位基因,平均每对引物扩增3.1个等位基因;12份野生大豆在分子水平上发生了一定的遗传变化,聚类结果可以看出,12份材料聚类分成了三大类,SSR分子标记与材料的地理来源并没有明显的相关性。野生大豆的亲缘关系与生长的地理环境、表型性状等具有一定的相关性,同一区域收集到的野生大豆也表现出遗传分化。     

利用RAPD标记鉴定大豆种质

本研究以５７个中国大豆祖先吕系及育成品种和１８个美国大豆祖先品系为ＤＮＡ样品来源，通过随机引物ＰＣＲ扩增基因组ＤＮＡ的多态性，探索利用ＲＡＰＤ标记鉴定和相关种质的可能性。研究结果表明，５０个１０摩尔随机引物共扩增可分辩产物２４６个，其中８２．４％的随机引物可产生多态性产物，所扩增产物的５４．４％至少在两个基因毒草境存在差异。上ＰＣＲ扩增产物分别以１和０记录存在与否。扩增产物间的成对比较可产生非相似     

Genetic diversity of wild and cultivated soybeans growing in China revealed by RAPD analysis

To evaluate the genetic diversity and to clarify the genetic relationships of wild and cultivated soybeans growing in China, 21 wild soybean accessions and 27 cultivated soybean landraces were analysed by using the random amplified polymorphic DNA method. The data show that wild soybean has a higher genetic variation than cultivated soybean, indicating that genetic variation has been reduced by domestication of wild soybean. Based on Nei's genetic similarity coefficient, all the accessions were classified into two major clusters, corresponding to wild and cultivated varieties of soybean. Furthermore, within each species, the accessions tend to form sub‐clusters that are in agreement with their geographical origins, demonstrating that an extensive geographical genetic differentiation exists in both species. For cultivated soybean, the varieties from the same geographical region but with different seasonal types were found to have closer genetic relationships than varieties from different geographical regions but with the same seasonal type. This result indicates that geographical differentiation plays a key role in the genetic differentiation of both wild and cultivated soybeans. Cultivated soybean varieties with different seasonal types in a region might have been established mainly from the local genotypes.     

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.     

中国野生大豆群体农艺加工性状与SSR关联分析和特异材料的遗传构成

选用204对SSR标记对全国野生大豆群体(174份代表性样本)的基因组扫描,采用TASSEL软件的GLM (general linear model)方法对百粒重、开花期、成熟期、干豆腐得率、干豆乳得率和耐淹性性状值关联分析,解析与性状关联位点的优异等位变异,鉴别出一批与农艺、加工性状关联的优异等位变异及携带优异等位变异的载体材料;进一步分析极值表型材料的遗传构成。结果表明: (1)累计51个位点(次)与性状关联,有些标记同时与2个或多个性状相关联,可能是性状相关的遗传基础;关联位点中累计16位点(次)与连锁分析定位的QTL一致;(2)与地方品种群体和育成品种群体的关联位点比较,发现野生群体关联位点只有少数与之相同,群体间育种性状的遗传结构有明显差异。(3)与多性状关联的位点其等位变异对不同性状的效应方向可相同可不同,如GMES5532a-A332对百粒重和耐淹性的相对死苗率都是增效效应,而GMES5532a-A344对百粒重是减效效应,对相对死苗率是增效效应;(4)极值表型材料间的遗传构成有很大差异。表型值大的材料携带较多增效效应大的位点等位变异,例如N23349的百粒重是9.08 g,含有4个增效效应较大的位点等位变异;表型值小的材料携带较多减效效应大的位点等位变异,如N23387的百粒重是0.75 g,含有4个减效效应较大的位点等位变异。关联作图得到的信息可以弥补连锁定位信息的不足,尤其是全基因组位点上复等位变异的信息为育种提供了亲本选配和后代等位条带辅助选择的依据。     

Seed quality attributes of food-grade soybeans from the U.S. and Asia

Diversity of food-grade soybeans is critical for utilization of genetic resources in cultivar development, germplasm enhancement, and end-product commercialization. The objective of this study was to assess seed quality attributes and phenotypic variability among 54 U.S. and 51 Asian food-grade cultivars and breeding lines. The results showed greater genetic diversity of protein content, calcium content, seed hardness, and seed size uniformity than other quality traits in both small- and large-seeded genotypes evaluated in this study. Among the small-seeded soybeans, the U.S. genotypes were more diverse and exhibited higher swell ratio and oil content but lower stone seed ratio and protein content than Asian accessions. Among the large-seeded accessions, U.S. genotypes had higher stone seed ratio and oil content but lower swell ratio and protein content, and were less diverse than Asian genotypes. The characterization of diverse food-grade soybeans will facilitate parent selection in specialty soybean breeding.     

Genome-wide genetic dissection of germplasm resources and implications for breeding by design in soybean

“Breeding by Design” as a concept described by Peleman and van der Voort aims to bring together superior alleles for all genes of agronomic importance from potential genetic resources. This might be achievable through high-resolution allele detection based on precise QTL (quantitative trait locus/loci) mapping of potential parental resources. The present paper reviews the works at the Chinese National Center for Soybean Improvement (NCSI) on exploration of QTL and their superior alleles of agronomic traits for genetic dissection of germplasm resources in soybeans towards practicing “Breeding by Design”. Among the major germplasm resources, i.e. released commercial cultivar (RC), farmers’ landrace (LR) and annual wild soybean accession (WS), the RC was recognized as the primary potential adapted parental sources, with a great number of new alleles (45.9%) having emerged and accumulated during the 90 years’ scientific breeding processes. A mapping strategy, i.e. a full model procedure (including additive (A), epistasis (AA), A × environment (E) and AA × E effects), scanning with QTLNetwork2.0 and followed by verification with other procedures, was suggested and used for the experimental data when the underlying genetic model was usually unknown. In total, 110 data sets of 81 agronomically important traits were analyzed for their QTL, with 14.5% of the data sets showing major QTL (contribution rate more than 10.0% for each QTL), 55.5% showing a few major QTL but more small QTL, and 30.0% having only small QTL. In addition to the detected QTL, the collective unmapped minor QTL sometimes accounted for more than 50% of the genetic variation in a number of traits. Integrated with linkage mapping, association mappings were conducted on germplasm populations and validated to be able to provide complete information on multiple QTL and their multiple alleles. Accordingly, the QTL and their alleles of agronomic traits for large samples of RC, LR and WS were identified and then the QTL-allele matrices were established. Based on which the parental materials can be chosen for complementary recombination among loci and alleles to make the crossing plans genetically optimized. This approach has provided a way towards breeding by design, but the accuracy will depend on the precision of the loci and allele matrices.