大豆核心种质和微核心种质的构建、验证与研究进展

我国作物种质资源长期库中保存大豆资源2.3万余份,数量居世界之首。然而,在大豆新品种培育中的利用率仅为1%左右,导致大豆育成品种的遗传基础趋于狭窄。主要原因是缺少对其重要经济性状的鉴定,尤其是缺少多年多点的评价,难以有目的选择有重要价值的育种亲本。为了加速大豆种质资源的评价并促进其利用,在国家基础研究项目(973)的连续资助下,开展了“大豆核心种质构建(1998-2003)”和“大豆微核心种质基因多样性(2004-2009)”研究,目的是浓缩大豆资源的遗传多样性,强化其表型和基因型鉴定,为发掘和利用大豆资源中的优异基因提供指导。本文在研究构建不同比例(占总体2%~5%)大豆核心种质和大豆微核心种质(占总体1%)的同时,介绍了核心种质补充和完善的研究进展。为了验证核心种质的代表性,在构建方法方面,从SSR位点、样本组成、取样比例、低频率等位变异4个方面对代表性进行了分析,并用随机抽样方法对核心种质代表性进行了检测和验证。文中还介绍了利用核心种质和微核心种质在新基因发掘、种质创新和育种利用方面的研究进展,尤其介绍了与育种单位密切合作,建立基于核心种质的种质创新与利用体系的研究成效。围绕遗传多样性、核心种质利用方式进行了讨论,指出大豆核心种质为性状鉴定、新基因发掘、新种质创造和新品种培育等理论研究和实际应用提供材料基础,具有潜在的应用前景。实践证明,大豆种质资源的系统研究与利用,将促进我国大豆种质资源由数量保存型向研究应用型转变。     

黄麻核心种质的遴选

遴选黄麻核心种质可为黄麻种质创新及新品种选育奠定基础。本研究以300份黄麻种质资源为基础,基于SSR分子标记和农艺性状考察,结合地理来源构建核心种质。结果表明,11个农艺性状变异系数变幅在13.06%～84.87%,表现出丰富的遗传多样性。按农艺性状聚类分析可划分为8个类群,按分子标记聚类可划分为10个类群。结合2个聚类分析、地理位置并按比例取样,建立一个由108份品种(系)组成的预选核心种质。采用44对SSR引物对其进行遗传差异分析,在遗传相似系数为0.65时,可把108份品种(系)分成圆果种和长果种两大类。根据遗传差异分析,剔除遗传相似系数大于或等于0.85的遗传冗余,获得84份品种(系)的核心种质,其中圆果种60份和长果种24份。比较84份核心种质与300份种质的农艺性状变异系数及Shannon-Wiener指数发现,两者之间相差不大,表明遴选的84份核心种质可以最大限度代表300份黄麻种质资源的遗传多样性加以利用和保存。     

中国绿豆种质资源初选核心种质构建

以国家作物种质资源数据库中5 072份国内绿豆资源为材料, 根据14个农艺性状, 利用地理来源(省)和性状群进行分组, 分别采用比例法、平方根法和多样性指数法确定取样数及聚类选择和随机选择2种个体选择法构建了13个不同的绿豆初选核心样本, 对不同的取样方法及总资源间进行了品种间平均相似性系数、性状符合度、遗传多样性指数和数量性状变异系数的比较。结果表明, 聚类选择取样优于随机取样, 按照性状群分组优于按省分组; 在聚类选择条件下采用多样性指数法确定取样数优于平方根法和比例法。最终确定按性状群分组, 利用多样性指数确定取样数, 聚类选择个体为绿豆核心种质构建的最佳方案。用此方案, 构建了包含719份绿豆种质的初选核心种质, 取样比例为14.2%, 性状符合度达100%。     

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.     

Mining Elite Sea-Island Cotton Germplasm Based on Phenotyping and SSR Markers

[Objective] An elite germplasm resource of sea-island cotton with outstanding traits was mined in order to accelerate the breeding process of new varieties. [Method] The core collections of sea-island cotton germplasm consisted of 178 accessions were used as experimental materials in this study. Analyses of variability and diversity were performed through detecting phenotypic data of six main breeding-targeted traits, including boll weight, boll number per plant, lint percentage, fiber length, fiber strength, and micronaire. The elite germplasm of sea-island cotton was selected according to 10% optimal sampling strategy based on the phenotypic value of each trait. The 120 pairs of polymorphic simple sequence repeat (SSR) primers were used to analyze the polymorphism of 178 accessions of the core collections. Then, we conducted the population structure and clustering analysis based on the genotyping results. According to the results of cluster analysis, the primary elite germplasm was further selected, and the final elite germplasm of sea-island cotton was identified. [Result] The results showed that there was a high variability and abundant genetic diversity in the 6 studied traits. In 178 accessions of sea-island cotton, 262 alleles were detected by 120 pairs of SSR primers, with an average of 2.18 loci. The average polymorphism information content (PIC) was 0.067 8-0.630 0, with an average of 0.296 0, showing moderate polymorphism. The cluster analysis showed that the core collection of sea-island cotton was divided into six groups. twenty-three elite germplasm resources of sea-island cotton were identified based on phenotypic value and cluster analysis of SSR markers. [Conclusion] The germplasm of sea-island cotton can be analyzed and evaluated based on the phenotyping and SSR markers, and then the elite germplasm of sea-island cotton can be identified. These results provided the material basis for the genetic breeding of sea-island cotton, as well as the important reference and basis for the mining and identification of crop elite germplasm.     

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.     

中国花生小核心种质与ICRISAT微核心种质的SSR遗传多样性比较

明确花生种质资源的遗传多样性和分布规律,对于发掘优良种质资源,选配优良亲本,拓宽育成品种的遗传基础具有重要意义。核心种质为种质资源的研究、评价和鉴定带来了方便。本研究从206对SSR引物中筛选26对引物对我国花生小核心种质和ICRISAT微核心种质共466份资源进行了遗传多样性分析,相似系数为0.49～0.99,鉴定出遗传差异最大的种质L2刚果(中国花生资源)与ICG12625(ICRISAT资源),相似系数为0.49。分析结果表明,多粒型花生的多态性信息量(0.761)和遗传多样性指数(0.97～1.11)均最大(平均相似系数最小,0.73～0.76),其次是普通型花生。中国花生种质资源与ICRISAT资源存在较大差异,尤其是ICRISAT的赤道型材料ICG12625,与中国花生资源的差异最大。相似系数和遗传多样性指数的分析结果均表明,我国花生种质资源的遗传多样性比ICRISAT资源丰富。     

苎麻核心种质构建方法

选用国家长沙苎麻圃的790份种质资源,在已有的25个性状数据的基础上,采用不同取样方法、不同系统聚类方法、不同遗传距离方法构建苎麻核心种质,用质量性状的多样性指数均值、表型保留比率均值和数量性状均值差异百分率、方差差异百分率、极差符合率和变异系数变化率等6个指标评价不同方法组合(取样方法、聚类方法、遗传距离)构建核心种质的优劣。选出合适的构建方法,构建苎麻核心种质。结果表明,不同的取样方法对质量性状和数量性状的遗传多样性影响不同,就质量性状的最大遗传多样性而言,选择优先取样+多次聚类随机取样方法比较适宜,而对数量性状的最大遗传多样性而言,选择优先取样+多次聚类变异度取样方法则较适宜。用优先取样+多次聚类随机取样方法取样时,采用最短距离法和重心法构建的核心种质最好,用优先取样+多次聚类变异度取样时,采用离差平方和法则是构建苎麻核心种质的最佳聚类方法。苎麻核心种质构建与质量性状的不同遗传距离无关,但数量性状以欧氏距离最佳。     

不同遗传距离聚类和抽样方法构建作物核心种质的比较

以棉花种质资源168个基因型，5个纤维性状[2.5%跨长(mm)、整齐度(%)、强度(gf/tex)、伸长度(%)、麦克隆值]数据为例，用混合线性模型分析方法无偏地预测基因型值，利用得到的基因型值，分别采用2种遗传距离(马氏距离和欧氏距离)和7种系统聚类方法(最短距离法、最长距离法、中间距离法、重心法、类平均法、加权配对算术平均法     

利用分子标记数据逐步聚类取样构建甘蔗杂交品种核心种质库

甘蔗杂交品种是商业品种选育的重要亲本资源,为了有效管理和评价这类资源,本研究使用SSR分子标记数据,依据不同相似性系数,采用逐步UPGMA聚类法对161份甘蔗杂交品种(136份来自前期构建的初级核心种质库和25份为新引入国家甘蔗种质资源圃的材料)构建核心种质库,以随机取样方法为对照。在核心种质库质量检测中,用Nei’s多样性指数、Shannon-Wiener多样性指数、总条带数、多态性条带数、多态性条带比例、变异系数符合率、极差符合率、方差差异百分率和均值差异百分率评价分析。结果表明,161份材料在20个SSR位点上具有丰富的多态性,获得294个条带,其中290个为多态性条带,平均多态性条带比例达98.64%;依据3种相似性系数(Jaccard、SM、Dice)和2种取样方法获得8个核心种质库,在核心种质库质量检测中Shannon-Wiener多样性指数、总条带数、多态性条带数表现出较高的检测效率,而其他指标相对较低,8个库中依据Jaccard或Dice相似性系数构建的核心种质库质量最优,该库由107份材料组成,Nei’s多样性指数(0.9785)和Shannon-Wiener多样性指数(4.1854)在P<0.05概率条件下与原库(分别为0.9801和4.4074)无显著差异,而且与原库的均值差异百分率为0 (<20.00%),极差符合率为94.32% (>80.00%),对原库分子和农艺性状遗传多样性都具有较好的代表性,可为后续甘蔗杂交品种资源的准确评价、优异基因发掘和开发利用提供重要的前期基础。     

中国花生核心种质的建立及与ICRISAT花生微核心种质的比较

以中国花生种质资源数据库中记录的6 390份花生资源为材料, 以其基本数据、特征数据和评价数据为信息, 采用分层分组聚类以及随机取样与必选资源相结合的方法, 构建了由576份资源组成的花生核心种质, 占基础收集品的9.01%。除出仁率外, 核心种质与基础收集品种间的其他14个性状平均值和多样性指数差异均不明显, 表明本研究建立的核心种质是有效的。与ICRISAT花生微核心种质的比较, 中国花生资源在龙生型和珍珠豆型方面具有优势, 叶片长、叶片宽、种子长、种子宽的遗传多样性丰富; 而ICRISAT花生资源在多粒型和普通型方面具有优势, 且植株高度和总分枝数的遗传多样性比中国花生资源丰富。     

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.     

山西普通菜豆初级核心种质农艺性状遗传多样性研究

本研究通过对152份山西省普通菜豆初级核心种质进行农艺性状鉴定及农艺性状多样性分析,旨在明确山西省普通菜豆种质资源的主要类型,各类型及性状的主要农艺性状特征及多样性特点。通过在田间进行农艺性状鉴定,同时利用表型数据,采用聚类分析、多样性分析和主成分分析法,确定了山西省普通菜豆种质资源的主要类型,各类型和各性状的农艺性状特征和多样性特点以及24个表型性状所属的主成分。结果表明山西普通菜豆初级核心种质可划分为5种类型。从各分组多样性来看,半蔓+蔓生-紫茎紫花型和蔓生-籽粒斑纹型多样性指数I最高,而直立型资源多样性指数I最低,从各性状多样性来看,质量性状多样性总体低于数量性状,但粒色则表现出明显丰富的多样性。通过主成分分析,将普通菜豆23个农艺性状归属为7个主成分,初步明确了各主成分代表的农艺性状;通过上述研究,明确了山西普通菜豆主要类型、多样性特征和各主成分代表的主要农艺性状,可为资源鉴定评价提供综合性状指标。     

Population structure and linkage disequilibrium of ICRISAT foxtail millet (Setaria italica (L.) P. Beauv.) core collection

Use of diverse germplasm is a key factor which allows high level of resolution due to extensive recombination in the history. Therefore, population used in association mapping should posses as many phenotypes as possible. One of the methods to obtain most of the phenotypes is to construct the core collection. The ICRISAT foxtail millet core collection consisting of 155 accessions was genotyped using 72 simple sequence repeat (SSR) markers to investigate the genetic diversity, population structure and linkage disequilibrium (LD). A high degree of molecular diversity among the accessions was found, with an average of 16.69 alleles per locus. STRUCTURE analyses classify the accessions into four subpopulations (SP) based on SSR allelic diversity. The Neighbor joining clustering and the principal coordinate analysis were in accordance with the racial classification. The distribution of molecular genetic variation among and within the four SP and three races showed high degree of variability within each group, and low level of genetic distance (GD) among the groups. LD decay of <40 cM of GD in foxtail millet core collection was observed, which suggests that it could be possible to achieve resolution down to the 40 cM level. From this investigation, it is evident that the foxtail millet core collection developed at ICRISAT is very diverse and could be a valuable resource for trait association mapping, crop breeding and germplasm management.     

The nabicol: A horticultural crop in northwestern Spain

Nabicol (B. napus L. var. pabularia) is a traditional crop in the Northwest of Iberian Peninsula (South of Galicia and North of Portugal) where it is grown during the winter season on small farms and gardens using traditional varieties. A collection of 36 populations of nabicol from Galicia (northwestern Spain) was evaluated during 2002 and 2003 in two locations and two growing seasons (spring/summer and autumn/spring) for 28 agronomic and morphological traits. The objectives of this study were to: (i) evaluate a collection of nabicol landraces from northwestern Spain, (ii) determine the suitability of this germplasm as a summer crop and (iii) study the genetic diversity among local populations. Significant differences were observed among populations for most traits. Genotype × environment interaction was significant for most of them. Spring/summer growing season could be recommended for growing nabicol but resistance to Lepidoptera pests attacking Brassica crops should be improved. Most populations had an agronomic value similar to the commercial variety. The most promising variety for horticultural use was MBG-BRS0063, which showed the highest yield. Morphological and agronomic data were subjected to cluster analysis and four groups were defined with a group clustering most populations. The low genetic diversity could be explained because populations were collected in close geographical areas and the selection made by farmers was always for a horticultural use. These results give information about the diversity and breeding value of the nabicol Spanish germplasm, which could be useful in breeding programs.     

黑龙江省大豆遗传结构及遗传多样性分析

利用22个表型性状和60个微卫星(simple sequence repeat, SSR)位点对黑龙江省140份代表性种质(78份地方品种和62份育成品种)进行分析, 根据UPGMA (unweighted pair group method with arithmetic mean)和Model-base对SSR数据进行遗传结构划分。结果表明, 参试品种可分为2大类群, 第II类群的各项多样性指标均高于第I类群, 2个类群遗传距离为0.2427;PCO结果显示这2个类群分布在不同区域, 这与地理来源和育成年代密切有关。依据品种类型分为育成品种和地方品种两组, 后者的各项多样性指标均高于前者, 两组间的遗传距离为0.1131。依据表型数据的PCO分析表明, 分布区域与品种类型有关, 与SSR结构分类的结果吻合度低, 两组品种主要在3个主成分的6个表型性状上有所不同。它们不是2个相对独立的遗传群体, 根据分子标记和表型分类各有特点;建议在种质遗传多样性研究中将分子数据和表型数据结合起来。     

中原牡丹品种初级核心种质构建与代表性检验

以400个中原牡丹品种的形态学和农艺学性状为基本数据,采用花型分组,组内按平方根比例策略确定取样量,并使用类平均聚类法取样,获得了中原牡丹品种初级核心种质,其取样比例占总体样本的30%.经检测,初级核心种质与总体种质的31个性状中仅花期1个性状的多样性指数达到了显著差异水平,初级核心种质多样性指数对总体种质的代表性达99.20%,表型保留比例达到了100%.表明,这个包含120个样本的初级核心种质能在表型性状上很好地代表总体种质的遗传多样性.     

甘薯核心种质构建方法研究

国家种质广州甘薯圃是我国目前唯一的室外营养体保存的国家种质资源圃,本研究以其中的1200份甘薯资源为材料,22个农艺性状为基础,采用因子分析和变量聚类两种方式对农艺性状进行降维处理,两步聚类、快速聚类和分层聚类3种聚类方法结合5个地理来源对资源分组,组内随机取样与必选资源相结合的方式构建核心种质,依照上述方案共构建出含15个甘薯核心种质的样本库。通过变异系数、标准差符合率和极差符合率评价15个构建方案的数量性状的代表性,多样性指数和表形频率方差评价质量性状的代表性。结果表明,以变量聚类进行降维、两步聚类结合地理来源分组、组内随机取样为甘薯构建核心种质的最佳方案,采用该方法构建的核心种质样本很好的保留了总资源的遗传多样性。     

基于SSR分子标记的闽楠(Phoebe bournei)核心种质的构建

为更好地发掘和利用现有闽楠种质资源,本研究利用7个SSR位点对江西和福建16个闽楠群体的237份材料进行基因型分析,采用逐步聚类(随机取样策略,位点优先取样策略)和模拟退火算法(等位基因数目最大化策略,遗传距离最大化策略)4种取样方法构建闽楠核心种质,并将各遗传多样性指标进行分析。结果表明:7个SSR位点共检测到50个等位基因,平均为7.143,平均有效等位基因为2.115,Shannon信息指数为0.778,观测杂合度为0.302,期望杂合度为0.442。基于逐步聚类方法构建的核心种质相较于基于模拟退火算法构建的核心种质各遗传参数指标都相对较高。对其进行t检验后,选择以基于逐步聚类位点优先取样策略在25%的取样比例下选取的种质为核心种质,其等位基因数、平均有效等位基因数、多态性位点百分率、Shannon多样性指数的保留率分别为初始种质的98%、104.92%、90.09%、97.94%。筛选出的59份核心种质材料能够较好地代表闽楠种质资源的遗传多样性,为闽楠的种质资源保存提供科学依据。