谷子种质资源表型及SSR遗传多样性分析

本研究对国内外124份谷子种质资源进行表型及SSR遗传多样性分析。表型性状分析表明,幼苗叶姿、穗颈2个质量性状变异较丰富;单株秆重、单株生物产量、单穗重3个数量性状变异系数较大,遗传多样性指数较高,表现出丰富的遗传多样性。表型性状聚类采用Ward法,在遗传距离7. 0处分为6大类群,各类群性状差异明显。SSR遗传多样性分析表明,52对引物在124份材料中共扩增特异性条带52条,平均遗传相似系数0. 834,变化范围为0. 615~0. 962。聚类分析表明,遗传相似系数为0. 746时,124份材料被分为4大类群,第Ⅰ、Ⅱ类群均来自中国河北;第Ⅲ类群来自哈萨克斯坦;第Ⅳ类群占参试材料的87. 90%,没有明显的地理聚类特征。但遗传相似系数为0. 843、0. 870时,进一步可分9个亚群、7个亚亚群,材料又基本按地区聚为一类。表型性状及SSR聚类分析均存在明显的地理聚类特征,结合表型性状鉴定出12份适宜新疆种植的谷子优异种质。     

燕麦种质资源主要农艺性状的遗传多样性分析

燕麦种质资源是燕麦育种的重要基础,对燕麦遗传多样性的研究不仅有助于种质资源的搜集、管理和利用,也有利于进行核心种质的研究。为了解不同地区燕麦种质资源在农艺性状上的遗传多样性,对74份皮、裸燕麦种质资源13个性状的遗传多样性进行了聚类分析与主成分分析。结果表明:各性状的遗传多样性指数较大,多样性指数最高的是主穗粒重,其次是千粒重和穗长;性状变异系数最大的是单株分蘖数,其后依次为单株粒重和主穗粒重,最小的为株高;根据品种间各性状的遗传差异,通过聚类分析将74份资源材料划分为5类,其中36份皮燕麦资源被分为2类,26份裸燕麦资源被分为2类,7份皮燕麦和5份裸燕麦被分为一类,其中,类群Ⅰ可作为高产育种目标的亲本,类群Ⅲ可作为粒型育种目标的亲本,类群Ⅳ、Ⅴ可作为株高和小穗等育种目标的亲本;8个数量性状主成分分析的结果表明,前4个主成分对变异的累计贡献率达86.27%,第一主成分反应产量,第二主成分反应粒型,第三、第四主成分分别反应分蘖数和株高。     

六棱大麦种质表型遗传多样性评价

为探究六棱大麦种质资源的遗传多样性,提高六棱大麦育种组合选配的针对性和育种效率。以来自国内外的89份六棱大麦种质为材料,考察了3个不同环境下参试材料的株高、穗下节间长、穗长、主穗粒数、单株穗数、千粒重、单株粒重、单株生物重等8个性状,综合参试材料3个不同环境的表现,通过变异系数及遗传多样性指数分析参试材料的遗传多样性,通过主成分分析、聚类分析等方法对参试六棱大麦种质进行了综合评价。结果表明:(1)六棱大麦的穗长、单株粒重2个性状变异较丰富,株型性状与穗粒数变异系数较小,六棱大麦育种的增益效应主要体现在穗长和粒重的适度增加;(2)六棱皮大麦与六棱裸大麦仅在千粒重性状上差异显著,六棱皮大麦各性状多样性指数普遍高于六棱裸大麦;(3)参试六棱大麦种质分为5类,其中第4类基本均为中国地方种质,分布广泛,但遗传距离较近;其他4类为国内外选育的种质,各具特性;(4)利用主成分二维排序分析筛选到以高产为基础,分别兼具矮秆、长穗且分蘖能力强、大粒及高生物重的4类优异种质;基于主成分构建了六棱大麦种质资源的综合评价方程。     

怒江干热河谷杧果种质资源的表型和AFLP遗传多样性分析

利用表型和AFLP标记,对怒江干热河谷57份杧果种质进行遗传多样性分析。表型性状分析结果表明:8个表型性状在不同的种质间表现出较大的差异,变异系数变化范围为16.98%~61.50%,多样性指数(H’)平均为3.975,其中单果重变异较大。AFLP分析结果显示:57份种质共产生1098条带,其中多态性条带为1032条,多态性比率为94.0%,相似系数在0.55~0.82之间。AFLP聚类分析结果及主成分分析结果均表明种质间具有复杂的遗传关系,且怒江干热河谷杧果种质的亲缘关系与地理分布没有明显的相关性。表型性状聚类和AFLP分子标记聚类分析的结果相对一致,均能较准确地将优势类群聚在一起,且表明57份杧果种质具有较丰富的遗传多样性。     

利用SSR标记研究茄子种质资源遗传多样性

本研究利用105个SSR分子标记分析了50份茄子种质资源遗传多样性。105对SSR引物中筛选出的20对多态性含量较高的引物,在50份茄子品种组成的群体中共检测出91个等位基因,平均每个基因位点检测到4.55个等位基因。PIC的变幅为0.202 1~0.735 6,平均为0.401 2。根据遗传距离并结合UPGMA聚类分析可将50份种质分为10个类群。SSR分子标记与品种资源性状聚类分析基本一致。     

405份CIMMYT引进小麦种质的遗传多样性分析

为了明确国际玉米改良中心(CIMMYT)引进普通小麦种质材料的遗传多样性特点,为其利用提供参考依据,本研究从均匀分布于小麦基因组的420对SSR引物中选择出条带清晰、多态性较好的62对引物对引自CIMMYT的405份普通小麦种质系进行遗传多样性检测。结果表明,62对SSR引物在405份CIMMYT材料中共检测到198个等位变异,每对引物检测到等位变异的数目为2~8个,平均每对SSR引物能够检测到3.19个等位变异。单个SSR引物的PIC值介于0.03~0.79之间,平均值0.48。405份CIMMYT材料A、B、D基因组之间多态性位点数和等位变异数相差不大,PIC平均值B基因组(0.53)A基因组(0.52)D基因组(0.39)。聚类分析结果显示,62对SSR引物能够将405份CIMMYT材料区分开来,在0.1285遗传距离处将供试材料分为24个类群,类型较为丰富,不同类群的材料在农艺性状和品质性状上存在差异。     

谷子主要育成品种在新疆的遗传多样性研究

以274份谷子种质资源为材料,利用聚类分析和主成分分析方法,对参试资源16个农艺性状的遗传多样性进行综合评价。结果表明,参试材料的11个数量性状的遗传多样性指数均大于2.000,在新疆表现出广泛的遗传多样性。基于种质资源间形态标记的遗传差异,将274份谷子种质资源聚类并划分为6大类群。第Ⅰ类群(105份材料)生育期较短,属早熟类型,但其他性状表现一般;第Ⅱ类群(19份材料)出苗-抽穗日数最小,全生育期最短,早熟性明显,穗下节间长度相对其他类群较长;第Ⅲ类群(10份材料)生育期较短,主穗长相对较长,其余性状表现均处于较低水平;第Ⅳ类群(58份材料)主穗长、单穗重在各类群中处于较高水平,生育期相对较短;第Ⅴ类群(26份材料)生育期最长,属晚熟类型,除主穗长度最小外其余性状均表现突出;第Ⅵ类群(56份材料)生育期相对较长,属中晚熟类型,株高较低,生物产量处于中等水平。9个数量性状的主成分分析结果表明,前3个主成分因子(单株秆重、株高、主穗直径)累计贡献率达70.41%,各主成分因子性状载荷值反映了育种中各性状的选择方向及潜力。综合评价谷子种质资源农艺性状,为新疆谷子资源收集、评价和利用提供一定的科学依据。     

河北省绿子叶黑豆种质资源表现型和ISSR标记遗传多样性分析

为了揭示河北省绿子叶黑豆种质资源的遗传多样性,为其研究利用提供理论根据,以46份原产河北省的绿子叶黑豆种质资源为试验材料,对其基于表型性状及ISSR标记鉴定结果进行了聚类分析,结果表明:7个ISSR引物共检测出60个等位变异,平均每个位点有8.6个等位变异,变幅为5～17个;ISSR引物的多态性信息量PIC变幅为0.721～0.927,平均0.820;利用表型性状和ISSR标记数据进行品种间遗传多样性分析,遗传相似系数变化范围分别为0.07～0.53和0.43～1.00,平均为0.284和0.704,遗传相似性变幅较大,河北省不同绿子叶黑豆品种间存在着丰富的遗传多样性。聚类结果显示,类群与品种来源地有关。     

谷子地方品种和育成品种的遗传多样性研究

对河南、河北和山东等地的482份谷子地方品种和近30年培育的谷子品种的11个形态性状和农艺性状进行种植观察,研究两类品种的遗传多样性。与地方品种相比,育成品种形态性状的多样性指数大多低于地方品种,育成品种的株高比地方品种显著降低,稳重、穗粒重和出谷率等性状显著提高。地方品种的广泛变异类型是现代谷子育种的重要遗传资源。     

应用SSR标记分析鹰嘴豆种质资源遗传多样性

种质资源的遗传多样性是育种工作的基础,本研究利用SSR标记对鹰嘴豆资源进行遗传多样性分析,旨在发掘鹰嘴豆资源中丰富的遗传变异。从48对鹰嘴豆SSR引物中筛选出18对核心多态性引物,对96份不同来源的鹰嘴豆种质资源进行SSR标记的遗传多样性分析。结果表明,18对SSR引物共检测到115个等位变异,占总检测位点的52.99%,每对SSR引物可检测出3~10个等位变异,平均6.39个;平均每个位点多态信息量(PIC)为0.8107,变化范围为0.6661~0.8984。Shannon's信息指数平均为1.4769,变化范围为0.0607~1.9584。PGMA聚类结果表明,在遗传相似系数0.59处,可将96份鹰嘴豆资源分为6个类群,类群Ⅰ包含9份资源,类群Ⅱ包含2份资源,类群Ⅲ包含28份资源,类群Ⅳ包含4份资源,类群Ⅴ包含40份资源,类群Ⅵ包含13份资源。本研究对我国鹰嘴豆种质资源的评价与利用、优异基因的挖掘、育种亲本材料的选择等提供科学依据。     

甜高粱种质资源在新疆的多样性表现及聚类分析

采用多样性指数、变异系数和聚类分析等方法,对国内外72份甜高粱种质资源24个性状进行遗传多样性研究。结果表明,14个质量性状中粒色的遗传多样性指数最高为1.6333,幼苗色和结实形式的遗传多样性指数最低为0,平均为0.7460;10个数量性状中穗长、茎粗、锤度、单穗粒重、单株秆重、出汁率、千粒重、株高、穗重、生育期都存在较大的变异,变异系数幅度为7.85%～53.01%,各性状多样性指数均较大,平均2.0061;穗长的多样性指数最大,为2.1383,生育期多样性指数最小,为1.7331,表明新疆现有甜高粱资源拥有丰富的遗传多样性。聚类分析将72份资源划分为四大类。     

Development of eSSR-Markers in Setaria italica and Their Applicability in Studying Genetic Diversity,Cross-Transferability and Comparative Mapping in Millet and Non-Millet Species

Foxtail millet ( Setaria italica L.) is a tractable experimental model crop for studying functional genomics of millets and bioenergy grasses. But the limited availability of genomic resources, particularly expressed sequence-based genic markers is significantly impeding its genetic improvement. Considering this, we attempted to develop EST-derived-SSR (eSSR) markers and utilize them in germplasm characterization, cross-genera transferability and in silico comparative mapping. From 66,027 foxtail millet EST sequences 24,828 non-redundant ESTs were deduced, representing ~16 Mb, which revealed 534 (~2%) eSSRs in 495 SSR containing ESTs at a frequency of 1/30 kb. A total of 447 pp were successfully designed, of which 327 were mapped physically onto nine chromosomes. About 106 selected primer pairs representing the foxtail millet genome showed high-level of cross-genera amplification at an average of ~88% in eight millets and four non-millet species. Broad range of genetic diversity (0.02–0.65) obtained in constructed phylogenetic tree using 40 eSSR markers demonstrated its utility in germplasm characterizations and phylogenetics. Comparative mapping of physically mapped eSSR markers showed considerable proportion of sequence-based orthology and syntenic relationship between foxtail millet chromosomes and sorghum (~68%), maize (~61%) and rice (~42%) chromosomes. Synteny analysis of eSSRs of foxtail millet, rice, maize and sorghum suggested the nested chromosome fusion frequently observed in grass genomes. Thus, for the first time we had generated large-scale eSSR markers in foxtail millet and demonstrated their utility in germplasm characterization, transferability, phylogenetics and comparative mapping studies in millets and bioenergy grass species.     

谷子β-胡萝卜素异构酶家族基因的表达与变异分析

株型是影响谷类作物产量的重要性状, 株型改良对提高作物产量具有重要意义。独脚金内酯(strigolactones, SLs)作为一种最新被鉴定的植物激素, 其通过抑制腋芽的伸长调控分枝/分蘖的形成。β-胡萝卜素异构酶(D27s)是SLs合成途径的关键酶, 通过对谷子(Setaria italica) β-胡萝卜素异构酶典型结构域Pfam:DUF4033进行分析, 鉴定到3个谷子D27s基因家族成员(Seita.8G168400、Seita.6G088800和Seita.3G050900)。蛋白质特性分析显示, 谷子D27s蛋白由271-277个氨基酸残基组成, 分子量为30.1-30.4 kDa, 等电点为5.85-9.31, 不稳定系数介于38.48-74.47之间, 且均定位于叶绿体; 系统进化分析发现, 谷子D27s家族成员位于3个不同进化分支; 顺式作用元件预测显示, SiD27-1 (Seita.8G168400)可能参与调控生物节律、生长素介导的生长发育以及干旱和低温等胁迫应答过程。基因表达分析显示, SiD27-1在谷子多分蘖材料中表达下调, 在低磷胁迫处理下, D27s基因均能产生不同程度的响应, 并且SiD27-1的响应较其它成员更快速。单倍型分析结果表明, SiD27-1的H001单倍型为优异单倍型, 对谷子的株高、抽穗期和产量改良具有重要应用价值。综上, 推测SiD27-1极可能在SLs合成中发挥关键作用并对谷子株型产生影响。研究结果为深入揭示D27s对谷子分蘖形成的调控机制奠定了基础, 也为谷子株型分子设计育种提供了优异的等位变异位点。     

Development and utilization of novel intron length polymorphic markers in foxtail millet (Setaria italica (L.) P. Beauv.)

Introns are noncoding sequences in a gene that are transcribed to precursor mRNA but spliced out during mRNA maturation and are abundant in eukaryotic genomes. The availability of codominant molecular markers and saturated genetic linkage maps have been limited in foxtail millet (Setaria italica (L.) P. Beauv.). Here, we describe the development of 98 novel intron length polymorphic (ILP) markers in foxtail millet using sequence information of the model plant rice. A total of 575 nonredundant expressed sequence tag (EST) sequences were obtained, of which 327 and 248 unique sequences were from dehydration- and salinity-stressed suppression subtractive hybridization libraries, respectively. The BLAST analysis of 98 EST sequences suggests a nearly defined function for about 64% of them, and they were grouped into 11 different functional categories. All 98 ILP primer pairs showed a high level of cross-species amplification in two millets and two nonmillets species ranging from 90% to 100%, with a mean of ～97%. The mean observed heterozygosity and Nei's average gene diversity 0.016 and 0.171, respectively, established the efficiency of the ILP markers for distinguishing the foxtail millet accessions. Based on 26 ILP markers, a reasonable dendrogram of 45 foxtail millet accessions was constructed, demonstrating the utility of ILP markers in germplasm characterizations and genomic relationships in millets and nonmillets species.     

Study of genetic diversity and relationships among accessions of foxtail millet [Setaria italica (L.) P. Beauv.] in Korea, China, and Pakistan using SSR markers

In this study, 28 simple sequence repeat (SSR) primer sets were used to analyze the genetic diversity, population structure, and genetic relationships among 37 accessions of foxtail millet from Korea, China and Pakistan. A total of 298 alleles were detected with an average allele number of 10.6 per locus among 37 foxtail millet accessions. The number of alleles per locus ranged from 2 (b226) to 20 (b236). Of the 298 alleles, 138 alleles (46.3%) were rare (frequency < 0.05), 152 alleles (51.0%) were detected at an intermediate frequency (range, 0.05?C0.50), and eight alleles (2.7%) were abundant (frequency > 0.50), respectively. The average gene diversity values were 0.652, 0.692, and 0.491 and polymorphic information content values were 0.621, 0.653, and 0.438, for accessions from Korea, China, and Pakistan, respectively. The accessions from China showed higher SSR diversity than those from Korea and Pakistan. A phylogenetic tree constructed using the un-weighted pair group methods with arithmetic mean algorithm revealed three major groups of accessions that were not congruent with geographical distribution patterns with a few exceptions. The lack of correlation between the accession clusters and their geographic location indicates that the diffusion of foxtail millet from China to Korea might have occurred through multiple routes. Our results provide support for the origin and diffusion route of foxtail millet in East Asia. This SSR-based assessment of genetic diversity, genetic relationships, and population structure among genetic resources of foxtail millet landraces will be valuable to foxtail millet breeding and genetic conservation programs in Korea.     

不同地区啤酒大麦品种农艺性状鉴定与分类研究

为筛选可用于云南啤酒大麦改良的优良种质材料,对107份不同来源啤酒大麦品种的农艺性状进行鉴定与分类研究.结果表明,云南啤酒大麦的主要特点是成熟期长,分蘖较强,植株高,旗叶面积大,穗长、穗粒数、穗粒重、千粒重、秸秆干重中等,穗草比较高;国外引进品种株型中等紧凑,叶片细长,穗粒重和千粒重较大,穗草比适中.从中筛选出19份性状优良的材料.通过聚类分析,可将供试材料划分成4个性状不同的类群,各类群的农艺性状差异明显,有利于育种目标材料的选择.     

谷子 MADS-box基因家族的鉴定和表达分析

MADS-box基因是真核生物中一类重要的转录因子,参与调控多项植物的生长发育过程。然而关于谷子穗发育的MADS-box基因研究比较少。本研究使用序列相似性检索,在Phytozome 13.0数据库中筛选并且鉴定出了68个谷子MADS家族成员,并对这些家族成员的物理化学性质、系统发育树、染色体定位、表达谱等进行了全面的分析。结果表明,谷子MADS家族成员在染色体上分布不均匀,可以分为5个亚族。通过组织特异性表达谱分析得到,多数MADS基因在穗中表达量要高于其他器官。此外利用转录组测序技术对发育初期的谷穗和成熟期的谷穗进行了转录组测序分析,筛选到数个与谷穗分生组织发育相关MADS-box基因。为进一步揭示MADS-box基因在谷子穗发育过程中的作用奠定了重要的基础。     

基于SSR标记的谷子遗传多样性研究

用21个分布在谷子9条染色体上的SSR标记,对120份来自于核心种质的谷子材料进行遗传多样性研究。21个标记共检查出305个等位变异,各标记检测出的等位变异数在3～26个之间,平均每个位点检测出的等位变异数为14.5个;21个位点的平均多态信息量(PIC)为0.809。基于21个SSR标记的分子鉴定,计算了120份材料间的遗传相似系数,其变化范围为0.8393～0.9672,平均值为0.8906。根据计算的遗传距离,对120份谷子材料进行UPGMA聚类,在遗传相似系数0.8865处这些材料被划分为4个类群,分类结果与这些谷子来源地生态类型总体上表现一致,分别为西北内陆类群、黄土高原内蒙古高原类群、华北平原类群以及华北平原近年育成种类群。     

谷子籽粒蛋白质、脂肪、千粒重的遗传变异

以来自华北夏谷区、东北春谷区、西北高原早熟春谷区、西北高原中晚熟春谷区等4个主要谷子生态区的270个品种为材料,包括育成品种220份和地方品种50份,测定了籽粒千粒重、蛋白质和脂肪含量。结果表明:270份谷子品种平均蛋白质含量(11.18±1.14)%,脂肪含量(4.00±0.42)%,千粒重(2.87±0.28)g。育成品种的蛋白质含量显著低于地方品种蛋白质含量,而育成品种和地方品种的脂肪含量、千粒重差异不显著;不同生态区品种之间蛋白质含量、千粒重差异显著,但脂肪含量差异不显著。西北早熟春谷区品种蛋白质含量平均最高、千粒重最大,华北夏谷区品种蛋白质含量平均最低、千粒重最小;蛋白质含量和千粒重显著正相关,蛋白质含量、千粒重和生育期显著负相关。本研究报道了近30年来我国谷子主产区谷子品种的蛋白质、脂肪含量和千粒重的差异,对于谷子品质育种具有借鉴意义;研究认为谷子籽粒的蛋白质、脂肪含量和千粒重应作为品质育种的重要指标加以重视。     

Exploring Potential of Pearl Millet Germplasm Association Panel for Association Mapping of Drought Tolerance Traits

A pearl millet inbred germplasm association panel (PMiGAP) comprising 250 inbred lines, representative of cultivated germplasm from Africa and Asia, elite improved open-pollinated cultivars, hybrid parental inbreds and inbred mapping population parents, was recently established. This study presents the first report of genetic diversity in PMiGAP and its exploitation for association mapping of drought tolerance traits. For diversity and genetic structure analysis, PMiGAP was genotyped with 37 SSR and CISP markers representing all seven linkage groups. For association analysis, it was phenotyped for yield and yield components and morpho-physiological traits under both well-watered and drought conditions, and genotyped with SNPs and InDels from seventeen genes underlying a major validated drought tolerance (DT) QTL. The average gene diversity in PMiGAP was 0.54. The STRUCTURE analysis revealed six subpopulations within PMiGAP. Significant associations were obtained for 22 SNPs and 3 InDels from 13 genes under different treatments. Seven SNPs associations from 5 genes were common under irrigated and one of the drought stress treatments. Most significantly, an important SNP in putative acetyl CoA carboxylase gene showed constitutive association with grain yield, grain harvest index and panicle yield under all treatments. An InDel in putative chlorophyll a/b binding protein gene was significantly associated with both stay-green and grain yield traits under drought stress. This can be used as a functional marker for selecting high yielding genotypes with ‘stay green’ phenotype under drought stress. The present study identified useful marker-trait associations of important agronomics traits under irrigated and drought stress conditions with genes underlying a major validated DT-QTL in pearl millet. Results suggest that PMiGAP is a useful panel for association mapping. Expression patterns of genes also shed light on some physiological mechanisms underlying pearl millet drought tolerance.