籼稻标记性状等基因系的构建

选用涉及水稻(Oryza sativa L．)全部12条染色体的、表现简单遗传且易于识别的形态标记材料27份,以早籼品种浙辐802为轮回亲本,经10余次回交,转育成一套籼型标记等基因系。在此基础上,对同一染色体上的标记进行聚合,育成了15份双标记等基因系。该套材料除所带标记性状外,生育期、株高、分蘖力和穗子大小等主要农艺性状与轮回亲本基本相仿。     

陆地棉染色体片段代换系（BC5F3和BC5F3︰4）产量和纤维品质性状表现的初步评价

本研究以海岛棉海1为供体亲本,陆地棉中棉所36为轮回亲本构建了一套棉花染色体片段代换系,通过对BC5F3和BC5F3∶4这两个世代材料表型数据的分析,结果显示,同年份中与轮回亲本比较,群体各性状的极差和遗传变异系数都比较大;1942个BC5F3单株群体中衣分最高达49.57%,超轮回亲本比例为71.78%,纤维断裂比强度最高为36.5cN/tex;658个BC5F3∶4株行群体中纤维上半部平均长度最大值32.25mm,超轮回亲本比例为47.57%,断裂比强度最高达到32.2cN/tex,超轮回亲本比例为40.27%;通过相关性分析,主要产量性状和纤维品质性状在两个世代间呈极显著正相关,衣分与纤维长度及强度在群体内呈极显著负相关。这些结果表明,通过高代回交后连续自交得到的染色体片段代换系群体中存在丰富的遗传变异,含有大量具有丰产、纤维品质优异的单株,两个世代材料具有较好的一致性与稳定性;产量和品质同步改良还存在一定难度。本研究为进一步的近等基因系分析、基因克隆、基因聚合效应分析等储备了大量的研究材料。     

水稻导入系群体的构建与保存

利用我国和国际水稻所的21个优良品种作轮回亲本、来源于24个国家和地区的188份品种资源作供体亲本,通过杂交、连续回交结合性状筛选,构建了近6万份具有轮回亲本遗传背景的近等基因导入系.通过初步的鉴定、整理、编目,共有41255份材料保存于上海市农业生物基因中心中期库,构建了专门的数据库,可实现全球范围的网络共享, 已分发利用了1200余份资源.     

覆盖野生稻基因组的染色体片段替换系的构建及其米质相关数量性状基因座位的鉴定

染色体片段替换系（CSSL）是基因组水平快速初步定位数量性状基因座位（QTL）的良好材料,而水稻的品质性状是多基因控制的数量性状,因此可用替换系鉴定控制水稻品质性状的QTL。本文用分子标记辅助选择技术（MAS）构建了由133个株系组成的以‘特青’（籼稻品种）为轮回亲本,以海南的一种普通野生稻为供体亲本,覆盖绝大部分野生稻基因组的染色体片段替换系。利用这套替换系,初步定位了控制稻米外观和理化品质性状的15个QTL,为今后水稻品质性状QTL的克隆以及稻米品质相关性状的改良提供了依据。     

小麦条锈菌鉴别寄主抗条锈病基因Yr9的微卫星标记

以含有Yr9的抗条锈病近等基因系Taichung29＊6／Yr9及其轮回亲本Taichung29为材料,用目的基因所在1B染色体上32对微卫星引物对其基因组DNA进行PCR扩增,发现引物Xgwm582在近等基因系与轮回亲本间可扩增出特异性DNA片段。经F2代分离群体177个抗、感单株检测证实,该片段位点与抗条锈病基因Yr9紧密连锁,遗传距离为3．7cM,确定Xgwm582可作为抗条锈病基因Yr9的标记。     

陆地棉HB红花性状特异SCAR分子标记的开发

利用Operon系列引物筛选到1个与HB红花性状基因连锁的RAPD标记OPA15^1160,对差异条带进行克隆与核苷酸测序,根据测序结果设计SCAR引物,在HB红花近等基因系及其白花轮回亲本中进行PCR扩增程序优化和鉴定,筛选出一对引物可稳定扩增出与HB红花性状基因连锁的特异片段,获得了与HB红花性状基因紧密连锁的SCAR标记HB^-330。利用具黄色花瓣紫红色基斑的海岛棉与粉红花瓣的红叶棉等种质材料以7LHB红花近等基因系与白花轮回亲本杂交的F1、BC1F1、F2群体,对该SCAR标记的特异性与准确性进行了鉴定与验证,在红花植株中扩增出了330bp大小的片段而在白花植株中未扩增出,证明该标记准确性高、重复性好。HB红花是通过远缘杂交转自野生二倍体比克氏棉的性状,已成功地应用于性状标记杂交棉育种。该SCAR标记不仅为HB红花标记杂交种的纯度鉴定提供了有效技术手段,也为新品种保护提供了技术支持,促进了红花性状杂交种的分子标记辅助育种进程。     

苏麦3号感赤霉病近等基因系的选育及分子标记

小麦品种苏麦3号高抗由禾谷镰刀菌（Fusarium graminearum Schw.)引起的小麦赤霉病,已成为全世界广泛应用的抗源。为更精确地研究苏麦3号的赤霉病抗性,实验用感病品种川980为供体亲本,苏麦3号为轮回亲本,通过不断回交和自交,把川980的赤霉病感病基因导入苏麦3号,创造了感赤霉病的苏麦3号近等基因系（S016）,并得到以下结果：（1）育成的苏麦3号感赤霉病近等基因系,经两年多点抗赤霉病性状鉴定,表明该近等基因系感赤霉病感病性稳定,而其他形态特征,特性与苏麦3号一致。（2）用RAPD,RFLP分析抗,感苏麦3号近等基因系之间的差异,RFLP分析发现该近等基因系在2D染色体上存在差异。RPAD的随机扩增产物OPH191400可能与近等基因系中的赤霉病感病基因非连锁。（3）用近等基因系验证已报道的与小麦抗霉病基因有关的分子标记,均未发现这些分子标记与导致该近等基因系抗性差异的基因有关。     

一个水稻长芒基因AWN-2的定位与克隆

本研究在以广西普通野生稻DP30为供体,籼稻品种9311为受体构建的染色体片段代换系中,筛选鉴定出1份稳定遗传的具有长芒表型的染色体片段代换系。利用该代换系与轮回亲本9311构建了BC5F2分离群体,对控制长芒表型的基因进行了遗传分析,发现该群体中长芒与短芒的单株分离比符合3:1的比例,表明该长芒性状受一对显性核基因控制。采用图位克隆法,将目的基因精细定位于水稻第4染色体分子标记M7和M8之间的12.14 kb范围内,该区域只有一个候选基因即LOC_Os04g43840,测序分析发现与栽培稻日本晴、9311相比,CSSL5的LOC_Os04g43840基因在5'UTR区有29个碱基的缺失和2个碱基的置换,因此,推测LOC_Os04g43840可能为该长芒候选基因。     

利用基于PCR的分子标记区分普通小麦-提莫菲维小麦渐渗系

采用定位于小麦2B染色体上的72对分子标记对含小麦抗白粉病基因Pm6的8份普通小麦（T.aestivum L.）-提莫菲维（T.timopheevii zhuk.）渐渗系材料进行分析, 通过分子标记标图确定8份材料中渗入的提莫菲维小麦染色体片段的大小, 同时结合连锁图谱对这些材料进行了遗传和物理标图。参考本研究所用的分子标记在染色体2B上的定位结果, Pm6基因被位于2B 染色体长臂近末端2BL-6区域, 提莫菲维小麦2G染色体渐渗片断长度由短到长排列顺序为: IGV1-465相似文献   

基于染色体置换系的野生稻抽穗期及紫色性状QTL的鉴定

利用一套以9311为背景的普通野生稻染色体片段置换系群体为研究材料,进行野生稻相关QTL的定位与基因的鉴定。在多年多点的抽穗期调查数据基础上,结合200多个分子标记引物的基因型鉴定结果,定位到11个与抽穗期相关的QTL;选取携带相关QTL导入片段的两个置换系进一步研究,发现2个抽穗期主效QTL均为已克隆基因的等位基因。利用叶鞘、稃尖、柱头颜色与9311差异显著的一个单片段置换系定位到来自野生稻的紫色性状基因Or C,初步鉴定与栽培稻的等位基因功能不同。这些结果再次表明染色体片段置换系是行之有效的QTL定位以及基因发掘的遗传群体。通过对抽穗期、紫色性状相关QTL的定位与基因的鉴定,为进一步的功能研究提供了基因资源。     

Association mapping for yield and grain quality traits in rice (Oryza sativa L.)

Association analysis was applied to a panel of accessions of Embrapa Rice Core Collection (ERiCC) with 86 SSR and field data from two experiments. A clear subdivision between lowland and upland accessions was apparent, thereby indicating the presence of population structure. Thirty-two accessions with admixed ancestry were identified through structure analysis, these being discarded from association analysis, thus leaving 210 accessions subdivided into two panels. The association of yield and grain-quality traits with SSR was undertaken with a mixed linear model, with markers and subpopulation as fixed factors, and kinship matrix as a random factor. Eight markers from the two appraised panels showed significant association with four different traits, although only one (RM190) maintained the marker-trait association across years and cultivation. The significant association detected between amylose content and RM190 was in agreement with previous QTL analyses in the literature. Herein, the feasibility of undertaking association analysis in conjunction with germplasm characterization was demonstrated, even when considering low marker density. The high linkage disequilibrium expected in rice lines and cultivars facilitates the detection of marker-trait associations for implementing marker assisted selection, and the mining of alleles related to important traits in germplasm.     

Association studies of dormancy and cooking quality traits in direct-seeded indica rice

Association analysis was applied to a panel of accessions of Assam rice (indica) using 98 SSR markers for dormancy-related traits and cooking quality. Analysis of population structure revealed 10 subgroups in the population. The mean r ² and D ^′ value for all intrachromosomal loci pairs was 0.24 and 0.51, respectively. Linkage disequilibrium between linked markers decreased with distance. Marker-trait associations were investigated using the unified mixed-model approach, considering both population structure (Q) and kinship (K). Genome-wide scanning, detected a total of seven significant marker-trait associations (P < 0.01), with the R ² values ranging from 12.0 to 18.0%. The significant marker associations were for grain dormancy (RM27 on chromosome 2), α-amylase activity (RM27 and RM234 on chromosomes 2 and 7, respectively), germination (RM27 and RM106 on chromosome 2), amylose (RM282 on chromosome 3) and grain length elongation ratio (RM142 on chromosome 4). The present study revealed the association of marker RM27 with traits like dormancy, α-amylase activity and germination. Simple correlation analysis of these traits revealed that these traits were positively correlated with each other and this marker may be useful for simultaneous improvement of these traits. The study indicates the presence of novel QTLs for a few traits under consideration. The study reveals association of traits like dormancy, α-amylase activity, germination, amylose content, grain length elongation ratio with SSR markers indicating the feasibility of undertaking association analysis in conjunction with germplasm characterization.     

Design of a Brassica rapa core collection for association mapping studies

A Brassica rapa collection of 239 accessions, based on two core collections representing different morphotypes from different geographical origins, is presented and its use for association mapping is illustrated for flowering time. We analyzed phenotypic variation of leaf and seed pod traits, plant architecture, and flowering time using data collected from three field experiments and evaluated the genetic diversity with a set of SSR markers. The Wageningen University and Research Centre (WUR) and the Vavilov Research Institute of Plant Industry (VIR) core collections had similar representations of most morphotypes, as illustrated by the phenotypic and genetic variation within these groups. The analysis of population structure revealed five subgroups in the collection, whereas previous studies of the WUR core collection indicated four subgroups; the fifth group identified consisted mainly of oil accessions from the VIR core collection, winter oils from Pakistan, and a number of other types. A very small group of summer oils is described, that is not related to other oil accessions. A candidate gene approach was chosen for association mapping of flowering time with a BrFLC1 biallelic CAPS marker and a BrFLC2 multiallelic SSR marker. The two markers were significantly associated with flowering time, but their effects were confined to certain morphotypes and (or) alleles. Based on these results, we discuss the optimal design for an association mapping population and the need to fix the heterogeneous accessions to facilitate phenotyping and genotyping.     

浙江省地方豇豆种质资源的鉴定与评价

“第三次全国农作物种质资源普查与收集行动”浙江项目组从全省32个县(市/区)共收集到豇豆地方资源76份.本研究通过对76份种质43个基本农艺性状的田间调查,发现76份种质包括30份长豇豆(Vigna unguiculata(L.)Walp.ssp.sesquipedialis)、46份普通豇豆(Vigna unguic...     

一个新的水稻叶形突变体（tll1）的遗传分析与精细定位

利用甲基磺酸乙酯（ethylmethane sulphonate,EMS）诱变粳稻品种日本晴获得了一个遗传稳定的叶形突变体thread-like leaf1（tll1）。该突变体在杭州表现为矮化、窄叶,极端时仅剩主脉,呈细丝状。将该突变体分别与籼稻品种南京6号、浙辐802和9311进行正反交配组,遗传分析表明该突变体性状由1对隐性单基因控制。通过SSR和STS分子标记对F2代分离群体进行遗传定位,将该基因初步定位在第12染色体SSR标记RM247和RM101之间。随后利用已公布的粳稻品种日本晴和籼稻品种9311的基因组序列,发展了7对有多态的STS标记,最终将该基因定位在FL13和FL14之间约94.3kb的区间内,为进一步克隆TLL1基因奠定了基础。     

Segregation distortion in Arabidopsis C24/Col-0 and Col-0/C24 recombinant inbred line populations is due to reduced fertility caused by epistatic interaction of two loci

A new large set of reciprocal recombinant inbred lines (RILs) was created between the Arabidopsis accessions Col-0 and C24 for quantitative trait mapping approaches, consisting of 209 Col-0 x C24 and 214 C24 x Col-0 F(7 )RI lines. Genotyping was performed using 110 evenly distributed framework single nucleotide polymorphism markers, yielding a genetic map of 425.70 cM, with an average interval of 3.87 cM. Segregation distortion (SD) was observed in several genomic regions during the construction of the genetic map. Linkage disequilibrium analysis revealed an association between a distorted region at the bottom of chromosome V and a non-distorted region on chromosome IV. A detailed analysis of the RILs for these two regions showed that an SD occurred when homozygous Col-0 alleles on chromosome IV coincided with homozygous C24 alleles at the bottom of chromosome V. Using nearly isogenic lines segregating for the distorted region we confirmed that this genotypic composition leads to reduced fertility and fitness.     

Comparison of genomic SSR and EST-SSR markers for estimating genetic diversity in cucumber

Thirteen genomic microsatellite (gSSR) and sixteen expressed sequence tag (EST)-SSR (eSSR) markers were compared to estimate genetic diversity among 29 cucumber (Cucumis sativus L.) accessions. gSSR markers detected mean 4.46 alleles with a mean polymorphic information content (PIC) of 0.664, against eSSR markers with mean 3.38 alleles and a mean PIC of 0.397. gSSRs amplified more null alleles than eSSRs. Genetic diversity within the accession set was estimated by construction of dendrograms using gSSR or eSSR data. There was a clear consistency between gSSR and eSSR trees in terms of positioning of most cucumber germplasms. gSSR markers could separate various types of cucumber germplasms on the whole, although clustering of some accessions was not based on their geographical origins in eSSR tree. eSSR markers identified an independent sub-cluster containing five accessions resistant to downy mildew, suggesting a probable relationship between eSSRs and disease-resistance trait in cucumber. The Mantel test between gSSR and eSSR matrices revealed a good fit correlation (r = 0.836). The general dendrogram constructed using the combined data of gSSRs and eSSRs was similar to those obtained separately with each marker.     

内蒙古78份葱属野生种表型遗传多样性分析

为了揭示内蒙古葱属野生种表型遗传多样性,采用方差分析、主成分分析、聚类分析等方法,对收集的78份野生种的种质资源14个表型性状进行了遗传多样性评价。结果表明:内蒙古葱属野生资源存在丰富的遗传多样性。(1)14个表型性状平均变异系数为43.0%,叶片宽的变异系数最大,为94.7%;种子厚的变异系数最小,为14.6%。14个表型性状在居群间的差异除叶片宽达到显著(P0.05)水平外,其余性状均表现差异不显著;(2)14个表型性状可归成为6个主成分因子,累计贡献率达到80.77%,最大程度上反映了所有78份葱属种质资源的表型特征,在前6个主成分包括的14个农艺性状中的株丛直径、叶片长、叶片宽、单株叶片数、花序长度、花序宽度和种子宽等性状是造成葱属种质资源表型差异的主要因素;(3)14个表型性状间存在显著或极显著的相关性;(4)采用欧氏距离系统聚类法将供试材料分为7大类,78份葱属种质资源基本上按种区分开,并和地理条件有一定的关系,种间和材料间表型差异很明显。本研究为内蒙古葱属资源的利用提供了科学依据。