黄淮冬麦区175个小麦品种的SSR多态性及其与株高、产量相关性状的关联分析

为了解小麦品种资源的遗传多样性, 筛选株高、产量相关性状相关标记的等位变异, 选用108对覆盖小麦各同源染色体且多态性高的SSR引物, 对黄淮麦区175个小麦品种进行分析。共检测到448个等位变异, 平均每个标记4.15个等位变异, 变化范围为2~14个;全部SSR位点的多态性信息含量(PIC)变化范围为0.075~0.869, 平均为0.561。聚类分析显示同一地区或同一育种单位育成的、具有共同亲本的品种多数聚为一类。关联分析表明, 与株高、产量相关性状显著关联(P<0.01)的标记有23个, 其中3个标记达到极显著(P<0.001)水平。标记wmc128(1B)和wmc236(3B)与小穗数极显著相关, 分别解释小穗数变异的10.5%和8.0%;标记Xgwm129(2B)与千粒重达到极显著相关, 可以解释千粒重变异的19.3%。     

中国小麦微核心种质及地方品种籽粒休眠特性的分子标记鉴定

为探索我国小麦微核心种质及地方品种籽粒休眠的遗传基础,利用已报道的4个3AS上的SSR标记(Xbarc57、Xbarc294、Xbarc310和Xbarc321)和1个3BL上的Viviparous-1基因标记Vp1-b2对107份我国小麦微核心种质及31份地方品种进行籽粒休眠的分子标记鉴定。结果表明,5个分子标记在试验材料中表现出丰富的等位变异,具有5～6种等位类型,与籽粒萌芽指数(GI)密切相关。根据一般线性模型分析结果,各位点的等位变异显著影响籽粒休眠,其中Vp1-b2和Xbarc294对籽粒休眠作用较其他标记大,可分别解释65.8%和61.2%的表型变异;其次是Xbarc310(56.3%)和Xbarc57(55.8%),最小的是Xbarc321(53.3%)。而5个标记联合可解释95.9%的性状变异,其次是Vp1-b2和Xbarc294的组合(89.1%),解释变异最小的标记组合是Vp1-b2和Xbarc321(79.4%)。5个分子标记即可解释籽粒休眠的绝大部分表型变异,说明我国小麦微核心种质及地方品种籽粒休眠特性受3AS和3BL上的2个主效基因控制。     

小麦抽穗期和开花期性状的全基因组关联分析

为揭示控制小麦抽穗期和开花期性状的遗传位点,利用均匀分布在21条染色体上的106对SSR标记,对205份小麦品种构成的自然群体进行关联分析。结果表明,在106对SSR标记中共检测到879个等位变异,主要等位变异频率的变化范围为0.175~0.986,平均为0.548;多态性信息指数的变化范围为0.028~0.900,平均为0.557。采用混合线性模型共检测到7个与抽穗期和开花期显著关联（P<0.01）的位点,其中1个存在极显著关联（P<0.001）。另外,Xgwm182（5D）在灌溉条件下与抽穗期显著关联,Xbarc42（3D）在雨养条件下分别与抽穗期和开花期显著关联,这两个位点与前人的研究结果一致。Xgwm102（2D）在灌溉条件下与开花期显著关联,而Xgwm124（1B）在雨养条件下与抽穗期显著关联。Xgwm130（7A）在雨养和灌溉条件下均与抽穗期相关联。研究结果为小麦抽穗期和开花期性状的分子标记辅助选择育种提供了信息。     

利用关联分析发掘小麦自然群体旗叶叶绿素含量的优异等位变异

为揭示小麦自然群体干旱胁迫条件下旗叶叶绿素含量的变化, 筛选相关标记的优异等位变异, 以262份小麦种质资源组成的自然群体为材料, 分别种植在北京的2个试验地点, 均设雨养和灌溉处理, 于开花期和灌浆期检测旗叶叶绿素含量。以分布于21条染色体的169个SSR标记检测所有材料的基因型, 利用STRUCTURE 2.3.2软件分析群体结构, 用TASSEL软件的MLM (mixed linear model)方法对小麦自然群体的旗叶叶绿素含量进行关联分析。在此基础上, 将携带某等位变异的所有材料表型均值与携带无效等位基因(null allele)材料表型均值比较, 估计等位变异的表型效应, 鉴别优异等位变异。共检测到2048个等位变异, 每位点2~37个等位变异, 平均12个。每位点的标记多态性信息量(PIC)为0.008~0.936, 平均0.628。在22个标记位点共检测出40个(次)与旗叶叶绿素含量极显著的关联, 其中11个标记位点有2次以上的关联, Xwmc419-1B和Xgwm501-2B分别有3次关联。在Xcfa2123-7A、Xgwm232- 1D和Xgwm429-2B位点分别检测到效应值大于4.0的等位变异。     

一个水分胁迫应答蛋白与小麦抗旱性的关系及其基因的定位

分析与小麦抗旱性密切相关的水分胁迫应答蛋白, 定位蛋白基因并挖掘与其连锁的分子标记对小麦抗旱分子辅助选择具有重要意义。在一年两点田间全生育期抗旱性鉴定基础上, 以-0.5 MPa PEG-6000模拟干旱胁迫处理小麦幼苗48 h, 并应用SDS-PAGE方法检测分子量约66.2 kD的水分胁迫应答蛋白, 分析其表达与小麦抗旱性的关系。在128个小麦品种(系)中, 检测出67份表达该蛋白, 另61份未表达该蛋白; 前者的平均抗旱指数为1.00, 而后者为0.80, 有极显著差异(P<0.01), 且各抗旱性等级中表达该蛋白的品种比例随着抗性等级的降低而减小。利用晋麦47×西农2208杂交后代230个F₃株系进行遗传分析, 发现该蛋白表达由1对显性基因控制; 目的基因与位于小麦5AS染色体上的5个SSR标记(Xgwm129、Xgwm304、Xbarc56、Xbarc117和Xbarc197)连锁, 位于邻近标记Xbarc56和Xbarc117之间, 遗传距离分别为2.2 cM和2.9 cM。用这两个紧密连锁标记检测128个小麦品种(系), 有67个品种(系)与晋麦47具有相同的标记位点, 其中86.6%的品种(系)(58/67)在水分胁迫后表达目标蛋白。该约66.2 kD的水分胁迫应答蛋白与小麦的抗旱性密切相关, 与其紧密连锁的分子标记可为小麦抗旱分子辅助选择提供依据。     

小麦骨干亲本繁6条锈病成株抗性特异位点及其在衍生品种中的遗传解析

基于已获得的控制小麦条锈病成株抗性“一致性”QTL区段80个SSR标记,结合小麦骨干亲本繁6及其衍生的39个后代小麦品种进行田间条锈病成株期抗性表型鉴定,揭示了骨干亲本繁6遗传物质及其成株抗性在其衍生品种的遗传规律。结果表明,骨干亲本繁6在条锈病条中31、32和33混合生理小种诱导下表现成株抗性,7个衍生后代品种表现全生育期抗性;用控制小麦条锈病成株抗性QTL区段的80个SSR标记对繁6及其后代衍生品种的其他亲本进行分子扫描,共发现9个来自繁6基因组的特异SSR标记,即Xwmc631、 Xgwm359、 Xwmc407、 Xgwm501、 Xgwm148、 Xgwm539、 Xgwm533、 Xgwm299和Xgwm639,其中,Xwmc631、 Xgwm359、 Xgwm501、 Xgwm299和Xgwm639在繁6衍生后代的4个子代中表现较高的遗传贡献率。以SSR标记与小麦条锈病成株抗性的关联分析发现6个SSR标记与小麦条锈病成株抗性显著相关,其中来自繁6的特异SSR等位变异Xgwm539-2D和Xgwm299-3B与严重度、反应型、普遍率、病情指数及病程曲线下面积(AUDPC)均具显著相关性,表明繁6的成株抗性及其控制遗传位点在其衍生后代品种选育过程中得到了很好的定向选择,并在西南麦区小麦条锈病抗性育种中发挥了重要作用。     

小麦株高相关性状与SNP标记全基因组关联分析

株高是影响小麦产量和控制倒伏的重要因素,研究小麦株高相关性状的遗传机制对高产育种具有指导意义。以205份中国冬麦区小麦品种(系)为材料,利用分布于小麦全基因组的24 355个单核苷酸多态性(SNP)标记对株高相关性状进行关联分析。共发现38个与株高相关性状显著关联(P0.0001)的SNP,分布在1B、2A、2B、3A、3B、3D、4A、4B、5A和6D染色体上。其中,11个位点至少在2个环境中稳定表达,可用于开发CAPS标记。同时,发掘了一批株高性状相关基因的优异等位变异,如降低株高的等位变异Bob White_c48009_52,平均降低株高12.9 cm;控制穗下节间长的等位变异BS00039422_51-C和IAAV1698-A,分别调控穗下节间长5.9 cm和6.6 cm。本研究发掘的控制小麦株高基因位点为在分子水平上研究小麦株高复杂性状提供了有价值的参考。     

我国小麦主推品种穗发芽抗性鉴定及相关分子标记的评价

为了培育抗穗发芽品种,对我国10个小麦主产省份的75个品种进行了穗发芽抗性鉴定,同时利用3个分子标记Vp1B3、Xbarc310和Barc294对上述品种进行了分子检测。结果表明,不同品种间穗发芽抗性存在明显差异。Vp1基因的等位变异与穗发芽抗性关系不密切,Vp1B3标记难以用于品种的穗发芽抗性筛选。主效QTL(QPhs-3AS)与种子休眠关系密切,Xbarc310和Barc294可作为穗发芽抗性选择的重要参考,且Barc294较Xbarc310的选择效果好。上述结果为抗穗发芽小麦品种的改良和推广提供了重要信息。     

一个普通小麦Trx超家族新基因TaNRX的克隆与抗旱相关标记开发

抗旱相关基因的挖掘和分子标记开发对选育抗旱小麦品种有重要意义。采用同源克隆、电子克隆、RACE技术和生物信息学分析手段,获得了普通小麦硫氧还蛋白(Trx)超家族一个新基因(TaNRX)的全长cDNA序列(GenBank登录号为KC890769),包含2015 bp,其中开放阅读框1734 bp;预测编码577个氨基酸,分子量为63.79 kD,含3个Trx活性功能区,其中2个存在典型的Cys-X1-X2-Cys结构,具有催化氧化还原反应的活性。TaNRX基因被定位在小麦的5B染色体短臂上,包含4个外显子和3个内含子。比较该基因在两组极端相对发芽率品种中的序列差异,发现第1个内含子差异明显。基于差异位点开发了4个显性互补标记,并用其检测150份小麦品种(系)。检测到TaNRX基因在普通小麦中至少存在2种与抗旱相关的等位变异,分别是TaNRX-a和TaNRX-b;TaNRX-a基因型的品种(系)平均相对发芽率显著高于TaNRX-b基因型(P0.01),说明开发的标记可被用于小麦抗旱性鉴定筛选。     

小麦骨干亲本“胜利麦/燕大1817”杂交组合后代衍生品种遗传构成解析

小麦地方品种燕大1817是我国小麦育种骨干亲本之一,胜利麦/燕大1817杂交组合是北部冬麦区小麦品种遗传改良的基础组合。利用随机分布于小麦全基因组21条染色体上的175对在胜利麦和燕大1817间具有多态性的SSR标记(每条染色体平均8.3对)分析了燕大1817和胜利麦对其38份后代衍生品种的遗传贡献率。结果表明,在全基因组水平上,燕大1817对其后代衍生品种贡献率为26.8%,胜利麦对其后代衍生品种贡献率为43.6%;在部分同源群水平上,燕大1817对其后代衍生品种A、B和D基因组的贡献率分别为25.9%、25.7和26.4%,胜利麦对其后代衍生品种A、B和D基因组的贡献率分别为46.1%、39.1%和44.0%。说明引进种质对我国北部冬麦区小麦品种遗传改良起了重要作用。在染色体水平上,胜利麦对其后代衍生品种的21条染色体贡献率在20.0%~63.3%间,其中对1A染色体贡献率仅有20.0%,对7A染色体贡献率高达63.3%。骨干亲本燕大1817对其后代衍生品种的21条染色体贡献率在7.5%~44.2%间,其中对2A染色体贡献率仅有7.5%,对7D染色体贡献率可达44.2%。骨干亲本燕大1817对后代衍生品种贡献率较高的基因组(单元型)区段有7个,分别是3A上的Xwmc11–Xcfa2262、7B上的Xbarc1073–Xwmc475、1AL上的Xgwm357–Xwmc312、7DS上的Xbarc305–Xwmc506、4AS上的 Xgwm165–Xgwm610、1B上的Xwmc419–Xwmc134和2D上的Xcfd56–Xbarc228,其中,3A染色体上的Xwmc11–Xcfa2262区段对衍生品种贡献率高达77.5%。而胜利麦对后代衍生品种贡献率较高的基因组(单元型)区段有8个,分别是6BS上的Xwmc105－Xwmc397、3D上的Xgdm72–Xgdm8、2DS上的Xgdm5–Xgwm455、7AL上的Xbarc121–Xgwm332、5DL上的Xgwm174–Xwmc161、5BL上的 Xgwm499–Xbarc308、5A上的Xbarc141–Xgwm291和4BL上的Xgwm66–Xgwm251,其中6BS上的Xwmc105–Xwmc397区段对衍生品种的贡献率最高,达71.3%。这些基因组(单元型)区段上存在许多与产量、抗病、抗逆和适应性等重要农艺性状相关的基因和QTL,对北部冬麦区小麦品种遗传改良可能起了重要作用。     

Phenotypic and marker-assisted evaluation of spring and winter wheat germplasm for resistance to fusarium head blight

Fusarium head blight (FHB) caused by Fusarium species, is among the most devastating wheat diseases, causing losses in numerous sectors of the grain industry through yield and quality reduction, and the accumulation of poisonous mycotoxins. A germplasm collection of spring and winter wheat, including nine reference cultivars, was tested for Type II FHB resistance and deoxynivalenol (DON) content. Genetic diversity was evaluated on the basis of Simple Sequence Repeat (SSR) markers linked to FHB resistance quantitative trait loci (QTLs) and Diversity Arrays Technology (DArT) markers. The allele size of the SSR markers linked to FHB resistance QTLs from known resistance sources was compared to a germplasm collection to determine the presence of these QTLs and to identify potentially novel sources of resistance. Forty-two accessions were identified as resistant or moderately resistant to Fusarium spread, and two also had very low DON concentrations. Genetic relationships among wheat accessions were generally consistent with their geographic distribution and pedigree. SSR analysis revealed that several resistant accessions carried up to four of the tested QTLs. Resistant and moderately resistant lines without any known QTLs are considered to be novel sources of resistance that could be used for further genetic studies.     

应用SSR分子标记分析国外种质对我国小麦品种的遗传贡献

利用363对SSR标记分析了在我国小麦生产和育种中发挥了重要作用的11份国外引进品种和33份选育品种的遗传组成,旨在揭示国外种质对我国小麦品种改良的遗传贡献,指导种质资源引进和利用。国外种质包含了选育品种所发现等位变异的76.3%。与不同时期小麦品种等位基因多样性比较发现,国外种质的平均等位变异数最多(3.92),20世纪60年代(2.86)和70年代(3.01)基本一致,80年代有所升高(3.46)。品种间遗传距离比较与品种等位基因多样性结果相吻合。比较引进和选育品种在SSR位点的等位变异频率变化,发现至少在33个SSR位点,国外种质等位变异在我国小麦育种中被优先选择(该等位变异在引进和选育品种的分布频率均高于70%),其中一些位点已知与产量、生育期和抗病等性状密切相关。表明引进品种在以上基因组区域对我国小麦品种具有非常高的遗传贡献。     

应用SSR分子标记分析国外种质对我国小麦品种的遗传贡献

利用363对SSR标记分析了在我国小麦生产和育种中发挥了重要作用的11份国外引进品种和33份选育品种的遗传组成,旨在揭示国外种质对我国小麦品种改良的遗传贡献,指导种质资源引进和利用。国外种质包含了选育品种所发现等位变异的76.3%。与不同时期小麦品种等位基因多样性比较发现,国外种质的平均等位变异数最多(3.92),20世纪60年代(2.86)和70年代(3.01)基本一致,80年代有所升高(3.46)。品种间遗传距离比较与品种等位基因多样性结果相吻合。比较引进和选育品种在SSR位点的等位变异频率变化,发现至少在33个SSR位点,国外种质等位变异在我国小麦育种中被优先选择(该等位变异在引进和选育品种的分布频率均高于70%),其中一些位点已知与产量、生育期和抗病等性状密切相关。表明引进品种在以上基因组区域对我国小麦品种具有非常高的遗传贡献。     

Changes in allelic frequency over time in European bread wheat (Triticum aestivum L.) varieties revealed using DArT and SSR markers

A collection of 189 bread wheat landraces and cultivars, primarily of European origin, released between 1886 and 2009, was analyzed using two DNA marker systems. A set of 76 SSR markers and ~7,000 DArT markers distributed across the wheat genome were employed in these analyses. All of the SSR markers were found to be polymorphic, whereas only 2,532 of the ~7,000 DArT markers were polymorphic. A Mantel test between the genetic distances calculated based on the SSR and DArT data showed a strong positive correlation between the two marker types, with a Pearson’s value (r) of 0.66. We assessed the genetic diversity and allelic frequencies among the accessions based on spring- versus winter-wheat type as well as between landraces and cultivars. We also analyzed the changes in genetic diversity and allelic frequencies in these samples over time. We observed separation based on both vernalization type and release date. Interestingly, we detected a decrease in genetic diversity in wheat accessions released over the period from 1960 to 1980. However, our results also showed that modern plant breeding have succeeded in maintaining genetic diversity in modern wheat cultivars. Studying allelic frequencies using SSR and DArT markers over time revealed changes in allelic frequencies for a number of markers that are known to be linked to important traits, which should be useful for genomic screening efforts. Monitoring changes in the frequency of molecular DNA markers over time in wheat cultivars may yield insight into alleles linked to important traits that have been the subject of positive or negative selection in the past and that may be useful for marker-assisted breeding programs in the future.     

Genetic diversity of resistance genes controlling fusarium head blight with simple sequence repeat markers in thirty-six wheat accessions from east asian origin

Summary Fusarium head blight (FHB) is a serious disease of wheat worldwide that may cause substantial yield and quality losses. Breeding for FHB-resistant cultivars is the most cost-effective approach to control FHB. The objective of the present study was to determine the relationship of resistance between new resistant sources and Sumai 3 using five simple sequence repeat (SSR) markers closely linked to the major QTL for FHB resistance on chromosome arms 3BS and 6BS. All five SSR markers were highly polymorphic between Sumai 3 (and its derivatives) and susceptible Canadian wheat lines. Most of the Sumai 3-derived Chinese wheat accessions and three Canadian FHB-resistant lines had all the Sumai 3 SSR marker alleles on chromosome arms 3BS and 6BS. The Chinese landrace Wangshuibai and two Japanese accessions Nobeokabozu and Nyu Bai had the same banding patterns as Sumai 3 for all five SSR marker alleles, and another Chinese landrace Fangshanmai had three of the five SSR markers in common with Sumai 3, and therefore most likely carries the same QTL as Sumai 3 on 3BS and 6BS. The Brazilian cultivar Frontana had no alleles in common with Sumai 3 on either QTL, and the Chinese landrace Hongheshang had only one of the five SSR markers in common with Sumai 3, therefore likely carrying resistance genes different from Sumai 3. The Italian cultivar Funo is not the donor of either the 3BS QTL or 6BS QTL. All five SSR seem to be effective candidates for marker-assisted selection to increase the level of resistance to FHB in wheat breeding programs.     

利用SSR标记进行优质冬小麦品种(系)的遗传多样性研究

遗传多样性研究对于作物育种具有重要意义. 选择分布于21条小麦染色体上的59对SSR引物对48个优质冬小麦新品种(系)进行了遗传多样性分析. 共检测出209个等位位点, 每对引物等位位点数在2～9之间, 平均为3.5个. 位点多态性信息含量PIC变幅为0.16～0.87, 平均0.56. 8个引物组合在一起可将全部品种区分开来, 48个品种可分为五类,     

Comparison of molecular markers and coefficients of parentage for the analysis of genetic diversity among spring bread wheat accessions

The comparison of different methods of estimating genetic diversity could define their usefulness in plant breeding and genetic improvement programs. This study evaluates and compares the genetic diversity of 70 spring wheat accessions representing a broad genetic pool based on molecular markers and parentage relationships. The sample was composed of 32 accessions from the International Maize and Wheat Improvement Center (CIMMYT) and 38 from other breeding programs worldwide. Eight AFLP-primer combinations and 37 pairs of SSR primers were used to characterize the accessions and the Coefficients of Parentage (COP) were calculated from registered pedigrees. The average genealogical (COP) similarity (0.09 with a range of 0.0–1.0) was low in comparison to similarity calculated using SSR markers (0.41 with a range of 0.15–0.88) and AFLP markers (0.70 with a range of 0.33–0.98). Correlation between the genealogical similarity matrix (excluding accessions with COPs = 0) and the matrices of genetic similarity based on molecular markers was 0.34≤r≤0.46 (p <0.05). It is concluded that AFLP and SSR markers are generally in agreement with estimates of diversity measured using COPs, especially when complete pedigree data are available. However, markers may provide a more correct estimate due to some unrealistic assumptions made when calculating COPs, such as absence of selection. Furthermore, both COP and marker distances indicate that CIMMYT accessions are different from the worldwide group of accessions. This revised version was published online in July 2006 with corrections to the Cover Date.     

基于菜心(Brassica campestris)基因组Survey测序开发多态性SSR标记

为解决菜心SSR标记数量不足、已开发的位点多态性差等问题,本研究利用高通量测序技术,对‘四九-19’和‘3T6’两份菜心材料进行基因组Survey测序,规模化开发多态性SSR标记。两个菜心材料分别获得55 649 657个和59 300 433个Clean reads,分开拼接组装得到430 483个和499 876个Contig。在两个材料的Contig中搜索到共有的SSR位点为30 696个,其中以二和三核苷酸重复基序最为丰富,占总SSR位点的67%。分析比较发现,3 652个(12%) SSR位点在两份测序材料间具有潜在多态性,随机挑选50个SSR位点进行PCR扩增验证,48对(96%)引物在4份菜心材料中扩增出清晰的条带,其中31对(62%)引物在两份测序样品间具有多态性,19对(38%)引物在另两份菜心材料间具有多态性。结果表明,利用基因组Survey测序能开发SSR标记和开发具有多态性的SSR标记,本研究开发的多态SSR标记将进一步为菜心分子标记的发展和应用提供基础。