小麦EST-SSR标记的开发和遗传作图

【目的】利用小麦EST序列数据库开发EST-SSR标记。【方法】对GenBank/dbEST注册的普通小麦EST序列(2006.4.18—2007.2.4)进行SSR查找,采用Primer5.0软件设计EST-SSR引物,选用3个小麦品种进行有效性检测,利用RIL群体和Mapmaker/Exp3.0软件进行遗传作图。【结果】在265362条普通小麦EST序列中,共发现6314个SSR,占整个EST数据库的2.38%。其中二核苷酸、三核苷酸重复序列最多,分别为2237(35.43%)和2084(33.01%)个。二核苷酸重复中,以GA/CT和AG/TC出现频率最高、分别占SSR总数的17.85%和10.37%,其次是CA/GT(4.07%)和AC/TG(2.53%);三核苷酸重复中,CAA/GTT(3.93%)、CGG/GCC(3.83%)、CGC/GCG(3.36%)、GGC/CCG(3.14%)、CTT/GAA(2.53%)、TGC/ACG(2.27%)以较高的频率出现。根据筛选得到的微卫星序列共设计了596个EST-SSR引物对,选择其中95分以上的194个合成。PCR检测表明,165个引物对(85%)可以扩增出稳定清晰的带型;在RIL群体中检测到21个EST-SSR引物26个位点有多态性,将其中的23个位点整合到已有的小麦遗传图谱上。【结论】开发了165个小麦EST-SSR新标记,EST序列是小麦SSR标记的重要来源。     

一个谷子新抗锈基因的AFLP标记

【目的】研究谷子抗源的抗锈遗传规律,寻找和定位与谷子抗锈基因连锁的分子标记,为谷子抗锈病基因的定位、克隆和抗病育种等研究奠定基础。【方法】用谷子锈菌单胞菌系93-5接种十里香和豫谷1号及杂交后代F1、F2进行抗锈鉴定,并根据鉴定结果构建抗、感基因池;利用AFLP技术筛选128对EcoRⅠ/MseⅠ引物组合,从中寻找和定位与谷子抗锈基因连锁的分子标记;根据AFLP分析结果进行抗锈基因连锁分析并进行SCAR标记转化。【结果】根据十里香×豫谷1号杂交后代F2群体(131株)抗感谷锈病分离比例,确定十里香抗锈性由显性单基因控制。筛选获得3个与谷子抗锈基因Rusi1(暂命名)连锁的AFLP分子标记,经计算标记与该抗锈基因的遗传距离分别为7.4、9.2和27.4cM。将3个标记片段回收、克隆和测序,成功地将AFLP标记E+CTT/M+TAC-256转化为SCAR标记。初步构建了谷子抗锈基因Rusi1的遗传连锁图谱。【结论】谷子十里香抗锈性由显性单基因控制,Rusi1是一个新发现的谷子抗锈基因。     

鲤遗传连锁图谱的构建

利用JoinMap 4.0软件包,以德国镜鲤选育系为祖父母所培育的自交F2群体的68个个体为作图群体,首次以新型分子标记单核苷酸多态性(Single Nucleotide Polymorphism,SNP)为主要作图标记,以微卫星(Simple Sequence Repeats,SSR)和表达序列标签(Expressed Sequence Tag,EST)为辅助标记,采用CP(CrossPollinators)模型构建鲤的遗传连锁图谱。构建的遗传连锁图谱含有560个标记(174个SSR标记、41个EST-SSR标记和345个SNP标记),分布在50个连锁群上,最大连锁群由60个标记组成,最小连锁群仅含2个标记,平均每个连锁群有11.2个标记。最大连锁群的图距为198.1厘摩(centimorgan,cM),最小的连锁群图距为1.5 cM,图谱总图距为3 295.92 cM,标记间平均间距为7.21 cM,图谱覆盖率为76.26%。构建的图谱为中等密度的遗传连锁图谱可为进一步的鲤相关经济性状的QTL定位研究和分子标记辅助选择育种(MAS)打下基础。研究亮点:首次以新型分子标记SNP为主要作图标记并以SSR标记为...     

SSR分子标记在苹果研究中的应用

SSR(simple sequence repeat)分子标记是以聚合酶链式反应(PCR)技术为基础的共显性分子标记,也称为微卫星标记(microsatellite markers),其具有多态性高、通用性好、易检测且操作简单等优点,已被较多应用于苹果的品种鉴定、遗传多样性分析以及遗传图谱构建和基因定位等方面。就以上方面对苹果中SSR分子标记的应用进行概述并提出SSR分子标记在未来苹果中的研究重点和方向。     

QTL Mapping for Dough Mixing Characteristics in a Recombinant Inbred Population Derived from a Waxy × Strong Gluten Wheat (Triticum aestivum L.)

Protein and starch are the most important traits in determining processing quality in wheat. In order to understand the genetic basis of the influence of Waxy protein (Wx) and high molecular weight gluten subunit (HMW-GS) on processing quality, 256 recombinant inbred lines (RILs) derived from the cross of waxy wheat Nuomai 1 and Gaocheng 8901 were used as mapping population. DArT (diversity arrays technology), SSR (simple sequence repeat), HMW-GS, and Wx markers were used to construct the molecular genetic linkage map. QTLs for mixing peak time (MPT), mixing peak value (MPV), mixing peak width (MPW), and mixing peak integral (MPI) of Mixograph parameters were evaluated in three different environments. The genetic map comprised 498 markers, including 479 DArT, 14 SSR, 2 HMW-GS, and 3 Wx protein markers, covering 4 229.7 cM with an average distance of 9.77 cM. These markers were identified on 21 chromosomes. Eighteen additive QTLs were detected in three different environments, which were distributed on chromosomes 1A, 1B, 1D, 4A, 6A, and 7D. QMPT-1D.1 and QMPT-1D.2 were close to the Glu-D1 marker accounting for 35.2, 22.22 and 36.57% of the phenotypic variance in three environments, respectively. QMPV-1D and QMPV-4A were detected in all environments, and QMPV-4A was the nearest to Wx-B1. One minor QTL, QMPI-1A, was detected under three environments with the genetic distances of 0.9 cM from the nearest marker Glu-A1, explaining from 5.31 to 6.67% of the phenotypic variance. Three pairs of epistatic QTLs were identified on chromosomes 2D and 4A. Therefore, this genetic map is very important and useful for quality trait related QTL mapping in wheat. In addition, the finding of several major QTLs, based on the genetic analyses, further suggested the importance of Glu-1 loci on dough mixing characteristics.     

主要水产养殖动物QTL定位的研究现状

数量性状是一个多世纪以来遗传学研究的主要对象之一,因为它是许多重要作物,畜牧和人类的重要性状。最早的工作是在孟德尔定律重新发现之前由Galton[1]开展的。但长期以来,开展数量性状研究主要借助于统计学手段。20世纪80年代,尤其是90年代以来,RFLP,RAPD,AFLP,微卫星,SNP等DN     

青海不同产区大黄的X-射线衍射图谱研究（摘要）

[目的]为区分不同产地大黄药材的质量及鉴别提供依据。[方法]将供试材料青海玉树掌叶大黄（样品1）、青海海南唐古特大黄（样品2）与青海果洛唐古特大黄（样品3）分别粉碎成粉末,过100目筛,即可供X射线衍射试验用。设定管压30 kV,管流20 mA,2θ扫描范围3°～90°,扫描速度0.06°/s,每种样时间0.5 s,获得3种不同产区大黄的X-射线衍射图谱。试验数据以晶面间距d与特征峰相对强度L/Io表示,记为d/（I/Io）。将试验数据导入中药指纹图谱相似度计算软件,经选峰,设定匹配模板,将谱峰自动匹配,然后设定标准模板,进行谱峰差异性评价和整体相似性评价。[结果]样品1中所含的化学成分与样品2相同,但峰的强度（I/Io）不同,表明相同成分在两种样品中的含量不相等。同样把样品3的图谱与其他两个样品进行比较,也会得到相同的衍射峰值。表明不同产区的大黄中各化学成分的含量有差异,但其衍射图谱及衍射峰值具有一定的指纹特征。[结论]X-射线衍射法是鉴别不同产区的大黄及其他中药材的一种快捷有效的方法。     

小麦RIL群体遗传连锁图谱的构建及其多态性分析

以普通小麦重组近交系(recombinant inbred lines,RIL)‘Q9086×陇鉴19’为作图群体,利用SSR标记构建小麦遗传连锁图谱.结果表明:通过选用2 187对SSR引物筛选出RIL群体双亲表现多态性的引物共405对,多态性频率为18.52%.不同类型SSR标记多态性频率从小到大依次为Xpsp(4.4%)相似文献   

Identification of Co-dominant SSR Markers Associated with Genes Controlling α′-and α-subunit-null β-conglycinin Phenotypes in Soybean (Glycine max (L.) Merr.)

Studies have shown that the three subunits of β-conglycinin are the main potential allergens of soybean sensitive patients.And β-conglycinin has adverse effects on nutrition and food processing.So solation and production of lines with lower β-conglycinin content has been the focus of recent soybean breeding projects.Soybean lines with deficiency in one or all subunits of β-conglycinin have been obtained.An effective and rapid system to identify such mutations will facilitate genetic manipulation of the β-conglycinin subunit composition.Here,two segregating F_2 populations were developed from crosses between Cgy-1/cgy-1 (CC),an α′-lacking line (Δα′),and DongNong 47 (DN47),a wild-type (Wt) Chinese soybean cultivar with normal globulin components,and Cgy-2/cgy-2 (CB),an α-lacking line (Δα),and DN47.These populations were used to estimate linkage among the cgy-1 (conferring α′-null) and cgy-2 (α-null) loci and simple sequence repeat (SSR) markers.Seven SSR markers (Sat_038,Satt243,Sat_307,Sat_109,Sat_231,Sat_108 and Sat_190) were determined to co-segregate with cgy-1,and six SSR markers (Satt650,Satt671,Sat_418,Sat_170,Satt292 and Sat_324) co-segregated with cgy-2.Linkage maps being composed of seven SSR markers and cgy-1 locus,and six SSR markers and the cgy-2 locus were then constructed.It assigned that the cgy-1 gene to chromosome 10 at a position between Sat_307 and Sat_231,and the cgy-2 gene to chromosome 20 at a position between Satt650 and Satt671.These markers should enable map-based cloning of the cgy-1 and cgy-2 genes.For different subunit-deficiency types[α′-null,α-null and (α′+α)-null types],the two sets of SSR markers could also detect of polymorphism between three normal cultivars and seven related mutant lines.The identification of these markers is great significance to the molecular marker-assisted breeding of soybean β-conglycinin subunits.