Microsatellite Marker Identification of a Triticum Aestivum -Aegilops Umbellulata Substitution Line with Powdery Mildew Resistance

Summary Aegilops umbellulata acc. Y39 and Triticum carthlicum acc. PS5, immune to many powdery mildew isolates, were crossed to make an amphidiploid line Am9. The powdery mildew resistance of Am9 was transferred to common wheat cultivar Laizhou953 by crossing and backcrossing. In this study, the origin of powdery mildew resistance in a BC₃F_4:5 population derived from a cross of Am9 and Laizhou953 was identified. Microsatellite markers analysis showed that markers Xgwm257, Xgwm296, and Xgwm319, co-segregated with the powdery mildew resistance, whereas markers Xgwm210, Xgwm388/140, Xgwm388/170 and Xgwm526 were related to susceptibility and linked to resistance in repulsion. Of three markers related to resistance, Xgwm257 and Xgwm319 were codominant, whereas Xgwm296 was dominant. All three markers were Ae. umbellulata-specific indicating that resistance in the test population originated from Ae. umbellulata acc. Y39. The chromosome location and mapping of these linked microsatellite markers, the chromosome numbers of derived BC₃F_4:6 families, and chromosome pairing in F₁ plants from a cross of a homozygous resistant BC₃F_4:5 plant and Laizhou953, showed that wheat chromosome 2B was substituted by Ae. umbellulata chromosome 2U. This is the first gene conferring powdery mildew resistance transferred to wheat from Ae. umbellulata, and it should be a novel resistance gene to powdery mildew. It was temporarily designated PmY39.The first two authors made equal contributions     

STMS markers for grain protein content and their validation using near-isogenic lines in bread wheat

The present study, undertaken as a continuation of an earlier study on quantitative trait loci (QTL) analysis of grain protein content (GPC) in bread wheat (Prasad et al. 1999), includes the following: (1) identification of an additional molecular marker associated with GPC; (2) development of near‐isogenic lines (NILs) for high GPC; and (3) the use of three sets of NILs (a total of 10 NILs) to validate the two available markers linked with QTL for GPC. A total of 114 sequence‐tagged microsatellite site (STMS) primer pairs (that were not used in the previous study) were used for detection of polymorphism between the two parents (PH132, with high GPC; WL711, with low GPC) of a mapping population of 100 recombinant inbred lines (RILs). A total of 95 primer pairs gave amplification products, of which only 30 detected reproducible polymorphism between the parental genotypes. Bulked segregant analysis was conducted using these 30 primers on two bulks (each comprising eight RILs) representing the two extremes of the normal distribution. A solitary primer pair (WMC415) showed association with GPC, which was further confirmed through selective genotyping. Subsequently, 100 RILs were genotyped. A single‐marker linear regression analysis showed significant association between the marker WMC415 and GPC, thus identifying a quantitative trait locus (designated as QGpcccsu‐5A1), which explained 6.21% of the variation for GPC among the RILs. The above STMS marker, together with the STMS marker (WMC41) identified earlier, explains approximately 25% of the variation for GPC. In order to conduct validation of the above two available markers, 10 NILs were developed for high GPC using two genotypes (WL711 and HD2329) with low GPC as recipient parents and another two genotypes (PH132 and PH133) with high GPC as donor parents. NIL 2233 (with 11.7% GPC), derived from HD2329, when tried with WMC41 gave a characteristic amplification profile similar to that of its donor parent PH132, and NIL 2215 (with 11.9% GPC) derived from WL711, when tried with WMC415 gave an amplification profile that resembled its donor parent PH133. The remaining eight NILs with high GPC gave patterns similar to those of their corresponding recipient parents with both the markers, suggesting that either the QTL, other than those associated with the above markers, were actually transferred from the donor parents and contributed to high GPC in these NILs or that recombination had occurred between the markers identified and the corresponding QTL. Thus, the marker validation conducted using NILs, while demonstrating the utility of these two microsatellite markers for use in marker‐assisted selection in plant breeding, also suggested that many more QTL exist that would need to be identified using closely linked molecular markers.     

Development and characterization of SSR markers in Chinese jujube (Ziziphus jujuba Mill.) and its related species

In order to study the extensively genetic diversities of more than 700 cultivars of Chinese jujube, it is necessary to utilize various informative DNA markers. SSR markers are highly polymorphic, co-dominant, locus-specific markers widely used in genetic studies, but less used in Chinese jujube because of no specific primers available. In this study, we used the approach of selectively amplified microsatellite (SAM) to develop SSR markers for Chinese jujube and its related species. Three cultivars (Dongzao, Dalilongzao and Jinsixiaozao) were selected to perform the approach of SAM with CT repeats. There were totally 25 primers obtained, of which we selected 16 primers available to detect the polymorphism in populations of 24 Chinese jujube cultivars, two wild jujube varieties and two Indian jujube cultivars. Based on these primers, genetic relationships of the 28 samples were constructed in a dendrogram according to the UPGMA cluster analysis. The samples were clustered into three main groups, including Chinese jujube, wild jujube and Indian jujube as expected. The 16 sequence-specific SSR primers could efficiently distinguish all the 24 cultivars of Chinese jujube, except for two cultivars, Jinsixiaozao and its ‘stoneless’ mutant, Wuhexiaozao. As a result, SAM was a very efficient method in targeted developing sequence-specific SSR primers in Chinese jujube. Furthermore, SAMs could also be used as high polymorphic molecular markers independently. The further study would focus on developing other oligonucleotide repeat types and applying more SSRs available in the genetic research of Chinese jujube.     

利用高分辨率熔解曲线（HRM）分析梨微卫星标记

 以西洋梨（Pyrus communis L.）‘Nain Vert’的一个矮生型实生系与‘茌梨’（Pyrus bretschneideri Rehd.）的F1杂交群体为试材,对分别源自梨和苹果基因组的4个微卫星（即SSRs）序列多态性进行了高分辨率熔解曲线（high resolution melting,HRM）分析。结果显示,在测试群体中,TsuENH042和Hi15a13存在两种分型,TsuENH081存在3种分型,而Hi08d09只有1种类型。因此,HRM分析技术可以作为筛查果树微卫星标记多态性或对其进行基因分型的有效手段在相关研究中加以应用。     

内蒙古地区西门塔尔牛亲子鉴定微卫星标记筛选

研究旨在筛选适合于我国肉牛亲子鉴定的微卫星标记,以内蒙古乌拉盖地区368头西门塔尔牛为实验材料,利用荧光引物PCR和ABI3730测序仪对12个微卫星标记进行基因型检测,应用Cervus3.0软件统计12个微卫星标记的遗传多态性。结果表明:其中4个微卫星标记扩增效率较低,被剔除;其他8个微卫星的多态性丰富,平均等位基因数为11.38,平均期望杂合度为0.6686,平均多态信息含量为0.6354,适合于我国肉牛亲子鉴定。     

鲁西牛群体遗传多样性与生长发育性状的微卫星标记研究

以120头鲁西牛为研究对象,选用分布于牛60条染色体的30对微卫星引物中多样性丰富的8对微卫星位点,探讨与鲁西牛生长发育性状相关联的分子遗传标记。本研究采用微卫星标记技术和最小二乘线性模型,检测了鲁西牛群体的遗传多样性和分析了与鲁西牛生长发育性状相关的标记效应。结果,共检测到34个等位基因,每个微卫星座位的等位基因数为3～6个,平均等位基因数为4.25,该群体的基因平均杂合度(H)为0.561 3,多态信息含量(PIC)为0.498 3。结果表明,7个微卫星座位的不同基因型之间的差异达到显著(P<0.05)或极显著水平(P<0.01)。影响相应性状的最有利基因型分别是ETH185座位的AE与体高,BM711座位的CC和BM1824座位的BB与体长,TGLA53座位的AD和BM711座位的CC与管围,BM1824座位的BB和DVGA55座位的AB与腹围,TGLA53座位的AC、ETH185座位的BD、BM711座位的AB、DVGA46座位的AC、DVGA44座位的BB和DVGA55座位的BB与体质量。而对生长不利的基因型有:DVGA46的AD型,DVGA44的AD型,DVGA55的AA型和ETH185的BC型。...     

利用微卫星DNA标记分析槟榔江水牛群体遗传特征

槟榔江水牛是近年在我国云南西部发现的第一个本土河流型水牛.为了揭示其群体遗传多样性、群体遗传组成、其与河流型和沼泽型水牛的遗传关系及沼泽型水牛对该水牛的基因渗入等重要遗传背景信息,本研究采用30个微卫星DNA标记对141份槟榔江水牛样品和对照群体24份河流型水牛(摩拉水牛)样品、41份沼泽型水牛(滇东南水牛)样品进行了检测分析.结果,在槟榔江水牛群体中共检测到253个等位基因,其中,54个为3个群体所共享的等位基因,58个为只在槟榔江水牛与摩拉水牛间共享的等位基因,73个为只在槟榔江水牛与滇东南水牛间共享的等位基因,其余68个等位基因为该群体所独有.槟榔江水牛平均等位基因数、平均表观杂合度、平均期望杂合度和多态信息含量等参数均显著高于对照组群体,分别为8.433 3、0.640 5、0.600 9和0.594 7.该水牛群体近交系数FIS值为0.061 9.槟榔江水牛与摩拉水牛间的DA遗传距离为最小(0.114 4),在NJ树上聚为一支;而滇东南水牛与槟榔江水牛和摩拉水牛间的遗传距离较大(分别为0.382 9和0.555 3),其在NJ树上单独聚为一支.群体遗传结构分析显示,槟榔江水牛遗传组分为河流型与沼泽型2种类型水牛混合的模式(当K=2时),整个群体中有相当数量的个体存在沼泽型水牛的遗传渗入(群体中沼泽型水牛遗传组分为0.067 2).结果揭示,槟榔江水牛遗传资源独特,群体遗传多样性丰富,但该群体杂合子缺乏,且存在一定沼泽型水牛的基因渗入.     

利用微卫星标记分析5个家兔品种内的遗传多样性

本研究利用微卫星标记分析了我国5个家兔品种内的遗传多样性。结果表明:在5个品种中,新西兰兔有效等位基因数最多,为6.545 5;加利福尼亚兔有效等位基因数最少,为3.000。比利时兔平均多态信息含量和平均杂合度最大,分别为0.570 4和0.823 3;加利福尼亚兔平均多态信息含量和平均杂合度最小,分别为0.499 7和0.589 7。     

安徽省3个地方鸡品种的遗传多样性分析

运用微卫星标记对淮南麻黄鸡、皖南三黄鸡和五华鸡3个地方品种遗传多样性进行了分析。在5个座位中,共检测到23个等位基因数,平均有效等位基因数为1.5396,杂合度为0.2920,多态信息含量为0.4156。3个地方品种中,淮南麻黄鸡遗传多样性最低,五华鸡遗传多样性最高。根据5个微卫星座位等位基因频率计算群体遗传一致度和标准遗传距离并构建系统发生树,五华鸡与皖南三黄鸡遗传关系最近,皖南三黄鸡与淮南麻黄鸡最远。研究结果与这些地方品种的地理分布、育成历史相一致,同时也佐证了微卫星标记是研究鸡群体遗传关系的一个有用工具。     

应用微卫星标记分析湘西黄牛与其他黄牛品种的亲缘关系

为了分析湘西黄牛与国内外其他品种的亲缘关系,利用8个微卫星标记对湘西黄牛进行了遗传多样性分析。结果发现湘西黄牛等位基因丰富,在57~72个之间;平均多态信息含量在0.8547~0.9024之间,为高度多态性;平均杂合度在0.8867~0.9248之间;各杂交牛之间都存在一些特有等位基因和缺失等位基因;通过Nei＇s聚类分析,其与引进种公牛的遗传距离较大,与国内4个优良肉牛品种相比,可以单独聚为一类。