中国南瓜海南农家品种间的遗传特异性分析和DNA指纹图谱构建

在收集中国南瓜海南农家品种的基础上,本研究应用ISSR和SRAP标记技术对28份海南农家品种间的遗传特异性进行了分析,并构建指纹图谱,为中国南瓜海南农家品种鉴定、评价、保护和利用提供科学依据。结果表明,所供试的品种间存在显著的遗传特异性,具有特殊的遗传基础或背景,所筛选的6个ISSR引物和11对SRAP引物共产生了10个特异标记和11条唯一缺失带;应用ISSR引物组合UBC807/UBC814/UBC844/UBC868和UBC808/UBC814/UBC844/UBC868,以及SRAP引物组合Me1/Em2 Me1/Em10 Me2/Em3和Me1/Em1 Me1/Em10 Me8/Em3分别绘制了四张28份中国南瓜海南农家品种的DNA指纹图谱,所构建的DNA指纹图谱直观、简单。ISSR标记和SRAP标记技术可有效应用于中国南瓜海南农家品种DNA指纹图谱的构建和遗传特异性鉴定。     

Genetic diversity and population structure of Butea monosperma (Lam.) Taub.- a potential medicinal legume tree

Three molecular marker systems, Random Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeats (ISSR) and Sequence-Related Amplified Polymorphism (SRAP) were employed to investigate the genetic structure and diversity among the 14 natural populations of Butea monosperma collected from different geographical regions of India. Detected by 17 RAPD, 15 ISSR and 11 SRAP primer combinations, the proportions of polymorphic bands were 84.2 %, 77.2 % and 91.9 %, respectively, and the mean Nei’s genetic distances among the populations were 0.13, 0.10 and 0.13, respectively. Partitioning of genetic variability by Analysis of molecular variance (AMOVA) revealed that the high genetic diversity was distributed within the populations. AMOVA also revealed that the coefficient of gene differentiation among populations based on F_ST was very high irrespective of markers used. The overall gene flow among populations (Nm) was very low. Cophenetic correlation coefficients of Nei’s distance values and clustering pattern by Mental test were statistically significant for all three marker systems used but poor fit for ISSR data than for RAPD, SRAP and combined data set of all three markers. For all markers, a high similarity in dendrogram topologies was obtained, although some differences were observed with ISSR. The dendrogram obtained by RAPD, SRAP and combined data set of all three markers reflect relationship of most of the populations according to their geographic distribution.     

生境片段化对千岛湖岛屿上黄足厚结猛蚁遗传多样性的影响

利用相关序列扩增多态性(SRAP)分子标记法,对千岛湖片段化生境中14个岛屿上的黄足厚结猛蚁(Pachycondyla luteipes)种群遗传结构和多样性进行研究。利用5对SRAP引物对42份材料的基因组进行扩增,共得到大小在50-800 bp之间的71个可重复位点,其中63个为多态性位点,多态性比率达88.73%。AMOVA分析结果显示,65.03%的遗传变异存在于种群间,34.97%的遗传变异来自种群内(P < 0.001)。利用PopGene Version 1.32软件对SRAP多态性数据进行分析,不同岛屿种群的多态位点比例和Nei's基因多样性指数变化范围分别介于35.21%-91.55%和0.2662-0.4905之间,平均值分别为58.25%和0.3729,其中多态位点比率最高的岛屿为面积最大的JSE岛。多态位点比例和Nei's基因多样性指数与岛屿面积、海拔均无显著相关性,但与隔离度呈显著正相关关系。种群间遗传分化指数介于0.0777-0.9328之间,平均值为0.4419,基因流值介于0.0360-5.9350之间,平均值为1.0451,种群间遗传分化程度较高,基因流较低。利用UPGMA聚类分析法对14岛屿上的42个个体进行遗传聚类分析,表明地理距离较近的个体和岛屿具有优先聚在一起的趋势。Mantel 检验表明黄足厚结猛蚁各种群间地理距离与遗传距离间存在显著相关性(r=0.7757,P < 0.01)。以上结果表明地理隔离是影响千岛湖黄足厚结猛蚁种群遗传结构和多样性的主要因素。     

利用SRAP标记分析贵州砂梨资源遗传多样性

采用SRAP标记技术,对贵州59份砂梨种质的遗传多样性进行分析。结果表明:(1)筛选的15对SRAP引物共扩增出151个位点,其中120个是多态性位点,多态百分率为79.47%。(2)POPGENE分析表明,贵州砂梨资源在物种水平上有效等位基因数(Ne)为1.355 9,Nei’s基因多样性指数(H)为0.216 9,Shannon’s信息指数(I)为0.336 2;在区域品种群水平Ne=1.261 1,H=0.155 5,I=0.235 2,以黔西南地区资源的遗传多样性最高,六盘水市最低。(3)聚类分析显示,在遗传相似系数0.75处,可将59份砂梨资源分为6个组,聚类分析结果具有一定的地理区域特性。研究认为,贵州砂梨种质具有较为丰富的遗传多样性,并表现出明显的地理区域特性;SRAP标记与ISSR标记在揭示贵州砂梨种质遗传多样性上具有高度的一致性,但SRAP标记能在更小的变异范围内揭示更多的遗传信息。     

海南岛油茶种质资源遗传多样性的SRAP分析

为了解海南岛油茶(Camellia oleifera)种质资源的遗传多样性,采用SRAP分子标记,对海南岛油茶主要分布区的31个居群进行了遗传多样性和亲缘关系分析。结果表明,海南岛油茶资源的遗传多样性低,物种水平的多态性百分率(PPB)为98.30%,Nei’s基因多样性(H)为0.222 8,Shannon信息指数(I)为0.353 8;居群水平的PPB=40.96%,观测等位基因数(Na)为1.409 6,有效等位基因数(Ne)为1.237 1, H=0.138 5, I=0.208 3,这与海南岛油茶丰富的表型多样性水平不一致。海南岛油茶资源遗传分化较大,居群间基因交流有限,不同居群间的遗传分化指数(Gst)为0.380,基因流(Nm)为0.813 91。遗传变异主要发生在居群内,有38.05%的变异存在居群间,61.95%存在于居群内。遗传距离为0.022 6～0.276 4,平均为0.107 7,居群间的亲缘关系较近。UPGMA聚类分析表明,在遗传距离为0.11处,可将31个油茶居群聚为6类,表现出明显的行政区域性,而与地理距离关系不大。因此,海南岛油茶资源遗传多样性低,亲缘关系近可能导致自交或近交不亲和,可能是海南油茶林分花量大而结实低的主要内在原因。     

SRAP技术在遗传的研究进展

SRAP是一种新型的DNA分子标记,具有简便、稳定、中等产率和容易得到选择条带序列的特点。SRAP利用独特的引物设计对开放读码框(ORFs)进行扩增,上游引物长17bp,对外显子进行特异扩增,下游引物长18bp,对内含子区域、启动子区域进行特异扩增,因个体不同及其物种的内含子、启动子与间隔长度不等而产生多态性。本文阐述SRAP的原理和操作流程,综述了SRAP标记目前在植物遗传图谱构建、遗传多样性、基因定位、基因克隆、杂种优势利用等方面的研究进展及应用前景。     

Physiological,Biochemical and Molecular Responses of Barley Seedlings to Aluminum Stress

Barley (Hordeum vulgare L.) is one of the most Aluminum (Al) sensitive cereal species. In this study, the physiological, biochemical, and molecular response of barley seedlings to Al treatment was examined to gain insight into Al response and tolerance mechanisms. The results showed that superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activity were inhibited to different degrees following Al exposure. The MDA content also significantly increased with increasing Al concentrations. SRAP results indicated significant differences between Al treatments and controls in terms of SRAP profile, and the genomic template stability (GTS) decreased with increasing Al concentration and duration. These integrative results help to elucidate the underlying mechanisms that the barley response to Al toxicity.     

西南地区乡土杨树遗传变异的SRAP分析

采用SRAP分子标记技术对西南地区11种乡土杨树共333份样本的遗传变异进行分析,7对引物组合共扩增出215条带,其中多态性条带158条,多态性条带百分率为73.49%,表明11种乡土杨树间存在广泛变异。AMOVA分析结果显示,种间遗传变异分量为10.84,占总变异的48.70%,遗传差异达极显著水平(P0.001)。种间的遗传相似系数变幅在0.8199~0.9607之间,平均遗传相似系数为0.8983。聚类结果表明,昌都杨和藏川杨之间的遗传差异最小,大叶杨和三脉青杨之间的遗传差异最大。本研究结果为西南地区乡土杨树基因资源的保护、开发和利用提供了一定的科学理论依据。     

A comparative analysis of genetic diversity in medicinal Chrysanthemum morifolium based on morphology, ISSR and SRAP markers

The diversity and genetic relationship among 29 populations of Chrysanthemum morifolium, one of Chrysanthemum indicum and one of Chrysanthemum nankingense from China were analyzed using morphological traits and molecular markers. Twenty morphological traits were scored as well as 182 ISSR marker-fragments, as amplified by 22 primers [the percentage of polymorphic bands (PPB): 81.87%], and 243 SRAP marker-fragments as generated by 26 primer pairs (PPB: 75.72%). Mantel’s test indicated significant correlation (r = 0.624) of morphological trait and SRAP. By contrast, the morphological trait showed low correlation with ISSR (r = 0.246). Cluster analysis showed groupings of the accessions according to all four methods correlated well with their geographic region of origin, and most populations from the south of China were classified into one cluster and most populations from the north of China were classified into another cluster. Finally, an appropriate strategy for conserving the C. morifolium germplasm was proposed.     

The linkage maps of Dendrobium species based on RAPD and SRAP markers

Dendrobium plants are used commonly as tonic herbs and health food in many Asian countries,especially in China.Here we report the genetic map construction of two Dendrobium species with a double pseudo-testcross strategy using random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) markers.A F1 mapping population of 90 individuals was developed from a cross between D.officinale and D.hercoglossum.A total of 307 markers,including 209 RAPD and 98 SRAP,were identified and used for genetic linkage group (LG) analysis.The D.officinale linkage map consisted of 11 major linkage groups and 3 doublets,which covered 629.4 cM by a total of 62 markers with an average locus distance of 11.2 cM between two adjacent markers.The D.hercoglossum linkage map contained 112 markers mapped on 15 major and 4 minor linkage groups,spanning a total length of 1,304.6 cM with an average distance of 11.6 cM between two adjacent markers.The maps constructed in this study covered 92.7% and 82.7% of the D.hercoglossum and D.officinale genomes respectively,providing an important basis for the mapping of horticultural and medicinal traits and for the application of marker-assisted selection in Dendrobium breeding program.