基于ISSR分子标记的褐马鸡亲缘关系分析

褐马鸡(Crossoptilon mantchuricum)是中国特有濒危鸟类,国家一级保护动物。为了保护褐马鸡种质资源,保障驯养繁育种群和再引入种群的遗传基因结构优化,采用ISSR分子标记技术,对山西庞泉沟国家自然保护区和太原动物园两个种群35个褐马鸡进行了亲缘关系分析。从20条ISSR引物中筛选出10条扩增条带清晰稳定、重复性好的引物,共扩增出65条DNA条带,其中77%呈多态性。两个种群各个体之间的Nei's无偏差遗传距离和遗传相似度分析表明:个体间遗传相似性的平均值分别为0.5061与0.7591,遗传距离的平均值分别为0.4939与0.2409。用组平均法对35个褐马鸡个体进行聚类分析,结果显示:大多数同一种群的个体首先聚在一起,显示出生态地理条件相似的种群亲缘关系较近。     

26个川茶花品种亲缘关系的ISSR分析

采用ISSR分子标记,对重庆南山植物园中26个品种的川茶花古树进行了ISSR遗传多样性分析。选用14条扩增带型清晰且重复性好的引物共获得166条谱带,其中140条呈多态性,多态性比例(PPB)为84.34％。根据Nei-Li遗传相似性系数在0.708处,UPGMA结果将26个品种分为5个类群,在0.726处可进一步分为10个亚类群;26个川茶花品种的遗传相似性系数介于0.615和0.913之间,平均值为0.704,表明品种间亲缘关系较近。其中牡丹茶与花牡丹之间遗传相似性系数最大,为0.913,表明牡丹茶可能为花牡丹的芽变品种。川渝地区山茶古树资源丰富,但在古树的保护上也面临诸多问题,建议采取相应的措施加以保护。     

基于ISSR分子标记的叶子花亲缘关系分析和指纹图谱构建

为分析品种遗传多样性和遗传距离并构建品种聚类图和指纹图谱,该研究从DNA模板浓度、引物浓度、退火温度和循环次数等方面优化了叶子花ISSR-PCR反应体系和反应程序,利用11个ISSR引物对131个叶子花品种进行PCR扩增,扩增产物经琼脂糖凝胶电泳检测.结果表明:优化的ISSR-PCR反应体系中DNA模板浓度为0.5 n...     

大戟属植物亲缘关系的ISSR分析

利用ISSR标记对9种大戟属植物的亲缘关系进行了分析.结果表明:筛选出的24条ISSR引物共扩增251条带,其中多态性条带248条,多态率为98.8%.9种大戟属材料的遗传相似系数在0.51～0.69之间,平均为0.62.利用UPGMA法将9种大戟属植物分别聚为3个类群,第一类群包含地锦亚属与一品红亚属的5个种及大戟亚属的绿玉树,第二、第三类群分别包含大戟亚属中的3个种.ISSR标记获得的聚类结果与根据形态特征获得的分类结果有很大的差异.     

基于SSR标记的草莓品种亲缘关系分析

摘要：用18对SSR引物对96份草莓栽培品种资源进行扩增。在18个SSR位点共获得184个条带,其中多态性条带为172个,多态性条带比率为93.5％,不同引物扩增的多态性条带数为6～19个,平均为9.56个。各位点PIC值在0.5726～0.8885之间,平均0.8057。NTSYS软件进行相似性系数计算,UPGMA法进行聚类分析结果显示,96份来源不同的草莓品种被混杂地聚在一起,草莓品种的亲缘关系与其来源并不存在明显的相关性,中国地方品种的聚类相对集中,中国地方品种与中国选育品种之间亲缘关系相对较远。欧美品种和日本品种的遗传基础要宽于中国选育品种和地方品种。     

不同地区麦冬遗传多样性的ISSR分析

利用ISSR分子标记技术对5个地区6个麦冬居群进行遗传多样性分析。选用10条扩增带型清晰且重复性好的引物进行扩增,共获得115条带,其中89条具有多态性,多态性比例为77.39%。当遗传相似系数(GS)为0.61时,可将6个麦冬居群分为2大类群。居群的GS平均值为0.643,表明供试居群之间存在较近的亲缘关系,且地域越近的其亲缘关系关系越近。研究表明ISSR分子标记技术能够很好地用于不同地域同种物种的亲缘关系分析。     

基于SSR分子标记的酸浆属植物亲缘关系研究

近年来, 由于营养价值高、果实可当水果食用且具有潜在的药用价值, 酸浆属(Physalis)植物在全球范围内受到了越来越多的关注。采用SSR分子标记技术对我国范围内主要分布的4种酸浆属植物的22份样品进行了亲缘关系研究, 结果表明, 20对SSR引物共扩增出118个位点, 其中107个(90.7%)为多态性位点; 平均种间遗传相似系数为0.501。UPGMA聚类和PCoA分析结果得出相似结论, 并且将供试酸浆属植物样品分为两大类。其中, 酸浆(P. alkekengi var. francheti)的所有样品与其它酸浆属植物的遗传距离最远, 单独聚为一类, 与前人的研究结果非常吻合。研究表明, SSR标记遗传信息丰富, 可以用于酸浆属植物的亲缘关系分析。研究结果为酸浆属种质资源保护提供了有效信息, 并且为酸浆属植物的分子辅助育种奠定了重要基础。     

基于ISSR方法的芦笋育种亲本间亲缘关系分析

为了解芦笋(Asparagus officinalis L.)育种亲本间的遗传关系,采用ISSR方法对19份芦笋育种亲本进行亲缘关系和遗传多样性分析。结果表明,18条引物共扩增出187条谱带,多态性比率达93.6%。UPGMA聚类分析表明,亲本的相似系数为0.56~0.80,在相似系数0.62处可将亲本分成3大类群,第Ⅰ类群为美国芦笋品种?绿龙?;第Ⅱ类群由荷兰品种?金林?人工培育的部分亲本和美国品种?格兰德?组成;第Ⅲ类群为其余亲本组成。同一品种不同个体间也可能存在较大的遗传距离,芦笋经倍性育种培育出的个体表现出丰富的遗传多样性。这为芦笋杂交育种中的亲本筛选提供了理论依据。     

基于枇杷转录组序列的SSR分子标记引物开发

为获得更多的枇杷SSR引物,对枇杷转录组测序得到的1 kb以上的11 798条Unigenes进行SSR位点搜索。结果在3515 条Unigenes（6.77%）中共获得4438个SSR位点,其中主要重复类型为双碱基重复和三碱基重复,二者占SSR总数的68.27%,而四、五、六碱基重复类型较少,仅占1.42%。对选出的SSR标记采用Primer3进行引物设计,得到7911 对SSR位点特异引物,可用于枇杷遗传多样性分析、分子标记辅助育种、育种群体的建立等研究。     

利用ISSR和SRAP标记分析细辛资源遗传多样性与亲缘关系

采用ISSR、SRAP分子标记对61份细辛资源进行遗传多样性与亲缘关系进行分析,结果表明:(1)ISSR标记平均每条引物可获得8.35个DNA片段,多态性比率为86.3%,SRAP标记平均每对引物可获得7.85个DNA片段,多态性比率为86.0%。(2)利用相同数量的引物,ISSR标记揭示的多态性略高于SRAP标记。(3)按照种质间相似系数得出聚类图,可将所有细辛资源分开,在依据ISSR标记聚类分析中,生物学上北细辛和汉城细辛的划分,其作用不如地域来源的效应。SRAP分子标记中,大部分资源的聚类与地域性有关,但有4份汉城细辛优先聚类,SRAP分子标记在揭示基因组差异方面有一定的优势。(4)2种分子标记的聚类图中,来自同一产地的北细辛和汉城细辛优先聚类,其亲缘关系更近。聚类图中未出现北细辛与汉城细辛分别聚类。分子标记分类与传统植物学分类不一致。     

白沙蒿不同地理种群遗传分化的ISSR分析

运用ISSR技术,对白沙蒿的5个种群进行遗传分化的分析。12个引物在108个个体中共扩增出222个位点,其中,多态位点为218个,多态位点百分率为98.20%,Shannon多样性指数为0.315 4,具有高的多态性。种群间的遗传分化系数（G_st）为0.076 7,与AMOVA分析的结果一致（Ф_st为7.96%）,表明绝大多数的遗传变异存在于种群内部。各种群间遗传一致度高达98%以上,遗传距离很小,基因流达3.008 2,均表现出白沙蒿各种群存在着广泛的基因交流,有着很小的遗传分化。     

Genetic Diversity of Kenaf (Hibiscus cannabinus) Evaluated by Inter-Simple Sequence Repeat (ISSR)

Understanding genetic diversity is very useful for scientific utilization for breeding. In this study, we estimated the genetic distances in a panel of 84 kenaf accessions collected from 26 countries and regions using ISSR markers. The results of UPGMA analysis showed that kenaf germplasm had abundant genetic variation, with genetic dissimilarity coefficients ranging from 0.01 to 0.62. The in-group dissimilarity coefficient (0.29) was observed in 84 kenaf accessions, and all the accessions could be divided into three groups: cultivars (L_1–1), relatively wild species (L_1–2 and L_1–3), and wild species (the others). Further in-group analysis in group L_1–1 (0.19) revealed that the kenaf cultivars could be divided into five subgroups with distinct regional characteristics. It is imperative that genes be exchanged among all kinds of tested varieties from different origins. The results provide a useful basis for kenaf germplasm research and breeding.     

Population genetics of Ageratina adenophora using inter-simple sequence repeat (ISSR) molecular markers in China

Abstract

Understanding distribution and diversity of invasive weeds is essential for the development of efficient control measures against it. In the present study, inter-simple sequence repeat (ISSR) markers were used to assess the biogeographic relationships among populations of the invasive Crofton weed (Ageratina adenophora (Spreng.)) during 2004–2006 in China. A total of 100 ISSR primers with di-, tri-, tetra- and penta-nucleotide repeats were screened, from which 20 polymorphic and informative primers were selected. Amplification of the 20 primers generated a total of 479 polymorphic bands among the 64 weed populations, and a high level of genetic diversity (H _E = 0.1541 ± 0.0193) was detected in A. adenophora. Neighbor-joining (NJ) cluster analysis based on genetic distances among populations grouped the populations according to their geographical origin, i.e. (1) populations of southwestern Guizhou, (2) populations of Liangshan city in Sichuan, (3) populations of western Guizhou, (4) Guangxi populations plus Chongqing populations, (5) populations of southern Yunnan, and (6) populations of Yangtze River Valleys in Sichuan plus populations of western Yunnan. A significant positive correlation between geographical and genetic distance was detected by the Mantel test (r = 0.183, p = 0.0012). Based on the divergence relationships revealed by ISSR markers, it was assumed that A. adenophora mainly dispersed through wind and water in China.     

Assessment of Genetic Diversities of Selected Laminaria (Laminariales,Phaeophyta) Gametophytes by Inter-Simple Sequence Repeat Analysis

Inter-simple sequence repeat (ISSR) analysis was used to assess genetic diversity among 10pairs of male and female Laminaria gametophytes. A total of 58 amplification loci was obtained from 10selected ISSR primers, of which 34 revealed polymorphism among the gametophytes. Genetic distances were calculated with the Dice coefficient ranging from 0.006 to 0.223. A dendrogram based on the unweighted pair-group method arithmetic (UPGMA) average showed that most male and female gametophytes of the same species were clustered together and that 10 pairs of gametophytes were divided into four groups. This was generally consistent with the taxonomic categories. The main group consisted of six pairs of gametophytes, which were selected from Laminaria japonica Aresch. by intensive inbreeding through artificial hybridization. One specific marker was cloned, but was not converted successfully into a sequence characterized amplified region (SCAR) marker. Our results demonstrate the feasibility of applying ISSR markers to evaluate Laminaria germplasm diversities.     

Assessment of Genetic Diversities of Selected Laminaria （Laminariales, Phaeophyta） Gametophytes by Inter-Simple Sequence Repeat Analysis

Inter-simple sequence repeat (ISSR) analysis was used to assess genetic diversity among 10 pairs of male and female Laminaria gametophytes. A total of 58 amplification loci was obtained from 10 selected ISSR primers, of which 34 revealed polymorphism among the gametophytes. Genetic distances were calculated with the Dice coefficient ranging from 0.006 to 0.223. A dendrogram based on the unweighted pair-group method arithmetic (UPGMA) average showed that most male and female gametophytes of the same species were clustered together and that 10 pairs of gametophytes were divided into four groups. This was generally consistent with the taxonomic categories. The main group consisted of six pairs of gametophytes, which were selected from Laminaria japonica Aresch. by intensive inbreeding through artificial hybridization. One specific marker was cloned, but was not converted successfully into a sequence characterized amplified region (SCAR) marker. Our results demonstrate the feasibility.of applying ISSR markers to evaluate Laminaria germplasm diversities.     

Genetic Diversity and Phylogenetic Relationships in the Genus Lycopersicon (Tourn.) Mill. as Revealed by Inter-Simple Sequence Repeat (ISSR) Analysis

Inter-simple sequence repeat (ISSR) analysis was for the first time used to study the genetic diversity and phylogenetic relationships in 54 wild accessions and cultivars of the genus Lycopersicon. Analysis involved 14 ISSR primers homologous to microsatellite repeats and containing additional selective anchor nucleotides. In total, 318 ISSR fragments were amplified for the wild and cultivated tomato genomes. The interspecific polymorphism revealed with the ISSR primers was 95.6%. Species-specific ISSR fragments were detected for each tomato species. The highest number (more than 20) of species-specific fragments were obtained for L. esculentum sensu lato, although the intraspecific variation of ISSR patterns was low. UPGMA cluster analysis was used to construct a dendrogram and to estimate the genetic distances between the species of the genus Lycopersicon; between populations ofL. peruvianum, L. pimpinellifolium, and L. esculentum; and between tomato cultivars. The ISSR-based phylogeny was generally consistent with Lycopersicon taxonomy based on morphological and molecular evidence, suggesting the applicability of ISSR analysis for genotyping and phylogenetic studies in tomato.     

应用ISSR-PCR分析蒙古栎种群的遗传多样性

本研究应用ISSR标记技术对东北地区的优势树种蒙古栎(Quercus mongolica)的25个种群遗传多样性进行了分析, 目的是为蒙古栎早期选择提供依据。从60条ISSR引物中筛选出10个特异性强、稳定性好的引物进行ISSR分析。共获得位点数71个, 其中多态位点数56个, 多态位点百分率为78.87%。PopGene分析结果表明: 种群的平均多态位点百分率为45.2%, Shannon表型多样性指数(I)平均值为0.25, 具有较高的遗传多样性, 种群间存在一定程度的基因流(N_m为1.3818)和遗传分化(Nei’s信息多样性指数平均值为0.1068, G_st平均值为0.2657), 种群内的基因多样度占总种群的73.43%, 种群间占26.57%, 表明蒙古栎种群的变异主要来源于种群内。结合聚类分析和地理变异规律把种群划分为两个大的种群组: 小兴安岭种群组和长白山种群组。以上结果可为栎属种质资源的保护和利用以及物种分化研究提供基础资料。     

森林生态系中球孢白僵菌遗传多样性的ISSR分析

应用ISSR分子标记对安徽大别山区的球孢白僵菌遗传多样性进行了研究。从33个引物中筛选出12个多态性高、稳定性好的ISSRs用于正式的扩增分析,在2个自然保护区、3个不同季节和3个不同海拔梯度采集的48个菌株中共扩增出84条带,其中73条为多态性条带,多态性为81％,平均每个引物扩增出7条（2～11）。群体的多态位点百分率（PPL）达81％,Nei’s基因多样性（H）为0.3187,Shannon信息指数（I）为 0.4782。居群间的基因分化系数较小（Gst）0.1028。以上结果表明：安徽大别山区球孢白僵菌有较高的遗传多样性, 居群间遗传变异较小,居群内表现出较高水平的遗传分化。     

40份玉米自交系的ISSR遗传多样性分析

利用ISSR分子标记技术对40份玉米自交系进行亲缘关系分析。从59条ISSR引物中筛选出10条重复性高、多态性好的引物,分别对全部供试材料进行扩增,共扩出90条清晰谱带,其中多态性条带81条,多态性比率为90.00%,表明供试材料基因组DNA的多态性较高。用 NTSYSpc-2.10软件中的 UPGMA进行聚类分析,40份玉米自交系的遗传相似系数变化范围在0.65~1.00之间,在此基础上构建聚类分析树状图,揭示了玉米各自交系间的亲缘关系。本研究为今后分子水平上玉米优良自交系的改良与选育提供了一定的参考依据。