Genetic variation in relation to adaptability of three mangrove species from the Indian Sundarbans assessed with RAPD and ISSR markers

Rich genetic polymorphism is important for plants to adapt to changes because it enables the plant to make anatomical,physiological and biochemical changes in response to abiotic stress.Geomorphologic characteristics,demographic interference and a cumulative decrease in freshwater influx in the Indian Sundarbans region have proved detrimental to some economically important plants.In this study,genetic polymorphism of three mangrove species,Xylocarpus granatum,Excoecaria agallocha,and Phoenix paludosa,was assessed using RAPD and ISSR molecular markers.X.granatum,already in distress in the Sundarbans,had the least genetic polymorphism,14.56%in the RAPD analysis and 12.92% in the ISSR.Relatively higher genetic polymorphism was recorded for the profusely growing E.agallocha and P.paludosa: 24.66 and26.4% in RAPD; 24.87 and 20.32% in ISSR analysis respectively.A UPGMA dendrogram constructed using the similarity matrix from RAPD,ISSR and combined datashowed that for X.granatum,the least and highest salinity zones clustered together,whereas for E.agallocha and P.paludosa,higher and lower salinity areas clustered in different clades.Nei's genetic diversity,calculated from RAPD and ISSR data,was also in accordance with 0.0637 and 0.0583 for X.granatum,respectively,much lower than0.0794 and 0.0818 for E.agallocha and 0.0799 and 0.0688 for P.paludosa.This opposing degree of polymorphism might be attributed to the profusely growing E.agallocha and P.paludosa and precarious status of X.granatum throughout the Indian Sundarbans.

A Genetic Linkage Map of Kenaf (Hibiscus cannabinus L.) Based on SRAP, ISSR and RAPD Markers

Kenaf (Hibiscus cannabinus L.) is one of the most economically important crops for non-wood fiber production. The objective of this study was to establish a genetic linkage map of kenaf with higher density of molecular markers. A semi-wild variety Ga42 and a cultivar Alain kenaf were used as parents to construct an F2 population consisting of 155 plants. The genetic linkage map comprising 134 marker loci was constructed, including 65 sequence-related amplified polymorphism (SRAP), 56 inter-simple sequence repeat (ISSR), and 13 randomly amplified polymorphic DNA (RAPD) markers. This map spans 2 108.9 cM and contains 20 linkage groups with an average marker density of 15.7 cM between the adjacent markers.

Assessing Genetic Structure and Relatedness of Jerusalem Artichoke (<i>Helianthus tuberosus</i>L.) Germplasm with RAPD, ISSR and SRAP Markers

Jerusalem artichoke (Helianthus tuberosus L.) is an old tuber crop with a recently renewed interest in multipurpose improvement, but little effort has been made to characterize its genetic resources. A study was conducted to assess genetic structure and genetic relatedness of 47 diverse Jerusalem artichoke accessions using RAPD, ISSR and SRAP markers. A total of 296 (87.1%) polymorphic bands were detected from 13 RAPD markers;92 (80%) from six ISSR primers;and 194 (88.6%) for nine combinations of SRAP primers. Five optimal clusters were inferred by the STRUCTURE program from the RAPD or ISSR data, while six optimal clusters were found from the SRAP data or combined marker data. Significant linear relationships between the distance matrices for all pairs of individual accessions were detected for all marker pairs and the estimated correlation coefficient was 0.40 for RAPD-ISSR, 0.53 for RAPD-SRAP, and 0.43 for ISSR-SRAP. Based on the combined data, the neighbor-joining clustering of the 47 accessions matched closely with those inferred from the STRUCTURE program. Three ancestral groups were observed for the Canadian germplasm. Most diverse germplasm harbored in the USA collection. These findings not only reveal the compatible patterns of genetic structure and relatedness inferred with three marker types, but also are useful for managing Jerusalem artichoke germplasm and utilizing diverse germplasm for genetic improvement.

Genetic analysis of selected Sargassum fusiforme (Harvey) Setchell (Sargassaceae, Phaeophyta) strains with RAPD and ISSR markers

Since the 1980s,Sargassum fusiforme has been cultivated in Zhejiang,South China,and nowadays it becomes one of the important commercial seaweeds in China.With traditions of eating habits in the East Asian countries,this brown alga is used as food,because it contained functional oligo/polysaccharides and chemical components,and was regarded playing roles in antioxidant activities and regulating immunology.Through over 15 years’selection,breeding and cultivation,we obtained three strains with good traits and testified their characters during the production,which included the cultivars with high yield and other two good characters,either all the selected strains were applied in the Sargassum production.To avoid confusion during the selection and nursery,it was preferred to establish one fingerprint for distinguishing the Sargassum cultivars from different strains.Random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR)methods were adopted to analyze the genetic diversities of the selected S.fusiforme strains.With that,one fingerprint with RAPD markers was constructed,and one sequence characterized amplifi ed region(SCAR)marker to S.fusiforme was obtained.It is indicated that the applied fingerprint could be valid in S.fusiforme genetic and germplasm justification,and will be positive to molecular marker assistance in its selection and cultivation.

基于ISSR和RAPD分子标记的15株糙皮侧耳遗传多样性分析

平菇(糙皮侧耳)是我国食用菌栽培主要品种之一,国内各地栽培品种较多,平菇资源的遗传多样性较丰富。为了明确平菇菌株间的亲缘关系及遗传距离,借助ISSR和RAPD两种分子标记技术,对收集到的国内主产区的15个平菇菌株进行DNA遗传多样性分析,为后期平菇适应性试验提供理论依据。通过ISSR和RAPD两种分子标记引物筛选试验,获得9对多肽引物,利用9对引物对15份平菇DNA样品进行多样性PCR扩增,并采用NTSYS-PC分析软件对供试菌株的遗传多样性进行聚类分析。结果显示,参试平菇菌株遗传距离在0.26~0.56;9对引物共扩增产生了102条清晰条带,平均每对引物扩增产生11.3条带;多态性条带90条,多态性条带比例为88%,平均每对引物组合扩增产生10条多态性条带;在遗传距离0.36的水平上,供试的15个平菇品种分为4个类群,P101和P121分别单独分为一个类群,P102、P112、P120、P122、P104、P125、P107、P113、P124归为一个类群,P117、P123、P118、P119归为一个类群。供试材料之间遗传差异大,遗传多样性较丰富,研究结果为本地区平菇产业发展及育种提供新的菌株。

Identification of Tremella fuciformis strains Using SRAP, ISSR and RAPD markers

Three types of molecular markers (SRAP, ISSR and RAPD) were used to identify four Tremella fuciformis strains, T6 (white), T7 (white), T8 (yellow) and T9 (light yellow). Twelve SRAP primer pairs, ten ISSR primers and eight RAPD primers were screened, and identification data obtained using the three molecular markers were consistent in that the four T. fuciformis strains were divided into three groups with T7 and T9 clustered together in a single group. Each RAPD primer generated a higher average number of polymorphic bands than either the SRAP or ISSR primers, and the average similarity between the four strains was 81.34%. SRAP markers reflected more genetic information compared with the two other markers, and the average similarity was 68.98%. Genetic information reflected by ISSR markers was intermediate between SRAP and RAPD, and the average similarity was 77.48%.

基于形态标记和ISSR分子标记的长瓣铁线莲遗传多样性分析

为快速区分鉴定市场上繁多的长瓣铁线莲(Clematis macropetala)品种,了解长瓣铁线莲间的遗传多样性水平及亲缘关系,本试验利用形态标记结合ISSR分子标记对25个长瓣铁线莲品种和1个野生种进行遗传多样性研究。结果表明:14个数量形状的变异系数在0.24%~2.32%之间;15个假质量性状的信息多样性指数H在0~2.29%之间,遗传多样性指数D在0~0.94%之间;在欧式距离为15时,可将29个表型性状划分为七类,在欧式距离为15时,可将26个长瓣铁线莲划分为五类。12条引物共扩增出144个条带,多态性条带占比为100%。利用引物UBC815、UBC824和UBC836构建26个长瓣铁线莲的指纹图谱,可快速区分鉴定26个长瓣铁线莲。UPGMA聚类结果显示,在遗传相似系数为0.68时,可将26个长瓣铁线莲划分为八大类。形态标记结合ISSR分子标记可有效鉴别长瓣铁线莲种质资源,以期为长瓣铁线莲种质资源收集保存、创制新品种等提供理论基础。

Genetic Diversity of Tunisian and Chinese Alfalfa (Medicago sativa L.) Revealed by RAPD and ISSR Markers

Medicago sativa L.) is one of the most important forage crops in the world. The genetic variability analysis of 19 alfalfa populations collected from three sites in South Tunisia (Gabes, Kebili, Tozeur) and 1 from North West China were carried out using Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers. Five RAPD primers amplified 44 bands of which 22 were polymorphic;and five ISSR primers amplified 51 bands of which 33 were polymorphic. The percentage of polymorphic bands detected by RAPD and ISSR was 50% and 64.7%, respectively. The resolving power (Rp) varied between 0.6 and 4.1 with an average of 2.02 for RAPD marker and between 0.7 and 6.5 with an average of 2.28 for ISSR marker. However the Average Informativeness band (AvIb) was ranged from 0.2 to 0.9 with an average of 0.5 in RAPD marker and from 0.29 to 0.7 with an average of 0.624 in ISSR marker. The RAPD marker revealed less within population genetic diversity than ISSR marker. Although Cluster (UPGMA) and Correspondence Factorial Analyses (CFA) indicate that populations’ clustering made independently both from the geographical origin.

Assessment of Genetic Variation in Soybean (<i>Glycine max</i>) Accessions from International Gene Pools Using RAPD Markers: Comparison with the ISSR System

Soybean (Glycine max) is one of the most important crops in the world in terms of total production and usage. It is also among the least diverse species. The main objectives of the present study were to 1) assess the level of genetic variation among soybean (G. max) accessions from different countries using Random Amplified Polymorphic DNA (RAPD) markers and 2) compare Inter Simple Sequence Repeats (ISSR) and RAPD marker systems in detecting polymorphic loci in soybeans (G. max). Genomic DNAs from 108 soybeans (G. max) accessions from 11 different gene pools were analyzed using several ISSR and RAPD primers. The average level of polymorphic loci detected with the RAPD primers was 35%. The soybean accessions from the China, Netherlands, and Canada gene pools were the least genetically variable with 25%, 26%, and 30% of polymorphic loci, respectively. Accessions from Hungary (43%) and France (48%) showed the highest level of polymorphism based on the RAPD analysis. Overall, RAPD data revealed that the accessions from different countries are closely related with 64% genetic distance values below 0.40. The levels of polymorphic loci detected with the RAPD and ISSR marker systems were in general moderate and similar even if they target different regions of the genome. A combination of different marker systems that include RAPD/ISSR, microsatellites (SSR), and SNPs should provide the most accurate information on genetic variation of soybean (G. max) accessions.

鸡血藤ISSR体系优化及引物筛选

以鸡血藤的入药部位藤茎为试验材料,采用改良CTAB法进行DNA提取,利用正交试验和单因素试验对ISSR-PCR反应体系进行优化和构建。结果表明,鸡血藤最佳ISSR-PCR反应体系为DNA模板60 ng,Taq酶2.00 U,Mg2+浓度1.75 mmol/L,dNTP浓度0.25 mmol/L,引物浓度0.20 μmol/L,最佳退火温度44.6℃。从100条ISSR引物筛选出20条适合鸡血藤的ISSR引物。

RAPD和ISSR标记检测黄麻属遗传多样性的比较研究

应用RAPD和ISSR标记,分别对来自非洲地区和中国的15份黄麻野生种(10个种),以及来自中国、印度、越南、日本等地的12份黄麻栽培品种基因组DNA的遗传多样性进行检测。结果表明:(1)参与分析的所有RAPD和ISSR引物都对DNA模板浓度的梯度变化不敏感,对比RAPD和ISSR在PCR反应中的稳定性,ISSR优于RAPD;(2)25个RAPD和ISSR引物分别扩增出329条和283条带,多态比率分别为89.36%和92.5%,显然,ISSR检测出的多态性条带的能力优于RAPD;(3)RAPD和ISSR标记检测同组供试材料种间或种内的遗传相似性系数(GS)范围,分别为0.48～0.98和0.33～0.97,ISSR检测出种间遗传多样性的分辨力较RAPD为高,但两者GS相关系数高达0.955;(4)RAPD和ISSR标记的分子聚类获得了趋势相近,但不完全相同的聚类树,两种标记均能准确地把原始黄麻野生种与栽培种中的长果种和圆果种及其近缘野生种聚在不同的种群中,分子分类结果与传统经典分类相符,并揭示出种间或种内基因型的遗传差异与亲缘关系,而ISSR比RAPD能检测到种间更高的遗传差异性;(5)两种标记均可揭示种间与种内的遗传多样性及其进化的亲缘关系,可为进一步开展黄麻分子辅助育种和起源与进化研究提供有价值的理论依据。

辣椒种质遗传多样性的RAPD和ISSR及其表型数据分析

用RAPDI、SSR分子标记及28个表型性状数据对辣椒属5个栽培种的13份材料进行了分析,结果表明:23条RAPD引物共扩增出209条带,平均每个引物扩增出9.09条,多态性位点比率为83.73%;16条ISSR引物共扩增出94条带,平均每个引物扩增出5.88条,多态性位点比率为79.79%.与RAPD相比,ISSR标记检测到的有效等位基因数(Ne)及Shannon多样性指数(I)、遗传离散度(Ht)和遗传分化系数(Gst)等遗传多样性参数都较大,多态性位点比例在亲缘关系较近的一年生辣椒(Capsicum annuum)种内较高,说明ISSR有更高的多态性检测效率,并且适合亲缘关系较近的种群间遗传多样性分析.基于RAPDI、SSR的聚类与基于表型数据的聚类之间存在极显著正相关,且都能将C.annuum与其它栽培种区分开来.

利用RAPD和ISSR分子标记分析地黄种质遗传多样性

用RAPD与ISSR技术对地黄的 8个品种和 2个脱毒品系进行了种质遗传多样性分析。分别从 80条RAPD引物和 4 4条ISSR引物中筛选出适合地黄种质分析的 17条RAPD引物和 10条ISSR引物用于RAPD和ISSR分析。 17条RAPD引物共扩增出 177条带 ,多态性位点数为 10 9;多态性位点比率为 6 1 5 8% ;平均多样性指数 (I)为0 3135 ;每个位点的有效等位基因数 (Ne)是 1 36 4 1;10条ISSR引物共扩增出 110条带 .多态性位点数为 79;多态性位点比率为 71 5 8% ;平均多样性指数 (I)为 0 35 77;每个位点的有效等位基因数 (Ne)是 1 4 0 37。基于扩增条带数据库建立了各自的Jaccard遗传相关系数矩阵 ,构建了相似的分子树状图 ,将 10个供试材料分为 2类 :一类群含组培 85 5、大田 85 5、组培 930 2、大田 930 2、金状元和金白 6个材料 ;另一类群含北京 1号、大红袍、地黄 910 4和野生地黄 4个材料。两种分子标记的分析结果呈极显著正相关 (r=0 6 4 9)。结果表明 ,RAPD与ISSR标记适合于地黄种质遗传多样性分析 。

用RAPD、ISSR和AFLP标记分析系谱关系明确的甘薯品种的亲缘关系

用RAPD、ISSR和AFLP标记对系谱关系明确的7个甘薯品种进行了亲缘关系分析。24个RAPD引物、14个ISSR引物和9对AFLP引物分别扩增出173、174和168条多态性带。3种分子标记在检测甘薯品种间遗传差异上相关程度高，其中RAPD与ISSR之间的相关系数最大为0．9328。用ISSR标记估计的品种间遗传距离为0．1286～1．0932，平均0．4883．大于其余2个标记的估计值。3种分子标记皆可揭示甘薯品种的亲缘关系，其中ISSR标记产生的聚类图与系谱图最吻合，认为ISSR标记更适于分析甘薯品种的亲缘关系。

苦瓜种质遗传多样性的RAPD和ISSR分析

采用RAPD和ISSR分子标记技术对38份苦瓜种质进行遗传多样性分析。结果表明：10个RAPD和10个ISSR51物分别扩增出93条和81条带，多态性比率分别为50．54％和61．29％；RAPD和ISSR标记检测供试材料的遗传相似性系数（Gs）范围，分别为0．287～1和0．221～1，ISSR（平均GS值0．672）检测多态性效果高于RAPD（平均G5值0．694）。RAPD标记聚类分析将供试种质分为3个类群6组，分类结果与苦瓜瓜瘤的表型分类比较相似；ISSR标记聚类分析将供试种质分为3个类群7组，ISSR标记划分类群与形态上以颜色分类比较接近。RAPD和ISSR标记的遗传相似性系数呈显著相关（r=0．550）。两个标记整合后聚类分析可检测到更大的遗传变异，结果与苦瓜的农艺性状分类和地理分布有一定的相关性。

正品龙胆遗传多样性的RAPD及ISSR分析

目的用RAPD和ISSR方法分析了4种正品龙胆的亲缘关系及遗传多样性,为正品龙胆的品种鉴定及栽培育种提供了分子生物学依据。方法优化RAPD及ISSR的PCR反应体系,对产物琼脂糖凝胶电泳结果进行数据统计。结果从100个RAPD引物和32个ISSR引物中分别筛选出10个RAPD引物和4个ISSR引物。通过遗传距离系数UPGMA聚类法分析数据,构建类聚关系树系图。结论RAPD及ISSR的PCR反应体系可以获得理想的实验结果,适用于龙胆品种遗传多样性及亲缘关系的分析。

耐旱苔藓植物DNA提取及优化RAPD、ISSR反应体系的建立

耐旱苔藓植物常常单个个体矮小、生物量低,如何从细小的单个个体中有效提取总DNA是进一步开展居群遗传多样性研究的关键。本研究以古尔班通古特沙漠广泛分布的刺叶墙藓(Tortula desertorum)为对象,使用快速提取法、2×CTAB法及DNeasy plant mini kit试剂盒提取法等3种方法对刺叶墙藓单个个体的总DNA进行提取。结果表明,2×CTAB法提取的DNA纯度高,凝胶电泳显示无明显降解现象,适宜作为PCR扩增的模板。利用所提取的单个个体DNA为模板,建立了优化的RAPDI、SSR反应体系。

果蔗种质资源的RAPD和ISSR分析

利用RAPD和ISSR分析 3 0份果蔗种质的遗传背景 ,筛选出几个扩增条带清晰、多态程度较高、稳定可靠的引物 (其中RAPD引物SS和ISSR引物WW可一次性准确地检测出黑皮果蔗、地方果蔗和自育杂交种间的区别 ) ,而且ISSR又比RAPD检测到更多的遗传多态性 (二者的多态性条带比分别为 61 .3 5 %和 5 6.8% )。

用RAPD和ISSR检测的橡胶树野生种质和栽培品种的遗传多样性(英文)

用19个RAPD引物和12个ISSR引物对14份野生橡胶树种质和我国的37份栽培品种进行了遗传多样性分析.RA PD 引物共产生132条带, 多态性带占88.6% ,相似系数变化范围在0.432-0.947,ISSR引物共产生101条带,多态性带占87.1% ,相似系数为0.505-0.941。平均基因杂合度分析表明野生种质比栽培品种具有较高的遗传多样性。根据UPGM A 法对51份材料进行聚类分析,结果表明,ISSR分析中所有材料可分为2类:第一类为野生种质,第二类为栽培品种;而RAPD分析中野生种质和栽培品种不能被分为明显的两大类。虽然ISSR和RAPD的聚类分析结果存在差异,但对两种方法进行的相关分析表明,他们之间仍存在极显著相关性,相关系数为0.574。品种PR107、热研217等一些栽培品种可以通过特异带在51份供试材料中被区分开。这些结果可以对橡胶树的育种工作起到一定的指导作用,同时RAPD和ISSR 技术也是进行橡胶树品种鉴定和遗传多样性研究的有效手段。

20份新疆紫花苜蓿种质RAPD和ISSR遗传多样性分析

【目的】采用RAPD和ISSR分子标记对新疆20份野生紫花苜蓿种质进行遗传多样性分析。【方法】用梯度PCR仪对RAPD引物和ISSR引物进行扩增筛选,筛选出多态性好的引物用于20份种质材料的研究。【结果】通过筛选分别得到6对RAPD和ISSR引物对20份新疆紫花苜蓿进行遗传多样性分析。6对RAPD引物共扩增出93个条带,多态性带91个,多态带百分率97.85%;平均每引物扩增出15.50条带,每引物平均扩增出的多态条带15.17。6对ISSR引物共扩增出157个条带,多态性带152个,多态带百分率96.82%;平均每引物扩增出26.17条带,每引物平均扩增出多态条带25.33。RAPD标记的Nei’s基因多样性变异范围为0.176 3~0.274 0,平均0.207 1;ISSR标记的Nei’s基因多样性范围为0.085 0~0.154 1,平均0.113 3。【结论】2种标记聚类分析结果均表明种质聚类与地理来源有一定的相关关系。