37份龙眼种质资源亲缘关系的ISSR分析

本研究利用ISSR技术对37份龙眼种质资源进行遗传多样性检测。研究结果表明,从100条ISSR引物中筛选出7条重复性好,条带清晰的引物对37份龙眼品种基因组DNA进行扩增,共扩增出54条带,其中43条具有多态性,比率为79．6％。不同龙眼品种间遗传相似系数变幅为0．69～0．97,平均达0．83,说明ISSR标记能够揭示材料间较高的遗传多样性。UPGMA聚类结果表明,ISSR标记能将37份龙眼品种完全区分开,并能将来源于中国、越南和泰国的37份龙眼品种分别聚类到中国、越南和泰国三大品种群,说明龙眼品种资源的亲缘关系与地理因素有关,三个国家的龙眼品种之间存在较大的遗传差异。本研究结果将为为龙眼品种资源的研究利用提供参考。     

用ISSR分析四川苎麻品种(系)间的遗传关系及雄性不育分子标记的建立

本文从100条ISSR引物中筛选出21条引物,对8个四川苎麻品种(系)间的遗传关系进行了分析。PCR扩增结果表明,21条引物在8份材料中共扩增出86条带,平均每条引物扩增出4.1条,其中多态性位点71个,各引物扩增出的位点数3~8个不等,平均每条引物可以检测到3.4个多态性位点。聚类分析和遗传距离分析结果表明,供试品种川7、川8与其亲本遗传距离较远。3个杂交品种(川7、川8、川9)均表现特有的偏父本遗传现象。此外,本研究用ISSR引物U835筛选到了1个雄性不育分子标记,并将其扩增产物克隆测序,结果表明该序列大小为658bp。根据该序列,此标记被转化成稳定的SCAR标记,可用于苎麻雄性不育分子标记辅助育种。     

14个南丰柑桔品种及品系间的ISSR遗传多样性分析

柑桔是江西省南丰县生物资源中十分有特色的种质资源。本研究利用ISSR分子标记对南丰柑桔品种的遗传多样性进行研究。从100个ISSR引物中筛选出17个多态引物,对14份柑桔样品进行ISSR-PCR分析。结果显示,共扩增出162条DNA条带,其中多态性条带为134条,占82.71%,其片段大小在200～2000bp之间。UPGMA聚类分析结果表明,以遗传相似系数为0.67为分界线,可以将14份供试柑桔样品分成三类:第一类包含以ss-28为代表的7个柑桔品种;第二类包含以杨小2,6为代表的3个柑桔品种第三类包括以早系97-1为代表的4个柑桔品种。本研究结果对于南丰蜜桔的辅助育种或品种(株系)苗木鉴别具有重要的参考价值。     

利用小麦SSR标记分析鸭茅种质资源的遗传多样性

使用小麦(Triticum aestivum)SSR引物和扩增程序,以中国春小麦(T. aestivum)品种为对照,利用SSR标记对来自国内外的45份鸭茅(Dactylis glomerata L.)种质资源进行遗传多样性研究.结果表明,20对引物扩增出295个条带,多态性条带为187条,多态性条带比率为61.15%,鸭茅种质资源的遗传相似系数范围为0.7848～0.9513.聚类分析和主成分分析将供试材料分为6大类,聚类结果不仅能反映鸭茅生态适应性特征与其生长发育状况及生产性能相关,还显示出国产鸭茅品种遗传基础较为狭窄.研究表明将小麦SSR引物用于检测鸭茅遗传多样性行之有效.     

大豆种质资源RAPD标记遗传多样性研究

为深入研究并充分利用野生大豆资源,本文利用RAPD分子标记对40份大豆材料加以分析,旨在从DNA分子水平上探索野生大豆、地方品种和育成品种之间的遗传多样性状况。结果表明:50个RAPD引物筛选出具有多态性且扩增条带清晰的引物38个,共检测出407条带,其中多态谱带309条,多态性程度为75.92%。每个引物可扩增出2～14条多态性带,平均产生多态性谱带8.1条;平均多样性指数为2.3377,变幅范围为0.5865～4.2133。遗传相似系数变幅范围为0.44～0.92,平均为0.75。野生大豆的多态比例(94.35%)、多样性指数(2.2336)分别高于育成品种(87.47%、1.7331)和地方品种(83.54%、1.6198)。遗传相似系数为野生大豆(0.6498)地方品种(0.7015)育成品种(0.7177),育成品种与地方品种间为0.6599,育成品种与野生大豆间为0.6487,地方品种与野生大豆间为0.6045。UPGMA聚类分析结果表明,40份大豆材料聚为6类,育成品种和地方品种各自聚为一类,野生大豆聚为4类。野生大豆特异等位基因数远远高于育成品种和地方品种二者的相加之和。本研究从分子水平上揭示了野生大豆与栽培大豆区别明显,宜作为一个独立的种,同时野生大豆变异幅度大,遗传基础广,是大豆育种实践中的优良基因资源。     

马铃薯遗传资源多样性的SRAP分析

采用SRAP分子标记，对44份马铃薯品种的遗传多样性进行了分析。结果显示, 随机抽取27对SRAP引物，对基因组DNA进行特异扩增，其中23对引物扩增出多态性条带，共获得104个多态性条带, 多态性引物比率达85.2%, 平均每对引物产生4.5个多态性条带，表明SRAP标记具有较高的多态性比率。44份种质资源的SRAP标记遗传距离为0.147-0.741。当遗传距离D=0.67时，将44份种质资源分为四大类群，包括一个复合类群和三个独立类群。其中复合类群可进一步分为7个亚群。从而在DNA水平上证明了所研究马铃薯种质资源具有丰富的遗传多样性。     

SSR和SRAP标记研究油菜杂交种骨干亲本的遗传多样性

用SSR和SRAP两种分子标记方法研究51份甘蓝型油菜杂交种亲本系的遗传多样性,并对两种分子标记研究结果进行比较。结果发现,在51份材料中,45对SSR引物共扩增出194条多态性条带,平均每对引物为4.3条,25对SRAP引物共扩增出197条多态性条带,平均每对引物为7.9条。UPGMA聚类分析表明,SSR和SRAP标记都可将51份亲本材料划分为五大类群,本所选育的玻里马细胞质雄性不育系（Polima CMS）的主要保持系和恢复系都聚在同一类群的不同亚群中。根据系谱资料分析发现,SRAP标记划分的类群与系谱资料更为接近,SRAP标记更适用于遗传关系较近材料的遗传多样性分析。SRAP标记揭示的亲本间遗传距离要大于SSR标记揭示的遗传距离。两种不同标记方法揭示出油菜亲本遗传多样性的差异主要是由不同的标记方法揭示的标记位点等位基因变异数不同造成的。     

小麦杂种优势群研究III普通小麦和斯卑尔脱小麦微卫星分子标记遗传差异的研究

选用１５份我国不同生态区的普通小麦品种（系）及９份不同国家的斯尔脱小麦品种（系）,利用微卫星分子标记对小麦种间,品种（系）间遗传差异进行了研究,探讨扩大杂交小麦育种亲本遗传基础的途径。所利用的１８对微卫星引物的２４份材料中均有搁增产物,共扩增出分子量小于５００ｂｐ的条带４９５条,其中４６８条带（占９６．７８％）具有多态性,平均每个引物可拉增出２６．６条多态性带。研究发现,９份斯卑尔脱小麦微卫星多态     

应用RAMP标记研究黑麦属遗传多样性*

对黑麦属(Secale L．)10个野生居群和11个栽培居群共21份材料进行了RAMP(random amplified microsatellite polymorphism)标记分析，结果表明，被测材料间RAMP标记多态性较高。80个：RAMP引物中，有41个引物(占50．5％)可扩增出清晰且具多态性的条带。这41个引物共扩增出445条带，其中428条(占95．9％)具有多态性，每个引物可扩增出3—19条多态性带，平均10．4条。RAMP标记遗传相似性系数(GS)变异范围为0．266-0．658，平均值为0．449。RAMP标记可将所有21份黑麦材料完全区分开，聚类结果与材料的地理分布有一定关系，但与黑麦属传统的系统分类体系存在明显差异。据此认为，RAMP标记可以有效地评价黑麦属植物的遗传多样性，并为其物种亲缘关系的界定提供信息。     

基于ISSR指纹的甘薯食用品种的遗传多样性分析

运用ISSR分子指纹技术检测甘薯基因组DNA的多态性,是鉴定甘薯遗传多样性和亲缘关系有效的方法之一。本研究选用6个ISSR标记对17份甘薯品种进行了DNA指纹扩增,共扩增出55条谱带,其中多态性谱带50条,多态性水平为90.91%;UBC825引物扩增出带型最多,有16种;根据Nei’s遗传距离计算获得的聚类树状图表明,在GS为0.625时,参试的17份甘薯品种分为4个组群;遗传相似性分析表明,顶芽菜用品种间遗传相似系数最高0.877,叶柄食用品种和烤薯型品种之内的遗传相似系数分别为0.778和0.740。结果表明甘薯主要食用间具有较好的遗传多样性,但同类型品种间却又有较高的遗传相似性,这类品种需要引进或创制新资源。     

菊属11个野生种和12个栽培品种遗传关系的ISSR分析

为进一步明确菊属野生种与栽培品种的遗传关系和多样性,本研究利用ISSR分子标记技术对菊属11个野生种和12个栽培品种之间的遗传关系进行比较分析。从75个ISSR引物中筛选出了14个引物,对供试材料的DNA进行扩增,共获得142条清晰可辨的谱带,多态位点比率为95.1%;菊属野生种的平均有效等位基因数（effective number of alleles,Ne）、平均Nei＇s基因多样性指数（Nei’s gene diversity,H）及Shannon信息指数（shannon’s information index,I）均高于栽培菊花,说明各野生种间基因差异比较显著,多态性强于栽培品种。UPGMA聚类结果表明：菊属野生种呈现由低倍向高倍进化的趋势;栽培菊花之间遗传关系复杂,大体可以推断出平瓣是菊花的基本瓣形;菊花脑与栽培菊花亲缘关系最近,小红菊、龙脑菊、若狭滨菊与栽培菊花关系亦较近,神农香菊与其它材料关系最远。本研究的结果表明菊属野生种与栽培品种之间遗传关系比较复杂,而ISSR分子标记技术可以较好地从分子水平上揭示出菊属植物间的遗传关系。     

利用ISSR揭示不同类型月季遗传多样性

本研究应用正交设计法对ISSR反应体系中的各个主要影响因子进行了优化筛选,确立了适合月季ISSR-PCR反应的最佳体系。结果表明,25μL的ISSR反应体系中各组分的最适浓度分别为：1×PCR缓冲液、1U Taq DNA聚合酶、800pmol／L 引物、0.16mmol／L dNTPs、Mg^2＋ 1.5mmol／L。筛选了33个ISSR引物,共得到了11个多态性比较高的ISSR引物,占所筛引物的33.33％。利用筛选出的11条ISSR引物对3种月季类型的23份月季材料进行遗传多样性分析,共扩增出477条DNA带,其中多态性位点有14个,平均每条引物可以检测到4.5个多态性位点。用NTSYS软件对样品进行了UPGMA聚类分析,聚类结果表明丰花月季基本能聚为一类,切花月季与藤本月季交叉聚在一起。这表明月季种质的遗传差异与其应用分类的相关性不紧密。     

Assessment of genetic diversity in Solanum trilobatum L., an important medicinal plant from South India using RAPD and ISSR markers

Solanum trilobatum L. is an Indian medicinal plant containing rich amount of steroidal glyco-alkoloids that can be used as precursor for commercial steroid production. Two efficient marker systems such as Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) were used for the first time to assess the genetic diversity across 14 S. trilobatum accessions obtained from five South Indian states. Twenty out of 60 RAPD primers generated 189 distinct bands of which 160 were polymorphic with an average of 8 polymorphic bands per primer. A maximum of up to 15 fragments were amplified with an average of 9.45 bands per primer and the amplicons varied in size between 100 and 3,000 bp. The percentage of polymorphism ranged from 55.5 to 100, with an average of 84.6. ISSR profiling using 7 out of 20 primers amplified 83 bands and the number of amplified fragments varied from 2 to 16 with a size range of 200–1,800 bp. Totally 72 polymorphic bands were obtained using 7 ISSR primers at an average of 10.28 polymorphic bands per primer. Polymorphism percentage varied from 50 to 100 among the selected accessions resulting in an average percentage of polymorphism of 86.7. The PIC values ranged from 0.49 to 0.93 for RAPD and 0.16 to 0.90 for ISSR primers. The study pointed out that ISSR markers were more efficient than RAPD markers in evaluating the degree of genetic variation in S. trilobatum. The UPGMA cluster analysis grouped all Tamil Nadu accessions in one cluster and other state accessions in another cluster. The Principal component analysis also substantiates this clustering pattern. Thus the phylogenetic relationship and a high genetic variation revealed in the present study could provide a baseline data for conservation and improvement of this plant in future. Also the molecular markers identified in this study will be helpful in authentication of this species to prevent adulteration in herbal medicine.     

Analysis of Genetic Diversity and Relationships in Waxy Rice (Oryza sativa L.) using AFLP and ISSR Markers

Opaque endosperm is the main phenotypic indicator for waxy rice, but other phenotypic and genotypic variation among waxy rice accessions has largely been ignored. Previous studies showed that wide diversity in starch physiochemical properties exists in both indica and japonica waxy rices, especially for starch gelatinization temperature (GT) which could be divided into a high- and a low-GT group. In the present study, amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat (ISSR) molecular markers were employed to examine genetic diversity and relationships of 56 waxy rice accessions. A total of 358 AFLP fragments were amplified with five primer combinations, showing a high level of polymorphism (78.3%). A total of 190 ISSR bands were generated with a single primer and a primer pair, showing a very high level of polymorphism (92.2%). The genetic distance matrices obtained from the two sets of markers were significantly correlated (r = 0.731, P = 0.004). The dendrogram generated with combined AFLP and ISSR markers could clearly differentiate the indica and japonica groups. Newly released varieties and breeding lines within each subspecies tended to be clustered together, whereas landraces were more distantly placed in the dendrogram. Only one AFLP band was found specific to the indica type, while no specific bands were found for starch GT. The implications for the conservation and breeding of waxy rice are discussed.     

基于ISSR分析28份香蕉种质的基因组DNA多样性

利用ISSR标记技术分析了28份香蕉种质的遗传多态性。从100个ISSR引物中筛选出8个多态性引物,共扩增出55条DNA带,其中46条为多态性带,占83.6%,平均每个引物扩增的DNA带数为6.88条。依相似系数0.73的水平,将香蕉28个品种划分为6大类。其中云南BB（BB）和东莞高把大蕉（ABB）在相似系数为0.94时,二者的亲缘关系较近。Pisang Ceylan（AAB）和FHIA-18（AAAB）相似系数水平接近为1,表明二者亲缘关系最近。本研究为香蕉遗传关系的建立及品种鉴定提供理论基础。     

Comparative Analysis of Genetic Diversity in Two Tunisian Collections of Fig Cultivars Based on Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats Fingerprints

This study characterized the genetic diversity of 18 Tunisian fig cultivars using random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR). Both random and ISSR primers tested generated a total of 116 RAPD and 47 ISSR markers. Considerable genetic variation was observed among fig cultivars sampled from two regional Tunisian collections with an average diversity of 4.57. RAPD and ISSR banding patterns and genetic distances values reflected the high level of diversity among the collections and lower variability between the two collections. The correlation between the RAPD and ISSR similarity matrices computed for the 153 pairwise comparisons among the 18 varieties was lower and significant. An analysis of molecular variance showed that 92% of the total genetic diversity resided within collections, whereas only 8% between collections. The results indicated that in the local fig germ plasm the information provided by RAPD and ISSR is not analogous, most likely as a consequence of the fact that the two classes of markers explore, at least in part, different portions of the genome.     

甜菜品种（系）的ISSR标记数字指纹图谱构建及聚类分析

摘要：本文针对来源于荷兰的4个引进甜菜品种和国内的6个甜菜品系（其中2个为一年生野生甜菜）进行了ISSR指纹图谱构建和聚类分析研究。筛选出稳定性高且多态性好的6个引物用于试验。利用筛选的6条引物ISSR-PCR 共扩增出51个条带, 其中多态性条带百分率为86.3%. 利用该6条引物ISSR-PCR建立的指纹图谱能将试验中的全部甜菜品种都鉴定区分开。只利用2条引物L1和UBC846 扩增的8个多态性条带构建了10个甜菜品种（系）的数字指纹识别码,该数字指纹图谱能完全区分10个甜菜品种（系）,结果显示ISSR 指纹图谱能非常有效的鉴定不同的甜菜品种。利用生物软件NTSYS-pc针对10个试验甜菜品种（系）的ISSR 扩增条带进行遗传相似性聚类分析,结果显示10个甜菜品种（系）的相似系数为0.43与0.83之间,平均为0.62。利用非加权组平均法（UPGMA）进行聚类分析,结果显示10个甜菜品种（系）聚类为2个组和3个亚组。UPGMA 聚类分析能清楚的显示10个甜菜群体间的遗传关系并且聚类结果与10个甜菜群体的特性一致, 说明ISSR标记能用于甜菜不同群体间遗传距离的评估。     

微卫星标记分析我国南方常规水稻品种的遗传差异

利用12个DNA微卫星标记对2005-2007年参加南方稻区国家水稻品种区域试验的23个长江中下游早籼、晚粳和华南早籼常规品种进行标记检测,结果共检测到等位基因47个,平均每个标记3.9个,变幅2～7个,多态性频率（FP）平均值0.632,变幅0.403～0.842。遗传差异分析表明：长江中下游早籼与长江中下游晚粳(平均遗传相似系数AGSC=0.036)、华南早籼与长江中下游晚粳（AGSC=0.125）品种间的遗传差异明显,长江中下游早籼与华南早籼（AGSC=0.449）品种间的遗传差异中等,华南早籼（AGSC=0.606）、长江中下游早籼（AGSC=0.682）、长江中下游晚粳（AGSC=0.761）品种间的遗传差异较小。     

Inter-Simple Sequence Repeat fingerprints to assess genetic diversity in Tunisian fig (Ficus carica L.) germplasm

The genetic diversity of 18 Tunisian fig cultivars was investigated at the DNA level using the Inter Simple Sequence Repeat (ISSR) associated with the Polymerase Chain Reaction (PCR). Using a set of primers, the most informative ones were selected that were characterized by an important Resolving power value of 29.6. A total of 47 discernible fragments were scored from samples, with a mean of 11.7 fragments per primer. The 90.4% of sample that were polymorphic were scored as molecular markers to examine the Tunisian fig germplasm polymorphism at DNA level. A large genetic diversity as related to ISSR patterns was found within the local Tunisian fig germplasm. An UPGMA dendrogram exhibits the unstructured variability in this crop. Moreover, the principal component analysis shows that the observed diversity was typically continuous. Our data provide a large number of ISSR markers that are useful in the fingerprinting of Ficus carica L. cultivars, and in the understanding of the genetic relationships among these accessions.