西瓜种质资源遗传多样性的SRAP分析

【研究目的】研究西瓜材料之间的亲缘关系及其分类,为进一步利用和创新材料、培育新的品种提供依据;【方法】采用SRAP技术对64份西瓜种质资源的遗传多样性进行了研究;【结果】从700多个引物组合中筛选出了51个条带清晰、多态性高的引物组合来分析供试材料,共产生431条扩增带,其中243条带具有多态性,多态性频率平均为56.4%,除去非洲野生型57号材料,其它材料间多态率为39.4%。将243条多态性SRAP条带,利用NTSYS软件计算了64个材料间的遗传相似系数,其变化范围为0.47~0.97,除非洲野生型材料57号等4个差异较大的材料,其它材料间变化范围仅为0.87~0.97;【结论】说明目前西瓜育种的大多数材料间同源性较高,遗传分化较小,其遗传基础非常狭窄。该研究结果对利用特殊种质、合理选择杂交亲本具有一定的参考价值。     

利用RAPD分子标记分析玉米种质遗传多样性

以RAPD分子标记技术对玉米的10个品种的全基因组DNA进行PCR扩增,再用Popgene32软件和SPSS 13.0软件分析扩增结果,研究10个玉米的种质资源遗传关系。结果表明：（1）所选20个引物可扩增出214条RAPD条带;（2）所选引物扩增条带的多态性比率为86.4%,RAPD分子标记扩增10个玉米品种间的相似性系数分别在0.316-0.654之间;（3）用RAPD-PCR扩增条带的分析结果建立了遗传相关系数矩阵、构建了分子树状图、可将10个玉米品种分为3个类群;（4）RAPD分子标记适合于构建玉米的DNA指纹图谱,进行品种鉴定和遗传分析。     

36个马铃薯品种的SSR分析

为明确36个马铃薯品种在DNA分子水平上的遗传差异,利用SSR分子标记技术对其多态性进行了分析。试验从90对SSR引物中筛选出适宜于36个马铃薯品种基因组DNA扩增的引物10对,PCR扩增获得301个SSR条带,多态性条带百分率为71.1%。引物STACCA3和SSR111扩增的SSR指纹图谱差异明显,可作为36个马铃薯品种间鉴别的分子依据。36个马铃薯品种间的遗传距离(GD)变幅为0.33～0.85,平均为0.54,其中,大于GD平均值的品种有20个,占供试品种的55.56%。以GD值0.57为基准,将36个马铃薯品种划分成7类,从DNA分子水平揭示出各供试品种间的亲缘关系。     

11个茶树品种遗传多样性的ISSR和TRAP比较分析

【研究目的】分析茶树种质资源的遗传多样性,比较ISSR和TRAP在茶树中的标记效率;【方法】用12个ISSR引物和18个TRAP引物组合分别对11个茶树品种的基因组DNA进行扩增;【结果】ISSR引物共产生61条带,多态性带占83．6％,遗传距离变化范围在0．231～0．707。TRAP引物共产生69条带,多态性带占79．7％,遗传距离变化范围为0．200～0．755。基于两种标记的聚类结果存在一定的差异,TRAP聚类结果能较好地体现供试品种的亲缘关系、地域特性和树型特点,与传统的基于形态特征建立的分类结果比较一致。在11个茶树品种间以及在亲缘关系比较近的黄旦、铁观音、毛蟹、白样观音4个品种间,ISSR的标记指数MI值都高于TRAP的MI值;【结论】ISSR具有更大的标记效率。也具有较高的多样性检测能力;TRAP比ISSR更适合于茶树特异种质资源和重要农艺性状的筛选。     

基于SSR标记新疆陆地棉的DNA指纹图谱构建及遗传多样性分析

【目的】利用Simple sequence repeats(SSR)标记构建新疆近40年间审定的120个陆地棉品种的DNA指纹图谱并进行遗传多样性分析。【方法】从586对候选引物中筛选得到78对多态性高、扩增稳定且均匀分布于棉花染色体上的引物,并用这78对引物构建120个陆地棉品种的DNA指纹图谱。【结果】78对核心引物在120个材料中检测到392个等位位点,其中多态性位点324个,多态性比率达82.7%。24个标记位点在17个品种上具有特征谱带。采用12对引物组合即可鉴定120个棉花品种。聚类分析显示,120个陆地棉品种遗传相似系数变化范围为0.50~0.96,平均为0.73,遗传相似系数偏高,表明新疆陆地棉品种的遗传基础较狭窄。【结论】引物组合法是构建DNA指纹图谱最有效的方法。遗传相似系数矩阵将120个陆地棉品种分为三大类群,与系谱来源较为吻合。     

龙眼种质资源的RAPD分析

利用RAPD技术对龙眼的95份种质资源和一份荔枝种质资源进行研究。从200条随机引物中筛选出18条多态性引物,共扩增出197条带,DNA扩增带的多态性为100%,这表明96份材料的遗传多样性和遗传资源的丰富性。利用SPSS13.0统计软件对RAPD表型数据矩阵进行分析。根据欧氏距离（ED）法计算各样品间的遗传距离,采用UPGMA（类平均法）,进行分析,建立亲缘关系聚类树状图。结果表明,RAPD标记能在分子水平揭示龙眼的遗传背景与亲缘关系     

甜菜SCoT核心引物的筛选

【研究目的】为了筛选出适合甜菜品种遗传多样性分析的SCoT引物,为SCoT引物得以在甜菜中进行深入应用奠定基础,本研究甜菜以8个来自不同国家的甜菜品种为材料对80条SCoT引物进行扩增【方法】扩增方法采用SCoT-PCR最优反应体系包含2.0 μL的10×PCR buffer（含Mg2+）、10 ng的DNA、0.5 U的Taq DNA聚合酶、10 μmol/L的引物2 μL以及0.1 μL的dNTPs (2.5 mmol/L each)。【结果】结果从80条SCoT引物中筛选出20条多态性高的引物,这20条引物最多扩增出16条多态性条带,最少扩增出2条多态性条带,扩增总条带155条,其中多态性条带为134条,多态性条带的百分比为86.5%。【结论】通过对核心引物的有效性验证,表明筛选出的20个核心引物非常适合用于甜菜指纹图谱的构建。     

西瓜种质资源的遗传多样性及亲缘关系的AFLP分析

利用AFLP指纹图谱分析技术对57个西瓜材料进行了基因组DNA多态性分析。结果表明,18对AFLP引物共产生709条DNA带,平均每个引物组合扩增出39.4条带,其中440条为多态性带,占总带数的61.4%;根据DNA谱带计算品种间遗传相似系数,其变化范围在0.24-0.99之间,平均相似系数为0.70,说明西瓜品种间遗传基础狭窄。用NTSYS软件进行聚类分析,57个西瓜品种被划分为2个类群。其中类群1包括1个野生西瓜,类群2均为栽培种。在类群2中引进的资源聚在一起,杂交种和双亲或其一聚在一起。此外,本研究表明,AFLP指纹分析技术具有很高的分辨率,能够揭示西瓜品种间的差异,是鉴定西瓜品种的有力工具。     

西瓜种质资源的遗传多样性及亲缘关系的AFLP分析

利用AFLP指纹图谱分析技术对57个西瓜材料进行了基因组DNA多态性分析.结果表明,18对AFLP引物共产生709条DNA带,平均每个引物组合扩增出39.4条带,其中440条为多态性带,占总带数的61.4%;根据DNA谱带计算品种间遗传相似系数,其变化范围在0.24～0.99之间,平均相似系数为0.70,说明西瓜品种间遗传基础狭窄.用NTSYS软件进行聚类分析,57个西瓜品种被划分为2个类群.其中类群1包括1个野生西瓜,类群2均为栽培种.在类群2中引进的资源聚在一起,杂交种和双亲或其一聚在一起.此外,本研究表明,AFLP指纹分析技术具有很高的分辨率,能够揭示西瓜品种间的差异,是鉴定西瓜品种的有力工具.     

不同花型牡丹品种亲缘关系的SRAP分析

为了从分子水平上揭示牡丹品种间的遗传差异,本研究用SRAP-PCR技术对20个牡丹品种进行基因组DNA多态性分析,并用UPGMA法构建系统发育树,结果表明,20对引物扩增出131条清晰、稳定的DNA条带,其中90条谱带表现多态性,多态性检测率为68.7%,表明受试牡丹品种之间含较丰富的遗传多态性。UPGMA聚类结果表明,部分花型相同的品种相聚,但部分花型不同的品种亦相聚。说明聚类结果与牡丹品种的花型间可能有一定的相关性,但并未有完全一致的关系。     

8份剑麻种质亲缘关系的ISSR和RAPD分析

为了揭示剑麻栽培品种的遗传多样性,利用ISSR和RAPD分子标记技术对8份剑麻种质的亲缘关系进行分析。结果表明,筛选后选用的8条ISSR引物和8条RAPD引物,分别产生了53条和66条扩增条带,其中多态性条带分别为44条和61条,多态性条带百分率分别为83.02%和92.42%。根据2种标记的扩增结果,用UPGMA法对8份剑麻种质进行聚类分析,供试材料之间具有较高的遗传多样性,其品种间遗传相似系数分别为0.59~0.80和0.52~0.76。2个标记的聚类结果基本一致,但有点差异,可将供试的8份剑麻种质划分为2类群,而且2个标记聚类结果呈显著相关性,相关系数为0.70。可见,剑麻种质资源的遗传多样性丰富。     

利用RAPD、ISSR标记分析国兰种质资源遗传多样性的研究

旨在分析不同地区国兰间的亲缘关系,为国兰资源的开发及新品种的选育提供分子水平的参考依据。利用RAPD和ISSR分子标记技术,对7种国兰的21个品种资源进行遗传多样性和亲缘关系分析。结果表明：39个RAPD和ISSR引物在供试材料中共扩增出96条带型清晰的谱带,其遗传中31条为多态性条带;通过UPGMA聚类分析表明,21个国兰品种资源间遗传距离在1.91~6.60之间,其中春兰资源的遗传距离在1.91~4.38之间,建兰资源的遗传距离在2.60~6.20之间,寒兰资源的遗传距离在2.20~5.80之间,墨兰资源的遗传距离在3.52~6.60之间,表现出了较高的遗传多样性。聚类分析结果与传统的形态学分类结果基本一致,也说明分子标记可以在分子水平反映遗传资源的遗传多样性,具有灵敏度高、结果真实可靠等优点。本研究结果显示国兰品种间的亲缘关系与地理位置分布相关,为兰属植物的分类及遗传多样性研究提供了一定的理论支撑。     

应用RAPD分析四川马铃薯地方品种与部分栽培品种间遗传关系

为探讨四川省马铃薯地方品种与栽培品种间遗传关系,评价其在遗传研究与育种利用价值,指导杂交亲本选配,对21份地方品种和10个栽培品种进行了RAPD分析。选取8份差异较大的材料对170条RAPD引物进行筛选,获得20条多态性高、条带清晰、稳定的RAPD引物。20条RAPD引物共扩增出191个位点,其中多态性位点172个;平均多态性位点比率90.1%,平均多态性信息量(PIC)为0.822;栽培品种、地方品种平均PIC分别为0.823和0.814。遗传距离与聚类分析表明：31份材料间平均遗传距离0.4274,变幅为0.1481~0.5852,地方品种间平均遗传距离和变幅均高于栽培品种间,地方品种与栽培品种间平均遗传距离为0.4496;表明地方品种中可能存在更广泛的遗传变异,具有较高的育种利用价值。大部分地方品种出乎意料与栽培品种聚在不同的类中,与供试栽培品种间亲缘关系较远,可能具有较高的杂交组配潜力;而N5-33、N5-38和N6-22可能分别源自于大西洋、疫不加与南湖塔(或米拉),值得深入对比研究。     

Analysis of loquat germplasm (Eriobotrya japonica Lindl) by RAPD molecular markers

The loquat’s adaptation to Spain has proved very successful. In the Valencia area, the crop has met with very good environmental conditions for its development. Many new cultivars have been selected by growers and a European loquat germplasm collection has been established in Valencia at IVIA. An efficient sampling as well as implementation of germplasm resources requires the accurate identification of plant material. Molecular markers offer an effective tool for cultivar fingerprinting, estimation of genetic similarity and relationships. In this study, as a tool for germplasm management, RAPD markers were tested. Thirty-six primers were used to screen 33 cultivars. Twenty-three primers proved polymorphic. These primers generated 29 polymorphic amplification fragments that were selected as markers. Twenty-two cultivars out of 33 were identified by unique combinations of RAPD markers. Four different combinations were shared by two or more cultivars each. Cluster analysis based on the similarity matrix obtained from Nei’s coefficient among cultivars showed groupings that agreed according to geographical and genetic origin. RAPD technology was useful in distinguishing those cultivars obtained through hybridization but could not be used to distinguish those obtained by selection of mutations. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genotyping safflower (Carthamus tinctorius) cultivars by DNA fingerprints

Summary Carthamus tinctorius (2n = 2x = 24) (family Asteraceae), commonly known as safflower, is widely cultivated in agricultural production systems of Asia, Europe, Australia and the Americas as a source of high-quality vegetable and industrial oil. India ranks first in the production of safflower oil. Fourteen cultivars, widely cultivated in various agro-climatic regions of India, have been fingerprinted by RAPD, ISSR, and AFLP markers utilizing 36, 21 primers, and 4 primer combinations, respectively. On an individual assay basis, AFLP has proven to be the best marker system as compared with the other two markers applied as assessed by high discriminating power (0.98), assay efficiency index (33.2), marker index (18.2), resolving power (40.62), and genotype index (0.856). Thirty-six RAPD and 21 SSR primers could differentiate a maximum of eight and four cultivars, respectively, whereas, two AFLP primer combinations could fingerprint all the 14 cultivars. To understand genetic relationships among these cultivars, Jaccard's similarity coefficient and UPGMA clustering algorithm were applied to the three marker data sets. Mean genetic similarities ranged from 0.689 (AFLP) to 0.952 (ISSR). Correlation coefficient comparisons between similarity matrices and co-phenetic matrices obtained with the three markers revealed that AFLP displayed no congruence vis-a-vis RAPD and ISSR data. However, strong correlation was observed between RAPD and ISSR marker systems. This paper reports the start of molecular biology programme targeting nuclear genome of safflower, a major world oilseed crop about whose genetics very little is known.     

温州盘菜地方品种鉴定与指纹图谱的分子标记分析

利用基因组DNA的RAPD、ISSR与SRAP等3种分子标记技术,以日本芜菁品种作为外类群,对来自于温州不同地区具有代表性的10个盘菜品种进行品种鉴定与遗传多样性分析。10个RAPD引物共产生多态性条带70条,多态率为71．7％;12个ISSR引物共产生142条清晰带,其中多态性条带70条,多态率为49．3％;8个SRAP引物组合共产生105条谱带,其中多态性谱带78条,多态性比率为74．3％,表明品种间存在较高的多态性。用单个引物NAURP299、NAUISR43以及SRAP引物组合mel／em2,都可以将11个品种完全区分开来。基于3种标记的聚类分析结果表明,11个材料可以分为3大类,一定程度上能够揭示品种之间园艺学性状的相似性及亲缘关系远近。     

Analyses of genetic diversity in Cuban rice varieties using isozyme, RAPD and AFLP markers

A survey of the genetic diversity among the major cuban rice cultivars was conducted using isozyme, RAPD and AFLP markers. Polymorphisms were detected for esterases, peroxidases, alcohol dehydrogenases and polyphenoloxidases systems; 21 RAPD primers and four AFLP primer combinations. Heterozygosity arithmetic mean value (H_av(p)), the effective multiplex ratio (EMR) and the marker index (MI), were calculated for isozyme, RAPD and AFLP markers. The mean value of genetic similarity among the different varieties was 0.92 for isozyme, 0.73 for RAPD and 0.58 for AFLP analyses. Thus, AFLP were able to detect polymorphisms with higher efficiency than RAPD (+15%) and isozyme (+34%). Data from the isozyme, RAPD and AFLP analyses were used to compute matrices of genetic similarities. The efficiency of the UPGMA for the estimation of genetic relatedness among varieties was supported by cophenetic correlation coefficients. The resulting values indicated that the distortion level for the estimated similarities was minimal. The correlation coefficients obtained by the Mantel matrix correspondence test, which was used to compare the cophenetic matrices for the different markers, showed that estimated values of genetic relationship given for isozyme and RAPD markers (r = 0.89), as well as for AFLP and RAPD markers (r = 0.82) were properly related. However, AFLP and isozyme data showed only moderate correlation (r = 0.63). Although the genetic variability found among the different cultivars was low, both RAPD and AFLP markers proved to be efficient tools in assessing the genetic diversity of rice genotypes. This revised version was published online in July 2006 with corrections to the Cover Date.     

利用RAPD标记鉴定大豆种质

本研究以５７个中国大豆祖先吕系及育成品种和１８个美国大豆祖先品系为ＤＮＡ样品来源，通过随机引物ＰＣＲ扩增基因组ＤＮＡ的多态性，探索利用ＲＡＰＤ标记鉴定和相关种质的可能性。研究结果表明，５０个１０摩尔随机引物共扩增可分辩产物２４６个，其中８２．４％的随机引物可产生多态性产物，所扩增产物的５４．４％至少在两个基因毒草境存在差异。上ＰＣＲ扩增产物分别以１和０记录存在与否。扩增产物间的成对比较可产生非相似     

Comparing RAPD and AFLPTM analysis in discrimination and estimation of genetic similarities among apple (Malus domestica Borkh.) cultivars

Forty-one apple (Malus × domestica Borkh.) cultivars were screened for RAPD (Random Amplified Polymorphic DNA) and AFLP(Amplified Fragment Length Polymorphism) markers. RAPD analysis was performed with 35 arbitrary 10-mer primers, selected from 60 primers tested (kits A, C and E, Operon Technologies, Inc.). Of a total of 362bands observed, 208 (57.5%) were polymorphic. Three-hundred-and-eighty-one AFLP fragments were obtained with 8primer combinations, of which 218 (57.2%) were polymorphic. Cultivars differentiated through mutation were included in this study and showed identical patterns when analysed with both RAPD and AFLP analysis. The estimated genetic relationships were correlated (r = 73.7%) between the analysis with the two different markers. UPGMA analysis was performed and dendrograms were constructed using either the data apart from each(RAPD and AFLP) method or combined in a single joint matrix. The relationships among the forty-one studied cultivars were basically consistent with the known lineage and geographic origins of the cultivars. The four Portuguese cultivars included in this study clustered together and diverged from the other cultivars. Apparently they constitute an independent genetic pool, which could be of interest for apple plant breeders. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cultivar identification and genetic relationship among selected breeding lines and cultivars in chickpea (Cicer arietinum L.)

Twenty two RAPD and 22 ISSR markers were evaluated for their potential use in determination of genetic relationships in chickpea (Cicer arietinum L.) cultivars and breeding lines. We were able to identify six chickpea cultivars/breeding lines by cultivar-specific markers. All of the cultivars tested displayed a different phenotype generated either by the RAPD or ISSR primers. Though ISSR primers generated less markers than RAPD primers, the ISSR primers produced higher levels of polymorphism (% of polymorphic markers per primer) than RAPD primers. A high level of within cultivar homogeneity was observed in chickpea. Cultivars/breeding lines originating from a common genetic background showed closer genetic relationship. Chickpea lines with similar seed type(kabuli or desi) had a tendency to cluster together. This revised version was published online in August 2006 with corrections to the Cover Date.