新型标记SRAP在棉花F2分离群体及遗传多样性评价中的适用性分析

将新型分子标记SRAP(Sequence-related Amplified Polymorphism)应用于棉花的遗传研究,并建立了完整的PCR反应体系,此体系稳定可靠、扩增效果好、可重复性强。采用30个SRAP引物组合对海岛棉品种“Pima90”和陆地棉品种“邯郸208”进行比较扩增,29个引物组合可以获得多态性扩增,显示了较高的多态性。对上述两个品种的F2群体进行检测,共产生149个多态性条带,平均每个组合产生5．14个,单引物组合最多可产生13个多态性条带。用SRAP标记对11份陆地棉材料进行遗传多样性检测,30个引物组合中15个组合有多态性,得到22个多态性条带,显示了较高的多态性比率。研究结果表明,SRAP标记可在棉花分子生物学领域中广泛应用。     

酵母菌SRAP-PCR反应体系的优化及遗传多样性分析

从8个正向引物和11个反向引物随机组合成的88个引物对中筛选出14个引物组合,对19株酵母菌进行PCR扩增。采用正交试验设计方法,对SRAP的影响因素进行研究,从Mg2+、dNTPs、DNA浓度、引物、TaqDNA聚合酶设计5因素4水平进行优化试验,以确定PCR电泳分析的最佳条件。14个引物组合共扩增出279条带,其中92.5%呈多态性,相似系数范围为0.60~1.00。     

红掌品种亲缘关系SRAP分析

利用相关序列扩增多态性（SRAP）分子标记,从100对引物组合中筛选出 26对多态性高、条带清晰的SRAP引物,对33个红掌品种进行遗传多样性和亲缘关系分析。结果如下：（1）26对引物共扩增出366条条带,其中有314条多态性条带,多态性比率为85.79%。引物组合产生的条带数在9～23之间,平均每对引物组合扩增出14.1条和12.1条多态性条带。（2）根据SRAP扩增结果,利用UPGMA法进行聚类分析,33份材料的遗传相似系数在0.55～0.94之间,在遗传相似系数0.786处可将33个红掌品种分为5个类群。结果表明,供试品种遗传多样性丰富,本研究为品种鉴定和杂交育种提供了参考信息。     

应用SRAP标记分析黄瓜的遗传差异

利用49对SRAP引物对4种不同类型28份黄瓜种质资源进行了遗传差异分析.结果表明,有35对引物扩增出多态性,在28份资源间共产生724条扩增带,平均每对引物组合产生20.69条;共检测出337个多态性位点,多态性比率为46.5%,每对引物平均为96.3个.利用NTSYS软件分析遗传相似系数,UPGMA方法聚类分析表明,28份资源可聚为两大类.试验结果表明SRAP标记位点多,重复性好,可以揭示不同类型黄瓜种质之间的遗传基础.     

大豆种质资源SRAP分子标记中的引物筛选

以113个大豆栽培品种和20个野生品种为材料,从288对引物组合中筛选出12对多态性丰富、条带清晰、可重复性好的SRAP引物组合。用筛选出的12对引物组合对大豆品种进行PCR扩增,获得了带型丰富和清晰可辨的DNA的PAGE指纹图谱;共扩增出251条谱带,其中多态性条带220条,多态性谱带比率为87.6%,平均每个引物扩增出18.3条谱带。结果显示,所筛选出的12对引物组合可以有效的应用于大豆种质资源的SRAP分析。     

辣椒31个优良自交系的亲本类群分析

以包含我国重要尖椒品种的亲本材料在内的31份优良自交系为材料, 利用SRAP标记和基因型值分析技术开展了辣椒自交系间遗传差异的分析与类群划分研究。结果表明: 在30个引物组合中, 27个引物组合可以 在自交系间扩增出多态性条带, 共扩增出310个多态性条带, 平均每个引物组合产生11.5个多态性条带, 显示出SRAP技术具有较强的分析效率; 基于SRAP标记和Yule相似系数对这些自交系进行的聚类分析中, 可以基本区分辣椒的2个变种(C. annuum var. grossum和C. annuum var. longum), 而且可以反映出自交系间的亲缘及系谱关系; 在相似系数为0.67处, 可将这31个自交系分为4个类群; 基于基因型值和标准Euclidean距离对这些自交系进行的聚类分析可成功地将辣椒的两个变种完全区分; 在遗传距离约4.5处, 可将这31个自交系分为4个类群; 自交系间基于SRAP标记与基因型值的遗传距离存在一定的相关性。     

利用SRAP标记分析河南小麦栽培品种的遗传多样性

利用小麦SRAP标记对22个河南省小麦品种进行了遗传多样性分析,10对引物组合扩增获得169个条带,其中70个条带具有多态性,多态条带百分率为41.42％,每对引物平均产生7个多态性条带。22个供试材料的带型按照条带的有,无分别记录为1,0后,采用Nei 72方法计算不同品种的遗传距离,利用NTSYS软件进行非加权成组法（UPGMA）进行了聚类分析。结果表明SRAP标记技术能较真实地反映小麦品种间的亲缘关系,可以用于小麦品种遗传多样性的研究。     

利用SRAP标记分析海南旱稻地方品种的遗传多样性及其分子身份证的构建

利用SRAP标记对来自海南省的28份旱稻种质进行了亲缘关系分析。从255对引物组合中筛选出46对扩增条带清晰、多态性高的引物组合,并选用其中的18对引物组合对28份材料进行SRAP扩增,共扩增出180个条带,其中多态性条带160条,多态性比率为87.75%,平均每对引物组合产生10个条带和8.9个多态性条带。并且,运用UPMGA方法进行的聚类分析结果表明,供试旱稻种质的遗传相似系数范围为0.687 0-0.921 2,在遗传相似系数0.687 0处分为两大类群,第I类群全部为海南山栏稻种质,第Ⅱ类群为海南地方旱稻坡压和本研究组自育旱稻品种。对海南旱稻地方品种的划分表明,山栏稻种质间亲缘关系较近,海南地方旱稻坡压可能与籼型水稻种质具有一定的遗传相似性。此外,还构建了28份海南省旱稻种质的SRAP标记分子身份证,可为山栏稻种质的遗传亲缘关系,种质鉴定及新品种选育等方面提供依据。     

茄子种质遗传多样性及群体结构的SRAP分析

采用SRAP标记法对183份茄子(Solanum melongena)种质资源的遗传关系和群体结构进行分析。结果表明, 33对多态性SRAP引物组合共扩增出215条清晰稳定的多态性条带, 平均每对引物组合产生7条多态性条带。183份茄子种质的遗传相似系数介于0.276-0.813之间, 平均值为0.623, 表明茄子种质资源间遗传背景存在一定的差异。在遗传距离为0.345 6处可将183份茄子种质分为4组。通过群体结构分析可将种质划分为4个群体, 不同群体间的界限十分明显, 且群体间的基因渗透较高。     

利用SRAP标记研究四川高原青稞育成品种的遗传多样性

利用SRAP(Sequence-related Amplified Polymorphism)分子标记技术, 对25份来自四川高原的青稞育成品种进行了遗传多样性研究。结果表明: 64对引物组合共检测出999条清晰条带, 62对可以获得多态性条带, 多态性引物组合占96.9%, 共产生225条多态性条带, 占总条带数的22.5%。64对引物组合共扩增出333种等位变异, 平均每个引物组合检测到5.20种等位变异。遗传多样性在0(me9/em14, me9/em15)~0.8928(me6/em18)之间, 平均为0.5126。聚类分析结果表明, 25份材料可分成A、B、C 3大类, 材料聚类与其来源地有明显的相关性。25份材料间的平均遗传距离较小(0.3240), 平均遗传多样性较低(0.5126), 遗传基础较为狭窄。     

Comparative Evaluation of RAPD and AFLP Based Genetic Diversity in Brinjal (Solanum melongena)

The present investigation was carried out with an objective of evaluating genetic diversity in brinjal (Solanum melongena) using DNA markers. A total of 38 brinjal accessions including one wild-species, Solanum sisymbrifolium were characterized using random amplified polymorphic DNA (RAP D) and amplified fragment length polymorphism (AFLP) techniques. Out of 45 primers employed to generate RAPD profiles, reproducible patterns were obtained with 32 primers and 30 (93.7%) of these detected polymorphism. A total of 149 bands were obtained, out of which 108 (72.4%) were polymorphic. AFLP analysis was carried out using four primer combinations. Each of these primers was highly polymorphic. Out of 253 fragments amplified from these four primer combinations, 237 (93.6%) were polymorphic. The extent of pair-wise similarity ranged from 0.264 to 0.946 with a mean of 0.787 in RAPD, in contrast to a range of 0.103 to 0.847 with a mean of 0.434 in AFLP. The wild species clustered separately from the brinjal genotypes. In the dendrogram constructed separately using RAPD and AFLP markers, the brinjal genotypes were grouped into clusters and sub-clusters, and the varieties released by IARI remained together on both the dendrograms. All the 30 RAPD primers in combination and each of the four primer pairs in AFLP could distinguish the brinjal accessions from each other. AFLP was thus found to be more efficient than RAPD in estimation of genetic diversity and differentiation of varieties in brinjal.     

A comparative analysis of genetic diversity in blackgram genotypes using RAPD and ISSR markers

Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to study the DNA polymorphism in elite blackgram genotypes. A total of 25 random and 16 ISSR primers were used. Amplification of genomic DNA of the 18 genotypes, using RAPD analysis, yielded 104 fragments that could be scored, of which 44 were polymorphic, with an average of 1.8 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from two (OPA-13) to nine (OPK-4) and varied in size from 200 bp to 2,500 bp. Percentage polymorphism ranged from 16.6% (OPK-7) to a maximum of 66.6% (OPE-5, OPH-2, and OPK-8), with an average of 42.7%. The 16 ISSR primers used in the study produced 101 bands across 18 genotypes, of which 55 were polymorphic. The number of amplified bands varied from two (ISSR 858) to ten (ISSR 810), with a size range of 200–2,200 bp. The average numbers of bands per primer and polymorphic bands per primer were 6.3 and 3.4, respectively. Percentage polymorphism ranged from 25% (ISSR 885) to 100% (ISSR 858), with an average percentage polymorphism of 57.5% across all the genotypes. The 3-anchored primers based on poly(GA) and poly(AG) motifs produced high average polymorphisms of 54.98% and 58.32%, respectively. ISSR markers were more efficient than the RAPD assay, as they detected 57.4% polymorphic DNA markers in Vigna mungo as compared to 42.7% for RAPD markers. The Mantel test between the two Jaccards similarity matrices gave r =0.32, showing low correlation between RAPD- and ISSR-based similarities. Clustering of genotypes within groups was not similar when RAPD and ISSR derived dendrogram were compared, whereas the pattern of clustering of the genotypes remained more or less the same in ISSR and combined data of RAPD and ISSR.     

Identification of cultivars and validation of genetic relationships in Mangifera indica L. using RAPD markers

Twenty-five accessions of mango were examined for random amplified polymorphic DNA (RAPD) genetic markers with 80 10-mer random primers. Of the 80 primers screened, 33 did not amplify, 19 were monomorphic, and 28 gave reproducible, polymorphic DNA amplification patterns. Eleven primers were selected from the 28 for the study. The number of bands generated was primer- and genotype-dependent, and ranged from 1 to 10. No primer gave unique banding patterns for each of the 25 accessions; however, ten different combinations of 2 primer banding patterns produced unique fingerprints for each accession. A maternal half-sib (MHS) family was included among the 25 accessions to see if genetic relationships could be detected. RAPD data were used to generate simple matching coefficients, which were analyzed phenetically and by means of principal coordinate analysis (PCA). The MHS clustered together in both the phenetic and the PCA while the randomly selected accessions were scattered with no apparent pattern. The uses of RAPD analysis for Mangifera germ plasm classification and clonal identification are discussed.     

随机扩增多态性DNA技术在鲍氏层孔菌菌株鉴别中的应用

用20个随机引物对7个不同来源的鲍氏层孔菌菌株进行了RAPD分析．结果表明120个随机引物中,有17个引物的扩增产物DNA条带表现出明显的多态性,不同引物对供试菌株扩增出现的DNA条带数目少则10条,多达33条．DNA片段从250bp到2000bp;采用17个引物对7个鲍氏层孔菌菌株共扩增出DNA片段带377条,不同引物扩增出的DNA片段谱带存在较大差异．采用UPGMA系统聚类法,将7个菌株聚类为两大类,能直观准确地揭示菌株间的差异并加以鉴别．     

冬枣×宁梨巨枣的子代SRAP鉴定及遗传多样性分析

应用SRAP分子标记方法对冬枣×宁梨巨枣的子代进行了分子鉴定及遗传多样性分析。采用构建基因池的方法对SRAP分子标记引物进行筛选,从88对引物中筛选出15对多态性好、主带清晰的引物,并对子代进行了真实性鉴定及多态性分析。结果表明:(1)15对引物共产生95个多态性条带,平均每对引物产生6.3个多态性条带,显示了较高的多态性比率。(2)80个子代中44个具有父本特征带,鉴定为真杂种。子代遗传多样性及UPGMA聚类分析表明,子代个体与亲本间的遗传相似系数在0.55~0.98之间,个体差异明显。该研究结果为枣树杂交育种提供了重要的分子证据。     

中国扁蓿豆遗传多样性的ISSR分析

利用ISSR标记,对来源于中国7个省(市)的44个扁蓿豆居群的亲缘关系进行了分析。从80条ISSR引物中筛选出多态性强、重复性好的16条引物,对44个扁蓿豆居群基因组DNA进行扩增,共扩增出133条谱带,平均每个引物扩增出8.31条带,其中多态性带115条,多态性位点百分率为87.08%。扁蓿豆种质间遗传相似系数变化范围在0.436～0.908之间。利用UPGMA聚类分析,44个扁蓿豆居群被划分为8个类群,与地理来源有较高的相关性。     

S-SAP分子标记开发及其在苹果芽变鉴别上的应用

S-SAP(特异序列扩增多态性,Ssequence specific amplification polymorphism)是一种基于反转录转座元件的广泛应用于生物遗传多样性研究的分子标记。本研究从34对引物中筛选出7对具有谱带清晰、多态性高的引物组合,成功地开发了苹果基因组的S-SAP分子标记。27个元帅系芽变品种中,共扩增出588条谱带,每对引物组合平均扩增出84条谱带,其中多态性谱带48条,多态性谱带占总扩增出谱带数的8.2%,遗传相似系数介于0.73～0.90之间。对15个苹果芽变品种进行S-SAP分析,相似系数在0.42～0.94之间,以相似系数0.80为阈值,可以区分各芽变品系。开发的S-SAP分子标记可以有效地将苹果芽变品种区分,并为苹果生物遗传多样性与系统进化、品种鉴定提供新方法。     

Molecular Characterization of <Emphasis Type="Italic">Cyclamen</Emphasis> Species Collected from Different Parts of Turkey by RAPD and SRAP Markers

The genus Cyclamen (family Myrsinaceae) contains about 20 species, most of which occur in the Mediterranean region. Turkey has critically important Cyclamen genetic resources. Molecular characterization of plant materials collected from different regions of Turkey in which Cyclamen species grow naturally, namely Adana, Antalya, Ayd?n, Mu?la, ?zmir, Denizli, Kahramanmara?, Osmaniye, Eski?ehir, Trabzon, and Rize provinces, was performed using RAPD and SRAP markers. DNA was successfully amplified by 30 RAPD primers and 14 SRAP primer pairs. Among the 470 bands generated by the RAPD primers, 467 were polymorphic. The number of bands detected by a single primer set ranged from 11 to 22 (average of 15.6). The percentage polymorphism was 99.3 % based on the RAPD data. In the SRAP analysis, a total of 216 bands were generated, showing 100 % polymorphism. The number of bands detected by a single primer set ranged from 9 to 22 (average of 15.4). All data were scored and UPGMA dendrograms were constructed with similar results in both marker systems, i.e., different species from nine provinces of Turkey were separated from each other in the dendrograms with the same species being clustered together.