枇杷栽培种的随机扩增DNA多态性(RAPD)研究

运用RAPD(Random Amplified Polylnorphic DNA)分析技术,对江苏省常绿果树研究中心16个枇杷(E．japonic Thunb Lindl)品种的基因组DNA进行了分析,从50个随机引物中筛选出19个引物,可从各个品种的基因组DNA扩增出谱带,总数为233个,其中多态性DNA片段176个,公共性DNA片段57个,表明这些枇杷品种间存在着丰富的遗传多样性。根据DNA片段的电泳结果,计算出品种间遗传相似系数及遗传距离,应用UPG—MA方法建立了品种遗传关系的系统树,将16个品种分成5类。从随机引物中筛选出2个随机引物能从各个品种的基因组DNA扩增出DNA片段,在16个枇杷品种中2个引物扩增出19个DNA片段,其中3个为公共,16个为多态性或单态性DNA片段。这些结果可为枇杷品种鉴定、分类、亲缘关系确立及品种选育中杂交组合亲本选配提供一定的依据。     

中国水仙与欧洲水仙品种RAPD指纹的研究

利用随机多态性DNA(RAPD)技术对中国水仙和欧洲红口水仙共6个品种进行了分析。选用了12个10bp引物共扩增出119条DNA片段。计算各样本间的相似性系数和遗传距离,并采用UPGMA法对遗传距离进行了聚类分析。结果表明：中国水仙3个品种亲缘关系十分密切,红口水仙3个品种亲缘关系较近,而中国水仙与红口水仙3个品种间的亲缘关系较远。建立的水仙RAPD技术体系可帮助育种亲本的选配及水仙品种的鉴定。     

RAPD分析在绢丝昆虫亲缘关系研究中的应用Ⅰ.蓖麻蚕品种间的遗传差异

用RAPD技术对蓖麻蚕基因组DNA进行多态性研究，分析了5个蓖麻蚕品种间的遗传差异。结果表明，所采用的40个随机引物中，有27个引物扩增谱带清晰且重复性较好，扩增总片段数达243个，单个引物的扩增片段数在4～17之间，平均为9条，片段大小在033～30kb之间。不同蓖麻蚕品种间的遗传距离(Ｄ)在00683～01603之间，根据Ｄ值，由UPGMA聚类分析软件绘制了它们的聚类分子树。     

甜樱桃品种及其砧木的RAPD分析

利用RAPD技术,从130个随机引物中筛选出46个引物,对欧洲甜樱桃、欧洲酸樱桃、马哈利樱桃和野生中国樱桃4个类型樱桃种,以及欧洲甜樱桃与中国樱桃的种间杂交种共15个品种的基因组遗传变异进行分析。结果表明,46个随机引物均得到了稳定可重复的RAPD图谱,扩增出的DNA条带大小在100~2625bp之间,多态性位点数517个,多态性位点百分率为98.85%,每个随机引物扩增出的多态性DNA条带数在4~23条。品种间Nei遗传距离在0.166~0.479之间,平均遗传距离0.329;甜樱桃新品种‘秦樱1号’与‘秦岭玛瑙’、‘CDR-1’等10个樱桃砧木之间的遗传距离在0.248~0.376,并且根据遗传距离可以相互区分,所分析的15个樱桃品种均扩增出了特有的DNA条带,每个樱桃特有标记带在2~17个之间,共扩增出149个特有标记,据此可以进行樱桃品种及砧木的RAPD鉴定。研究认为利用RAPD技术可以在分子水平上对甜樱桃品种及其砧木进行快速鉴定。     

10种冬青属植物遗传多样性RAPD和AFLPs分析

采用RAPD和AFLP技术,对10种冬青属植物基因组进行DNA片段扩增,以研究该属种间遗传多样性.结果表明:在RAPD分析中,通过对100种10个碱基随机引物的筛选,发现11种引物能得到多态性较高扩增产物,11种引物共扩增出301条多态性条带,多态率为98.63%.在AFLP分析中,3对选择性引物组合均扩增出了丰富的多态性片段.利用RAPD和AFLP技术分析,结果按UPGMA类平均法进行聚类,聚类结果显示冬青和代茶冬青,木姜冬青和浙江冬青以及光枝刺缘冬青与毛枝三花冬青之间的亲缘关系最近.     

用RAPD技术探讨中国枣的种下划分

利用随机扩增多态性DNA(RAPD)技术对14个枣Ziziphus jujuba 品种和1个野生种——泰山酸枣Z．spinosus的遗传变异进行了研究。从120个10-碱基随机引物中筛选出37个多态性引物用于正式扩增,共扩增出429条DNA带,其中多态性带214条,占49．88％。根据DNA扩增结果计算了品种及类型间遗传距离,并用UPGMA构建了聚类树状图。分析结果表明：龙爪枣 Z．jujuba var．tortuosa、葫芦枣 Z. jujuba var．lageniformis、无核枣 Z.jujuba var．anucleatus 等几个变种内的遗传距离大于变种间遗传距离,认为枣的变种划分是不自然的,宜并入其原变种;枣种下不宜设变种,对枣种下的众多品种,应根据品种间的遗传关系,直接划分品种群。     

大麻性别的RAPD和SCAR分子标记

利用随机扩增多态性DNA(randomamplifiedpolymorphicDNA,RAPD)技术获得与大麻性别连锁的分子标记.将10株雄性大麻或10株雌性大麻的单个DNA样品等量混合分别组成雄性或雌性DNA池(DNApool),以提供具有相同遗传背景的雌、雄性DNA样品.每个随机引物分别用三个不同的循环程序进行PCR扩增.在30个随机引物中,用引物401扩增得到一条约2.5kb雄性多态性片段.对该片段进行了克隆和序列分析,并根据序列分析结果将上述RAPD分子标记转化为重复性和特异性更好的SCAR(sequencecharacterizedamplifiedregions)分子标记.     

大麻性别的RAPD和SCAR分子标记

利用随机扩增多态性DNA（random amplified polymorphic DNA,RAPD)技术获得与大麻性别连锁的分子标记,将10株雄性大麻或10株雌性麻的单个DNA样品等量混合分别组成雄性或雌性DNA池（DNApool),以提供具有相同遗传背景的雄,雄性DNA样品。每个随机引物分别用三个不同的循环程序进行PCR扩增,在30个随机引物中,用引物S401扩增得到一条约2．5kb雄性多态性片段,对该片段进行了克隆和序列分析 ,并根据序列分析结果将上述RAPD分子标记转化为重复性和特异性更好的SCAR（Sequence characterized amplified regions)分子标记。     

草莓DNA分子标记及其应用

综述了限制性长度多态性(RFLP)、随机扩增多态性DNA(RAPD)、扩增片段长度多态性(AFLP)、简单重复序列(SSR)等不同类型分子标记在草莓指纹图谱构建、品种鉴别、遗传多样性、进化、遗传作图以及相关性状的标记等方面的应用,分析了草莓分子标记研究中的关键问题,提出了今后研究方向。     

苜蓿遗传多样性的取样数目——RAPD和SSR群体标记法

紫花苜蓿为异花授粉植物,其DNA多态性研究的取样策略与遗传多样性分析直接相关.选取了4个苜蓿品种陇东(地方品种)、中兰1号(育成品种)、牧歌(Graze)和金皇后(Queen)(引进品种),设置了取样数目分别为10、20、40和60个单株进行DNA混合,利用RAPD(random amplification polymorphic DNA,随机扩增长度多态性DNA)和SSR(simple sequence repeat,简单重复序列)标记分别进行了遗传多样性分析,结果发现40和60个单株DNA混合样的聚类结果一致,表明10和20个单株组成的群体太小,随着苜蓿群体取样数目的增加,遗传多样性分析的准确性也随之增加,但是分析成本也相应提高.鉴于此,利用RAPD和SSR标记分析苜蓿遗传多样性时采用40个单株的DNA混合样是较适宜的群体大小.     

松花型花椰菜主要品种鉴定的分子标记分析

利用RAPD、ISSR和SRAP 3种分子标记对我国南方地区松花型花椰菜主栽品种进行鉴定,分析了品种间的遗传多样性。3种标记共产生370条扩增带,238条为多态性条带,其多态率为64.32%。其中只有SRAP标记的引物m e1/em1可将20个品种全部鉴别。遗传相似系数分析表明,松花型花椰菜品种之间的亲缘关系较近,遗传背景比较狭窄。聚类分析表明品种间的亲缘关系与熟性、地理分布相关。研究表明,分子标记能有效地应用于花椰菜品种鉴定,且综合多种分子标记分析品种间的遗传多样性将更加准确可靠。     

Assessment of genetic diversity among 16 promising cultivars of ginger using cytological and molecular markers

Ginger (Zingiber officinale Roscoe) is an economically important plant, valued all over the world. The existing variation among 16 promising cultivars as observed through differential rhizome yield (181.9 to 477.3 g) was proved to have a genetic basis using different genetic markers such as karyotype, 4C nuclear DNA content and random amplified polymorphic DNA (RAPD). The karyotypic analysis revealed a differential distribution of A, B, C, D and E type of chromosomes among different cultivars as represented by different karyotype formulas. A significant variation of 4C DNA content was recorded in ginger at an intraspecific level with values ranging from 17.1 to 24.3 pg. RAPD analysis revealed a differential polymorphism of DNA showing a number of polymorphic bands ranging from 26 to 70 among 16 cultivars. The RAPD primers OPC02, OPA02, OPD20 and OPN06 showing strong resolving power were able to distinguish all 16 cultivars. The extent of genetic diversity among these cultivars was computed through parameters of gene diversity, sum of allele numbers per locus and Shannon's information indices. Cluster analysis, Nei's genetic similarity and genetic distances, distribution of cultivars into special distance classes and principal coordinate analysis and the analysis of molecular variance suggested a conspicuous genetic diversity among different cultivars studied. The genetic variation thus detected among promising cultivars of ginger has significance for ginger improvement programs.     

Analysis of genetic diversity in cowpea (Vigna unguiculata L.Walp.) cultivars with random amplified polymorphic DNA markers

The genetic diversity among ten Indian cultivars of cowpea was analyzed using 18 sets of RAPD markers. A total of 181 bands with an average of 15 bands per primer were obtained. Out of 181 bands, 148 showed polymorphism (81.7%). The variation in genetic diversity among these cultivars ranged from 0.1742 to 0.4054. Cluster analysis based on Jaccard’s similarity coefficient using UPGMA with high bootstrap values revealed two distinct clusters I and II comprised of two and seven cultivars, respectively. Cluster II was further differentiated into various subclusters. Cultivar IC-9883 was found to be unique based on its altogether distinct position in the dendrogram and two-dimensional space projections.     

Application of RAPD markers for characterization of γ-ray-induced rose mutants and assessment of genetic diversity

Six parent and their 12 gamma ray-induced somatic flower colour mutants of garden rose were characterized to discriminate the mutants from their respective parents and understanding the genetic diversity using Random amplification of polymorphic DNA (RAPD) markers. Out of 20 primers screened, 14 primers yielded completely identical fragments patterns. The other 7 primers gave highly polymorphic banding patterns among the radiomutants. All the cultivars were identified by using only 7 primers. Moreover, individual mutants were also distinguished by unique RAPD marker bands. Based on the presence or absence of the 48 polymorphic bands, the genetic variations within and among the 18 cultivars were measured. Genetic distance between all 18 cultivars varied from 0.40 to 0.91, as revealed by Jaccard’s coefficient matrix. A dendrogram was constructed based on the similarity matrix using the Neighbor Joining Tree method showed three main clusters. The present RAPD analysis can be used not only for estimating genetic diversity present in gamma ray-induced mutants but also for correct identification of mutant/new varieties for their legal protection under plant variety rights.     

Relationships among <Emphasis Type="Italic">Leymus</Emphasis> species assessed by RAPD markers

The DNA genetic diversity of 40 accessions of genus Leymus was analyzed by random amplified polymorphic DNA (RAPD) markers. A total of 352 products were amplified by 34 10-mer arbitrary primers, among which 337 products (95.74 %) were found to be polymorphic. 5–14 polymorphic bands were amplified by each polymorphic primer, with an average of 9.91 bands. The data of 352 RAPD bands were used to generate Jaccard’s similarity coefficients and to construct a dendrogram by means of UPGMA. Great genetic diversity in genus Leymus was observed, the genetic diversity among the different species more abundant than that of the different accessions, and the different accessions in a species or the species from the same areas were clustered together.     

Comparative analysis of genetic relationships in barley based on RFLP and RAPD markers

Genetic relationships have seldom been analyzed with different types of molecular markers in order to compare the information provided by each marker class. We investigated genetic relationships among nine barley cultivars using separate cluster analyses based on restriction fragment length polymorphisms (RFLPs) and random amplified polymorphic DNAs (RAPDs). Genomic DNA restricted with three enzymes and hybridized with 68 probes revealed 415 RFLPs (74.2% of all bands). Among the 128 primers used for RAPD analysis, 100 provided a reproducible profile, 89 of which revealed 202 polymorphic and 561 monomorphic bands (26.5 and 73.5%, respectively). A nonrandom distribution of 62 RAPDs with a tendency to cluster near centromeric regions was produced when these RAPDs were mapped using 76 doubled-haploid lines derived from a cross between two of the nine cultivars. The correlation between the RFLP and RAPD similarity matrices computed for the 36 pairwise comparisons among the nine cultivars was equal to 0.83. The dendrograms obtained by cluster analyses of the RFLP and RAPD data differed. These results indicate that in barley the information provided by RFLPs and RAPDs is not equivalent, most likely as a consequence of the fact that the two marker classes explore, at least in part, different portions of the genome.     

中国茶树初选核心种质遗传多样性的RAPD分析

以中国茶树初选核心种质中的69份种质为实验材料,采用改良的SDS法提取它们的基因组DNA,并运用优化的RAPD 分析体系对基因组DNA进行分子标记遗传差异研究。从50个随机引物中筛选出32个扩增效果好的引物,对全部试验材料进 行了RAPD扩增共得到348条有效带,其中多态性带为328条(占94.3%),它们之间的遗传距离为0.223～0.723。研究结果表 明中国茶树初选核心种质的遗传结构、遗传多样性和遗传距离基本上能较好的代表中国的茶树种质资源。同时,指出结合形 态标记和DNA分子标记是构建茶树核心种质较好的选择。     

利用RAPD标记分析大麦种质资源的遗传多样性

利用RAPD标记对19份西藏近缘野生大麦材料、33份我国不同省市的地方品种以及8份国外引进大麦品种共60份大麦种质资源的遗传多样性进行检测.结果表明材料间遗传差异明显.32个RAPD引物中,有25个引物(占78.13%)可扩增出清晰且具多态性的条带,另外7个引物能扩增出1～3条清晰但无多态性的条带.每个引物可扩增出1～8条多态性带,平均为3.72条.32个引物共产生119条DNA片段,其中87条具有多态性,多态性比率(PPB)为73.11%,平均多态信息量(PIC)为0.434;每个位点平均有效等位基因数(Ne)为2.304;材料间遗传相似系数GS变化范围为0.757～0.981,平均值为0.871.19份来源于西藏的近缘野生大麦材料间GS值变幅为0.818～0.969,平均为0.892;33份我国栽培大麦地方品种间的GS值变化范围为0.783～0.981,平均为0.879;8份分别来自8个国家的栽培大麦品种间的GS值变幅为0.820～0.956,平均为0.882.根据RAPD标记分析的结果,对60份大麦种质资源进行聚类分析,在平均GS值0.871水平上60份大麦材料可聚为5类,聚类结果能在一定程度上反应材料的地理分布关系,但某些相同地理来源的材料也较分散地分布在整个聚类树中.本研究从分子水平上进一步证明了我国栽培大麦丰富的遗传多样性,是世界栽培大麦的遗传多样性中心之一.     

利用RAPD分析披碱草属、鹅观草属和猬草属模式种的亲缘关系

利用随机扩增多态性DNA（RAPD）技术对小麦族披碱草属、鹅观草属和猬草属3个属的模式种进行了基因组DNA多态性分析。42个引物产物的290条谱带中,257条（88.62%)表现出多态性,说明披碱草属、鹅观草属和猬草属3个属的模式种间具有丰富的遗传多样性。利用290个RAPD标记,计算材料间Nei氏遗传相似性系和遗传距离,在NTSYS程序中利用UPGMA进行聚类。结果表明,Elymus sibiricus种不同居群间的遗传差异较小,遗传距离在0.097-0.180之间。E.sibiricus,Roegneria caucasica和Hystrix patula的种间遗传差异明显,遗传距离在0.458-0.605之间。H.patula与E.sibiricus的亲缘关系较近。R.caucasica与E.sibiricus的亲缘关系较远。