应用RAPD技术研究柿品种间的亲缘关系

采用随机扩增多态性ＤＮＡ技术对原产中国,日本和韩斩部分柿品种间的亲缘关系进行了初步探讨。结果表明：（１）１５个供试品种ＯＰＡ－０６,ＯＰＡ－０８,ＯＰＡ－１９三个引物扩增后,品种间和引物间的谱带类型各不相同;（２）经６个随机引物扩增出的１５供试品种的谱带类型和相似指数的分析结果显示,“富有”和“次郎”,“平核无”和”仓光“,”磨盘柿“和”杵头柿“ 可具有较近的亲缘关系。     

‘黄金冠’桃及其品系亲缘关系的RAPD分析

利用RAPD技术对黄桃新品种‘黄金冠’及其它6个品系的亲缘关系进行了研究。从80个随机引物中筛选出15个重复性好、条带清晰的多态性引物对桃品系进行了RAPD扩增,共扩增出84条谱带,其中38条表现多态性,多态性比率为45.2%。利用NTSYS软件和UPG-MA聚类法对扩增结果进行了品种间相似系数的计算及聚类分析。结果表明:相似系数为0.78~0.86,黄桃19和其它桃品系亲缘关系最远,‘黄金冠’和锦黄2号的亲缘关系最近。     

部分中国原产柑柿种质的分子系统学研究

采用２５个随机引物对中日两国原产完全甜柿种质的ＲＡＰＤ分析结果表明：（１）供试中国甜柿和日本甜柿是相对独立分化的两上居群，中国甜柿的遗传多样性更为丰富；（２）在供试中国甜柿中，‘罗田甜柿’的５个单株间的遗传组成的差异较小，但与其它湖北甜柿和湘西甜柿的差异较大；（３）‘铜盆柿’与湖北甜柿间的相似指数较高，说明二者有某种亲缘关系；（４）湘西甜柿与供试日本甜柿间的亲缘关系较近，说明其可能来自日本或是与日     

部分中国原产甜柿种质的分子系统学研究

采用２５ 个随机引物对中日两国原产完全甜柿种质的ＲＡＰＤ 分析结果表明： （１）供试中国甜柿和日本甜柿是相对独立分化的两个居群， 中国甜柿的遗传多样性更为丰富； （２）在供试中国甜柿中， ‘罗田甜柿’的５ 个单株间的遗传组成的差异较小， 但与其它湖北甜柿和湘西甜柿间的差异较大； （３） ‘铜盆柿’与湖北甜柿间的相似指数较高， 说明二者有某种亲缘关系； （４） 湘西甜柿与供试日本甜柿间的亲缘关系较近， 说明其可能来自日本或是与日本甜柿的起源有关。     

浙江部分柿属植物鉴定及亲缘关系的SSR分析

应用SSR对浙江省分布的部分柿属植物(包括柿、浙江柿和金枣柿在内的3种46个基因型)进行了种质鉴定和亲缘关系分析。从已公开序列的20对柿属植物SSR引物中筛选出扩增谱带清晰和多态性丰富的引物11对;应用建立的技术体系,可将同物异名和同名异物的试材得以系统整理,一般认为是同一品种的兰溪大红柿与永康方柿,玉环长柿与永嘉长柿指纹图谱各有不同,应为不同种质。聚类分析结果表明,金枣柿(Diospyros sp.)与其他供试试材间的亲缘关系较远,支持其可能为一新种的观点;浙江柿(D.glaucifolia Metc.)与金枣柿的亲缘关系较远但与柿(D.kaki Thunb.)较近;部分供试日本柿种质与浙江省柿种质的亲缘关系较近。研究结果可为浙江省柿属植物种质资源保护和利用提供一定的科学依据。     

基于SSR标记技术的南丰柑橘种质资源亲缘关系研究

【目的】从分子水平探究南丰柑橘种质间的亲缘关系。【方法】应用SSR技术对28份南丰柑橘种质进行基因组的多态性分析。【结果】从91对引物中筛选得到13对SSR引物,可区分全部供试材料。共扩增得到64个等位基因,平均每个位点4.92个等位基因。以遗传相似系数0.18为界,金柑属金柑单独聚为1类,柑橘属27份供试样品聚为1类。以相似系数0.59为阈值,27份柑橘属供试种质分为5个类群,其中南丰蜜橘品种群(包含小果系、大果系、桂花蒂系、早熟系)所有20份供试种质与‘蜜广’聚为一类,‘小叶广’‘火广’‘火橘’聚为一类,‘红广’‘红橘’‘本地早橘’各为一类。【结论】南丰广橘品种群中‘蜜广’与南丰蜜橘品种群有较近的亲缘关系,而‘红广’与亲本南丰蜜橘和‘红橘’的亲缘关系均较远。‘小叶广’‘火广’和‘火橘’亲缘关系较近。     

菊花18个品种的RAPD分析

 采用RAPD 技术分析了18 个菊花品种DNA 的多态性。从80 个10 碱基随机引物中筛选出多态性频率高的3 个引物。扩增的多态性片段在600 bp～1300 bp 之间。检测出两个品种特有的分子标记, ‘大红托桂’有OPD15 (1200 bp) ,‘玉翎管’缺失OPA17 (1100 bp) 。瓣型一致的品种间基因型相似系数较高。     

几种玉兰亚属植物的RAPD亲缘关系分析

 以玉兰亚属的20个材料(含种和品种)为研究对象，用RAPD技术对其亲缘关系进行分析。从55个随机引物中筛选出15个，对所有供试材料进行扩增，共获得274条DNA谱带，其中262条为多态性带。建立的基于RAPD结果的亲缘关系树形图显示玉兰亚属的基因背景非常复杂，类群划分较为困难，说明中国的玉兰亚属种质资源非常丰富。     

不同山楂品种亲缘关系的RAPD分析

为探讨山楂品种间的亲缘关系,采用RAPD技术对20个不同品种的山楂材料进行了基因组DNA多态性分析。从120个引物中筛选出15个10bp的随机引物对所选山楂品种的DNA样品进行PCR扩增,共得到216条谱带,177条表现多态性,多态性比率达81.9%,其中包含27条特异性谱带,揭示了山楂植物丰富的遗传多样性。且利用NTSYS软件和UPGMA法对扩增结果进行了品种间相似系数的计算及聚类分析,结果表明相似系数在0.71～0.87,实生楂与其他山楂品种亲缘关系较远。     

大花蕙兰遗传多样性及亲缘关系的AFLP分析

对来源于日本、韩国和美国的42个大花蕙兰品种和两个国产兰属原生种进行了遗传多样性和亲缘关系的AFLP分析, 9对多态性引物在50～500 bp内共扩增出1 597条带, 其中多态性带1 565条, 多态性比率9810%。单引物对扩增的带数156～193条, 平均每对引物扩增带数177条。42个品种具特征带或缺失带。大花蕙兰品种间的遗传多样性丰富, 品种间的相似系数0.3399 ～0.8223, 平均相似系数0.5783。UPGMA聚类结果将供试品种分为4大类, 与根据花枝类型或花径大小、花色等形态指标分类的结果相吻合, 同一产地来源甚至同一育种公司选育出的品种能基本上聚类在一起, 反映出了品种间的亲缘关系。     

Molecular typing of rose cultivars using RAPDs

Summary

A rapid molecular typing method is described in this work. RAPD amplification products are very dependent upon various factors such as source of taq DNA polymerase, thermocycling programmes and DNA concentrations. This issue has been consequently been examined to establish a suitable experimental protocol. Subsequently, genomic DNA from 25 cultivars of rose were amplified using RAPD techniques with twenty 10-mer primers (Operon Kit A). The data obtained reveal no variability within cultivars and a high degree of variation between cultivars. With the patterns obtained with two of the primers (OPA-11 and OPA-17) all the rose cultivars were unequivocally identified. The results suggest that RAPD profiles provide a simple and efficient way to identify rose cultivars.     

Analysis of genetic diversity among Indian short-day onion (Allium cepa L.) cultivars using RAPD markers.

Summary

Randomly amplified polymorphic DNA (RAPD) markers were used to estimate genetic diversity among 24 cultivars of short-day onions. Total genomic DNA was extracted and subjected to RAPD analysis using 90 arbitrary 10-mer primers. Of these, 15 primers were selected which yielded 137 bands, 91.24% of which were polymorphic. None of the primers produced a unique banding pattern for each cultivar. RAPD data were used to calculate a Squared-Euclidian Distance matrix which revealed a minimum genetic distance between cultivars ‘AFLR-722’ and ‘PBR-140’, and a maximum genetic distance between cultivars ‘PBR-139’ and ‘A.Kalyan’, and ‘MS-48’ and ‘A.Kalyan’. Based on the distance matrix, cluster analysis was done using a minimum variance algorithm.The dendrogram thus generated, based on Ward’s method, grouped the 24 onion cultivars into two major clusters. The first cluster consisted of cultivars from the northern region, and the second of cultivars from the southern region of India. The present study shows that there is high diversity among the onion cultivars selected and indicates the potential of RAPD markers for identification and maintenance of onion germplasm for crop improvement purposes.     

15个樱桃品种的RAPD分析

利用RAPD技术,用104条随机引物对甜樱桃的14个品种和中国樱桃的1个品种进行遗传多样性分析,其中有14条引物的多态性较好。用任意一条能出现扩增带的引物,能明显区分开中国樱桃和甜樱桃,RAPD标记能够准确地进行种间的区分;而用一个引物或两个引物的组合只能鉴定出甜樱桃的一个品种。聚类结果显示,中国樱桃和甜樱桃的遗传距离最远;黄色果肉的养老和其他红色果肉的品种遗传距离较远。RAPD分析基本能够反映甜樱桃品种间的遗传多样性,但效果不理想,鉴定品种较为困难。     

Genetic diversity and phylogenetic relationships among and within species of oriental cymbidiums based on RAPD analysis

The classifications of oriental cymbidiums that are native to Korea, China, Taiwan and Japan were examined by molecular analysis. A total of 21 cymbidiums, 15 species including three Cymbidium gyokuchin, 4 Cymbidium kanran and 2 Cymbidium goeringii cultivars, were analyzed by using randomly amplified polymorphic DNA (RAPD) to determine the interspecific and intraspecific relationships. Twenty-two primers were used in the RAPD analysis to distinguish, by comparing differences in DNA banding patterns, all species and cultivars. Similarity values ranged from 0.501 for Cymbidium aloifoilum and C. kanran to 0.935 for Cymbidium ensifolium and Cymbidium marginatum with analysis of total bands score. Phylogenetic tree analysis of the RAPD results from the 21 cymbidiums identified specific groupings. The cymbidiums could be divided into two clusters based upon ecological traits. One trait was temperate zone preference, with each cymbidium preferring either an Asian or subtropical temperate zone. The group that comprised the subtropical cymbidiums was C. aloifoilum, Cymbidium insigne and Cymbidium lowianum. Additionally, we found that Cymbidium lancifolium and Cymbidium aspidistrifolium could be separated based on different flowering physiology and unique leaf form. The groups identified by morphological, physiological and ecological characteristics were in full agreement with those determined by RAPD analysis. The phylogenetic tree derived from the RAPD results was similar to that of the traditional classification. The data acquired from this study could be used for identification and classification of other orchid genera and oriental cymbidium.     

Genetic characterisation of oak seedlings, epicormic, crown and micropropagated shoots from mature trees by RAPD and microsatellite PCR

DNA was isolated from seedlings of Quercus robur, collected from a single provenance, and from epicormic, crown shoots and in vitro shoots from a single tree of Q. petraea using a CTAB method of extraction. DNA was obtained in sufficient quantity and purity, from 13 out of 30 seedlings, and from all isolations from epicormic and in vitro shoots (2.5–10.0 μg/g fresh/ weight). Smearing was minimised at a primer concentration of 0.12 μM with Taq polymerase at 0.5 unit/reaction. Nine primers produced 142 bands, 28 of which were polymorphic. A similarity index showed that 11 seedlings were closely related with high coefficients (0.85–0.90), but each could be identified from another using only 9 primers (OPA-02 and -05, OPG-04 and -05, OPE-01, -02, -03, -08, -09). DNA was isolated from crown, epicormic and in vitro leaves originating from a single 150-yr old tree of Q. petraea and analysed by randomly amplified polymorphic DNA (RAPD) and microsatellites. With each primer, a characteristic RAPD pattern was obtained, and it was common to all six epicormic shoots derived from different parts of a single branch of this tree; also to the shoots from the crown of the same tree with OPE1 OPA-05, OPA-08, OPA-01, OPA-02, OPA-04, OPA-05, OPG-02, OPG-10, OPE-12. Similarly, the RAPD pattern obtained from shoot cultures in vitro, derived from individual nodes of epicormic shoots produced by six different branch segments, were uniform for each of 15 primers. This work was repeated using microsatellite PCR. Three microsatellite loci AG16, AG 1/2 and AG 1/5 were amplified by PCR. It showed a uniformity of these microsatellite loci in shoots from the crown of the tree, and from epicormic shoots cultures derived from six different sections of branch.     

RAPD技术在石榴品种分类上的应用

以55个石榴栽培品种为试验材料,利用15条多态性好的随机引物进行RAPD分析,共扩增出125条带,其中多态性条带92条,多态性百分率73.6%,说明品种间有变异。应用TFPGA软件计算55个石榴品种间的Nei’s遗传距离,并用UPGMA法构建聚类图,将55个石榴品种分为4个类群,从DNA水平上揭示了石榴品种之间的亲缘关系。结果表明,采用UPGMA法聚类的结果与形态上的分类存在着一定的差异,即与根据花色和果味进行的分类没有相关性而与瓣型进行的分类有一定的相关性。     

茄子耐盐种质亲缘关系的RAPD分析

利用RAPD标记对14份耐盐茄子种质资源进行遗传多样性分析。结果表明：从100个引物中筛选出10个多态性较好的引物,共扩增出102条带,多态性条带所占的比例为77.45%。聚类分析结果显示：14份耐盐茄子材料的遗传相似系数在0.316～0.975之间|在遗传相似系数0.47处可将14份耐盐茄子材料分为3大类,大部分种质资源（85.71%）都聚在第3类（栽培种）,表明收集到的茄子耐盐种质亲缘关系较近。     

DNA fingerprinting and genetic diversity analysis of late-bolting radish cultivars with RAPD,ISSR and SRAP markers

Three molecular marker systems, RAPD (random amplified polymorphic DNA), ISSR (inter-simple sequence repeat) and SRAP (sequence-related amplified polymorphism), were employed for identification and genetic diversity analysis of 35 elite late-bolting radish cultivars. Detected by 35 RAPD primers, 22 ISSR primers and 17 SRAP primer combinations, the proportions of polymorphic bands were 85.44%, 85.2% and 85.41%, respectively, and the mean genetic similarity coefficients between pairs of genotypes were 0.781, 0.787 and 0.764, respectively. Each of the three molecular marker systems can identify all the cultivars. Five sets of three-RAPD primers, 3 sets of three-ISSR primers and 16 sets of three-SRAP primer combinations were able to distinguish all the cultivars. A linear relationship was observed between Resolving power (Rp) of a primer and its ability to distinguish genotypes. The 35 cultivars were clustered into three major groups based on the RAPD, ISSR and marker combination data with UPGMA, which are in high accordance with their own origins and main characteristics. The results demonstrated that these three marker systems could be useful for identification and genetic diversity analysis of radish cultivars.     

豆瓣菜种质资源遗传多样性的RAPD和ISSR分析

以8个豆瓣菜的品种为试材，用筛选出的79个RAPD引物和34个ISSR引物对这8个品种的基因组DNA进行扩增，分别扩增出361条和179条谱带，每个引物扩增出的带在3～10条之间，平均每个引物扩增出约5条带。根据所得的条带进行聚类分析，两种标记产生的聚类图存在一些差异，但它们都可以较好地将8个品种按亲缘关系的远近划分为3个不同的类群。Mantel测试得出相关系数r=0．58155，表明RAPD和ISSR两种分子标记技术的相关度很低。     

河南17 个桂花品种的RAPD 分析

 采用改良CTAB 法提取河南17 个桂花品种的基因组DNA , 利用RAPD 技术对其进行鉴定和分类研究。从100 个10 bp 的随机引物种筛选出15 个扩增效果较好的引物进行扩增, 共产生121 条带, 其中87 条为多态性带。根据扩增结果进行聚类分析, 得出反映各品种间亲缘关系的树状图。各个品种群内的不同品种间的亲缘关系接近, 而不同品种群间亲缘关系较远。RAPD 对基因组的分析结果与传统分类学的结果基本相符。