芥菜种质资源的RAPD和ISSR分析

利用RAPD和ISSR标记对28份芥菜种质资源进行遗传多样性分析,20个RAPD引物共扩增出162条清晰的谱带,其中140条显示多态性,平均每个引物扩增出7.0条多态性谱带.18个ISSR引物共扩增出143条清晰的谱带,其中多态性谱带124条,平均每个引物扩增出6.9条多态性谱带.基于RAPD和ISSR两种标记,利用UPGMA分别构建了28份芥菜资源的聚类树状图.结果表明,RAPD和ISSR分别聚类以及两种标记混合聚类均将28份芥菜种质分为3类:第Ⅰ类群中包括了15份种质;第Ⅱ类群中包括了9份种质;第Ⅲ类群中包括了4份种质.     

马尾松种质资源遗传多样性的RAPD分析

利用18条RAPD引物对35份马尾松种质资源进行了DNA扩增试验,共产生153条RAPD标记条带,其中多态性带137条,多态性百分率为89.5%,平均每个引物检测到的条带数为8.5条。结合聚类分析,进行马尾松种质资源遗传多样性研究,结果表明RAPD可以准确分析研究马尾松种质资源遗传多样性。     

31份野生树莓资源的RAPD分析

用RAPD技术分析了31份野生树莓资源的亲缘关系,从100个A系列随机引物中选出了15个多态性高、分辨能力强的引物。将这15个引物应用于31份野生树莓材料,共扩增出182条谱带,其中170条为多态性标记(占93.41%),平均每个引物扩增多态性谱带12.13条,表明不同树莓资源基因型间存在着极为丰富的遗传多样性。基于RAPD的扩增结果建立UPGMA聚类分析树状图,31份野生树莓资源在遗传距离0.5100处被划分为4个类群。从分子水平研究了树莓属资源的亲缘关系,为树莓种质资源的保存和利用及其杂交亲本的选配提供了依据。     

云南25份石榴资源的RAPD分析

用RAPD标记技术分析了25份云南地方石榴材料(品种或类型)的亲缘关系,从128个随机引物中筛选出12个重复性好、条带清晰的多态性引物,并用这些引物对25份云南石榴材料进行RAPD扩增,共扩增出110条谱带,平均每个引物9.17条。多态性程度达到71.8%。用Statistic分析软件计算出的遗传距离为0.027～0.342,对25份材料进行了UPGA法聚类,亲缘关系树状图显示,25份云南石榴材料的遗传背景较复杂,难以划分类群。研究结果与生产上根据风味、花色、皮色、子粒颜色、核软硬程度等某一性状及栽培目的的分类方法不一致。     

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。     

基于ISSR和SRAP标记的69份红毛丹种质资源DNA指纹图谱构建

利用筛选出的多态性高、谱带清晰的8条ISSR引物和18对SRAP引物分别对海南省保存的69份红毛丹种质资源进行PCR扩增,8条ISSR引物共检测到425条多态性条带,其中品种(系)特异条带47个,平均位点多态性信息含量(PIC值)为0.928,可鉴别的种质资源数25~67份;18对SRAP引物共检测到894条多态性条带,其中品种(系)特异条带69个,平均位点多态性信息含量(PIC值)为0.936,可区分的种质资源数为17~63份。综合各项指标,利用ISSR16和ISSR5引物21个条带用于构建红毛丹种质DNA指纹图谱,该图谱可有效区分69份红毛丹种质资源。     

我国南方长茄种质资源的ISSR标记分析

从分子水平用ISSR标记法对南方长茄资源的遗传多样性进行分析,从100个ISSR引物中共筛选出12个多态性明显、条带清晰、反应稳定的引物,对57个样品DNA共扩增出116条谱带,平均每个引物扩增出9·67条带,其中多态性位点84个(71%)。品种间遗传相似系数在0·51~0·98之间,表明茄子栽培种内品种间的遗传基础相对较狭窄。利用UPGMA聚类分析,能将57个南方长茄品种划分为6个类群,类群的划分与地方来源没有很大的关系。     

苦瓜种质资源ISSR遗传多态性分析

利用ISSR分子标记对30份苦瓜种质进行遗传多态性分析,从23个引物中筛选出13条重复性好,条带清晰的引物进行PCR扩增,共扩增出79条带,其中58个为多态性条带,占总带数的73.4%,每个引物扩增的条带数为4~10条,平均6.07条。对ISSR结果进行聚类分析,结果表明,30份苦瓜种质间的遗传距离为0.025~0.520。在遗传距离为0.21处可将30份苦瓜种质划分成6个品种群,其亲缘关系与种质地理分布和植物学性状特征等有一定关联。     

不同产地人参种质资源RAPD和SSR分析

以15个产地人参种质资源为试材,采用RAPD和SSR分子标记技术对其遗传多样性进行分析。结果表明:2种分子标记均能揭示不同地区人参种质间的遗传多样性。共筛选出11条RAPD随机引物,平均每条引物可扩增出3~17条DNA片段,共扩增出104条清晰条带,多态性条带100条,多态性百分率为96.15%,揭示供试材料间遗传相似系数(GS)为0.505 3~0.989 5,平均值为0.760 4。筛选出5对SSR引物,平均每对引物可扩增出1~8条DNA片段,共扩增出38条清晰条带,多态性条带35条,多态性百分率为92.11%,揭示供试材料间遗传相似系数(GS)为0.400 0~0.960 0,平均值为0.742 1。RAPD和SSR标记的聚类分析在分类上稍有差异,但总体趋势一致,RAPD、SSR及RAPD+SSR均将样品聚为三大类,均能揭示它们之间的亲缘关系,为人参种质资源的收集与利用奠定了基础。     

利用ISSR分子标记对39份莲藕种质资源的遗传多样性分析

采用ISSR分子标记技术对39份莲藕品种进行遗传多样性分析。结果表明:8个ISSR引物共扩增出89条带,其中有55条多态带,平均每个引物扩增的多态性带数为6.88条,多态性比率平均为61.8%;通过遗传相似系数和聚类分析,能将39份莲藕品种完全区分开;说明ISSR标记技术能较好地从分子水平揭示出莲藕品种的遗传多样性。     

Assessment of genetic diversity in cashew germplasm using RAPD and ISSR markers

Diversity and genetic relationship in 100 cashew germplasm accessions were analyzed by using RAPD and ISSR markers. Using 10 selected RAPD primers 60 bands were generated, of which 51 bands were polymorphic (85%), and with 10 selected ISSR primers 67 amplified bands were observed with 58 polymorphic bands (86.6%). Though both kinds of markers discriminated the accessions effectively, analysis of combined data of markers (RAPD + ISSR) resulted in better distinction of accessions. By combining markers, a total of 127 bands were detected, of which 109 bands (85.8%) were polymorphic and produced on an average of 5.45 polymorphic bands per primer. Primers with high polymorphic information content and marker index were identified for discriminating accessions. High percentage of polymorphism (>85%) observed with different markers indicated high level of genetic variation existing among the accessions. Genetic relationship estimated using similarity co-efficient (Jaccard’s) values between different pair of accessions varied from 0.43 to 0.94 in RAPD, 0.38 to 0.89 in ISSR and 0.43 to 0.87 with combined markers suggested a diversity (dissimilarity) ranging from 6 to 57%, 11 to 62% and 13 to 57% respectively and the diversity skewed around 50% indicated moderate diversity. The cluster analysis with UPGMA method separated the accessions broadly into 13 clusters and in that three into smaller clusters. Some correspondence between the molecular groupings and the morphological clusters were observed. Among the accessions, NRC-142 and NRC-12 were highly divergent and NRC-231 and NRC-232 were genetically similar.     

Genetic diversity of ash gourd [Benincasa hispida (Thunb.) Cogn.] inbred lines based on RAPD and ISSR markers and their hybrid performance

Ten inbred lines of ash gourd [Benincasa hispida (Thunb.) Cogn.] were crossed to produce 45 F₁ hybrids (without reciprocal) which were evaluated along with the parents for 20 growth- and yield-related traits, in a replicated field trial. High level of heterosis was observed among the hybrids for most of the traits examined, including yield. These inbred lines were analysed by using 42 RAPD primers those produced 282 DNA marker bands. A total of 130 RAPD markers were obtained with a mean of 3.1 per primer, which in combination discriminated all the inbreds from each other. Pair-wise genetic distance measurements ranged from 0.07 to 0.31, suggesting a wide genetic diversity for these inbreds. These inberds were also analysed with five ISSR primers of which four were informative. Twenty-six ISSR marker bands were generated of which 11 were polymorphic with an average of 2.80 per primers. The percentage of polymorphic bands produced were higher in ISSR markers (>80%) than generated through RAPD markers (46%). Although the results indicated significant positive correlations of genetic distance with hybrid performance and heterosis, the RAPD based genetic distance measures and use of limited ISSR markers in this present study could not effectively predict hybrid performance in this crop. The genetic variation among ash gourd inbred lines examined, herein, defined a marker array (combined ISSR and RAPD) for the development of a standard reference for further genetic analyses, and the selection of potential parents for predicting hybrid performance and heterosis.     

Genetic diversity analysis in date palm (Phoenix dactylifera L.): a comparative assessment using ISSR and RAPD marker assays

SUMMARY

A comparative study was conducted to evaluate genetic diversity in 45 genotypes of date palm (Phoenix dactylifera L.), including both male and female plants, employing RAPD and ISSR marker systems. The data were analysed to calculate the total number of bands, the number of polymorphic bands, the percentage polymorphism, the average number of bands per primer, the effective multiplex ratio (EMR), the polymorphic information content (PIC), the marker index (MI), and genetic similarity coefficients. The 37 RAPD and 53 ISSR primers used generated 363 and 608 scorable amplified products, respectively, of which 95.0% and 90.9% were polymorphic. The ISSR markers produced more information than the RAPD markers due to their higher EMR and MI values. Jaccard similarity values among male plants, female plants, and between all male and all female plants varied between 0.72 – 0.80. The results indicate the effectiveness of these two marker systems for demonstrating genetic relationships among date palm genotypes.     

羊蹄甲属3种园艺树种分子鉴定及亲缘关系的ISSR分析

 利用ISSR-PCR方法对香港羊蹄甲属(Bauhinia) 中最常见3种园艺树种: 红花羊蹄甲(B. purpurea) 、宫粉羊蹄甲(B. variegate) 和洋紫荆(B. blakeana) 的遗传变异进行了研究, 从74个ISSR引物中筛选出9个多态性引物用于正式扩增, 共扩增出80条DNA带, 平均每个引物扩增的DNA带的数目为8.88条, 多态性条带数目为55条, 占总条带数的68.8%。其中, 有8个引物扩增出差异条带和特异条带,并分别能准确地鉴定羊蹄甲属3个园艺树种。本研究的结果表明, ISSR - PCR的DNA电泳谱带在检测香港羊蹄甲(B. blakeana) 品种的真实性方面非常有用。同时, 通过对3个种扩增条带的叠加性进行分析, 证实了洋紫荆杂交起源的父母本为红花羊蹄甲和宫粉羊蹄甲。     

ISSR．PCR技术在桂花品种分类研究中的应用

 利用ISSR．PCR方法对桂花的19个品种进行了基因组多态性分析，从74个ISSR引物中筛选出13个多态性引物用于正式扩增，共扩增出9o条DNA片段，其中多态性DNA条带79条，占总扩增片段的87．8％ ，平均每个引物扩增的DNA带的数目为6．92条。根据ISSR扩增结果，应用RAPDistance软件进行Nei相似性系数和遗传距离计算，利用UPGMA法构建聚类树状图。聚类分析的结果把供试桂花的l9个品种分为8个大类，并对4个品种群的遗传关系和种下分类系统进行了探讨。     

香榧品种遗传变异与品种鉴定的ISSR分析

利用ISSR方法对香榧本地9个实生品种和8个优良无性系的遗传变异和品种间的遗传关系进行了分析,从50个随机引物中筛选出12条多态性引物用于正式扩增,共扩增出104个DNA片段,其中88个片段为多态性扩增片段,占总扩增片段的84.6％。根据ISSR引物的扩增图谱,确定部分供试品种的特异条带。基于Nei's遗传距离分析,利用UPGMA法构建了分子树状图。聚类结果把供试的香榧9个实生品种和8个优良无性系分为两大组,第一组包括大圆榧、米榧、旋纹榧、獠牙榧、芝麻榧、落霜榧、象牙榧,第二组包括小圆榧、茄榧和细榧优良无性系。     

Comparative analysis of genetic diversity in Indian bitter gourd (Momordica charantia L.) using RAPD and ISSR markers for developing crop improvement strategies

A genetic analysis of 38 diverse Indian bitter gourd (Momordicacharantia var. charantia, and var. muricata) accessions was performed using 29 RAPD and 15 ISSR markers. RAPD primers yielded 208 amplicons of which 76 (36.5%) were polymorphic providing an average of 2.6 amplicons per primer. RAPD amplicons per primer ranged from 3 (OPE-19, OPW-09) to 15 (OPW-05), and varied in size from 200 bp to 3000 bp. Fifteen ISSR primers provided a total of 125 bands of which 94 (74.7%) were polymorphic. Polymorphic ISSR markers ranged from 0 (UBC-841) to 12 (UBC-890) providing a mean of 6.3 amplicons per primer that ranged in size from 150 bp to 2700 bp. Nevertheless, the concordance among bitter gourd accession groupings after cluster analysis was relatively high (r = 0.77), indicating that RAPD- and ISSR-based diversity assessments in this germplasm array were generally consistent. The M.charantia var. charantia (domesticated) and var. muricata (wild, free-living) accessions examined were genetically distinct, and these differences provided for the development of strategies for genetic analyses and crop improvement in this species.     

SRAP、ISSR和RAPD分子标记技术在银耳菌株鉴别上的应用

Three types of molecular markers (SRAP, ISSR and RAPD) were used to identify four Tremella fuciformis strains, T6 (white), T7 (white), T8 (yellow) and T9 (light yellow). Twelve SRAP primer pairs, ten ISSR primers and eight RAPD primers were screened, and identification data obtained using the three molecular markers were consistent in that the four T. fuciformis strains were divided into three groups with T7 and T9 clustered together in a single group. Each RAPD primer generated a higher average number of polymorphic bands than either the SRAP or ISSR primers, and the average similarity between the four strains was 81.34%. SRAP markers reflected more genetic information compared with the two other markers, and the average similarity was 68.98%. Genetic information reflected by ISSR markers was intermediate between SRAP and RAPD, and the average similarity was 77.48%.     

Molecular analysis of genetic stability in micropropagated apple rootstock MM106

Random amplified polymorphic DNA (RAPD) markers were used to assess the genetic stability of 10 micropropagated plants regenerated through axillary buds of clonal apple (Malus pumila Mill.) rootstock MM106. Eleven random decamer primers were successfully used to analyse genomic DNA from mother plants and in vitro plant material. A total of 129 scorable fragments were amplified with an average of 11.73 bands per primer. Among them, 99 were monomorphic and 30 were polymorphic with 23.2% polymorphism. Among these 30, 12 were found monomorphic across seven plants and parent. Three plants could be regarded as off-types. Our results show that RAPD markers could be used to detect the genetic similarities and dissimilarities in micropropagated material.