大葱部分种质资源亲缘关系的RAPD分析

从36个大葱品种成株幼嫩叶片中提取基因组DNA,筛选出10个扩增较稳定的RAPD引物,进行PCR扩增,共扩增出89条带,其中多态性条带65条,多态率为73.03 %,应用UPGMA进行聚类分析,36个大葱品种间的遗传距离在1.732～4.727之间,在遗传距离3.883处,可将36份材料划分成3个复组合和1个独立组,同时建立了稳定的大葱品种RAPD标记体系。     

RAPD在枇杷品种鉴定中的应用

运用RAPD(RandomAmplifiedPolymorphicDNA,即随机扩增多态性DNA)分析技术,对16个枇杷品种的基因组DNA进行RAPD分析,从几个随机引物中筛选出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,表明不同枇杷品种基因型间存在着极为丰富的遗传多样性,扩增的DNA指纹图谱可将16个品种一一区分,为枇杷品种鉴定提供了新的方法,同时也为分子标记辅助育种提供了依据。     

基于SSR标记的黄桃种质资源亲缘关系分析

利用SSR标记技术对12个黄桃品种（系）进行遗传多样性检测.结果发现,SSR标记能够揭示材料间的遗传多样性,从蔷薇科苹果属35对SSR引物中筛选出8对扩增稳定的特异性引物,构建其指纹图谱.8对SSR引物共扩增出78条DNA片段,其中44条具有多态性,多态性比率为54.6％,材料间的遗传相似系数在0.64 ～0.88之间,平均为0.759.通过聚类,从分子水平对黄桃品种（系）的遗传关系进行分析,并对12份黄桃种质材料进行了SSR标记分类,为黄桃种质资源的研究利用提供参考.     

ISSR和RAPD分子标记技术在不同君子兰品种遗传多样性上的应用

采用ISSR标记和RAPD标记技术研究了15个君子兰品种的遗传多样性。结果表明:从55条寡聚核苷酸引物中筛选出10条简单重复序列引物,共扩增出65条带,平均每条引物扩增出6.5条带,其中5.4条多态性带,多态性比率为81.95%;从55条寡聚核苷酸引物中筛选出11条多态性引物,共扩增出47条带,平均每条引物扩增出4.27条带,其中32条多态性带,多态性比率为67.88%。POPGEN软件计算出君子兰品种间遗传距离为0.1123~0.6330,平均值为0.3263。基于遗传相似系数的UPGMA聚类分析将15个品种明显聚为二组,国兰系列品种为一组,日本兰系列品种组成另一组。     

山楂（Crataegus pinnatifida Bge.)遗传多样性的RAPD和ISSR标记分析

 利用RAPD和ISSR标记对35份山楂（Crataegus pinnatifida Bge.）资源进行了DNA多态性分析。12个RAPD引物共扩增出110条清晰的谱带,其中89条显示多态性,平均每个引物扩增出7.4条多态性谱带。13个ISSR引物共扩增出110条清晰的谱带,其中94条显示多态性,平均每个引物扩增出7.2条多态性谱带。基于RAPD和ISSR标记,利用UPGMA分别构建了35份山楂资源的聚类树状图。距离系数分别为0～0.62（RAPD）和0～0.64（ISSR）,表明山楂具有较高的遗传多样性。     

利用RAPD标记鉴定和区分梅花42个宫粉型品种

 利用RAPD标记，对形态特征相似的42个宫粉型梅花品种的基因组DNA进行随机引物PCR扩增。从120个引物中筛选出21个引物，共扩增出96条谱带，其中多态性谱带68条，占扩增谱带数的70．8％ ，品种问相似系数范围为0．737～0．961。在21个引物中，7个引物扩增的谱带可区分27个品种，7个引物相互组合可区分其余的l5个品种。梅花品种RAPD指纹图谱的建立可以为品种知识产权保护提供依据。     

白菜抗霜霉病基因的RAPD标记

研究了与抗霜霉病基因紧密连锁的RAPD分子标记, 利用白菜抗病品种014和感病品种010杂交的F₂群体144个单株为材料, 通过人工接种鉴定, 采用高抗单株和高感单株分别构建抗、感病池, 利用BSA法筛选了560条RAPD引物, 其中引物AY12在抗感病池中扩增出多态性片段AY12₁₂₃₈ , 通过F₂单株验证后证明AY12₁₂₃₈与抗霜霉病基因紧密连锁, 其遗传距离为6.7 cM。     

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.     

利用RAPD和SCAR标记鉴定草莓品种

 利用RAPD和SCAR标记对32份草莓材料进行了鉴定, 结果表明: 8个RAPD引物共扩增出85个标记, 其中71个为多态性标记, 多态性比率为83。5%。25份草莓材料的RAPD图谱差异较大, 易于区分。利用具有多态性的RAPD标记, 对28份草莓试材的亲缘关系进行分析, 初步鉴定出同名异物和同物异名品种。两个RAPD标记被转化为片段长度分别为378 bp和214 bp的显性SCAR标记, 其多态性与相应RAPD标记一致。利用这两个SCAR标记对草莓品种进行了初步鉴定。     

石斛属植物遗传多样性RAPD分析

应用随机扩增多态性DNA(RAPD)技术,对石斛属植物16个种进行了基因组DNA多态性分析.从82个随机引物中筛选出20个多态性好的引物进行RAPD扩增,扩增DNA片段数据用UPGMA聚类法构建系统发育树.20条引物共扩增出142条带,多态性带118条,约占总数的83.1%.聚类结果显示RAPD分子标记构建的系统发育树与经典分类系统一致.RAPD标记可为石斛属植物的系统分类研究提供分子生物学依据.     

Identification of RAPD markers linked to nut weight and plant stature in cashew

Cashew is an important edible nut crop of tropics. Bulk segregant analysis (BSA) was carried out on DNA bulks constituted from F₂ population and germplasm in order to link or associate molecular markers with economic characters. In all 458 RAPD, 31 ISSR and 21 pairs of SSR primers were used and identified polymorphic markers between parents. Though screening F₂ bulks with these markers identified markers polymorphic between the bulks but none could be validated with the individuals of their bulks. Hence screening with germplasm bulks was carried out and could identify four RAPD markers polymorphic between the bulks for nut weight and plant stature and also between the individuals of their bulks. Of the four, three markers were associated with nut weight amplifying at 775, 475, 275, bp region in primers OPN 14, UBC 184 and UBC 185 respectively. Out of these three, two markers were specific to low nut weight and one marker was specific to high nut weight and their bands were present in greater frequency (50–77.8% and 75%) of individuals constituting the respective bulks. Similarly, the another marker UBC 185₂₇₅ was detected which was specific to low plant stature and was present in 66.7% and 10% individuals constituting short and tall bulks respectively. Markers identified with bulks and with the individuals of bulks were validated further with more individuals of F₂ and germplasm.     

Molecular assessment of polymorphism among local Jordanian genotypes of the common fig (Ficus carica L.)

This research was conducted to assess polymorphism among local genotypes of common fig available in Jordan (one of countries of origin). Leaf samples were collected in spring for DNA isolation from 20 different local genotypes (5 cultivars and 15 landraces). Two more wild types and one foreign cultivar were included. The genotypes were assessed using the randomly amplified polymorphic DNA (RAPD) technique. Six of the 19 screened primers showed reproducible polymeric profiles. Out of 62 amplified bands, 48 were polymorphic (77%). They generated 1104 data entries (591 for present and 513 for absent bands). After determining Jaccard similarity index, some genotypes showed high genetic similarity (90% between F20 and F22), while other were less similar (3–18% between F11 and all other genotypes). Moreover, the primers were evaluated for their discriminating power, where primer RAPD06 showed the weakest power (0.431), while highest values of 0.989 and 0.996 were achieved for primers RAPD02 and RAPD13, respectively.     

30份莲种质资源的RAPD遗传多样性分析

采用RAPD分子标记技术对30份莲(Nelumbo nueifera Gaertn.)种质资源的遗传多样性进行鉴定.筛选出14对有效引物进行PCR扩增,共产生2 146条带,其中有1 516条为多态性条带,各对引物的平均多态性条带为108.29条,扩增条带的多态率达70.64％.对所得到的30份莲种质资源的RAPD图谱...     

中国野生葡萄抗霜霉病基因RAPD标记的筛选

以抗病的中国野生华东葡萄(Vitis pseudoreticulata)白河-35-1(Baihe-35-1)与感病的欧洲葡萄佳利酿(Carignane)杂交F2代为试材,从520条随机引物中筛选出21条可以扩增多态性DNA片断的随机引物,从中获得了6个抗霜霉病基因的RAPD标记,对这些标记回收、克隆、测序后,将得到的6个标记分别记录为:S294-369,S202-527,S382-615,S221-677,S416-1224和S390-725。通过分析这6个RAPD标记在F2代植株中的表现,证明这些标记都与霜霉病抗性的基因有连锁关系。这将为葡萄抗霜霉病育种的分子标记辅助选择提供分子依据。     

桃花粉可育基因连锁的RAPD 标记与SCAR标记的转化

 桃品种以重阳红(花粉不育) ×大久保(花粉可育) 的52株F₁代群体为试材, 应用RAPD技术, 结合集群分组分析法(Bulked Segregate Analysis, BSA) 构建花粉可育/不育基因池, 利用180个随机引物, 筛选在花粉可育基因池中稳定扩增的一个RAPD标记OPW03-900。经过重复性验证和群体单株验证, 该标记仅在花粉可育个体(重组型除去) 中出现, 与桃花粉可育/不育位点的连锁距离为5.80cM。将该特异性片段回收、克隆、测序, 并设计一对特异SCAR引物, 再对F1代个体进行PCR扩增, 发现该特异带的位点及重组型个体的数目与RAPD扩增结果一致, 片段大小为906 bp, 命名为SCW03-906,表明RAPD标记已成功转化为与桃花粉可育基因连锁的SCAR标记。该序列已被GenBank收录, 登录号为DQ659676。     

中国野生葡萄果实抗白腐病基因的分子标记

 以感病×抗病的葡萄种间杂交组合白玉霓×塘尾的F₁ 代17 个单株及塘尾自交一代16 个单株为试材, 采用BSA 法和RAPD 技术, 通过对155 个随机引物的筛选, 获得了一个与中国野生葡萄抗白腐病基因连锁的RAPD 标记OPP09-760 , 并在中国野生葡萄8 个种的32 个株系及欧洲葡萄16 个品种中得到验证。进一步将该DNA 片段克隆并测序, 根据两端序列, 设计了一对长26 bp 的特异引物分别扩增供试材料, 获得了与该RAPD 标记相同大小的DNA 片段, 成功地将RAPD 标记转化为SCAR 标记, 可以用作抗白腐病基因的分子辅助选择的依据。     

Evaluation of genetic diversity among Iranian soft-seed pomegranate accessions by fruit characteristics and RAPD markers

Soft-seedness in pomegranate is a desirable trait for fresh consumption of this valuable fruit. At the main Iran pomegranate collection, 21 pomegranate accessions gathered from different parts of Iran are registered as soft-seed genotypes. The aim of this research was to study these soft-seed pomegranate accessions using fruit morphopomological traits and DNA markers to reveal their relatedness. Thirty-six fruit characteristics were measured in these accessions together with applying 29 random decamer primers already reported to be polymorphic on pomegranate. Factor analysis on mean values of fruit characteristics determined 10 main factors and applied for grouping of the accessions using Ward's method. Also 14 of the random primers showed good amplification and polymorphism on these samples, and a total of 43 RAPD markers were produced. Estimates of genetic similarity, using Jaccard's similarity coefficient, ranged from 0.13 to 1.0 using the RAPD data. Grouping based on the fruit traits compared with that based on RAPD data did not produce a significant correlation (r = −0.36). Morphometric measurements and sensory evaluation confirmed that some accessions are hard or semi-hard seeded. This study showed that information based on fruit characteristics and RAPD markers are complementary for genetic discrimination in soft-seed pomegranate accessions. This might be due to the high level of similarity between soft-seed pomegranate accessions.     

应用RAPD技术评价丁香品种间遗传关系

采用随机扩增多态ＤＮＡ（ＲＡＰＤ）技术分析了６个丁香（Ｓａｒｉｎａａ）品种间的遗传关系。用１７个单引物共扩增出８３个ＤＮＡ片段，其中清晰可辨、重复的片段６２个，占总数的７４．６％。被检测出品种特有标记，可作为区别不同品种的重要性状。品种ｆｌｏｒａ和ＮａｕｄＮｏｔｃｕｔｔ间相似系数高达９８．１％，证明它们是同一品种不同无性系。通过品种间相似系数比较，确认品种Ｒｏｃｈｅｓｔｅｒ是从品种Ｆｌｏｒａ选育而来；品种Ｍａｉｄｅｎ’ｓＢｌｕｓｈ是由朝鲜丁香（Ｓｙｒｉｎｇａｏｂｉａｔａｖａｒ．ｄｉｌａｔａｔａ）和欧丁香品种ＭｒｓＭａｒｒｙＢｉｃｋｌｅ杂交而来。     

Genetic diversity analysis of ginger (Zingiber officinale Rosc.) germplasm based on RAPD and ISSR markers

A global collection of ginger germplasm consisting of 46 accessions were characterized using two types of molecular markers, RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter Simple Sequence Repeats). UPGMA dendrograms constructed based on three similarity coefficients, i.e., Jaccard's, Sorensen–Dice and Simple Matching using the combined RAPD and ISSR markers placed the accessions in four similar clusters in all the three dendrograms revealing the congruence of clustering patterns among the similarity coefficients and a rather less genetic distance among the accessions. Improved varieties/cultivars are grouped together with primitive types. Moreover, in the clustering pattern of the accessions, a geographical bias was also evident implying that germplasm collected from nearby locations especially with vernacular identity may not be genetically distinct. The clustering of the accessions was largely independent of its agronomic features.     

Application of RAPD and SI-typing techniques to confirm genetic integrity of radish (Raphanus sativus L.) varieties

The random amplified polymorphic DNA (RAPD) and self-incompatibility (SI) typing techniques were applied to settle a lawsuit between a Seed Company and a farmer concerning seed impurity in radish (Raphanus sativus L.). After sowing “Halra Woldong” (HW) a winter radish variety, two morphologically different radish plants grew. Genomic DNA from each of the two types were prepared and analyzed using ninety-six RAPD markers and four variety-specific markers were found. The polymorphic RAPD band patterns were compared with those of the twenty Korean radish varieties. The genetic integrity of the two varieties in question was identified and further confirmed by SI-typing of S-locus glycoprotein (SLG) and S-locus receptor kinase (SRK) genes.