中国部分兰花品种RAPD分析

用随机引物扩增多态DNA（RAPD）分析技术，对中国兰花迸行品种鉴定和分类研究。使用10种20个碱基的引物，对19个中国兰花品种的基因组DNA进行扩增，其产生83条扩增带，其中80条大多态性带。每个兰花品种都有其特有的扩增带可与其它品种区别开来。这种特异的条带可作为一个品种的分子标记应用于品种鉴定和分类RAPD对基因组的分析结果与传统分类学的结果基本相符。各个种内的不同品种问亲缘关系接近，而不同种间差异较丸RAPD技术为中国兰花品种的鉴定和分类提供了新的途径     

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。     

桃梅李杏四种主要核果类果树RAPD指纹图谱初探

采用 10个随机引物对桃、李、梅、杏 4种核果类果树的代表品种进行RAPD扩增 ,获得了几个树种的RAPD指纹图谱。结果表明 :应用单一引物可区别核果类果树的不同种乃至品种 ;10个引物的扩增产物聚类结果能很好地反映种间亲缘关系 ,梅和杏的亲缘关系最近 ,桃次之 ,李最远 ,从而在基因水平上支持梅和杏为同一亚属 ,李、桃各为一亚属的分类方案     

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。     

‘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。     

38个石榴品种的RAPD遗传关系分析

以山东省枣庄市峄城区中国石榴种质资源圃内38个石榴品种为试材,用20条RAPD引物对其进行扩增,根据扩增结果进行UPGMA聚类分析和主坐标分析,研究了38份材料的亲缘关系。结果表明:20条RAPD引物在38份材料中共产生146个位点,多态性位点为109,多态性比率为74.6%;38个品种间遗传相似系数变化范围为0.45~1.00;在相似系数为0.58处,将38个品种可以分为四大类,得出品种间的亲缘关系与形态学和地理分布没有明显相关性;38份材料可以分为两大类,主坐标分析结果与UPGMA的聚类结果存在一定差异。     

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。     

辽宁地区五种野生软枣猕猴桃RAPD遗传多样性分析

以CTAB法提取的5种野生软枣猕猴桃基因组DNA为模板,应用RAPD方法随机扩增多态性DNA,分析其亲缘关系。结果表明:5种软枣猕猴桃均表现出各自不同的多态性RAPD标记,5条单个引物分别扩增出一般为4～6条RAPD条带,其分子量约在250～2000bp之间。依据这些多态性标记计算其种间遗传相似性,建立了亲缘关系树形图。     

十五种柑桔种质资源的RAPD分析

对柑桔15个材料（其中包含14个种，1个为品种）进行了RAPD分析，从结果看，各材料之间具有不同的遗传距离，供试材料聚为8个类群，分别为柠檬，柠檬群；香橼，小香橼群；柚，红心柚群；宜昌橙，香圆群；香橙，大翼橙群；红桔，温州蜜柑群；枳登，富民枳群；枳群。采用RAPD技术可作为品种鉴别，亲缘关系和分类的手段。对所得结果进行了讨论。     

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（RAPD）技术对原产中国、日本和韩国的部分柿品种间的亲缘关系进行了初步探讨。结果表明：（1）15个供试品种经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个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’间可能具有较近的亲缘关系。     

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。     

桂花SCoT标记体系的建立及其在遗传多样性分析中的应用

以‘籽银桂’桂花叶片DNA为材料,以2 × Es TaqMasterMix 预混液为基本体系,针对影响SCoT-PCR反应的退火温度、模板DNA用量、引物浓度等因素进行优化,建立了适于桂花SCoT分子标记的反应体系。12 μL反应体系含有30 ng模板DNA,引物浓度为0.4 μmol ? L-1,不同的引物退火温度分别为48、49.9、54.3或56 ℃。利用该体系对12个桂花品种进行分析,扩增条带清晰,扩增产物在150 ~ 2 200 bp之间。12 条引物共扩增出244条带,其中多态性条带238条,多态性比率为97.47%。在相似系数0.57水平处,‘九龙桂’形成第Ⅰ组,其余11个品种形成第Ⅱ组。第Ⅱ组中‘柳叶桂’、‘香云’、‘早籽黄’、‘大花金桂’、‘桂冠籽金桂’、‘鹅黄’、‘籽金桂’和‘醉云’8个花色不完全相同的品种聚在一起;‘金桂’、‘早银桂’和‘籽银桂’3个不同花色的品种聚在一起,说明品种间的亲缘关系与花色不完全相关,这与其它分子标记结果一致。     

13个阿月浑子品种的RAPD分析

为探明引进的阿月浑子品种间的亲缘关系,采用RAPD 标记技术对引入的12 个国外品种和‘新疆优株’进行了分析。用筛选的21 条10 bp 随机引物进行PCR 扩增可扩增出137 个位点,其中多态位点122 个,多态位点比率89.05%;品种间遗传距离在0. 2015 ~ 0.5163 之间,表明各品种间存在一定的遗传差异。UPGMA 聚类分析表明,13 个阿月浑子品种在遗传距离0.40 处可划分为3 个类群,第1 类包括 ‘Kerman’、‘Larnaka’、‘M-38’、‘M-P3’、‘Ashoury’、‘N’、‘Joley’、‘Aegina’、‘Avidon’、‘B’和‘Peter’11 个品种;第2 类为‘Xinjiang select tree’品种;第3 类为‘Mateur’品种。     

部分桂花栽培品种的AFLP分析

 从22对AFLP引物中筛选出6对用来检测22个桂花品种和木犀属3个种的多态性位点,共检测到171个位点,其中多态性位点104个,占60.8%。利用DPSv 3.11软件计算25个样品之间的Nei遗传距离,并按照非加权算术平均数聚类方法（UPGMA）,构建树状分支图。AFLP分析结果表明：桂花花色较深的品种之间和花色较浅的品种之间分别存在着较近的亲缘关系,而花色深浅不同的两类品种之间亲缘关系较远。从聚类图上看,22个桂花品种中最后聚在一起的分别是3个银桂品种（遗传距离0.19处）和3个四季桂品种（遗传距离0.21处）,说明四季桂类和银桂类中的部分品种与金桂品种群和丹桂品种群有较远的亲缘关系。从分类上看,AFLP分析结果与传统的以形态特征为基础的分类结果并不完全一致。     

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.     

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。     

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。     

Characterization of white sapote (Casimiroa edulis Llave & Lex.) germplasm using floral morphology,RAPD and AFLP markers

Floral morphology, random amplified polymorphic DNA (RAPD), and amplified fragment length polymorphism (AFLP) were used to characterize and verify genetic diversity within a white sapote cultivar collection and to develop molecular markers for germplasm identification. On the basis of floral morphology, the cultivars were classified into three types: type I included 23 cultivars with large ovaries and small anthers; type II included 13 cultivars with small ovaries and large anthers; and type III included one cultivar, named ‘Maltby’, with a large ovary and large anthers. DNA was isolated from 39 cultivars of white sapote and subjected to RAPD and AFLP analysis using 24 and 7 primers, respectively. One hundred and sixty-eight RAPD and 286 AFLP bands were used to assess genetic characterization among white sapote. Sixty percent of the RAPD and 77% of the AFLP amplification products were polymorphic among accessions. RAPD or AFLP markers differentiated all white sapote cultivars effectively. Moreover, each flower type was characterized as specially associated with two RAPD bands. UPGMA dendrograms based on RAPD and AFLP data, showed the majority of the cultivars from flower type I and flower type II clustering together. Finally 101 RAPD markers and 220 AFLP markers were used to construct a neighbor-joining dendrogram. This showed that the 37 cultivars could be classified into six distinct clusters, between which the similarity coefficient was as low as 0.00–0.55, even though the cultivars were morphologically very similar. The remaining two cultivars namely ‘Smathers’ and ‘Maltby’ were found genetically very distant from the other cultivars in RAPD, AFLP or combined RAPD and AFLP based dendrograms. The results suggested that the level of genetic variation among white sapote cultivars is diverse and the morphological and molecular data may lead to representation of the cultivar relationships as well as flower type discrimination.     

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.