应用RAPD 标记和细胞质基因组PCR-RFLP技术研究大花蕙兰的遗传多样性

 利用RAPD、叶绿体和线粒体基因组PCR-RFLP标记系统评价了大花蕙兰20个品种的遗传多样性。在50个RAPD引物分析中, 有36个引物(72.0% ) 能揭示材料间的多态性, 材料间遗传相似系数为0.503～0.765, 平均0.598, 根据遗传相似系数进行聚类分析表明, RAPD标记能将所有材料区分开。在7个叶绿体基因组( cpDNA) 的PCR-RFLP分析中, 6个标记(87.5% ) 可扩增出1至多条清晰的谱带; 扩增产物经7种限制性内切酶消化后, 6个标记的19种引物/酶组合共检测到53条DNA片段, 其中多态性片段有37条, 占69.8%; 材料间遗传相似系数变化范围为0.571～0.949, 平均值为0.766。在8个线粒体基因组(mtDNA) 的PCR-RFLP标记分析中, 只有3个(37.5%) 标记能得到1条清晰的谱带; 利用7种限制性内切酶对3个标记的扩增产物消化后, 在10种标记/酶组合中, 共检测到33条酶切片段, 其中21条(63.6%) 具有多态性; 遗传相似系数为0.634～1.000, 平均0.829。这些结果表明, RAPD标记揭示的大花蕙兰遗传多样性最高, 其次为cpDNA PCR-RFLP标记, 而mtDNA PCR-RFLP标记揭示的遗传多样性最低。     

十二份杧果种质亲缘关系的SRAP分析

以12份杧果种质为试材,利用SRAP分子标记技术对其进行遗传多样性研究。结果表明:从36对引物中筛选出22对多态性好的引物,共扩增出251条DNA条带,其中多态性谱带181条,平均每对引物扩增得到15.08条多态性谱带,多态性比率为72.11%。UPGMA聚类表明,所有供试品种(系)之间的亲缘关系都比较近,相似系数在0.77~0.89;如果以相似系数0.78为标准,可将供试的12个品种(系)分为3类。表明利用SRAP技术能较好的区分杧果的亲缘关系;SRAP分子标记适合杧果的DNA遗传多样性分析,可作为杧果种质鉴定依据之一。     

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。     

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。     

甘肃42份枣种质资源遗传多样性的ISSR分析

利用ISSR分子标记技术对甘肃省42份枣种质资源的遗传多样性进行分析。结果表明:筛选出8条引物共扩增出76条DNA谱带,其中多态性谱带70条,多态性位点比率92.0%;按照42份枣种质间的遗传距离构建UPGMA聚类图,42份枣种质被划分为4个大类、10个亚类,遗传多样性处于较高水平;从聚类结果看,原产自甘肃省的枣品种遗传背景相似,主要根据原产地进行聚类,目的性品种交换现象较少。另外,对6个未知身份的枣品种进行了初步鉴定。     

仁用杏品种SRAP遗传多样性分析及指纹检索系统的开发

利用SRAP标记对24份仁用杏品种进行了遗传多样性分析。15对引物共扩增出280条带,其中241条为多态性谱带,多态性比率为85.34%;每个引物组合扩增谱带数在13 ~ 24条之间,平均为18.7条;多态性信息含量（PIC）在0.28 ~ 0.65之间,平均为0.51。基于引物Me4-Em4扩增的多态性谱带,构建的指纹检索系统可以区分24个仁用杏品种。根据SRAP扩增结果,利用UPGMA法构建树状聚类图,在相似系数为0.70处可将24个仁用杏品种分为4组,聚类结果与形态分类基本一致。     

中国北方引种的部分越橘品种的RAPD分析

利用RAPD技术对我国北方引种的15份越橘(Vaccinium spp.)栽培品种进行了遗传多样性分析。从70条随机引物中筛选出15条引物,共扩增出94个位点,其中多态性位点83个(88.3%)。材料间遗传相似系数变化为0.5426～0.8936,表明各品种间具有一定的遗传差异。UPGMA聚类分析结果可将供试材料分为2大类、5个亚类。聚类结果与各品种的杂交系谱相对应,从分子水平揭示了品种间的亲缘关系。     

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。     

萱草部分野生种和栽培品种亲缘关系的AFLP分析

 借助AFLP标记对35份萱草野生种和栽培品种进行亲缘关系研究, 结果表明, 7对引物组合对萱草共扩增出条带380条, 其中多态性条带357条, 平均多态性达到93.39% , 单对引物扩增条带19～85条, 平均每对引物组合扩增多态性条带51条。7对引物扩增出的多态性条带均超过90.0%。种质资源相似系数为0.3822～0.9656, 平均相似系数为0.7039。UPGMA聚类结果将供试材料分为3类, 即早花、中花和晚花类, 同一产地的品种基本能聚在一起。AFLP标记技术能较好地从分子水平揭示萱草种质资源的亲缘关系。     

海南椰子栽培品种的SSR标记分析

应用简单重复序列（SSR）标记方法,对海南的11个椰子栽培品种进行了遗传多样性分析。选取30对引物用于PCR扩增,有23对引物扩增出有效多态性片段136条,平均多态性百分率为95.77%,每对引物扩增出的带数2~12条不等,平均为6.17条,每个SSR位点的多态信息量（PIC）变化于0.173~0.896之间,平均为0.561。11份材料之间遗传相似系数变化范围0.061~0.861,说明海南椰子栽培品种之间存在丰富的遗传多样性。UPGMA聚类分析结果表明,11个椰子栽培品种分为四个类群和两个亚群。SSR标记反映出的品种间亲缘关系与形态学研究的分类结果并不完全吻合。     

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。     

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.     

Identification of 57 sweet orange cultivars using AFLP markers

Summary

Sweet orange (Citrus sinensis) represents an important group of Citrus fruit; however, the identification of sweet orange cultivars during vegetative growth can be difficult. A study on the genetic identification of sweet orange cultivars may be significant for the sweet orange nursery industry, for cultivar-rights protection, and is important for the genetic evaluation and conservation of these orange cultivars. In this study, amplified fragment length polymorphism (AFLP) markers were used to genotype 57 sweet orange cultivars. Ten PCR primer pairs generated 629 unique AFLP bands, with a size range of 50 ? 500 bp. Seventy-four bands (11.8%) were polymorphic. On average, each primer pair produced 62.9 fragments, with 7.4 polymorphic fragments. A dendrogram of the 57 sweet orange cultivars was constructed based on an UPGMA analysis using Jaccard?s coefficients of similarity. This provided a clear comparison of the genetic variation between cultivars and an ability to identify them. From Jaccard?s coefficients, 56 of the 57 cultivars examined were genetically close, with coefficient values ≥ 0.985. ?Variegated Navel? was less closely-related, with a much lower coefficient value (0.94). Among the 57 cultivars, 28 sub-groups, some consisting of only one cultivar, could be separated by their AFLP fingerprints. Compared to ISSR and SSR markers, AFLP seemed to be the preferential marker technique for the identification of sweet orange cultivars.     

SRAP在葱栽培品种遗传多样性研究中的适用性分析

为评价SRAP技术对葱品种进行鉴定和遗传关系分析的适用性, 对20个葱栽培品种的表型特征进行了观察记载, 利用256个SRAP引物组合对其进行了遗传多样性研究。结果表明: (1) 256个SRAP引物组合中有161个引物组合产生多态性条带, 占所用引物组合数的62.9%。161个引物组合共产生336条多态性条带, 不同引物组合产生的多态性条带数为1～6个, 平均2.1个。20个葱栽培品种遗传相似系数变幅为0.464～0.938, 平均0.703。(2) 依据SRAP进行聚类分析的分类结果与依据表型特征分类的结果一致。上述结果说明SRAP标记可以在葱栽培品种的鉴定和遗传多样性研究中应用。     

Genetic relationships of gladiolus cultivars inferred from fluorescence based AFLP markers

Gladiolus is one of the important commercial flowers with a large number of cultivars. However, genetic relationships among its genotypes have not been reported. This study analyzed genetic relatedness of 54 gladiolus cultivars using amplified fragment length polymorphism (AFLP) markers. A total of 24 AFLP primer pairs with three samples were initially screened, from which 9 primer sets that showed clear scorable and highly polymorphic bands were selected for AFLP reactions. Fluorescence-labeled amplification products were subjected to electrophoresis and then analyzed using an automated sequencer. A dendrogram was constructed by the unweighted pair group method using the arithmetic average (UPGMA). The number of AFLP fragments generated per primer set ranged from 10 to 151 with fragment sizes varying from 50 to 450 bp. A total of 660 AFLP fragments were detected, of which 658 (99.70%) were polymorphic. All the primers except E-AGG/M-CTA displayed 100% polymorphism. All cultivars were clearly differentiated by their AFLP profiles. The AFLP data were compared with previously obtained RAPD data and combined to generate a common dendrogram. The first cluster was dominated with indigenously bred cultivars while the second was dominated with exotic cultivars. This shows that most of the exotic cultivars as well as indigenous cultivars are closely related with each other. However, two indigenous cultivars viz., Pusa Suhagin and Pusa Archana share genetic similarity with exotic cultivars. Among the genotypes selected for the investigation, Pusa Gunjan was identified as the most distinct genotype. The AFLP markers developed will help future Gladiolus cultivar identification, germplasm conservation and new cultivar development. The assessed genetic relationships among gladiolus cultivars may enhance the efficiency of breeding program by selecting desirable parents with reduced breeding cycle.     

云南芋种质资源遗传多样性的AFLP分析

 采用荧光标记引物的AFLP分子标记技术, 用筛选出的“3 + 2”引物组合, 对48份云南芋种进行遗传多样性分析, 3对引物共扩增出184个DNA位点, 平均每对引物可检测出56.3个多态性位点,多态性位点高达91.8% , 云南芋种质资源在DNA分子水平上表现出极为丰富的遗传多样性。聚类分析表明: 野生种质和栽培种质的亲缘关系较远, 栽培种质基于AFLP标记的分类结果与形态性状基本一致, 少数材料差异较大。     

部分板栗品种遗传多样性的AFLP分析

利用荧光标记AFLP技术,采用7对M+3和E+3引物组合对30份板栗和日本栗栽培品种进行了总基因组DNA水平上的多态性检测,共获得962条可统计的条带,其中852条呈多态性,多态性带百分率达89％。揭示了板栗丰富的遗传多样性。7组引物在30个品种中检测到数目不等的品种特异带型,对供试板栗品种具有一定的鉴别价值。7对引物能将30个板栗和日本栗品种完全区分开。聚类分析结果表明,多数来源地相同的板栗品种资源表现出较为密切的亲缘关系。     

黄瓜种质资源遗传多样性及其亲缘关系的AFLP分析

 采用AFLP分子标记技术，对中国黄瓜种质资源遗传多样性及其与外来种质的关系进行了分析， 结果表明8对AFLP引物在70份黄瓜种质中共扩增出425条带，多态性带的比例为66％。供试黄瓜种质的平 均期望杂合度为0．376，中国种质的平均期望杂合度为0．387，明显高于国外种质的n 291。西双版纳黄瓜和印度野生黄瓜具有一些栽培种质没有的特异位点，中国栽培种质的特异位点多于外来栽培种质，后者也有一些中国栽培种质没有的特异位点。聚类分析将70份种质分为三大组群，即西双版纳黄瓜(Cucumis sativus L.vaF．xishuangbannanesis Qi et Yuan)组群，e sativus var．hardwickii野生黄瓜组群和栽培黄瓜组群。西双版纳黄瓜与栽培黄瓜的距离最远，与野生黄瓜次之。按一定的遗传距离可以将中国和外来栽培种质分开。大多数 华南型和华北型种质归属于不同的亚组。这些结果有助于有目的地利用这些变异拓宽育种材料的遗传背景。     

红肉猕猴桃种质资源果实性状及AFLP 遗传多样性分析

 对中国红肉猕猴桃种质资源进行收集和调查,并对其进行果实性状变异分析和AFLP 遗传多 样性及遗传关系分析。结果表明,红肉猕猴桃野生资源主要分布于湖南省、湖北省、河南省、江西省、 四川省和陕西省等地,共采集到52 份野生资源和2 份品种资源（包括软枣猕猴桃红肉类型、中华猕猴桃 红肉类型和美味猕猴桃红肉类型）。红肉猕猴桃种质资源在果实性状和DNA 分子水平上都存在丰富的变 异和较高的遗传多样性水平,4 对AFLP 引物共扩增出259 个多态性位点,多态性位点百分率为90.56%, Nei’s 基因多样性和Shannon’s 信息指数分别为0.318 和0.477;资源间遗传相似性系数介于0.568 ~ 0.883 之间,平均为0.714。聚类分析和主坐标分析将54 份资源划分为4 个组,软枣猕猴桃红肉类型单独聚为 一类;中华猕猴桃和美味猕猴桃红肉类型亲缘关系较近且有按地理来源优先聚类的趋势。果实性状数据 和AFLP 数据之间具有极显著的相关性,二者可结合用于红肉猕猴桃资源评价和保护利用工作中。