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。     

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。     

福建省主要杨桃品种遗传多样性分析

采用RAPD技术对17份杨桃品种资源进行遗传多样性分析.结果表明,13条随机引物共扩增出80条RAPD带,其中77条为多态性带,平均多态位点5.92条,多态率达97.19%,表明杨桃遗传多样性的复杂性.利用UPGMA构建了杨桃的亲缘关系树状图,遗传距离为0～2.16093,当入=2.08时,17份杨桃品种可以划分为3个...     

24个大果沙棘品种的RAPD分析

为了解沙棘种质资源的遗传多样性和品种间的亲缘关系,以24个大果沙棘品种为材料,利用随机扩增多态性DNA(RAPD)方法进行沙棘的遗传多样性分析。从120个随机引物中筛选出多态性丰富的18个引物用于扩增反应,在24个大果沙棘品种中检测到171个RAPD位点,其中多态性DNA带112条,多态位点百分比为65.5%,表明大果沙棘品种具有丰富的遗传多样性;在RAPD分析的基础上,对大果沙棘品种进行了聚类分析,结果表明,以相似系数0.58处作为分界,可将供试的24份大果沙棘种质分为4类。     

番茄种质资源遗传多样性的RAPD分析

利用RAPD分子标记技术,对63份番茄种质资源进行了遗传多样性研究。结果表明:从180个RAPD引物当中,筛选出22条稳定性好、多态性强的RAPD引物,共扩增出多态性带207条,多态性条带比率为86.96%,63份番茄栽培品种或品系间的相似系数介于0.28～0.99之间,说明RAPD能很好的对番茄野生种、近缘野生种和栽培种进行多态性分析。采用UPGMA进行聚类分析,得到与生物学分类地位基本一致的结果。     

燕山板栗种质资源遗传多样性的RAPD分析

为研究燕山板栗的遗传多样性,采用随机扩增多态DNA(randomamplifiedpolymorphicDNA,RAPD)技术对36份燕山板栗种质进行了分析。分析了燕山板栗的遗传丰富度,并对包括36个燕山板栗品种和8份外来板栗品种在内的44份板栗种质进行聚类分析。结果表明,RAPD能有效地区分品种间的差异,用16个随机引物经PCR扩增共得到132个片段,其中有83个多态性片段,占62.9%;不同遗传位点之间遗传多样度最大可达0.444,最小值为0.096,平均多样度为0.187;UPGMA法聚类,将44份板栗种质聚成4个大的类群,36份燕山板栗可分为3个大的类群,外来种质聚为一类。燕山板栗明显不同于外来品种。在RAPD图谱中,找到了19个品种(类型)的特异性标记和标记组合,可作为品种(类型)分子鉴别的依据。     

厚皮甜瓜遗传多样性的SSR分析

采用61对SSR特异引物对46份厚皮甜瓜材料进行分析,其中52对扩增出条带,47对引物具有多样性,扩增带分子量在150～1500bp,187条谱带中154条谱带具有多态性,多态率占82.35%,平均每个引物可扩增出3.60条带。同时,对材料网纹、果形、单果质量、有无条带、果皮颜色、果肉颜色、肉质、种子、有无麝香味、裂叶、是否自落和花性型等12个性状进行了田间调查,经过phylip软件和NTSYS软件分析后,计算出材料间遗传距离,并做出SSR分子标记和性状2个聚类图。结果显示,46份材料的遗传距离在0.0188～1.2119,而单一性状气味、果皮颜色、果肉颜色和表面网纹的遗传距离都小于0.1,十分接近;2种聚类分析结果存在较大差异,这与供试材料的亲缘关系十分复杂,大部分材料来源相近,具有相同的亲本,和回交亲本存在一定关系。     

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。     

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。     

杧果主要品种遗传多态性的AFLP标记研究

 采用扩增片段长度多态性AFLP标记对国内外31个杧果主要品种进行分类与亲缘关系研究。结果表明: 筛选出的14对引物组合在31份杧果种质中共扩增出了1 761条带, 其中多态性带的比例为97%。平均每对引物产生125.8条带和121.6条多态性带, 总的多态性带率为97%; 基于AFLP标记, 以0.48的相似系数为阈值, 所有供试　果材料被分为7大组群, 第一组群内以0.52为阈值, 又可分为6组。与形态标记相比, 基于AFLP标记的杧果分类体系更能反映杧果品种间的亲缘关系。     

A PCR-based assessment of genetic diversity,and parentage analysis among commercial mango cultivars and hybrids

Summary

Three different PCR methods [Random Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeats (ISSR), and Directed Amplification of Minisatellite DNA (DAMD)] were used to analyse genetic diversity and parentage among 20 mango cultivars, including 18 landraces and two hybrids (‘Amrapali’ and ‘Mallika’). These hybrids together with a third hybrid (‘Ratna’), and an out-group species (Mangifera sylvatica) were also analysed for parentage. Fifteen, seven and four primers were used to amplify a total of 158, 69 and 59 distinct DNA fragments by RAPD, ISSR and DAMD, respectively. Of these, approx. 85%, 64% and 90% were polymorphic, respectively. Genetic distances between pairs of mango cultivars were measured separately by each method and depicted graphically as a Neighbor Joining (NJ) tree. The three methods revealed different groupings of cultivars and hybrids. A NJ tree based on the cumulative data from all methods correlated well with the parentage of the mango hybrids, and the grouping of cultivars on a regional basis. Genetic markers likely to be associated with important agronomic traits were identified by further analysing the hybrids, with their respective parents, using all three methods. On the basis of the highest number of polymorphic bands observed (90%), DAMD was judged to be the best method with which to analyse mango germplasm.     

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.     

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。     

Assessment of genetic diversity among mango (Mangifera indica L.) genotypes using RAPD markers

Knowledge about the extent of genetic diversity/relatedness in mango germplasm is vital for developing coherent strategies for future gains in productivity. The genetic diversity/relatedness among mango cultivars/genotypes developed in Pakistan has not been investigated previously. We have assessed the genetic diversity among 25 mango genotypes/cultivars using randomly amplified polymorphic DNA (RAPD). Sixty random ten-mer primers were surveyed, out of which 45 yielded amplicons in all the genotypes. Genetic similarity between genotypes/cultivars was in the range of 64–89% with an average of 74%. Similarly, the genetic relatedness among all variants derived from a mango cultivar Chaunsa was in the range of 81.18–88.63%. These coefficients were utilized to construct a dendrogram using the unweighted pair group of arithmetic means (UPGMA). The genotypes were grouped into three (A, B, C) clusters. Generally, genotypes originating from Pakistan were grouped in cluster ‘A’ while cluster ‘B’ primarily composed of southern India as well as Florida cultivars. Kensington Pride was the most distantly related genotype which grouped with Maya and Yakta, forming a distinct cluster ‘C’.     

SSR标记及其在葡萄上的应用

SSR(simple sequence repeat)作为第2代基于PCR的一种新型DNA分子遗传标记,数量丰富、多态性高、重复性好、分布于整个基因组、特异位点扩增、发生频率高、共显性遗传。对SSR引物开发,SSR标记在葡萄品种鉴别、系谱重建、遗传图谱构建、商业品种保护等方面的研究进展进行了综述,并指出了未来的研究方向。     

Genetic relatedness of sweet cherry (Prunus avium L.) cultivars from Ukraine determined by microsatellite markers

Microsatellite (SSR) markers were used to characterise 23 sweet cherry cultivars of Ukrainian, and four cultivars of non-Ukrainian, origin. To assess their genetic diversity and relatedness, 11 pairs of primers were applied to microsatellite loci, resulting in amplification of 66 SSR alleles. The mean value of the number of different alleles, and the polymorphic index content, amount to 7.333 and 0.700, respectively, demonstrating a significant genetic diversity of the investigated sweet cherry cultivars. Four highly polymorphic SSR loci (EMPAS02, EMPAS06, PceGA34, UDP98-412), which belong to the list recommended by the European Cooperative Program for Plant Genetic Resources, can be used as a minimum genetic marker set for identification of the majority of the studied cultivars; however, for successful discrimination of the most similar cultivars, more markers, located on all chromosomes of sweet cherry, appear to be necessary. Application of unweighted variable-group method using averages clustering allowed elucidation of the relatedness among the sweet cherry varieties, and showed that the Ukrainian cultivars combine genetic material of local, western European, and probably Caucasian origin; however, the origin of several cultivars still remains unclear, and should be studied additionally.     

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。     

基于表型和SRAP 标记的切花菊品种遗传多样性分析

 利用45 个表型性状和SRAP 标记分析56 个切花菊品种的遗传多样性。表型变异分析结果 表明：21 个性状表现出品种内一致性高及品种间特异性强;主成分分析发现,主成分贡献值较大的性状 有花序直径、花序类型和瓣型等花部性状,其次是叶部和茎杆性状,说明所选用的切花菊品种在分类时 应以花部性状为主,叶部和茎杆性状为辅;表型性状基于遗传距离UPGMA 聚类,将56 个切花菊品种分 为平瓣类、匙瓣类、桂瓣类、匙瓣-平瓣类和管瓣类,聚类结果大致按照花径-瓣型-花型分类。14 对 SRAP 引物组合扩增56 个切花菊品种的DNA,共扩增出454 条带,其中多态性带423 条,占扩增带数的 93.17%,多态性含量PIC 值在0.72 ~ 0.89 之间,平均为0.82,说明切花菊品种在分子水平上具有丰富的 遗传多样性。基于SRAP 标记的UPGMA 聚类分析显示：品种间遗传相似系数在0.64 ~ 0.97 之间,将56 个切花菊品种分为平瓣类-匙瓣、桂瓣类和管瓣类,聚类结果大致按照花径-瓣型-花型分类。Mantel 检验相关性系数为0.682,两种聚类结果有相似之处,均能很好体现试验切花菊品种间的遗传关系。     

与苹果果皮红色性状相关的RAPD分子标记的筛选

用355条10bp随机引物来筛选与苹果果实红色性状连锁的分子标记。通过分离群体分组分析,在果实红色与非红色2个对比基因池间进行了扩增筛选,初步选出49个多态性引物,进一步对这些多态性引物进行群体上的分析发现S519可以扩增出大约1000bp左右的一条与红色性状相关的DNA特异带。对本研究的73个来自3个不同杂交组合的F1后代个体分析表明,该标记判断果实红色性状的符合率为76.7%。鉴于获得的多态性标记对果皮颜色预测的准确率不高的现状,我们采用了双标记组合分析方案,即用S5191000分别与以往获得的3个与果色性状相关的标记(S12891200、S12351000和S21071200)配对组合进行判断,从而使得标记对目标性状预测的正确性大为提高。另外,对总共获得的4个RAPD标记在9个栽培品种上进行了分析,大多数品种的表现与理论一致。