利用RAPD技术进行枣树的品种鉴定

用试剂盒法提取了5种枣树品种的总DNA,使用RAPD技术对其进行了品种鉴定.选出12种扩增较稳定的引物,来扩增5种枣树品种总DNA,共扩增出34条带,每种引物扩增出1～5条带;多态性条带共2条,多态性比例为58.82%.利用DPS软件所做的聚类分析结果表明:5个品种间可聚成2类:优系99-1(1号)、婆枣(3号)、赞皇大臻(2号)可聚为A类;优系97-4(4号)、梨枣(5号)可聚为B类,同一类品种间体现了一定的相关性.     

14个八仙花品种亲缘关系的ISSR分析

利用ISSR技术对14个八仙花品种进行了亲缘关系的研究。从49个引物(引物选自加拿大哥伦比亚大学UBC公司公布的100条引物)中筛选了8个条带清晰、多态性高、重复性好的引物,对14个八仙花品种DNA进行扩增,共扩增出478条带(平均每个引物扩增59.75条DNA片段),多态性条带420条。用POPGENE分析数据得到14个品种的遗传距离为0.000 2～0.087 4,遗传相似性系数为0.916 3～1.000 0;利用NTSYSpc 2.1聚类分析,在遗传相似性系数约为0.38处将14个八仙花品种明显分为两大类,其中花园蕾丝、绿色夏天和圣代草莓被聚为第一类,其余11个品种被聚为第二类。     

微卫星分子标记鉴定黑龙江主要水稻品种及遗传多样性的研究

选用分布于水稻12条染色体上的75对SSR引物,对25个水稻品种进行DNA扩增,筛选到特异扩增条带的SSR引物25对。共检测出71条多态性条带,用UPGMA聚类分析结果分析,遗传相似性系数为0.54处可分成2类,在第一类中遗传相似系数在0.73~0.94之间,除了松粳3号和龙粳14不能区分之外,其它品种均能区别,表明近缘品种之间的检出率较低。     

莲藕品种DNA多态性的初步研究

利用RAPD技术对12个莲藕品种进行了DNA多态性分析。从85个随机引物中筛选出12个引物扩增基因组DNA,共获得128条带,其中106条为多态性标记,每个引物平均提供10．7个标记信息。由UPMGA方法得到的聚类分析结果表明了12个品种间的亲缘关系,聚类结果与它们的系谱关系比较吻合。     

应用AFLP技术检测莲藕遗传多样性的初步研究

利用AFLP技术对12个莲藕品种进行了DNA多态性分析。从10对引物中筛选出2对引物应用于扩增基因组DNA,共获得72条带,其中64条为多态性标记,每个引物平均提供36个标记信息。由UPMGA方法进行聚类分析结果显示了12个品种间的亲缘关系,聚类结果与它们的系谱关系非常吻合。     

惠州引种马铃薯品种的ISSR分子鉴定

采用ISSR分子标记技术对7个惠州市引种的马铃薯品种进行遗传多样性研究.从30条ISSR引物中共筛选出3条引物,对7个马铃薯品种基因组DNA进行有效扩增,共扩增出25条带,其中多态性带20条,多态性比率为80%.用NTSYS-pc22.10 e软件计算各马铃薯品种阃的Jaccard遗传相似系数,7个马铃薯品种之间的遗传相似性系数界于0.35～0.85,平均遗传相似性系数为0.693.利用UPGMA进行的系统聚类分析表明,7个马铃薯可划分成2组,广引1号独立聚为一类,剩下6个马铃薯聚为另一大类.试验结果说明,引种的马铃薯品种间亲缘关系较近,遗传基础狭窄,有必要进一步扩大引种的数量和范围.     

SRAP标记技术在花生种子纯度鉴定中的应用

研究了分属3个市场型的10个中国花生栽培种的序列相关扩增多态性(SRAP)。结果表明, SARP技术可以揭示花生栽培种的遗传差异。在所试验的74对SRAP引物中,2对只获得了单态性条 带,其余72对总共产生了1,812条带,其中1035条(57．12％)为多态性带,每对引物组合平均获得25．16 条带、14．38条多态性带。在这73对引物组合中,总带数和多态性条带的数目分别为7-63和2-44 条。10个花生栽培种的简单匹配相似系数为0．6967-0．9171不等,根据SRAP指纹图谱进行聚类分析 可将这10个品种分为5类。用64对引物筛选作图亲本24-3与B4,获得329条多态性条带。此项研究 为花生品种鉴定和遗传研究提供了新的DNA分子标记技术手段。     

枣树不同品种遗传多样性的RAPD分析

为了验证现有枣树材料的亲缘关系,本研究利用RAPD标记对21种枣树品种的遗传多样性进行了分析。结果表明：从36个RAPD引物中,筛选出20个可扩增出清晰且多态性高的条带的引物用于RAPD-PCR扩增,共获得185条DNA片段,其中173条具有多态性,多态率达93.51%,平均每个引物可以扩增出9.25条。利用DPS软件建立了21种枣树品种的聚类分析树状图,聚类分析结果显示：在相似系数82.48处所有枣树品种被聚为一类,在65.98处21种枣树品种被分为两大类群。     

基于ISSR标记分析豌豆农家品种遗传多样性

为掌握内蒙古部分地区种植的农家豌豆品种的亲缘关系,本研究通过ISSR分子标记技术,对来自内蒙古自治区部分地区及甘肃省张掖市的29份豌豆农家品种的遗传多样性进行分析。结果显示,从19个ISSR引物中筛选出多态性明显、条带清晰、反应稳定的引物共7个。29份DNA材料共扩增出118条条带,其中116条为多态性条带,平均每个引物扩增出的条带数为16.9条,多态性比率为98.31%。Shannon多样性指数平均为0.505 9,每个位点的有效等位基因数为1.569 6,品种间遗传相似系数变幅为0.370 4～0.928 6,表明本研究的29个豌豆农家品种具有丰富的遗传多样性。利用UPGMA聚类分析,以遗传相似系数0.69为界限,将29份材料划分为5类,聚类结果与豌豆种群地域性分布规律关联性较高,表明ISSR分子标记技术的遗传多样性分析能够将地理来源相近的居群进行聚类划分,其研究结果可运用于豌豆种质资源保护及育种工作中。     

菠萝17份种质的ISSR分析

利用ISSR分子标记技术对17份菠萝种质进行了基因组DNA多态性分析。从62个ISSR引物中筛选出6个多态性引物用于PCR扩增，共扩增出70条DNA条带，其中多态性条带44条，所占比例为62.86%，平均每个引物扩增的DNA条带的数目为11.67条。6个品种具有特异性的扩增带，可作为种质鉴定的依据。根据ISSR扩增结果建立的UPGMA聚类图，在距离系数约0.1处可把供试的17份菠萝种质分为3大类。同时对供试品种的遗传关系进行了探讨     

Genetic diversity in soybean genotypes from north‐eastern China and identification of candidate markers associated with maturity rating

Random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to estimate the genetic relationships among 101 soybean cultivars developed in north‐eastern China. Fifty‐three fragments of the 100 RAPD markers and 35 SSR markers tested were polymorphic across the 101 soybean cultivars. Similarity values among these soybean cultivars ranged from 45.2% to 100% for RAPD data, and ranged from 36.1% to 100% for SSR data. The similarity matrices for SSR data and RAPD data were moderately correlated (r = 0.31, P < 0.05). Cluster analyses indicated that the cultivars released from the same seed company were mostly grouped together. A principal component analysis, based on the combined RAPD and SSR data, yielded a good separation of soybean varieties with different maturity ratings [represented by soybean Heat Unit (HU)]. The varieties with HU < 2200 were well separated from those with HU > 2200. Four RAPD markers and eight SSR markers were significantly associated with the maturity ratings of soybean.     

白菜种质遗传多样性与亲缘关系的ISSR标记分析

摘要：从分子水平用ISSR标记法对白菜遗传多样性进行分析,从100个ISSR引物中共筛选出11个多态性明显、条带清晰、反应稳定的引物,对65个样品DNA共扩增出107条谱带,平均每个引物扩增出9.72条带,其中多态性位点102个(93.5% )。种间遗传相似系数在遗传距离在0.40～0.65 之间,表明白菜栽培种内品种间的遗传基础相对较宽,存在较大的遗传变异性。利用UPGMA聚类分析表明：能将65个白菜地方品种划分为四大类。由ISSR标记聚类结果所表现出的大多种质之间的亲缘关系与其来源地有较大的相关性,但也有地理差别很大的白菜资源遗传关系较近的情况。本研究还表明,ISSR 标记比RAPD 标记具有更高的稳定性,在植物遗传多样性的分子标记或克隆研究中,可优先使用ISSR 标记。     

A comparative analysis of the genetic relationships between rye cultivars using RFLP and RAPD markers

Summary The genetic similarities of eight closely related rye cultivars were estimated using two molecular marking techniques: restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD). Cultivars were evaluated for variation by 11 random cDNA and genomic clones used in combination with four restriction enzymes and 40 decamer primers. A total of 53 polymorphic RFLP fragments and 94 polymorphic RAPD fragments were observed. Based on the presence/absence of fragments, two genetic similarity matrices were calculated which were then used in cluster analysis. Differences between pair of cultivars were observed in RFLP and RAPD dendrograms. RFLP analysis produced estimates of genetic relationships more in accordance with the partially known pedigree of the cultivars than did RAPD analysis. The use of bulk samples of DNA in these analyses affected the sensitivity of RAPD assays more strongly. Dendrograms which took into account all fragments produced, either by RFLP or RAPD, reflected better the relationships between cultivars than did dendrograms based on only one type of marker. This reflects the importance of the number of markers used in determining the genetic relationships between genotypes.     

Genetic relationships in cultivars of hop, Humulus lupulus L., determined by RAPD analysis

Random amplified polymorphic DNA (RAPD) was used to evaluate the genetic variability and relationship of 65 hop cultivars from all the major hop-growing regions in the world. Twenty-eight selected random primers used in the RAPD reaction generated an average of 38.6%) polymorphic fragments, which was sufficient to produce 47 different RAPD profiles among the cultivars examined. The level of genetic variability was much higher than previously reported. Genetic similarity was estimated and UPGMA cluster analysis was performed using the RAPD data. Cluster analysis separated the cultivars into genetically related RAPD groups which were compared with pedigree data and grouping of the hop cultivars by essential oil type. The RAPD groups, strongly supported by pedigree data, gave more precise information on the level and distribution of genetic variability within hop cultivars than characterization by essential oils. Cultivars were divided into American and European groups, supporting the distinction between two geo-graphically distinct hop germplasms. Five genetically distinct groups revealed differences within the European germplasm, reflecting past hop breeding practices which have been adopted in different regions. The use of RAPD markers for hop germplasm characterization and genetic diversity study is discussed.     

部分烟草种质遗传多样性与亲缘关系的ISSR标记分析

烟草遗传资源多样性与亲缘关系研究，是烟草遗传育种与起源演化研究的重要基础，本文首次应用ISSR标记，对烟草属（Nicotiana）4个种30份材料的遗传多样性进行分析。从70个ISSR引物中共筛选出16个多态性明显、条带清晰、反应稳定的引物，对30个样品DNA共扩增出309条谱带，平均每个引物扩增出19.31条带，多态性条带比率（PPB）达93.20%。种间遗传相似系数在0.26~0.96之间，表现出丰富的遗传多态性。系统聚类结果显示，N. glutinosa、N. suaveolens、N. gossei 3个野生种间存在较大的遗传差异，遗传相似系数在0.29~0.52之间；27份栽培品种种内遗传相似性相对较高，在0.54~0.96之间，显示出栽培种内的遗传基础相对比较狭窄，但其中白肋21、台烟7号与其他供试材料有较大的遗传差异。ISSR聚类分析表明，当L1取值为D = 0.475时，可将3个野生种与27份烟草栽培品种明显区分开，反映出种间的遗传差异；当L2取值为D = 0.776时，可将30份材料分为2个大类、3个小类和6个独立的个类，较好地揭示了烟草属种间或栽培种品种类型间的遗传多样性与亲缘关系，可为烟草遗传育种和遗传连锁图谱构建的杂交亲本选择提供科学依据。本研究还表明ISSR标记比RAPD标记具有更高的稳定性，在植物遗传多样性的分子标记或克隆研究中，可优先使用ISSR标记。     

Comparing RAPD and AFLPTM analysis in discrimination and estimation of genetic similarities among apple (Malus domestica Borkh.) cultivars

Forty-one apple (Malus × domestica Borkh.) cultivars were screened for RAPD (Random Amplified Polymorphic DNA) and AFLP(Amplified Fragment Length Polymorphism) markers. RAPD analysis was performed with 35 arbitrary 10-mer primers, selected from 60 primers tested (kits A, C and E, Operon Technologies, Inc.). Of a total of 362bands observed, 208 (57.5%) were polymorphic. Three-hundred-and-eighty-one AFLP fragments were obtained with 8primer combinations, of which 218 (57.2%) were polymorphic. Cultivars differentiated through mutation were included in this study and showed identical patterns when analysed with both RAPD and AFLP analysis. The estimated genetic relationships were correlated (r = 73.7%) between the analysis with the two different markers. UPGMA analysis was performed and dendrograms were constructed using either the data apart from each(RAPD and AFLP) method or combined in a single joint matrix. The relationships among the forty-one studied cultivars were basically consistent with the known lineage and geographic origins of the cultivars. The four Portuguese cultivars included in this study clustered together and diverged from the other cultivars. Apparently they constitute an independent genetic pool, which could be of interest for apple plant breeders. This revised version was published online in July 2006 with corrections to the Cover Date.     

部分蔷薇（Rosa multiflora）属植物的RAPD分析

蔷薇作为园林观赏植物,在中国城市绿化中应用较为广泛,但目前应用的蔷薇品种较为混杂,尚无系统研究。本研究采用RAPD技术对部分蔷薇属植物进行亲缘关系的分析,旨在进一步明确蔷薇种、品种,为引种育种以及材料选择提供参考。对蔷薇属植物的2个种、1个变种、20个品种进行了RAPD分析,16个引物对供试蔷薇材料PCR扩增获得175条谱带,其中多态性条带151条（86.3%）表现出丰富的遗传多态性。且不同花色系蔷薇属植物间的遗传多态性不同,白色系75.16%,粉红色系73.46%,黄色系37.50%。利用UPGMA法构建树状聚类图,蔷薇属植物的2个种、1个变种、20个品种在阈值为0.667处被分划为两大类群,这两大类群间的遗传距离较近,黄蔷薇与黄刺枚单独被划分为一个类群,与上述两类群间的遗传距离较远。聚类结果显示出了蔷薇的演化趋势：白色、单瓣是蔷薇较为原始的一个性状,花径大、重瓣性高是现代蔷薇栽培品种的性状,而部分粉红色半重瓣蔷薇则是从野生种到现代栽培品种的中间过渡类型。     

Random amplified polymorphic DNA variation among German and Dutch asparagus cultivars

DNA polymorphism among nine cultivars of Asparagus officinalis L. was measured using random amplified polymorphic DNA (RAPD). Of 69 reproducible amplification products from 12 arbitrary decamer primers, 49 RAPD markers were polymorphic and could be used to distinguish six German and three Dutch asparagus cultivars. Even with very small sample sizes, genetic similarity measurements based on the RAPD data allowed accurate grouping of the nine cultivars into distinct clusters, with the exception of two individuals which clustered to closely related varieties. Two German cultivars showed high genetic similarity and were distinct from the remaining German varieties. The German and Dutch cultivars were clearly separated by a relatively large genetic distance.     

Radiomutants of chrysanthemum (Dendranthema grandiflora Tzvelev) of the Lady group: RAPD analysis of the genetic diversity

Random amplified polymorphic DNA (RAPD) markers were used to study the molecular characterization of 10 new radiomutants of chrysanthemum. The original cultivar ‘Richmond’ differed in genetic distance from its Lady group mutants. The analysis of genetic similarity indices revealed low diversity within the radiomutants. The dendrogram obtained after cluster analysis separated the new cultivars as a group that differed from the original cultivar ‘Richmond’. The Lady group cultivars, derived from one original cultivar by radiomutation, could be distinguished from each other by using RAPD markers of only a single primer or sets of two or three primers. Polymerase chain reaction analysis proved the efficiency of the RAPD method for DNA fingerprinting of the original cultivar ‘Richmond’ and its new radiomutants.     

利用RAPD标记鉴定大豆种质

本研究以５７个中国大豆祖先吕系及育成品种和１８个美国大豆祖先品系为ＤＮＡ样品来源，通过随机引物ＰＣＲ扩增基因组ＤＮＡ的多态性，探索利用ＲＡＰＤ标记鉴定和相关种质的可能性。研究结果表明，５０个１０摩尔随机引物共扩增可分辩产物２４６个，其中８２．４％的随机引物可产生多态性产物，所扩增产物的５４．４％至少在两个基因毒草境存在差异。上ＰＣＲ扩增产物分别以１和０记录存在与否。扩增产物间的成对比较可产生非相似