应用RAPD技术对不同基因组组合生物型遗传多态性的研究(英文)

利用ＲＡＰＤ技术对不同基因组合的鱼类进行了基因组指纹图谱构建,在ＤＮＡ水平上对基因组成分进行了分析,探讨了其遗传多态性。ＲＡＰＤ结果发现,在２６个随机引物扩增的产物中,平均每个个体观察到约１４２个ＲＡＰＤ标记,单个引物获得的标记平均为５．４。其中４个引物扩增的图谱可将不同的生物型区分开：Ｓ－２６引物的扩增图谱（Ｆｉｇ．１）可将红鲫（ＲＡ）与其它组合区分开,还可将鲤鲫杂种一倍体（ＣＡ）与鲫鲤杂种三倍体（ＣＡＡ）和人工复合三倍体鲤（ＣＣＡ）区分开;Ｓ－８引物（Ｆｉｇ．２）可区分开红鲤（ＲＣ）和镜鲤（ＭＣ）;Ｓ－４５引物（Ｆｉｇ．３）可区分开ＲＣ和ＣＡ;Ｓ－２２引物则可区分开ＣＡＡ和ＣＣＡ。六种生物型均存在基因组特异性的图谱即各自独特的“诊断性”图谱,作者由此建立了详细的分子标记检索表（Ｔａｂｌｅ１）。通过对ＲＡＰＤ图谱的量化分析,利用ＵＰＧＭＡ构建了不同生物型的遗传关系树图;反映了鲤鲫及各种组合生物型之间的遗传相似关系：ＲＣ和ＭＣ属同一种系,聚为一族;ＣＡＡ和ＣＡ基因组类型相同,聚为一族;ＣＣＡ虽自成一体,但可与ＣＡＡ和ＣＡ聚为一族;而ＲＡ与其它组合遗传距离较远,自成一族。ＲＡＰＤ的结果也表明各种生物型内个体间     

RAPD标记构建家蚕分子连锁图

以大造、Ｃ１０８及其Ｆ２群体构建了１个家蚕的ＲＡＰＤ连锁框架图,该图含ＲＡＰＤ标记位点１８２个,来自大造的１０３个标记分属前２３个连锁群,来自Ｃ１０８的７９个标记分属后１６个连锁群,覆盖基因组的总长度为１１４８．３ｃＭ（ｃｅｎｔｉｍｏｒｇａｎ）,它能与本室用同一群体构建的ＳＡＤＦ图谱相整合,亦可与相应的ＲＦＬＰ图谱相互补充。     

RAPD标记构建家蚕分子链锁图

以大造、Ｃ１０８及其Ｆ２群体构建了１个家蚕的ＲＡＰＤ连锁框架图,该图含ＲＡＰＤ标记位点１８２个,来自大造的１０３个标记分属前２３个连锁群,来自Ｃ１０８的７９个标记分属后１６个连锁群,覆盖基因组的总长度为１１４８．３ｃＭ（ｃｅｎｔｉｍｏｒｇａｎ）,它能与本室用同一群体构建的ＳＡＤＦ图谱相整合,亦可与相应的ＲＦＬＰ图谱相互补充。     

应用RAPD技术对不同基因组组合生物型遗传多态性的研究

利用RAPD技术对不同基因组合的鱼类进行了基因组指纹图谱构建,在DNA水平上对基因组成分进行了分析,探讨陈春遗传多态性。PARD结果发现,在26个随机引物扩增的产物中,平均每个个体观察到约142个RAPD标记,单个的扩增图谱（Fig.1)可将红鲫（RA）与其它组合区分开,还可将鲫鲫杂种二倍体（CA)鲫鲤杂种三倍体（CAA）和人工复合三倍体鲤（CCA）区分开;S－8引物（Fig.2)可区分开红鲤（RC）和镜锂（MC）;S－45引物（Fig.3)可区分开RC和CA;S－22引物则可区分开CAA和CCA。六种生物型均存在基因组特异性的图谱即各种独特的“诊断性”图谱,作者由此建立了详细的分子标记检索表（Table1）。通过对RAPD图谱的量化分析。利用UPGMA构建了不同生物型的遗传关系树图,反映了鲤鲫及各种组合生物型之间的遗传相似关系:RC的MC属同一种系,聚为一族;CAA和CA基因组类型相同,聚为一族;CCA虽自成一体,但可与各种生物型内个体间的遗传相似率均大于群体间的（P＜0．05）。此外,本研究还对基因剂量效应对PAPD扩增图谱的影响进行了探讨。综合以上结果,RAPD技术有简捷,灵魂、花费少等特点、在鱼类品系鉴定和遗传多态性研究方面具有血型血清学和蛋白质多态性等其它技术无可比拟的优势。     

RAPD标记构建水稻分子连锁图

利用随机扩增多态性ＤＮＡ（ＲＡＰＤ）,在一个水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）的双单倍体（ＤＨ）群体中发展分子标记,仅用５２ 个ＲＡＰＤ标记建成了一个水稻ＲＡＰＤ分子连锁图。该图覆盖基因组的总长度为８９８．４ ｃＭ （ｃｅｎｔｉｍ ｏｒｇａｎ）,标记间的平均间距为１７．３ ｃＭ,它能与用同一群体构成的ＲＦＬＰ图谱互相补充     

水稻DH群体的分子连锁图谱及基因组分析

利用扩大的籼粳杂交来源（窄叶青８号×京系１７）的水稻（ＯｒｙｚａｓａｔｉｖａＬ．）加倍单倍体（ＤＨ）群体,构建了包含４４４个位点的分子连锁图谱,覆盖水稻基因组１９６２ｃＭ（ｃｅｎｔｉＭｏｒｇｏｎ）,标记间的平均图距小于５ｃＭ。此图谱包括２７６个ＲＦＬＰ标记、３４个ＲＡＰＤ标记、８９个微卫星标记、１０个ＡＦＬＰ标记、２６个端粒重复相关序列（ＴＡＳ）标记以及９个同工酶标记。该遗传图谱与其它的水稻高密度遗传图谱具有较高的可比性,并有自己的特点,适于进行各种持续性的遗传学研究     

中国白菜RAPD分子遗传图谱的构建

以芜菁（Ｂｒａｓｓｉｃａ ｃａｍｐｅｓｔｒｉｓ Ｌ．ｓｓｐ．ｒａｐｉｆｅｒａ Ｍｅｔｚｇ）和结球白菜（Ｂ．ｃａｒｎｐｅｓｔｒｉｓ Ｌ．ｓｓｐ ．Ｐｌｋｉｎｅｎｓｉｓ（Ｌｏｕｒ．）Ｏｉｓｓｏｎ）杂交的Ｆ２群体为试材,采用ＲＡＰＤ标记,用８４个１０核苷酸随机引物构建了白菜的ＲＡＰＤ遗传图谱。该图谱覆盖基因组的１６３２．４ｃＭ（ｃｅｒｔｉ Ｍｏｒｇａｎ）标记间的平均间隔为１６．５ｃＭ。其中最长的连锁群为     

大豆灰斑病菌生理小种的RAPD标记

运用随机扩增多态性ＤＮＡ（ＲａｎｄｏｍａｍｐｌｉｆｉｅｄｐｏｌｙｍｏｒｐｈｉｃＤＮＡ,ＲＡＰＤ）技术对发生于中国东北的大豆灰斑病菌（Ｃｅｒｃｏｓｐｏｒｉｄｉｕｍｓｏｊｉｍｕｍ）的１０个生理小种进行基因组ＤＮＡ多态性分析,用１３个１０－核苷酸随机引物获得了１１１个ＲＡＰＤ标记,其中８６．５％具有多态性,通过聚类分析确定了供试小种间的亲缘关系,试验证明,ＲＡＰＤ技术分析大豆灰斑病菌遗传传变异可提供大量     

新疆风毛菊属二新种

新疆风毛菊属二新种沈观冕（中国科学院新疆生物土壤沙漠研究所乌鲁木齐８３００１１）ＴＷＯＮＥＷＳＰＥＣＩＥＳＯＦＳＡＵＳＳＵＲＥＡＤＣ．（ＣＯＭＰＯＳＩＴＡＥ）ＦＲＯＭＸＩＮＪＩＡＮＧ,ＣＨＩＮＡＳＨＥＮＫｕａｎＭｉａｎ（ＸｉｎｊｉａｎｇＩｎｓｔｉｔ...     

花生种子高纯度DNA的提取（简报）

花生种子高纯度ＤＮＡ的提取（简报）王艳,何军贤,傅家瑞（中山大学生命科学学院,广州５０２７５）关键词花生;种子;ＤＮＡ;十六烷基三甲基溴化铵ＲＡＰＩＤＡＮＤＥＦＦＩＣＩＥＮＴＰＵＲＩＦＩＣＡＴＩＯＮＯＦＤＮＡＦＲＯＭＰＥＡＮＵＴ（ＡＲＡＣＨＩＳＨＹＰ...     

RAPD analysis reveals genetic variability among sexual and Apomictic Paspalum dilatatum poiret biotypes

Paspalum dilatatum is a valuable forage grass in the subtropics. This species consists of several sexual (tetraploid) and apomict (penta- and hexaploid) biotypes. It has been proposed that the presence of a genome of unknown origin, the X genome, is responsible for apomixis in penta- and hexaploid biotypes. Here we evaluated the utility of random amplified polymorphic DNA (RAPD) markers for discriminating sexual and apomictic P. dilatatum biotypes. DNA samples from nine accessions, including P. intermedium, P. juergensii, and P. dilatatum (ssp. flavescens, and the common and Uruguayan biotypes) were analyzed with 86 RAPD primers. Three hundred sixty-two fragments were scored and genetic similarity estimates revealed that the penta- and hexaploid biotypes were highly similar (S(D) > or = 0.913). Forty RAPDs were unique to the penta- and hexaploid biotypes. Overall RAPD markers were useful for assessing genetic variation among closely related P. dilatatum genotypes as well as generating putative X genome markers.     

锦鲤4个人工雌核发育家系的微卫星标记研究

利用Crooijmans et al.(1997)分离的包含CA重复单元的普通鲤鱼（Cyprinus carpio L.)的8个微卫星DNA标记,对从锦鲤（Cyprinus carpio L.)的红白,大正和昭和3个不同品系中所获得的4个不同人工雌核发育家系的20尾个体进行PCR扩增。电泳结果表明,8对引物在20尾个体中均能重复稳定地扩增出相应的同源序列。随引物不同,各等位基因数为1－11个,大小在68－264bp。在MFW4,MFW7,MFW19,MFW20,MFW23和MFW24 6个微卫星的扩增结果中,20尾个体的扩增图谱呈现了高度的遗传多态性,不同雌核发育家系内个体的遗传异质性也较大。其中大正（TaS)和红白1（RW1）的个体不仅花色分化显著,而且个体间的平均遗传距离分别高达0．28。通过对微卫星等位基因和基因型分析发现,由于锦鲤品系中的每一个体是通过不断地杂交选育而获得,基因组来源复杂,基因高度杂合。因此,只进行1代的人工雌核发育,其家系内仅部分个体的部分座位出现纯合。所获得的人工雌核发育锦鲤为后续的色素遗传调控机制研究提供了必要的实验材料;同时,所鉴定的微卫星分子标记为进行锦鲤的分子标记育种的基因组作图提供了理想的工具。     

Microsatellite and RAPD polymorphisms in Ontario corn hybrids are related to the commercial sources and maturity ratings

Random-amplified polymorphic DNA (RAPD) and microsatellite markers were used to estimate the genetic relationships among 37 Ontario corn hybrids. Almost all (95%) of the 160 RAPD fragments and all of the 79 microsatellite alleles were polymorphic across the 37 hybrids. Similarity values among the hybrids ranged from 31% to 86% when based on the RAPD data. The similarities based on microsatellite markers ranged from 12% to 77%. The genetic diversity revealed by microsatellite marker analysis was higher than that obtained from RAPD analysis. The similarity matrices for the microsatellite data and the RAPD data were moderately correlated (0.43). Cluster analyses based on either type of marker showed that most of the hybrids from the same company were closely related to each other. Both dendrograms clustered similar pairs or groups of hybrids. A principal component analysis, based on the combined RAPD and microsatellite data, yielded a good separation of the hybrids with Ontario Corn Heat Unit (OCHU) values <2800 from those with OCHU values >2800. Seventeen RAPD markers and 5 microsatellite markers were significantly associated with the OCHU ratings of the hybrids.     

Intravarietal heterogeneity of durum wheat is an important component of the species biodiversity

Sixty-four durum wheat varieties of the domestic breeding (USSR and Russia) were studied for herogeneity using various genetic markers: storage proteins (gliadins), RAPD, and microsatellite (SSR) markers. About a third of the studied varieties (24) were shown to be heterogeneous at the protein markers. These varieties contained from two to six biotypes. Using the molecular markers, the biotypes were found to differ not only in the gliadin-coding genes as determined with the protein markers, but also in other chromosome regions. Moreover, using SSR markers, some additional subbiotypes were detected within the biotypes defined with the gliadin markers. Thus, the intravarietal durum wheat heterogeneity is an important component of general biodiversity of the species.     

Intravarietal heterogeneity of durum wheat is an important component of species biodiversity

Sixty-four durum wheat varieties of the domestic breeding (USSR and Russia) were studied for heterogeneity using various genetic markers: storage proteins (gliadins), RAPD, and microsatellite (SSR) markers. About a third of the studied varieties (24) were shown to be heterogeneous at the protein markers. These varieties contained from two to six biotypes. Using the molecular markers, the biotypes were found to differ not only in the gliadin-coding genes as determined with the protein markers, but also in other chromosome regions. Moreover, using SSR markers, some additional subbiotypes were detected within the biotypes defined with the gliadin markers. Thus, the intravarietal durum wheat heterogeneity is an important component of general species biodiversity.     

Analysis of tetraploid Elymus species using wheat microsatellite markers and RAPD markers.

An analysis of Amplification fragment polymorphism of DNA from 27 accessions of 19 tetraploid Elymus species was carried out using 18 wheat microsatellite (WMS) primer pairs and 10 decamer primers. Ten WMS primer pairs produced multiple polymorphism on all accessions tested. Two independent phenograms, one based on WMS-PCR and one on RAPDs, separated the 19 tetraploid species into two main groups, viz., the SH genome species group and the SY genome species group. The results coincide with the genomic classification of these species and hence support previous studies showing that Elymus is not a monophyletic genus. The assays indicated that accessions within a species cluster together, which concurs with the morphological classification. Interspecific and intraspecific polymorphisms were detected by the WMS-PCR and RAPD analyses. Variation was observed among accessions of Elymus caninus. The WMS-PCR detected a much higher level of polymorphism than the RAPD analysis. WMSs seem to be more efficient markers than RAPD markers for studying the population diversity of Elymus species. The potential of cross-species amplification of microsatellite markers as an additional source for genetic analysis and applications in Elymus is discussed in the context of these results.     

Pathogenic and molecular characterisations of pigeonpea wilt pathogen Fusarium udum

Thirty two pathogenic isolates of Fusarium udum from different pigeonpea growing areas in India were studied for pathogenic and molecular variability. Pathogenic variability was tested on 12 pigeonpea differential genotypes, which revealed prevalence of five variants in F. udum. The amount of genetic variation was evaluated by Polymerase Chain Reaction (PCR) amplification with 20 random amplified polymorphic DNA (RAPD) markers and nine microsatellite markers. All amplifications revealed scorable polymorphisms among the isolates, and a total of 137 polymorphic fragments were scored for the RAPD markers and 16 alleles for the simple sequence repeat (SSR) markers. RAPD primers showed 86% polymorphism. Genetic similarity was calculated using Jaccard's similarity coefficient and cluster analysis was used to generate a dendrogram showing relationships between them. Isolates could be grouped into three subpopulations based on molecular analysis. Results indicated that there is high genetic variability among a subpopulation of F. udum as identified by RAPD and SSR markers and pathogenicity on differential genotypes.     

Using RAPD for estimating genetic polymorphism in and phylogenetic relationships among species of the genus Lycopersicon (Tourn.) Mill

RAPD genome analysis of 53 species and cultivars of the genus Lycopersicon (Tourn.) Mill. revealed their high genetic polymorphism (Tourn.) Mill., based on which their phylogenetic relationships were inferred. In total, 248 polymorphic DNA fragments were amplified. Intraspecific polymorphism was maximum (79%) in L. peruvianum and minimum (9%) in L. parviflorum. In general, genome divergence among cross-pollinating tomato species was substantially higher than in self-pollinating species. An UPGMA dendrogram constructed from the RAPD patterns was consisted with the Lycopersicon phylogeny inferred from the molecular data of RFLP, ISSR, and microsatellite analyses and with a classification based on morphological characters. The relationships of taxa within the genus Lycopersicon are discussed.     

A genetic map of melon (Cucumis melo L.) based on amplified fragment length polymorphism (AFLP) markers

 Genetic maps facilitate the study of genome structure and evolution, and the identification of monogenic traits or Mendelian components of quantitative traits. We evaluated 228 RAPD, microsatellite and AFLP markers for linkage analysis in melon (Cucumis melo L.) varieties MR-1 (resistant to Fusarium wilt, powdery and downy mildews) and Ananas Yokneum (AY; susceptible to these diseases) and constructed a detailed genetic map. The mapping population consisted of 66 backcross progenies derived from AY×(MR-1×AY). Despite a relatively low level of polymorphism in the species, AFLP markers were found to be more efficient in mapping the melon genome than RAPD or microsatellite markers. The map contains 197 AFLPs, six RAPDs and one microsatellite marker assigned to 14 major and six minor linkage groups, and covers 1942 cM with the average distance between adjacent markers of approximately 10 cM. The maximum distance allowed between markers is 27.5 cM. About 11% of the intervals (20 out of 173) are over 20 cM (but less than 27.5 cM). The map has immediate utility for identifying markers linked to disease resistance genes that are suitable for marker-assisted breeding. The use of microsatellite markers for integration with other maps is also discussed. Received: 12 March 1997 / Accepted: 20 May 1997