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。     

赞皇大枣的群体遗传多样性评价

为了探明三倍体赞皇大枣的遗传基础,利用22个多态性引物对赞皇县及周边县区的50个赞皇大枣类型、26个酸枣类型、8个枣品种等3个群体进行了RAPD扩增,共扩增285个位点。遗传多样性分析表明:赞皇大枣群体的多态性位点百分率为58％,小于枣品种群体(66％)和酸枣群体(89％);赞皇大枣群体共扩增出205种带型,而酸枣群体扩增出416种带型;赞皇大枣群体内平均遗传距离为0．087,小于枣品种群体(0．181)和酸枣群体(0．254)。说明:赞皇大枣的遗传多样性小于枣品种群体,酸枣群体有较丰富的遗传背景。     

甘蓝型油菜―萝卜附加系中萝卜染色体的(dp)RAPD标记

在已掌握甘蓝型油菜―萝卜附加系染色体构成,并构建萝卜分子连锁图谱的基础上,以甘蓝型油菜―萝卜附加系为材料,利用(dp)RAPD分子标记技术,筛选附加系中外源萝卜9条染色体(A～I)的分子标记,为进一步研究萝卜染色体与连锁群对应关系奠定基础.试验筛选了576个随机引物(组合),有413个能从附加系中扩增出萝卜染色体的(dp)RAPD标记,平均有效引物(组合)占71.7%,其中RAPD反应有效引物率67.7%,dpRAPD反应有效引物率73.2%.试验共得到899个萝卜染色体(dp)RAPD标记,包括354个RAPD标记和545个dpRAPD标记,覆盖萝卜整个基因组的全部9条染色体,不同染色体的标记数目变化范围在41～160,其中最多的是染色体C,最少的是染色体I.     

野蔷薇的DNA提取和RAPD反应体系的建立

以野蔷薇(Rosa multiflora)的冬末初春芽和夏初叶片为材料,研究了DNA的提取方法,并对影响随机扩增多态DNA(RAPD)的反应因素进行了优化.建立了适合野蔷薇RAPD的反应体系和反应程序,在20μL反应体系中,25ng模板DNA,2.0mmol/LMgCl2,0.2mmol/LdNTPs,1UTaqDNA聚合酶,0.1μmol/L引物.扩增程序为:94℃预变性4min,然后94℃变性45s,37℃退火1min,72℃延伸1min,37个循环,最后72℃延伸5min,4℃保存.     

紫花桔梗和白花桔梗的RAPD指纹图谱鉴定研究

用RAPD方法对紫花桔梗和白花桔梗进行了指纹图谱的研究，以期为桔梗花色的鉴定提供分子依据。采用BSA法从200个随机引物中筛选出具有多态性的7个引物。结果引物OPG02在检测的紫花桔梗个体中均能各自扩增出一条580bp左右的特异带，而在白花桔梗中则未见。表明该RAPD标记具有紫花桔梗特异性，可用于鉴别紫花桔梗和白花桔梗。     

Molecular based assessment of genetic diversity within Barbary fig (Opuntia ficus indica (L.) Mill.) in Tunisia

In this work, we report for the first time on the analysis of genetic diversity within a set of 36 Tunisian Opuntia ficus indica (L.) Mill. ecotypes using RAPD markers.     

用RAPD标记分析高羊茅的遗传多样性

采用RAPD分子标记技术对从国外引进的15个高羊茅品种的遗传多样性进行了研究。从60个随机引物中筛选出12个有效引物,它们共扩增出85条DNA带,其中59条为多态性带,占69.41%,平均每个引物扩增出多态性带4.92条;利用NTSYS-PC软件计算出的不同品种间Jaccard遗传相似系数(GS)变化范围较大,为0.373~0.932;根据得到的遗传相似性矩阵进行非加权组法(UPGMA)聚类分析,建立高羊茅品种的分子系统树状图;以相似系数0.68为标准,可将所有品种分为3类,品种翠丽和贝克各自聚为一类,其余13个品种聚成一类。     

Pathogenicity and RAPD analysis of Phytophthora nicotianae pathogenic to pepper in Tunisia

Nine isolates of Phtophthora nicotianae were isolated from infected pepper plants. Their pathogenicity was studied in Capsicum annuum in comparison with P. nicotianae isolates from tomato and tobacco. The pathogenicity test showed that pepper isolates of P. nicotianae are adapted to their host. Banding patterns obtained by RAPD analysis with six oligonucleotide primers revealed polymorphism that grouped the isolates independently of the plant host. The polygenic dendrogram showed that pepper isolates were more similar to tomato isolates than to tobacco isolates. The RAPD bands of 1300 and 1500 bp, detected with primers OPD-01 and OPD-10, respectively, appeared specific to the most pathogenic pepper isolates. The OPK-08-1950 seems specific to the isolates of P. nicotianae from tomato. These results suggest that host specified might occur in P. nicotianae and that may be due to interspecific hybridization events resulting in novel pathogenic behavior.     

Cloning of DNA fragments specific to the pathotype and race of <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Japanese isolates of Verticillium dahliae, a causal agent of wilt disease in many plants, are classifiable into pathotypes based on their pathogenicity. Because these pathotypes are morphologically indistinguishable, establishing a rapid identification method is very important for the control of this pathogen in Japan. For cloning DNA fragments that are useful for identification and specific detection of V. dahliae pathotypes, we performed random amplified polymorphic DNA (RAPD) analyses using various isolates. One polymerase chain reaction (PCR) product, E10-U48, was specific to isolates pathogenic to sweet pepper. The other product, B68-TV, was specific to race 1 of isolates pathogenic to tomato. The specificity of these sequences was confirmed by genomic Southern hybridization. Further analyses revealed that the region peripheral to B68-TV obtained from the genomic DNA library includes the sequence specific to all isolates pathogenic to tomato (races 1 and 2). Moreover, sequence tagged site (STS) primers designed from B68-TV and its peripheral region showed race-specific and pathotype-specific amplification in a PCR assay. The probes and primers obtained in this study are likely to be useful tools for the identification and specific detection of pathotypes and races of V. dahliae. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB095266.     

中国主栽双孢蘑菇菌株的DNA多态性

本文以中国双孢蘑菇不同主栽产区的9个具代表性的双孢蘑菇菌株为材料,用20个随机引物对其进行RAPD分析。结果表明,20个随机引物中,有15个随机引物的扩增产物DNA片段表现出明显的多态性,其中每个引物对不同菌株扩增出的DNA片段数目各不相同,而且不同引物扩增出的DNA带谱也不同,差异较大。这15个引物对供试菌株总共扩增出168条DNA片段,其中最大的片段可达6.77kb,最小的DNA片段仪有0.31kb。同时,比较了供试菌株两两间的遗传相似性,发现它们之间的变化不大,其相似系数从0.6891到1.0000,而平均相似系数仅为0.8500,表明供试菌株之间的亲缘关系较近,遗传变异不丰富。另外,采用系统聚类法中的类平均法,对供试菌株相似系数两两间进行聚类分析并生成树状图谱,直观准确地揭示了它们之间的差异。当相似系数为77％时,所有供试菌株都被聚为一大类,说明我国主栽双孢蘑菇菌株的遗传基础较狭窄,可能来源于同一品系,这为进一步开展双孢蘑菇菌种的遗传改良提供了理论依据。