【作者】 周娜；

【导师】 柳李旺；

【作者基本信息】 南京农业大学 ， 蔬菜遗传育种与生物技术， 2014， 硕士

【摘要】 萝卜(Raphanus sativus L.)世界上广泛栽培的蔬菜品种之一。我国萝卜种质资源丰富,是起源地之一。深入研究和合理利用种质资源对开展萝卜育种工作尤为重要。目前生产上,要求新品种丰产、广适、多抗及商品性好、品质优,这些目标的实现,与掌握的种质资源数量和质量密切相关。为了种质创新以及更加有效的利用优势资源,可以采用多种方法和手段对萝卜种质资源的遗传多样性进行系统研究。目前对于种质资源遗传多样性的研究方法颇多,优缺点共存。其中,分子标记不受季节、环境条件等外界因素影响,能直接反映研究材料分子水平差异。所以利用分子标记方法研究育种材料间的遗传差异,为萝卜杂交育种中亲本选配和后代纯合性鉴定提供可信度更高的理论依据,也为萝卜基因库的构建提供技术保障。本研究运用ISSR, RAPD及RAMP三种分子标记,结合系统聚类分析,对来源于国内外的32份萝卜种质的亲缘关系及遗传多样性进行分析研究。1选用35个ISSR随机引物,从中筛选出多态性较好的引物8个,对32份萝卜种质进行ISSR分析,扩增共产生条带82条,其中69条多态性带,多态性比率为84.1%,平均每个引物产生多态性条带8.6条；系统聚类分析将32份种质分为4大类群,第一大类群由28个品种组成,又分为3个亚组,第二大类群由MTHS单独组成,第三大类群由LYDHP和LSHLB组成,红皮红心的YZLB单独聚为第四大类群。2选用随机引物48个,筛选出多态性较好的6个,对来源不同的32份萝卜种质差异性进行RAPD标记分析,扩增产生总带数58条,多态性带为50条,多态性比率为86.2%,平均每个引物产生多态性带8.3条；系统聚类分析将供试材料分为4大类组,第一大组包括16份材料,又可分为4个亚组,第二大组11份材料,又可分为3个亚组,第三大组包括4份材料,以绿皮萝卜为主。3选用96对RAMP引物,筛选出8对引物组合对32份萝卜种质遗传多样性进行了RAMP分析,结果扩增产生总条带122条,多态性带113条,多态性比率92.6%,平均每对引物产生多态性条带高达14.1条；系统聚类分析将供试材料分为3个组,第一组包括29份材料,第一组由QTQ和FLXX组成,第三组由MTHS材料单独构成。4综合三种标记结果进行聚类分析表明,单个标记与综合聚类结果不完全一致,但是都与形态标记分类基本一致。都能将MTHS材料单独聚类；将来源于重庆周边地区的CBL类型、FT类萝卜基本聚在同一大类,亲缘关系表现很近；将JG与BJSG、根形长柱形的白皮萝卜JSNBZ和ZDC及YCLBF、LSHLB和LYDHP等都在一定相似系数处相聚,表明萝卜根形与聚类关系非常密切。本研究中综合三种标记分析聚类结果与RAMP标记分析结果最接近；综合三种标记分析结果进行聚类分析能更加直观的反映出供试材料间的亲缘关系,与主要园艺学性状的一致性比基于单一标记分析的结果更高。更多还原

【Abstract】 Radish (Raphanus sativus L.) is one of the important vegetables cultivated worldwide. China is one of area of radish origin and the germplasm resource very abundant, intensive study and rational utilization Is particularly important for breeding work of radish germplasm resource.On the preduction at the present, requirement of new fertility, good fitness, (and commodity, quality Requirement of new cultivar fertility, Widely suitable, multi-resistant and Good commercial,good quality, but the realization of the target depends on the mastery of the radish germplasm resources quantity and quality and the reasonable use. The study on the genetic diversity of radish can benefit the conservation,classification,identification,genetic enhancement and effective utilization of the radish germplasm resource. Molecular marker which can detect the genetic diversity at DNA level,is a powerful tool to study genetic diversity. Using molecular marker technology research and breeding materials of genetic diversity will provide radish classification, origin, genetic relationship with the basis of molecular level, And can be used as a selection, the progeny of radish genetic breeding to determine a rapid analysis method, At the same time for the construction of core radish germplasm repository to provide reliable technical support. This study adopted the ISSR markers, RAPD markers and the RAMP markers, Combining with the system cluster analysis, Of 32 radish germplasm genetic diversity and genetic relationship were studied.1. From 35 primers screened eight good polymorphic random primers, the ISSR analysis of 32 radish germplasm, amplification produced 82 bands, including polymorphism with 69, polymorphism percentage is 84.1%, each primer an average of 8.6 bands polymorphic primers. System clustering analysis 32 germplasm can be divided into four groups, the first big groups consists of 28 species, is divided into three subgroups, second largest groups of MTHS alone, third largest groups composed of LYDHP and LSHLB two materials, red one red hearts of YZLB alone as the fourth largest groups.2. From 48 primers screened 6 good polymorphic random primers, the RAPD analysis of 32 radish germplasm, a total of 58 stripe amplification, polymorphism of 50, polymorphism percentage is 86.2%, each primer an average of 8.3 bands polymorphic primers; System clustering analysis were used to material group is divided into four categories, the first big group including 16 material, and can be divided into four subgroups, the second largest group of 11 copies of materials, and can be divided into three subgroups, the third largest group includes four material, give priority to with green radish, The fourth largest group consists of MTHS separate from Japan.3. From 96 pairs of primers screened eight pairs of primers, of 32 radish germplasm are analyzed in the RAMP, amplification produced 122 bands,113 polymorphism bands, polymorphism percentage is 92.6%, average each pair of primers produced 14.1 polymorphic bands; System clustering analysis will be divided into three groups, the germplasms in the first group includes 29 of material, the first group consists of QTQ and FLXX 2 material composition, the third group consists of MTHS material separately.4. Comprehensive results of three kinds of marker cluster analysis showed that a single tag and comprehensive clustering sequence is not completely consistent, but they are almost the same classification and morphological markers.MTHS material can be separated clustering; CBL, FT type radish coming from the surrounding area chongqing can be basicly clustered same groups, their genetic relationship is very close; JG and BJSG, JSNBZ and ZDC and YCLBF, LSHLB and LYDHP are all be clustered together at a certain similarity coefficient, the shape of the root is showed very close relationship with clustering. Three comprehensive clustering tags, more intuitive to reflect the relationship among the germplasms, its consistency with horticulture traits is higher than any single marker. And in this study integrated clustering analysis results to the RAMP mark the closest.更多还原