【作者】 郑鹭；

【导师】 祁建民；

【作者基本信息】 福建农林大学 ， 生物化学与分子生物学， 2007， 硕士

【摘要】 蓖麻是重要的油料作物。为深化蓖麻种质资源研究，本研究以国内外引进的81份蓖麻品种为材料，在完成蓖麻供试材料DNA提取的基础上，对81份材料进行了ISSR和SRAP分子标记，用筛选出的多态性较好的20个ISSR引物和20对SRAP引物组合对其进行PCR扩增和标记分析。按Nei-Li的方法计算其相似系数分别对81份材料进行分子聚类分析，其中对有栽培利用价值的75份材料的6个产量和品质性状（含油率）进行主成分分析。结果表明：（1）在75份蓖麻品种6个产量与含油率品质性状主成分分析结果表明：在前三个主成分中，第一主成分为产量构成因子，贡献率为58.41％；第二主成分为含油率构成因子，贡献率为13.87％。第三主成分为株高构成因子，贡献率为11.17％。三个主成分累计贡献率达83.45％，均有互为独立的性状构成因子。其主成分构成因素合理，能够较客观地揭示蓖麻种质基因型主成分构成性状向量特性及其相对重要性。（2）从品种主成分在欧氏距离系统聚类图可知，当横切线取值D=2.49时，可把75份品种分成L1-1、L1-2、L1-3和L1-4四类。这4类间的品种农艺性状有一定的差异。L1-1性状特征表现出：植株较矮，含油率较低，而亩产量中等偏上，单株果穗数较多。L1-2中的品种表现出来的一致性状特征是：株高中等，果穗较长，含油率、产量以及单株蒴果数都较高。L1-3中品种的性状特征是：植株较高（除了闻喜大粒蓖麻外）、单株有效穗较多、产量中等、含油率适中。L1-4中含有占大部分的59个蓖麻品种，这些品种的产量以及含油率都较L1-1和L1-2要低，品质一般。（3）利用75份品种前两个主成份二维聚类分析可见：当在横坐标1处作竖切线，在竖坐标0处作横切线，可把75个品种散布图分划成Ⅰ、Ⅱ、Ⅲ和Ⅳ4个象限，其中第Ⅰ象限中的11个品种为性状优良和较高含油率的蓖麻种质资源。在第Ⅱ象限中共有8个品种，这些品种都是高产中低含油率的种质资源。在第Ⅲ象限中，有33个品种为高含油率的种质资源。而在第Ⅳ象限中的其他多数品种是产量和含油率较低的品种资源。从蓖麻育种的目标以及对蓖麻优异品质的评价来看，第1象限中的品种可以直接应用于市场推广应用。（4）对81份供试材料提取的基因组DNA，筛选出多态性较好的20个ISSR引物共扩增出303条带，多态条带计252条，多态条带比率（PPB）为83.17％，遗传相似系数变幅范围在0.45685-0.94631；筛选出多态性较好的20个SRAP引物共扩增出263条带，多态条带计214条，多态条带比率（PPB）为81.37％，遗传相关系数变幅范围在0.32558-0.92973。（5）从ISSR和SRAP两种标记的分子聚合聚类结果表明：当在遗传相异系数为33.5％处作切割线L1时，81份材料可以分为L1-2、L1-3、L-4三个小类群以及由64份品种构成的大类群L1-1。其中L1-2小类群由6个分别来源于山西、辽宁、山东以及中国农科院的品种组成。L1-3小亚群包含有2个福建品种和2个山西品种。L1-4小亚类含有7个同源于山西的品种。L1-1大类群中包含有64个品种，由国内外各地栽培品种组成。当在遗传相异系数为30％时，对L1-1作切割线L2时可将其分为一个大亚类群L2-1和两个小亚类群L2-2和L2-3。其中L2-2含有7个品种，L2-3含有4个品种，这两个小亚类群共11个品种全都是山西省的地方品种。当对大亚类群L2-1细分时，又可将其分为两类群Ⅰ和Ⅱ。Ⅰ类里有24个品种，都为国内品种，其中福建品种4份，山西品种20份。Ⅱ类里有28份品种，这一类的品种来源地较复杂。来源地有山西、安徽、辽宁、云南和法国等地。（6）从81份供试材料ISSR和SRAP以及两者聚合所得的数据构建的分子聚类树状图可知，两种标记都可较好的把不同地域来源的材料区分开来，大多数来源相同的品种都能够聚在一起，展现了蓖麻品种资源丰富的遗传多样性。虽然ISSR和SRAP的聚类分析结果存在差异，但对两种方法进行的相关性分析表明，他们之间仍存在极显著相关性，相关系数为0.6769，达极显著标准(r_0.01=0.0452)。在对两种标记复合聚类与单一聚类的比较当中，SRAP标记聚类结果与之比较靠近，因此SRAP在品种资源的遗传多样性及亲缘关系研究中有更强的可靠性。综上所述：本研究对深化蓖麻种质资源遗传多样性与亲缘关系，以及在遗传育种的利用研究都可起到重要的指导作用，研究表明ISSR和SRAP两种分子标记是蓖麻品种鉴定和遗传多样性研究的有效手段。更多还原

【Abstract】 Ricinus Communis is an important oil plant. In order to further research on the Ricinus Communis resources. The research did the ISSR and SRAP markers to 81 materials in the base of extract their DNA. Then evaluated the genetic diversity and relationships of 81 Ricinus Communis varieties according to ISSR and SRAP markers by amplified with 20 ISSR primers and 20 SRAP primers. And principal components (PC) of six yield and quality characters in 75 Rininus Communis varieties which have cultivation value were analysed. The main results showed as follows:(1) The results of the Principal components (PC) of six yield and quality characters in 75 Rincinus Communis varieties are as follow: the first three PCs, which might be regarded as yield component factor (58.41%), oilyield factor (13.87%) and plant height factor (11. 17%). The cumulative contribution of the three factors was 83.45% among the varieties. It indicated that three factors were independence to each other. And it can post the vector character and the importance of component factor to Rincinus Communis core germ plasm.(2) From the graph of hierarchical cluster diagram of 75 Rincinus accessions, it shows: at the level of D=2. 49, all varieties were clustered into four different agriculture character groups, L1-1、L1-2、L1-3 and L1-4. The agriculture characters of varieties of L1—1 are: low, low fruit yield and middle oil yield. L1-2’ s are: middling tall, long fruit, high oil、capsule、and fruit yield. L1-3’s are: more tall、more capsule and middle oil and fruit yield. In the Ll-4, it has 59 Rincinus Commnuis varieties which oil and fruit yield are lower than the varieties of L1—1 and L1-2.(3) Base on the graph of D-sorting fig. based on the first and second principle components of 75 Rincinus Commnuis cultivars, it shows that: when makes the vertical tangent in x-coordinate 1, and does in vertical tangent in coordinates 0, it may divide 75 varieties dispersion patterns I, II, III and IV 4 quadrants. There are 11 varieties in the first quadrant, all of them are fine and the higher oil extraction rate. There are 8 varieties in the second quadrant, these varieties all are in the highproduction and the low oil rate. In the third quadrant, there have 33 varieties, all of them are have high oil extraction rate. But in the fourth quadrant, most of it has lower output and lower oil extraction rate. Looked from the castor-oil plant breeding goal as well as to the castor-oil plant outstanding quality appraisal that, in the first quadrant variety may directly be supposed to use in the market introduction application.(4) 303 DNA bands were amplified with 20 ISSR primers, of which 252 (83. 17%) were polymorphic. The similitude soefficient are age form 0.45685 to 0.94631. And 263 DNA bands were amplified with 20 SRAP primers, of which(81. 37%) were polymorphic. The similitude soefficient are age form 0.32558 to 0.92973.(5) Form two UPGMA cluster based on the aggregation between ISSR and SRAP, we will know that: at the level of D= 33.5, 81 materials may divide into four part, L1-1、L1-2、L1-3 and L1-4. L1-2 is made form six varieties, which originates from Shanxi, Liaoning, Shandong. L1—3 contains two Fujian varieties and two Shanxi varieties. The Ll-4 includes seven varieties, all of them are form Shanxi. At the level of D=0. 30, L1-1 can be divided into two three part: L2-1、L2-2 and L2-3. L2-2 includes seven varieties, L2-3 includes four varieties, all 11 varieties of two part are from Shanxi. When subdivides to L2-1, it also may divide into I and II. There 24 varieties in I, all of them are the domestic variety. In II kind, it has 28 varieties, the origination of these varieties is more complex. The origin has places such as Shanxi, Anhui, Liaoning, Yunnan and France.(6) Form three UPGMA cluster based on ISSR、SRAP and their aggregation, we will know that , both of two markers can division different clime varieties and exhibit they resourceful heredity diversity. Though there have some different between two markers, they exist best notability relativity with each other (0.6769) by relativity analyzed. When compare to their aggregation result, SRAP is close up to it than ISSR. So SRAP is doughty reliability to the study of varieties genetic diversity and relationships.The result of this research must have some guidance function for the breeding work of Rincinus Commnuis, at the same time it shows that ISSR and SRAP are availability to be used for identifing Rincinus Communis varieties and for researching their genetic diversity.更多还原