应用SSR和SRAP标记分析番木瓜种质资源遗传多样性及其亲缘关系

发布时间：2018-03-18 08:35

本文选题：SSR　切入点：SRAP　出处：《福建农林大学》2017年硕士论文　论文类型：学位论文

【摘要】：木瓜(Carica papaya L.)也称番木瓜,是世界上主要的热带水果作物之一,在世界各地大部分地区广泛种植和消费。据研究,番木瓜产量高且具有营养学和药用功效,因此成为世界范围内具有重用应用前景的水果。过去几年,全球木瓜产量大幅增加,已经成为发展中国家重要的出口创汇农产品,已成为亚洲和拉丁美洲成千上万人的经济支柱,因此选育高产优质的木瓜品种具有重要意义。过去,由于木瓜在生长发育的幼苗期难以分辨出雌雄株,加上木瓜病害及品种间遗传多样性狭窄等因素的影响,严重阻碍了木瓜的育种进程。近年来,随着新的研究技术不断进步和发展,这些问题逐步得到了成功的解决。然而,对木瓜的遗传多样性研究的报道却很少。分子标记技术在木瓜上日益广泛的应用为木瓜育种和生产的长足进展铺平了道路。目前,简单重复序列(SSRs)标记由于具有多态性信息含量高和操作简便等优点,已成为木瓜遗传多样性研究上应用最多的分子标记方法。另一方面,序列相关扩增扩增多态性标记(SRAP)是一种在木瓜研究上未得到广泛应用的新型分子标记技术。对于中国和尼日利亚木瓜遗传来说,研究主要集中在抗性品种的选育、性别相关标记的鉴定和形态多样性等方面,而关于中国和尼日利亚木瓜品种的遗传多样性则缺乏相关信息。本研究的目的弥补这一空白,增加木瓜研究可用的遗传资源及方法。对收集于中国和尼日利亚这两个不同地理区域木瓜品种的遗传关系进行了研究,并与已有的形态学数据进行了比较。首先利用54个分子标记(SSR和SRAP)分析了在野外收集到的60份(其中40份来自中国,16份来自尼日利亚)木瓜种质资源的遗传多样性及亲缘关系,利用木瓜鲜叶提取DNA后,根据不同分子标记的方法对其进行了扩增。利用SDS-PAGE电泳技术对扩增产物进行检测分析,通过NTSYSpc(2.10e版)软件确定了多态性信息含量值,并计算了遗传多样性,将木瓜种质资源分为三组,分别是中国组、尼日利亚组和中国-尼日利亚组。SSR标记共扩增到34个,29个和31个等位基因,平均每个位点等位基因数分别是3.78,3.22和3.44,平均多态信息量分别为0.76,0.73和0.61,尼日利亚和中国的木瓜种质资源等位基因数在每个分组中的变化范围是1-6个。其中标记SSR02和SSR21具有最高的多态性信息含量值,因此这两个标记最适合用于分析中国和尼日利亚木瓜种质资源遗传多样性,而SSR49获得的多态性信息含量值最低。SRAP分子标记分别检测到88个,25个和67个等位基因,平均每个位点的等位基因数分别是1.96,1.47和1.76。每个分组的多态性信息量分别是0.49,0.37和0.47。M9E1和M2E16两个标记扩增到的多态性信息含量最多,表明这两个引物组合是可用于同类分析的高多态性信息量引物。利用SSR-SRAP结合方法同时扩增来自中国和尼日利亚的木瓜种质资源,可获得121个,54个和98个等位基因,平均每个位点的等位基因数分别是4.4个,2.07个和2个,多态性信息量分别是0.52,0.5和0.5。利用SSR-SRAP综合引物和SSR引物在每个位点至少可以检测到一个等位基因,可有效区分木瓜种质资源,而SRAP标记每个位点上检测到的等位基因数在不同区组间有所变化。SSR标记获得的平均多态性信息量最高,因此表明SSR标记比SSR-SRAP综合标记或单独使用SRAP引物可获得更多的等位基因多态性含量,这也解释了为什么SSR标记或许是目前可用于木瓜研究的最佳选择。SRAP-SSR、SSR、SRAP扩增结果的聚类分析表明,木瓜种质资源具有丰富遗传多态性,其遗传相似系数(GS)分别是0.65、0.59和0.66。根据地理来源不同可将每大类木瓜种质资源再分为两个亚类,表明中国和尼日利亚的木瓜具有较高的遗传差异性。而且在第一大类中,尼日利亚木瓜资源根据地理来源被分为不同的亚类,说明不同地理来源的木瓜具有遗传多样性。综上所述,SSR标记更适合于进行木瓜种质资源亲缘关系和遗传多样性研究。
[Abstract]:Papaya (Carica papaya L.) also known as papaya, is one of the major tropical fruit crops in the world, widely cultivated in most areas around the world and consumption. According to the research, and has a high yield of papaya nutrition and medicinal effect, so the application prospect of the fruit become reusable worldwide. Over the past few years, the global papaya production increased significantly in developing countries, has become an important export of agricultural products, has become a pillar of tens of thousands of people in Asia and Latin America economy, therefore improving the yield and quality of papaya varieties has important significance. In the past, due to the growth and development of papaya in the seedling stage is difficult to distinguish between male and female plants, coupled with the impact of papaya diseases and genetic diversity among the varieties of factors such as stenosis the seriously hampered the breeding process of papaya. In recent years, along with the new technology of continuous progress and development, these problems have been successfully step by step The solution. However, the genetic diversity of papaya has rarely been reported. The application of molecular marker technique has been more and more widely in the papaya paved the way for the rapid progress of papaya breeding and production. At present, simple sequence repeat (SSRs) markers because of the merits of polymorphism information content is high and the operation is simple, has become papaya study on the genetic diversity of the most widely used on the molecular marker method. On the other hand, sequence related amplified polymorphism amplification (SRAP) is a new type has not been widely used in papaya research on molecular marker technology for China and Nigeria papaya genetic breeding research, mainly concentrated in the resistant varieties, sex related markers the identification and morphological diversity, and genetic diversity of Chinese and Nigeria papaya varieties of the lack of relevant information. The purpose of this study is to fill this gap, increase Study on genetic resources and methods available. The genetic relationship of papaya collected in China and Nigeria these two different geographical regions of papaya varieties were studied, and compared with the morphological data which have existed. Firstly, 54 molecular markers (SSR and SRAP) were analyzed in 60 samples collected (40 of which were from China, 16 from Nigeria) papaya germplasm resources of genetic diversity and genetic relationship of DNA extraction with fresh papaya leaves, according to the different methods of molecular markers were amplified. The amplified products of detected and analyzed by SDS-PAGE electrophoresis, through NTSYSpc (2.10e version) software to determine the polymorphism information content the value of genetic diversity and calculation, papaya germplasm resources were divided into three groups, China group, Nigeria group and Nigeria group China -.SSR markers were amplified to 34, 29 and 31 alleles Because, the average number of alleles per locus is 3.78,3.22 and 3.44 respectively, the average polymorphism information content were 0.76,0.73 and 0.61, the variation range of papaya germplasm resources. The number of alleles in Nigeria and Chinese in each group is 1-6. The markers SSR02 and SSR21 polymorphic information content of the highest value, so the two the most suitable for the analysis of genetic markers Chinese and Nigeria papaya germplasm resources diversity, polymorphism information content and SSR49 value obtained the lowest.SRAP molecular markers were detected in 88, 25 and 67 alleles per loci alleles are polymorphic information content 1.96,1.47 and 1.76. each packet are the most polymorphic information content of 0.49,0.37 and 0.47.M9E1 and M2E16 two markers amplified, indicating that these two primer combinations can be used for high polymorphism information content were similar analysis At the same time. By using the SSR-SRAP method combined with amplification from Chinese and papaya germplasm resources in Nigeria, available in 121, 54 and 98 alleles, the average number of alleles per locus were 4.4, 2.07 and 2, the polymorphism information content were 0.52,0.5 and 0.5. with SSR-SRAP primers and SSR primers in each locus can be detected at least one allele, which can effectively distinguish papaya germplasm resources, the average polymorphic information content and the number of alleles per locus SRAP markers detected in different regions vary between groups of.SSR markers is the highest, it showed that SSR marker than SSR-SRAP markers using SRAP alone or combined primers can obtain more allelic polymorphism content, which also explains why the SSR marker is perhaps the best choice available for.SRAP-SSR, papaya SSR for study, clustering analysis of SRAP results of the table Ming, papaya germplasm had rich genetic diversity, the genetic similarity coefficient (GS) are 0.65,0.59 and 0.66. according to the different geographical origin to each of the main categories of papaya germplasm resources were divided into two subtypes, showed genetic differences China and Nigeria papaya has high. And in the first category, Nigeria papaya according to the geographical origin resources are divided into different sub categories, indicating that the different geographical origin of the papaya has genetic diversity. To sum up, the SSR marker was more suitable for papaya germplasm genetic relationship and genetic diversity research.

【学位授予单位】：福建农林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S667.9

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