本文選題：SSR　切入點(diǎn)：SRAP　出處：《福建農(nóng)林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：木瓜(Carica papaya L.)也稱番木瓜,是世界上主要的熱帶水果作物之一,在世界各地大部分地區(qū)廣泛種植和消費(fèi)。據(jù)研究,番木瓜產(chǎn)量高且具有營養(yǎng)學(xué)和藥用功效,因此成為世界范圍內(nèi)具有重用應(yīng)用前景的水果。過去幾年,全球木瓜產(chǎn)量大幅增加,已經(jīng)成為發(fā)展中國家重要的出口創(chuàng)匯農(nóng)產(chǎn)品,已成為亞洲和拉丁美洲成千上萬人的經(jīng)濟(jì)支柱,因此選育高產(chǎn)優(yōu)質(zhì)的木瓜品種具有重要意義。過去,由于木瓜在生長(zhǎng)發(fā)育的幼苗期難以分辨出雌雄株,加上木瓜病害及品種間遺傳多樣性狹窄等因素的影響,嚴(yán)重阻礙了木瓜的育種進(jìn)程。近年來,隨著新的研究技術(shù)不斷進(jìn)步和發(fā)展,這些問題逐步得到了成功的解決。然而,對(duì)木瓜的遺傳多樣性研究的報(bào)道卻很少。分子標(biāo)記技術(shù)在木瓜上日益廣泛的應(yīng)用為木瓜育種和生產(chǎn)的長(zhǎng)足進(jìn)展鋪平了道路。目前,簡(jiǎn)單重復(fù)序列(SSRs)標(biāo)記由于具有多態(tài)性信息含量高和操作簡(jiǎn)便等優(yōu)點(diǎn),已成為木瓜遺傳多樣性研究上應(yīng)用最多的分子標(biāo)記方法。另一方面,序列相關(guān)擴(kuò)增擴(kuò)增多態(tài)性標(biāo)記(SRAP)是一種在木瓜研究上未得到廣泛應(yīng)用的新型分子標(biāo)記技術(shù)。對(duì)于中國和尼日利亞木瓜遺傳來說,研究主要集中在抗性品種的選育、性別相關(guān)標(biāo)記的鑒定和形態(tài)多樣性等方面,而關(guān)于中國和尼日利亞木瓜品種的遺傳多樣性則缺乏相關(guān)信息。本研究的目的彌補(bǔ)這一空白,增加木瓜研究可用的遺傳資源及方法。對(duì)收集于中國和尼日利亞這兩個(gè)不同地理區(qū)域木瓜品種的遺傳關(guān)系進(jìn)行了研究,并與已有的形態(tài)學(xué)數(shù)據(jù)進(jìn)行了比較。首先利用54個(gè)分子標(biāo)記(SSR和SRAP)分析了在野外收集到的60份(其中40份來自中國,16份來自尼日利亞)木瓜種質(zhì)資源的遺傳多樣性及親緣關(guān)系,利用木瓜鮮葉提取DNA后,根據(jù)不同分子標(biāo)記的方法對(duì)其進(jìn)行了擴(kuò)增。利用SDS-PAGE電泳技術(shù)對(duì)擴(kuò)增產(chǎn)物進(jìn)行檢測(cè)分析,通過NTSYSpc(2.10e版)軟件確定了多態(tài)性信息含量值,并計(jì)算了遺傳多樣性,將木瓜種質(zhì)資源分為三組,分別是中國組、尼日利亞組和中國-尼日利亞組。SSR標(biāo)記共擴(kuò)增到34個(gè),29個(gè)和31個(gè)等位基因,平均每個(gè)位點(diǎn)等位基因數(shù)分別是3.78,3.22和3.44,平均多態(tài)信息量分別為0.76,0.73和0.61,尼日利亞和中國的木瓜種質(zhì)資源等位基因數(shù)在每個(gè)分組中的變化范圍是1-6個(gè)。其中標(biāo)記SSR02和SSR21具有最高的多態(tài)性信息含量值,因此這兩個(gè)標(biāo)記最適合用于分析中國和尼日利亞木瓜種質(zhì)資源遺傳多樣性,而SSR49獲得的多態(tài)性信息含量值最低。SRAP分子標(biāo)記分別檢測(cè)到88個(gè),25個(gè)和67個(gè)等位基因,平均每個(gè)位點(diǎn)的等位基因數(shù)分別是1.96,1.47和1.76。每個(gè)分組的多態(tài)性信息量分別是0.49,0.37和0.47。M9E1和M2E16兩個(gè)標(biāo)記擴(kuò)增到的多態(tài)性信息含量最多,表明這兩個(gè)引物組合是可用于同類分析的高多態(tài)性信息量引物。利用SSR-SRAP結(jié)合方法同時(shí)擴(kuò)增來自中國和尼日利亞的木瓜種質(zhì)資源,可獲得121個(gè),54個(gè)和98個(gè)等位基因,平均每個(gè)位點(diǎn)的等位基因數(shù)分別是4.4個(gè),2.07個(gè)和2個(gè),多態(tài)性信息量分別是0.52,0.5和0.5。利用SSR-SRAP綜合引物和SSR引物在每個(gè)位點(diǎn)至少可以檢測(cè)到一個(gè)等位基因,可有效區(qū)分木瓜種質(zhì)資源,而SRAP標(biāo)記每個(gè)位點(diǎn)上檢測(cè)到的等位基因數(shù)在不同區(qū)組間有所變化。SSR標(biāo)記獲得的平均多態(tài)性信息量最高,因此表明SSR標(biāo)記比SSR-SRAP綜合標(biāo)記或單獨(dú)使用SRAP引物可獲得更多的等位基因多態(tài)性含量,這也解釋了為什么SSR標(biāo)記或許是目前可用于木瓜研究的最佳選擇。SRAP-SSR、SSR、SRAP擴(kuò)增結(jié)果的聚類分析表明,木瓜種質(zhì)資源具有豐富遺傳多態(tài)性,其遺傳相似系數(shù)(GS)分別是0.65、0.59和0.66。根據(jù)地理來源不同可將每大類木瓜種質(zhì)資源再分為兩個(gè)亞類,表明中國和尼日利亞的木瓜具有較高的遺傳差異性。而且在第一大類中,尼日利亞木瓜資源根據(jù)地理來源被分為不同的亞類,說明不同地理來源的木瓜具有遺傳多樣性。綜上所述,SSR標(biāo)記更適合于進(jìn)行木瓜種質(zhì)資源親緣關(guān)系和遺傳多樣性研究。
[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.

【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S667.9

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Ze-You Li;Yong Wang;Wen-Tao Shen;Peng Zhou;;Content determination of benzyl glucosinolate and anti—cancer activity of its hydrolysis product in Carica papaya L.[J];Asian Pacific Journal of Tropical Medicine;2012年03期

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本文編號(hào)：1628832