本文選題：甘藍(lán)型油菜 + SSR分子標(biāo)記　； 參考：《華中農(nóng)業(yè)大學(xué)》2013年碩士論文

【摘要】：我國是世界上最大的油菜生產(chǎn)國,也是世界上第一個(gè)成功大面積利用油菜雜種優(yōu)勢的國家。油菜雜交種由于具有產(chǎn)量高、抗逆性強(qiáng)和適應(yīng)性廣等優(yōu)點(diǎn),深受種子企業(yè)的親睞和廣大農(nóng)民歡迎。但隨著市場商業(yè)化的不斷推進(jìn),不法經(jīng)營者的人為摻假或以次充好以及混雜或以不正當(dāng)手段獲取育種家的親本用于生產(chǎn)雜交種,損害了農(nóng)民、企業(yè)和育種家的合法利益。因此,迫切需要建立對雜交油菜品種及其親本進(jìn)行真實(shí)性和純度鑒定的技術(shù)體系,為親本材料的知識產(chǎn)權(quán)保護(hù)和維護(hù)農(nóng)民的合法權(quán)益提供技術(shù)支撐。 SSR標(biāo)記由于具有共顯性、穩(wěn)定性好、遺傳方式簡單以及能檢測多個(gè)等位位點(diǎn)等優(yōu)點(diǎn),被廣泛應(yīng)用于玉米、水稻等主要農(nóng)作物品種的純度和真實(shí)性鑒定。本研究利用SSR標(biāo)記對本實(shí)驗(yàn)室的120份甘藍(lán)型油菜種質(zhì)資源進(jìn)行分析,篩選出一批多態(tài)性豐富的SSR引物,同時(shí)構(gòu)建了骨干親本SSR-DNA的指紋圖譜,為油菜雜交種純度鑒定及其親本的真實(shí)性提供了可靠的參考依據(jù)。主要結(jié)果如下： 1、從120份材料中選取6個(gè)不同遺傳背景的材料所組成小樣本(包括2個(gè)不育系、2個(gè)恢復(fù)系和2個(gè)常規(guī)品系),對本實(shí)驗(yàn)室擁有的1938對SSR引物進(jìn)行篩選,共得到多態(tài)性引物543對,經(jīng)過比較選擇出帶型清晰、穩(wěn)定的SSR引物173對。 2、用173對引物對120份甘藍(lán)型油菜材料進(jìn)行分析,共擴(kuò)增出646條帶,其中多態(tài)性條帶512條,多態(tài)性比率為80%。一對SSR引物擴(kuò)增多態(tài)性帶數(shù)為2-8個(gè),平均為3.8個(gè)；信息多態(tài)量PIC值變化范圍0.05-0.92,平均為0.61。 3、利用512條多態(tài)性條帶對120份材料進(jìn)行UMPGMA聚類分析,以遺傳相似系數(shù)0.61為閥值,將120份材料分為A、B兩個(gè)大類。A類共100份材料,在相似系數(shù)0.69處劃分4個(gè)亞類,其中大部分不育系材料聚在第一亞類和第二亞類,說明不育系材料遺傳變異相對較�。欢�42份恢復(fù)系在兩大類中均有分布,表明恢復(fù)系材料遺傳變異比較豐富。 4、根據(jù)173對引物在其中32份核心親本中的擴(kuò)增結(jié)果,對有條帶出現(xiàn)賦值為“1”,無條帶出現(xiàn)賦值為“0”,構(gòu)建了32份甘藍(lán)型油菜核心親本的標(biāo)準(zhǔn)數(shù)字化指紋圖譜。同時(shí)初步設(shè)計(jì)親本材料圖譜快速查詢計(jì)算機(jī)數(shù)據(jù)庫程序。
[Abstract]:China is the largest rapeseed producer in the world and the first country in the world to successfully utilize heterosis in large area. Rapeseed hybrids are popular among seed enterprises and farmers because of their high yield, strong resistance and wide adaptability. But with the development of commercialization of the market, the illegal operators' artificial adulteration or subdued, mixed or improper means to obtain the parents of breeders is used to produce hybrids, which is harmful to the legitimate interests of farmers, enterprises and breeders. Therefore, it is urgent to establish a technical system to identify the authenticity and purity of hybrid rape varieties and their parents so as to provide technical support for protecting the intellectual property rights of parents and safeguarding the legitimate rights and interests of farmers. Because of its advantages of co-dominance, good stability, simple genetic mode and the ability to detect multiple alleles, SSR markers have been widely used to identify the purity and authenticity of major crop varieties such as maize and rice. In this study, 120 Brassica napus germplasm resources in our laboratory were analyzed by SSR markers, and a batch of polymorphic SSR primers were screened, and the fingerprints of the backbone parent SSR-DNA were constructed. It provides a reliable reference for the purity identification of rapeseed hybrids and the authenticity of their parents. The main results are as follows: 1. A total of 1938 pairs of SSR primers were screened from 1938 pairs of polymorphic primers (including 2 male sterile lines, 2 restorer lines and 2 conventional lines) from 6 materials of different genetic backgrounds, and a total of 543 pairs of polymorphic primers were obtained. After comparison, 173 pairs of SSR primers with clear band and stable pattern were selected. 2, 120 rape (Brassica napus L.) materials were analyzed with 173 pairs of primers, and 646 bands were amplified, of which 512 were polymorphic bands, with a polymorphism ratio of 80000. The number of polymorphic bands amplified by a pair of SSR primers ranged from 2 to 8, with an average of 3.8, and the range of PIC values of information polymorphism was 0.05-0.92, with an average of 0.61. 3. Using 512-polymorphic bands, 120 materials were analyzed by UMPGMA cluster analysis. The genetic similarity coefficient was 0.61 as the threshold value, and 120 materials were divided into two categories. A group of 100 materials were divided into four subgroups at the similarity coefficient of 0.69. Most of the male sterile lines were clustered in the first and second subclasses, indicating that the genetic variation of the male sterile lines was relatively small, while 42 restorer lines were distributed in the two major categories, indicating that the restorer lines had relatively rich genetic variation. 4. According to the amplification results of 173 pairs of primers in 32 core parents, the standard digital fingerprint of 32 core parents of Brassica napus was constructed. At the same time, the computer database program of parent material atlas was designed.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：S565.4

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本文編號：1976657