白菜型油菜花色遗传及花色基因精细定位

发布时间：2018-03-11 01:09

本文选题：白菜型油菜　切入点：白花　出处：《华中农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：白菜型油菜是我国三大油菜类型之一,也是芸薹属中代表性的栽培作物。植物的花色主要是由花瓣中含有的植物色素决定,为了解析白菜型油菜花色性状的分子机制,了解芸薹属中花色的不同调控途径,揭示芸薹属花色进化的模式,本课题利用白菜型油菜黄花品种P3246和白花品种W01杂交后代的F2群体进行花色基因的初步定位和精细定位。主要的研究结果如下:1.花瓣和叶片类胡萝卜素含量的测定采用高效液相色谱法(HPLC)对P3246和W01的叶片和花瓣进行了类胡萝卜素含量的测定,不论是在叶片还是花瓣中两个材料间类胡萝卜素的组成成分没有差异,而各成分的含量均有差异。特别是在花瓣中,含量最高的两种成分是紫黄质和叶黄素,含量最高的紫黄质在P3246中含量高达3009.86μg/g,占花瓣中类胡萝卜素总量的86.6%,在W01中含量为242.06μg/g,占花瓣中类胡萝卜素总量的72.0%,P3246中类胡萝卜素的总量为3475.60μg/g,W01中类胡萝卜素的总量为336.38μg/g,前者是后者的10.3倍。2.花色遗传规律的分析将白菜型油菜黄花品种P3246(P1)和白花品种W01(P2)杂交,构建F2分离群体,共获得3104个F2单株。在盛花期进行花色性状考察,其中黄花单株2321株,白花单株783株,经卡方检验花色表型分离比符合3:1的理论值(X_c~2=0.073,X_(0.05,1)~2=3.84,P0.05),确定白菜型油菜白花性状受单个隐性基因控制。3.花色基因的定位采用集团分离分析法(BSA)对142个InDel标记进行筛选,获得4个与花色基因连锁的InDel标记,初步将花色基因定位在A02染色体。从对应的A02染色体区段选择50个InDel标记,另外根据该区段参考基因组序列新开发了106个SSR标记,对亲本和Bulk池进行筛选获得15个多态性SSR和InDel标记,采用BSA分析将花色基因初步定位在标记A02-1142和A02-1144之间,根据标记与花色基因间的交换单株数确定遗传距离为1.5cM。用该区段9个多态SNP标记和1个多态性InDel标记对F2群体2330单株进行基因型分析,最终将花色基因定位在SNP标记A02_SNP-378和A02_SNP-1100之间,遗传距离为0.39cM,对应白菜参考基因组A02染色体约162kb的物理区间内,其中SNP标记A02_SNP-389与白花基因共分离。4.目标区段候选基因的分析在白菜参考基因组V1.5的162kb目标区段物理序列上,共发现25个注释基因。结合同源拟南芥基因序列注释,对这25个基因的功能进行了初步分析。
[Abstract]:Brassica campestris (Brassica campestris) is one of the three rapeseed types in China, and it is also a representative cultivated crop in Brassica. The flower color of Brassica campestris is mainly determined by the pigment contained in the petals, in order to analyze the molecular mechanism of the color character of Brassica campestris. To understand the different regulation ways of flower color in Brassica, and to reveal the pattern of flower color evolution in Brassica. In this study, the primary mapping and fine mapping of flower color gene were carried out by using F2 population of Chinese cabbage yellow flower variety P3246 and white flower variety W01. The main results were as follows: 1. Determination of carotenoid content in petals and leaves. The content of carotenoid in leaves and petals of P3246 and W01 was determined by high performance liquid chromatography (HPLC). There was no difference in the composition of carotenoids between the two materials in leaves or petals, but the contents of each component were different, especially in petals, the two components with the highest content were violaxanthin and lutein. The highest content of purple lutein in P3246 was as high as 3009.86 渭 g / g, which accounted for 86.6% of the total carotenoid in petals, 242.06 渭 g / g in W01, and 72.0 渭 g / g in total carotenoid in P3246. The total amount of carotenoid in P3246 was 3475.60 渭 g / g W01. 336.38 渭 g / g, the former is 10.3 times as much as the latter. The genetic law of flower color was analyzed. The Chinese cabbage (Brassica napus L. yellow flower variety P3246P1) and the white flower variety W01P2) were crossed. A total of 3104 F2 plants were obtained by constructing F _ 2 segregated population. The characteristics of flower color were investigated at the flowering stage, including 2321 single yellow flowers and 783 white flower plants. The phenotypic segregation ratio of flower and color of Brassica campestris was determined to be in accordance with the theoretical value of 3: 1. The phenotypic segregation ratio of flower and color of Brassica campestris was in accordance with the theoretical value of 3: 1. The phenotypic segregation ratio of flower and color of Brassica campestris was determined to be in accordance with the theoretical value of 3: 1. The phenotypic segregation ratio of flower and color of Brassica campestris was determined to be controlled by a single recessive gene. Four InDel markers linked to the floral color gene were obtained, and the floral color gene was preliminarily located on the A02 chromosome. 50 InDel markers were selected from the corresponding A02 chromosome segment, and 106 new SSR markers were developed according to the reference genomic sequence of the A02 chromosome. Fifteen polymorphic SSR and InDel markers were obtained from parent and Bulk pool screening. Using BSA analysis, the floral and color genes were preliminarily located between markers A02-1142 and A02-1144. The genetic distance was 1.5 cm based on the number of individual plants exchanged between the marker and the floral and color genes. The genotypes of 2330 plants in F2 population were analyzed by 9 polymorphic SNP markers and 1 polymorphic InDel marker. Finally, the flower color gene was located between the SNP markers A02SNP-378 and A02SNP-1100, and the genetic distance was 0.39 cm, corresponding to the physical interval of the reference genome A02 chromosome of Chinese cabbage, about 162kb. Among them, A02SNP-389 and A02SNP-389 were coisolated with the white flower gene. 4. Analysis of candidate genes in target region of Chinese cabbage reference genome V1.5, a total of 25 annotated genes were found on the physical sequence of 162kb target region of Chinese cabbage reference genome V1.5, which combined with homologous Arabidopsis gene sequence annotation. The function of the 25 genes was preliminarily analyzed.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S565.4

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