本文選題：白菜型油菜　切入點：白花　出處：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：白菜型油菜是我國三大油菜類型之一,也是蕓薹屬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決定,為了解析白菜型油菜花色性狀的分子機制,了解蕓薹屬中花色的不同調(diào)控途徑,揭示蕓薹屬花色進化的模式,本課題利用白菜型油菜黃花品種P3246和白花品種W01雜交后代的F2群體進行花色基因的初步定位和精細定位。主要的研究結(jié)果如下:1.花瓣和葉片類胡蘿卜素含量的測定采用高效液相色譜法(HPLC)對P3246和W01的葉片和花瓣進行了類胡蘿卜素含量的測定,不論是在葉片還是花瓣中兩個材料間類胡蘿卜素的組成成分沒有差異,而各成分的含量均有差異。特別是在花瓣中,含量最高的兩種成分是紫黃質(zhì)和葉黃素,含量最高的紫黃質(zhì)在P3246中含量高達3009.86μg/g,占花瓣中類胡蘿卜素總量的86.6%,在W01中含量為242.06μg/g,占花瓣中類胡蘿卜素總量的72.0%,P3246中類胡蘿卜素的總量為3475.60μg/g,W01中類胡蘿卜素的總量為336.38μg/g,前者是后者的10.3倍。2.花色遺傳規(guī)律的分析將白菜型油菜黃花品種P3246(P1)和白花品種W01(P2)雜交,構(gòu)建F2分離群體,共獲得3104個F2單株。在盛花期進行花色性狀考察,其中黃花單株2321株,白花單株783株,經(jīng)卡方檢驗花色表型分離比符合3:1的理論值(X_c~2=0.073,X_(0.05,1)~2=3.84,P0.05),確定白菜型油菜白花性狀受單個隱性基因控制。3.花色基因的定位采用集團分離分析法(BSA)對142個InDel標記進行篩選,獲得4個與花色基因連鎖的InDel標記,初步將花色基因定位在A02染色體。從對應(yīng)的A02染色體區(qū)段選擇50個InDel標記,另外根據(jù)該區(qū)段參考基因組序列新開發(fā)了106個SSR標記,對親本和Bulk池進行篩選獲得15個多態(tài)性SSR和InDel標記,采用BSA分析將花色基因初步定位在標記A02-1142和A02-1144之間,根據(jù)標記與花色基因間的交換單株數(shù)確定遺傳距離為1.5cM。用該區(qū)段9個多態(tài)SNP標記和1個多態(tài)性InDel標記對F2群體2330單株進行基因型分析,最終將花色基因定位在SNP標記A02_SNP-378和A02_SNP-1100之間,遺傳距離為0.39cM,對應(yīng)白菜參考基因組A02染色體約162kb的物理區(qū)間內(nèi),其中SNP標記A02_SNP-389與白花基因共分離。4.目標區(qū)段候選基因的分析在白菜參考基因組V1.5的162kb目標區(qū)段物理序列上,共發(fā)現(xiàn)25個注釋基因。結(jié)合同源擬南芥基因序列注釋,對這25個基因的功能進行了初步分析。
[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.
【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】：2017
【分類號】：S565.4

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