红苞凤梨嵌合体中白化细胞失绿分子机理研究

项目名称： 红苞凤梨嵌合体中白化细胞失绿分子机理研究

项目编号： No.31300585

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 马均

作者单位： 四川农业大学

项目金额： 21万元

中文摘要： 红苞凤梨（Ananas bracteatus）嵌合体品种叶由绿色正常组织与白化组织相嵌而成，具金边、金心等多种类型，且花果艳丽多彩，观赏期长达3～4月，为重要的新型观赏植物。但其嵌合性状在繁殖过程中易发生变异甚至消失，导致大量再生植株因丧失观赏价值而被淘汰。因此，深入研究嵌合体中白化细胞形成的分子机理对嵌合品种育种和繁殖有着重要意义。本申请项目将在进行了嵌合体离体培养下变异规律和组织切片等研究基础上，以组织培养获得的全绿、全白植株作为典型材料，对嵌合体白化细胞失绿突变分子机理进行研究。利用AFLP分子标记进行绿、白两种类型细胞基因组DNA差异研究，寻找导致失绿的基因组DNA差异；通过RNA-seq转录组和表达谱测序筛选绿、白细胞间差异表达基因；通过RACE技术获得差异表达基因全长，采用real time qPCR检测差异基因的表达模式，RNAi基因沉默以验证功能，弄清细胞失绿的分子机理。

中文关键词： 红苞凤梨；白化机理；叶绿素合成；基因功能；

英文摘要： Variegated chimera varieties of Ananas bracteatus have been planted as important landscape plants for its ornamental values of color leaf and fruit. The leaf of it was composed of green normal tissues and abino tissues. However the variegated chimera character of Bromeliad is unstable. Many of the regenerated plants got by in vitro tissue culture lost its ornamental value for the changes of leaf color. It is very important and meaningful to lucubrate the characteristics and mechnism of chimera for its breeding and propagation. Based on the preliminary studies of the variation and microexamination of the chimera, the complete green or white plants derived from in vitro culture were used in the further study of mutation molecular mechnism of abino cells in the aaplied project. Research in the diversity of genomic DNA between normal cells and abino cells was carried out by AFLP molecular marker technique to discover the mutation of genomic DNA which results in chlorosis. Differentially expressed genes was screened by RNA-seq technique to identify the mRNA diversity between normal cells and albino cells. The full length of differentially expressed genes was isolated by RACE technique, expression patterns of it was detected by real time qPCR.The function of it was verified by RNAi.

英文关键词： Ananas comosus var. bracteatus；albino mechanism；chlorophyll biosynthesis；gene function；