云南药用野生稻高光效基因的筛选及功能分析

项目名称： 云南药用野生稻高光效基因的筛选及功能分析

项目编号： No.31460054

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 柯学

作者单位： 云南省农业科学院

项目金额： 50万元

中文摘要： 培育高光效水稻是提高水稻产量和解决粮食需求的有效途径，高光效基因发掘在其中尤为重要。药用野生稻(O. Officinalis)具备高光效特性：强生长势、20倍以上于栽培稻的生物量积累和较高的净光合速率。我们发现云南药用野生稻的羧化效率、光饱和点均为栽培稻的2倍以上。前期，我们构建了云南药用野生稻基因组BIBAC文库，转入栽培稻日本晴建立了渗入系库，获得了净光合速率高于日本晴的渗入系T1代株系。本项目拟加代T1代株系到T2代，测定T2代株系的生物学性状，筛选出高光效T2代株系；利用基因组重测序和RNA-Seq技术分析渗入系T2代株系中来自云南药用野生稻基因组的DNA和基因表达特征，结合筛选BIBAC克隆测序，预测候选基因；通过基因沉默和过表达途径验证和分析候选基因功能，从中鉴定出云南药用野生稻高光效基因。这不仅对揭示云南药用野生稻高光效机理有科学意义，而且在培育高光效水稻材料方面有实践意义。

中文关键词： 高光效；渗入系；BIBAC文库；基因筛选；功能分析

英文摘要： Breeding high photosynthetic efficiency of rice is the way to improve the yield and solve huge demand for food. Wild rice Oryza Officinalis Wall. evolved of some excellent traits during long-term of natural selection such as a strong growth potential with more than 20 times that of cultivated varieties, a great accumulation of biomass, and a higher net photosynthetic rate. And our result also indicated that the carboxylation efficiency and light saturation point of O.Officinalis of Yunnan was 2 times more than that of cultivated varieties. Our past research, a genomic BIBAC library of O. Officinalis of Yunnan was been constructed and used to transform Nipponbare for introgression lines. And among of the introgression lines, some T1 generations with higher net photosynthetic rate had been identified. This study, based on the T2 generations of introgression lines with high photosynthetic efficiency trait that screened out by measuring saveral biological characters, is to predict high photosynthetic efficiency candidate genes from T2 generations using genomic resequencing and RNA-Seq technology, that is, genomic resequencing can find out the differential DNA sequences from O. Officinalis when compared with Nipponbare genome, and then according to these DNA sequences, probes are designed to screen and sequence BIBAC clones to obtain the intact target DNA sequences; RNA-Seq technology can find out the differentially expressed genes between T2 generations of introgression lines and Nipponbare, witch is helpful to screen candidate genes from the target DNA. Ultimately, after transformating candidate genes into introgression lines and Nipponbare to identify their functions by RNAi and overexpression technics rerepectively, the high photosynthetic efficiency genes of O. Officinalis of Yunnan are determined. This will, to some extent, reveal the genetic basis of strong growth potential and great biomass accumulation traits of O. Officinalis, find out genes that can be applied to improve the photosynthesis and yield potential of cultivated varieties, which is important theoretical significance and application value.

英文关键词： high photosynthetic efficiency;introgression lines;BIBAC library;gene screening;functional analysis