应用SSH技术鉴定枳抗CTV相关基因  

Identification of CTV Resistance-Related Genes in Poncirus trifoliata L.Using Suppression Substractive Hybridization

作　　者：程春振[1,2,3] 贝学军[1,2,3] 阳佳位[1,2] 吴波[1,2,3] 朱世平[1,2] 张永艳[3] 钟云[3] 曾继吾[3] 

机构地区：[1]西南大学园艺园林学院 [2]西南大学柑桔研究所,重庆400716 [3]广东省农业科学院果树研究所,广州510640

出　　处：《西南大学学报（自然科学版）》2013年第12期35-41,共7页Journal of Southwest University(Natural Science Edition)

基　　金：国家国际科技合作项目(2012DFA30610);农业部公益性行业(农业)科研专项(201203075-07);“十二五”农村领域国家科技计划课题(2011AA100205);广东省科技计划项目(2012A020200016,2012B091100169)基金资助

摘　　要：应用SSH技术构建了CTV诱导的枳的正向和反向cDNA文库,经菌液PCR检测,从两个文库中分别随机挑选了100个阳性克隆测序,共获得169条表达序列标签(ESTs).用每条EST序列对应的克里曼丁基因组预测转录序列进行GO功能注释,去除在两个文库中均有出现的ESTs后,共得到与已知基因具有较高相似度,涉及39个特异性表达基因的EST 83条,并对4个特异性表达基因进行了实时定量PCR验证.结果表明:CTV侵染之后,枳启动了相关抗逆防御机制来减少伤害,木质素代谢、RNA干扰、赤霉素、生长素以及光敏色素调控可能在抗CTV中发挥了一定作用.Two suppression subtractive hybridization libraries were constructed using CTV infected and mock-infected Poncirus trifoliata L.leaves.One hundred positive clones randomly picked from each library were sequenced,and 169ESTs were obtained.Harvest citrus C52and Clementine genome were applied to annotate these ESTs directly according to the relative unigene or gene locus.After removing ESTs detected in both libraries,a total of 83ESTs involving 39differentially expressed genes（DEGs）were gained.To validate the gene expression patterns,gene expressions of four DEGs were identified using realtime reverse transcription PCR（RT-PCR）.The results indicated that resistance of P.trifoliatato CTV disease may be induced by CTV infection and,additionally,lignin metabolism,RNA interfering,GA, auxin and phytochrome regulation probably play key roles in CTV resistance of this species.

关 键 词：枳 抑制性差减文库 柑桔衰退病 表达序列标签 

分 类 号：S512.1[农业科学—作物学]