‘无籽’瓯柑CsLTP和CsLOX基因的克隆与表达分析  

Cloning and expression analysis of CsLTP and CsLOX in seedless Ougan (Citrus suavissima 'Seedless')

作　　者：张敏[1] 刘志辉[1] 宋雪恩 朱咪咪[1] 党婉誉 颜福花 张迟[1,4] 

机构地区：[1]浙江农林大学亚热带森林培育国家重点实验室培育基地,浙江临安311300 [2]浙江省平湖市乍浦镇农技水利服务中心,浙江平湖314200 [3]浙江省丽水市林业科学研究院,浙江丽水323000 [4]浙江农林大学浙江省农产品品质改良技术研究重点实验室,浙江临安311300

出　　处：《浙江农林大学学报》2014年第6期823-830,共8页Journal of Zhejiang A&F University

基　　金：国家自然科学基金资助项目(31000897);浙江省自然科学基金资助项目(Y3090494);浙江农林大学科研发展基金资助项目(2007FR106)

摘　　要：采用同源序列法,结合cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术,克隆获得‘无籽’瓯柑Citrus suavissima‘Seedless’脂转移酶基因CsLTP和脂氧合酶基因CsLOX的全长cDNA序列;CsLTP cDNA全长667 bp,1个开放阅读框,编码215个氨基酸,与脐橙C.sinensis,粗柠檬C.jambhiri的脂转移蛋白的相似性分别为95%和60%;CsLOX cDNA全长2 915 bp,1个开放阅读框,编码895个氨基酸,与粗柠檬、美洲黑杨Populus deltoids的相似性分别为99%和51%。荧光定量聚合酶链式反应(PCR)表明:CsLTP和CsLOX的表达量均在‘无籽’瓯柑花粉粒形成期开始显著下降,并显著低于普通瓯柑Citrus suavissima。‘无籽’瓯柑花粉发育过程中油脂的转运及氧化的异常,可能成为‘无籽’瓯柑花粉败育的原因之一。To gain new insight into the male sterile characters of seedless Ougan,homologous cloning was undertaken for clones of full length cDNAs of CsLTP （lipid transfer protein） and CsLOX （lipoxygenase） genes of seedless Ougan （Citrus suavissima ‘Seedless’）.Then,bioinformatic analyses were performed online to predict the characteristics of encoded proteins.The expression model of CsLTP and CsLOX with pollen development was analyzed through real time polymerase chain reaction （PCR）,and statistic analysis was performed by Duncan＇s multiple comparison test.The cDNA of CsLTP was 667 bp in length encoding 215 amino acids,which was similar to LTPs of Citrus sinensis （95％） and Citrus jambhiri （60％）.The cDNA of CsLOX was 2 915 bp in length encoding 895 amino acids,which was similar to LOXs of C.jambhiri （99％） and Populus （51％）.Compared to seedy Ougan,the mRNA transcripts of CsLTP and CsLOX in seedless Ougan decreased noticeably （P＜0.05） after the pollen formation stage leading to aberrant lipid transportation and oxygenation in seedless Ougan.Thus,the turbulence of lipid metabolism was related to pollen abortion in seedless Ougan.

关 键 词：园艺学 '无籽’瓯柑 花粉败育 CsLTP CsLOX 荧光定量PCR 

分 类 号：S603[农业科学—园艺学]