红肉蜜柚PDR型转运蛋白基因的发掘及CmPDR11-2表达与耐草甘膦初步分析  

Discovery of PDR Transporter Gene in ‘Hongrou miyou' and Preliminary Analysis of CmPDR11-2 Expressed with Glyphosate Tolerance

作　　者：宋敏娜[1,2] 姚婷婷[1,2] 潘腾飞 蒲小龙[1,2] 张迪[1,2] 谢冬梅 潘东明 SONG Mirma;YAO Tingting;PAN Tengfei;PU Xiaolong;ZHANG Di;XIE Dongmei;PAN Dongming(College of Horticulture,Fujian Agricultural and Forestry University,Fuzhou,Fujian 350002,China;Institute of Postharvest Science and Technology of Horticultural Products,Fujian Agricultural and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学园艺学院,福建福州350002 [2]福建农林大学园艺产品贮藏保鲜研究所,福建福州350002

出　　处：《热带作物学报》2018年第9期1758-1765,共8页Chinese Journal of Tropical Crops

基　　金：福建省科技计划项目(No.2016S0022);福建农林大学创新基金项目(No.KFA17354A)

摘　　要：基于红肉蜜柚RNA-seq数据库,筛选出50个ABC转运蛋白家族基因,其中包含11条PDR型基因。对11条柚PDR基因进行命名和生物信息学分析。同源性分析结果表明:CmPDR11-2编码的氨基酸序列与拟南芥中转运百草枯除草剂的AtPDR11基因同源性最高,达69.25%,利用RT-PCR技术克隆得到CmPDR11-2的ORF序列。该序列长度为4 371 bp,编码一个含有1 456个氨基酸的蛋白质,分子量165.138 03 ku,该蛋白属于稳定的亲水性蛋白,无信号肽,具有12个跨膜结构,定位于细胞膜。实时荧光定量PCR结果表明,草甘膦除草剂胁迫处理条件下,1~15 d处理组柚叶的CmPDR11-2基因相对表达量呈整体上升趋势,且均高于对照组,反应迅速且持久,这表明CmPDR11-2基因表达与红肉蜜柚耐草甘膦有关。Based on the RNA-seq database of ‘Hongrou miyou＇50 ABC transporter family genes were screened, including 11 PDR type genes. The nomenclature and bioinformatics analysis of 11 pomelo PDR genes were carried out. Homology analysis showed that the amino acid sequence encoded by CmPDR11-2 had the highest homology（69.25%） with the AtPDR11 gene that transports paraquat herbicides in Arabidopsis. The ORF of PDR gene was named as CmPDR11-2, which was a full-length of 4 371 bp, encoding a 1 456 amino acids with molecular weight 165.138 03 ku. The protein was stably hydrophilic, without signal peptide, with 12 transmembrane structure located in cell membrane. The results of real-time fluorescence quantitative PCR analysis showed that under the condition of glyphosate herbicide stress, the relative expression of CmPDR11-2 in the leaves treated with glyphosate for 1–15 days showed an overall upward trend, both of which were higher than the control, and the reaction was rapid and durable. This indicated that the expression of CmPDR11-2 gene was related to ‘Hong rou mi you＇ glyphosate tolerance.

关 键 词：红肉蜜柚 PDR CmPDR11-2 克隆 实时荧光定量PCR 

分 类 号：S682[农业科学—观赏园艺]