番荔枝GA20氧化酶基因的克隆与表达分析  

Cloning and Expression Analysis of GA20-Oxidase Gene from Sugar Apple(Annona squamosa)

作　　者：刘锴栋 袁长春 黎海利 刘金祥 陈燕 莫亿伟[2] 李华富 

机构地区：[1]岭南师范学院生命科学与技术学院,广东湛江524048 [2]绍兴文理学院生命科学学院,浙江绍兴312000

出　　处：《植物生理学报》2015年第10期1697-1705,共9页Plant Physiology Journal

基　　金：广东省科技计划项目(2013B020304008);国家自然科学基金项目(31201586);广东省教育厅科技创新项目(2013KJCX0124);湛江市热带特色资源植物技术开发重点实验室项目(2014A06008);湛江师范学院科研创新团队资助项目(2013CXTD05)

摘　　要：本研究利用同源克隆和RACE-PCR的方法获得番荔枝GA20ox基因的全长c DNA序列,命名为As GA20ox,Gen Bank登录号为KR676623。序列分析表明,克隆获得的番荔枝As GA20ox基因编码区长为1 257 bp,编码418个氨基酸。序列比对显示与油棕和海枣等GA20ox的相似度分别为71%和70%。构建类似蛋白系统进化树显示,番荔枝As GA20ox与簇毛麦、香蕉等分子进化距离较近。预测As GA20ox蛋白定位在细胞外基质和过氧化物酶体中,不具备信号肽。蛋白质分子量为47.2k Da,等电点为6.21。实时定量RT-PCR结果表明,在不同的组织、不同的器官中,As GA20ox的表达量存在差异,其中在花蕾期I、未成熟的种子、结果枝的幼嫩茎段和雌蕊中的表达量相对较高。A full-length c DNA sequence of homologous GA20 ox gene was cloned by employing homology gene cloning and RACE-PCR from sugar apple（Annona squamosa L.）, which was named as As GA20ox（GenBank accession KR676623）. Sequence analysis showed that the As GA20 ox gene has a 1 257 bp open reading frame（ORF） encoding 418 amino acids. Sequence alignment displayed that it was a similarity of 71% and 70% with GA20 ox of Elaeis guineensis and Phoenix dactylifera, respectively. The phylogenetic tree showed that sugar apple As GA20 ox and Dasypyrum villosum GA20 ox are closest in molecular evolution distance, followed by Musa acuminata subsp. Malaccensis GA20 ox. It was speculated that As GA20 ox protein is located in the extracellular matrix and peroxisomal. It do not have a signal peptide. The putative protein molecular weight was 47.2 k Da and its theoretical isoelectric point was 6.21. The real-time PCR results suggested that As GA20 ox gene showed a high transcription level in flower buds I, immature seeds, young stems of fruiting branches and pistils. The results laid a solid foundation for further research on As GA20 ox protein structure and the relationship between the gene and plant flower development.

关 键 词：番荔枝 GA20ox 基因克隆 表达模式 

分 类 号：Q943.2[生物学—植物学] S667.9[农业科学—果树学]