甘蓝型油菜磷脂二酰甘油酰基转移酶(BnPDAT1)cDNA的克隆和功能鉴定  

Cloning and Characterization of Phospholipids:Diacylglycerol Acyltransferase(BnPDAT1) cDNA from Brassica napus L.

作　　者：谭太龙[1] 冯韬[1] 罗海燕[1] 彭烨[1] 刘睿洋[1] 官春云[1] 

机构地区：[1]湖南农业大学农学院/国家油料作物改良中心湖南分中心,湖南长沙410128

出　　处：《作物学报》2016年第5期658-666,共9页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31401419);湖南省教育厅优秀青年项目(13B051);国家高技术研究发展计划(863计划)项目(2011AA10A104;2012AA101107);湖南省作物种质资源与创新利用重点实验室科学基金开放项目(11KFXM08)资助~~

摘　　要：磷脂二酰甘油酰基转移酶（phospholipids：diacylglycerol acyltransferase,PDAT1）是植物三酰甘油（triacylglycerol,TAG）合成的关键酶。本文在甘蓝型油菜湘油15号c DNA中克隆到3个PDAT1全长编码序列（coding sequence,CDS）,经比对分别定位于A02、A10、C09染色体,分别命名为Bn PDAT1-A02、Bn PDAT1-A10和Bn PDAT1-C09,其序列长分别为1998、2002和2005 bp,各自编码665、666、667个氨基酸。预测Bn PDAT1基因编码蛋白定位于细胞质膜,具有典型的PDAT1保守结构域。多序列比对和进化分析表明,Bn PDAT1基因编码蛋白与甘蓝、拟南芥、亚麻芥PDAT1蛋白具有较高的同源性。酵母互补实验证实,该基因编码蛋白具有PDAT1酶活性。Bn PDAT1基因在湘油15号中的表达现先上升后降低趋势,在开花后25~30 d达最大值,但3个拷贝的表达变化规律存在差异。Phospholipids：diacylglycerol acyltransferase（PDAT1） is a key enzyme in triacylglycerol（TAG） biosynthesis of plants. In this study, three novel PDAT1 coding sequences（CDSs） were isolated from c DNA of Brassica napus L. cv. Xiangyou 15 seeds, which were mapped to the chromosomes A02, A10, and C09, and designated as Bn PDAT1-A02, Bn PDAT1-A10, and Bn PDAT1-C09, respectively. Three Bn PDAT1 CDSs were 1998, 2002, and 2005 bp in length and encoded predicted proteins with 665, 666, and 667 amino acid residues, respectively. Bn PDAT1 proteins were predicted to be located on the cell membrane and have a typical PDAT1 conserved domain. Multiple sequence alignments and phylogenetic analysis showed that the deduced amino acid sequences of Bn PDAT1 were highly homologous to previously reported PDAT1 in Brassica oleracea, Arabidopsis thalian, and Eruca sativa. Furthermore, the catalytic enzyme activity of the cloned Bn PDAT1 genes was confirmed by the yeast complementary experiment. The expression level of Bn PDAT1 s increased gradually in seed development and reached the maximum from 25 to 30 days after flowering. However, three Bn PDAT1 copies were also found to be different in expression pattern.

关 键 词：甘蓝型油菜 磷脂二酰甘油酰基转移酶 基因克隆 酵母互补实验 

分 类 号：S565.4[农业科学—作物学]