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磷脂:二酰甘油酰基转移酶(PDAT)能够催化磷脂酰胆碱(PC)sn-2位的酰基转移至二酰甘油(DAG)上,形成三酰甘油(TAG)和溶血磷脂酰胆碱,这是另外一条不同于Kennedy途径的三酰甘油合成方式,故明确油茶(Camellia oleifera)PDAT基因的功能对阐明油茶种子油脂合成机制具有重要意义。本研究通过cDNA末端快速扩增(RACE)技术克隆油茶CoPDAT基因的全长cDNA序列,该序列长2773 bp,开放阅读框为2022 bp,编码673个氨基酸。亚细胞定位分析确定CoPDAT基因所编码蛋白质定位于内质网上。实时荧光定量PCR分析结果表明,CoPDAT在油茶不同组织/器官中均有表达,在种子中不同发育时期的相对表达量呈“上升-下降-上升-下降-上升”的表达特征,在花后42周表达量最高。CoPDAT基因与油茶种子油脂含量的灰色关联度值为0.8356,表明该基因与油茶种子油脂的合成积累关系密切。本研究为阐明CoPDAT基因在油茶油脂合成过程中的功能以及为油茶的分子遗传改良提供参考。
Phospholipid: Diacylglycerol acyltransferase (PDAT) catalyzes the transfer of acyl groups at the sn-2 position of phosphatidylcholine (PC) to diacylglycerols (DAGs) to form triacylglycerols (TAGs) and lysophosphatidylcholines, This is another triacylglycerol synthesis that is different from the Kennedy pathway. Therefore, it is of great significance to elucidate the function of the PDAT gene of Camellia oleifera to elucidate the mechanism of oil synthesis in Camellia oleifera seed oil. In this study, we cloned the full-length cDNA of CoPDAT from Camellia oleifera by rapid amplification of cDNA ends (RACE), which is 2773 bp in length and 2022 bp in open reading frame (ORF) encoding 673 amino acids. Subcellular localization analysis confirmed that the protein encoded by the CoPDAT gene was localized on the endoplasmic reticulum. Real-time quantitative PCR analysis showed that CoPDAT was expressed in different tissues and organs of Camellia oleifera. The relative expression of CoPDAT at different developmental stages was “up-down-up-down-up” 42 weeks after the highest expression. The gray correlation coefficient between CoPDAT gene and oil content of Camellia oleifera seeds was 0.8356, which indicated that the gene was closely related to the synthesis of Camellia oleifera seed oil. This study was designed to elucidate the function of CoPDAT gene in the synthesis of Camellia oleifera oil and to provide reference for molecular genetic improvement of Camellia oleifera.