木薯AGPase大亚基MeAGPL3基因启动子调控区域鉴定  

Promoter Regulatory Domain Identification of AGPase Large Subunit Gene Me AGPL3 in Cassava

作　　者：马平安 陈新[3] 沈佳琦 张帅兵[1] 胡元森[1] 王文泉 Ma Ping'an;Chen Xin;Shen Jiaqi;Zhang Shuaibing;Hu Yuansen;Wang Wenquan(College of Biological Engineering,Henan University of Technology,Zhengzhou,450001;College of Tropical Crops,Hainan University,Haikou,570228;Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101)

机构地区：[1]河南工业大学生物工程学院,郑州450001 [2]海南大学热带作物学院,海口570228 [3]中国热带农业科学院热带生物技术研究所,海口571101

出　　处：《分子植物育种》2021年第21期7122-7131,共10页Molecular Plant Breeding

基　　金：河南省高等学校重点科研项目(19A180016);河南省属高校基本科研业务费专项(2018QNJH15);河南工业大学高层次人才基金项目(2018BS017);中国科技部科技合作计划项目(KY201701005)共同资助。

摘　　要：木薯是重要的热带作物,具有高淀粉积累等特点,但其高效积累机制尚不明确。为研究腺苷二磷酸葡萄糖焦磷酸化酶(AGPase)大亚基对木薯淀粉积累的调控作用,本研究通过RT-PCR技术对木薯AGPase5个大亚基基因(MeAGPL1~MeAGPL5)进行筛选,发现MeAGPL3基因在木薯块根中高表达且受100μmol/L外源ABA正向调控;克隆了该基因2455 bp 5’侧翼序列,利用该序列及4个不同长度的5’端截短片段分别与β-葡萄糖醛酸酶(GUS)基因融合构建表达载体,转化本氏烟草,通过检测各转基因烟草GUS蛋白活性,发现-1至-1711 bp序列具有最佳的启动活性。利用100μmol/L外源ABA处理各转基因烟草株系,分析处理前后GUS蛋白活性变化,发现ABA信号响应元件分布于-1711至-2150区域。本研究为从转录调控角度阐释木薯淀粉高效积累机制提供理论支撑。Cassava(Manihot esculenta Crantz) is the staple crop and has a high starch content in its storage root.However, the inner regulatory mechanism of starch accumulation is unclear. In order to study the regulation of AGPase large subunit on starch accumulation in cassava, five AGPase large subunit genes( MeAGPL1 to MeAGPL5) were screened by RT-PCR. It was found that MeAGPL3 gene was highly expressed in cassava root tubers and positively regulated by 100 μmol/L exogenous ABA. The study cloned 2 455 bp MeAGPL3 gene 5’ flanking sequence, constructed five 5’ truncated vectors which fused with the GUS gene and transformed into tobacco. GUS staining and quantitative determination in transgenic tobacco showed that the core promoter region of MeAGPL3 gene was-1 to-1 711 bp. After 100 μmol/L exogenous ABA treatment, the GUS protein activity of transgenic tobacco lines before and after treatment was examined. It was found that ABA signal response elements were distributed in the region of-1 711 to-2 150. This study provides theoretical support to explain the mechanism of cassava starch accumulation from the perspective of transcriptional regulation.

关 键 词：腺苷二磷酸葡萄糖焦磷酸化酶 MeAGPL3 启动子 5’端截短 GUS蛋白 

分 类 号：S533[农业科学—作物学]