烟草科技
2016-03-29 星期二
烟草农学:2020 ,9:1-5
翟妞, 郑庆霞, 刘萍萍, 徐国云, 张慧, 李晶, 陈千思, 武明珠, 周会娜. 烟草查尔酮异构酶基因2(NtCHI2)功能分析[J]. 烟草科技, 2020 (9): 1-5.
ZHAI Niu, ZHENG Qingxia, LIU Pingping, XU Guoyun, ZHANG Hui, LI Jing, CHEN Qiansi, WU Mingzhu, ZHOU Huina. Functional analysis of chalcone isomerase gene 2 (NtCHI2) in tobacco[J]. Tobacco Science & Technology, 2020 (9): 1-5.

烟草查尔酮异构酶基因2(NtCHI2)功能分析
翟妞1, 郑庆霞1, 刘萍萍1, 徐国云1, 张慧1, 李晶2, 陈千思1, 武明珠1, 周会娜1
1. 中国烟草总公司郑州烟草研究院, 郑州高新技术产业开发区枫杨街2号 450001;
2. 云南中烟工业有限责任公司技术中心, 昆明市五华区科医路41号 650106
摘要:
烟草查尔酮异构酶(Chalcone isomerase,CHI)是黄酮类代谢的一个关键酶,烟草中有两个基因拷贝(NtCHI1NtCHI2),相关研究多集中在NtCHI1基因,对NtCHI2的研究较少。为研究烟草NtCHI2的功能,主要以NtCHI2的过表达和干扰植株叶片为材料,进行了实时定量PCR和CHI活性分析以及芸香苷含量检测,同时进行了NtCHI2体外表达蛋白的酶活分析。结果表明:NtCHI2体外表达蛋白具有查尔酮异构酶活性;过表达株系中NtCHI2的表达量升高5~75倍,但CHI酶活性和芸香苷含量基本没有变化;干扰株系中NtCHI2的表达量降至70%,CHI酶活性和芸香苷含量分别降低50%和90%。烟草NtCHI2参与芸香苷的合成代谢调控,在一定程度上NtCHI2基因的表达量和芸香苷的含量正相关。
关键词:    烟草    查尔酮异构酶基因(NtCHI2)    查尔酮异构酶活性    芸香苷   
Functional analysis of chalcone isomerase gene 2 (NtCHI2) in tobacco
ZHAI Niu1, ZHENG Qingxia1, LIU Pingping1, XU Guoyun1, ZHANG Hui1, LI Jing2, CHEN Qiansi1, WU Mingzhu1, ZHOU Huina1
1. Zhengzhou Tobacco Research Institute of CNTC, Zhengzhou 450001, China;
2. Technology Center, China Tobacco Yunnan Industrial Co., Ltd., Kunming 650106, China
Abstract:
Chalcone isomerase (CHI) is one of the key enzymes in flavonoid metabolism. In tobacco, two copies of CHI genes, NtCHI1 and NtCHI2, are identified, while little is known on the function of NtCHI2 compared to that of NtCHI1. To study the function of NtCHI2, its rutin content, CHI activity, and NtCHI2 expression level were analyzed for NtCHI2-OE strains and NtCHI2-RNAi strains. We also examined the CHI activity of recombinant NtCHI2 expressed in vitro. The results showed that NtCHI2 had a CHI activity. In the NtCHI2-OE strains, the NtCHI2 expression level was up-regulated by 5-75 folds, however no significant changes in CHI activity and rutin content were detected. In NtCHI2-RNAi strains, the NtCHI2 expression level was down-regulated to 70%, and correspondingly, CHI activity and rutin content decreased by 50% and 90% respectively. NtCHI2 was involved in the metabolic regulation of rutin, and the expression of NtCHI2 was positively correlated with the rutin content to some extent.
Key words:    Tobacco    Chalcone isomerase gene (NtCHI2)    Chalcone isomerase (CHI) activity    Rutin   
收稿日期: 2020-01-14     修回日期: 2020-03-17
DOI: 10.16135/j.issn1002-0861.2020.0027
基金项目: 郑州烟草研究院院长科技发展基金项目“烟草荧光细胞分选技术建立及在多酚代谢研究中的应用”(902018CA0250);国家烟草专卖局烟草基因组重大专项项目“典型香型烟草品质形成的代谢特征研究”[110201401012(JY-12)]和云南中烟工业有限责任公司科技项目“烟草苗期代谢缺陷研究及在基因育种筛选中的应用”(2018JC06)。
通讯作者: 周会娜,E-mail:joanzhn@163.com     Email:joanzhn@163.com
作者简介: 翟妞(1981-),博士,副研究员,主要从事烟草代谢研究。E-mail:zhainiu2005@163.com
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