烟草倍半萜合酶基因NtTPS21的克隆及功能鉴定  

Cloning and Functional Characterization of Sesquiterpene Synthase Gene NtTPS21 in Tobacco

作　　者：杨晴 刘少华 杨长青 耿锐梅[1] 张钊 刘东阳[4] 杨爱国[1] 李依婷[1] YANG Qing;LIU Shaohua;YANG Changqing;GENG Ruimei;ZHANG Zhao;LIU Dongyang;YANG Aiguo;LI Yiting(Institute of Tobacco Research of CAAS,Qingdao 266101,China;Graduate School of Chinese Academy of Agricultural Sciences,Beijing 100081,China;Qujing Tobacco Company of Yunnan Province,Qujing,Yunnan 655000,China;Liangshan Tobacco Company of Sichuan Province,Liangshan,Sichuan 615000,China)

机构地区：[1]中国农业科学院烟草研究所,青岛266101 [2]中国农业科学院研究生院,北京100081 [3]云南省烟草公司曲靖市公司,云南曲靖655000 [4]四川省烟草公司凉山州公司,四川凉山615000

出　　处：《中国烟草科学》2022年第3期39-46,共8页Chinese Tobacco Science

基　　金：中国农业科学院科技创新工程(ASTIP-TRIC01);中央级公益性科研院所基本科研业务费专项(1610232020001);中国烟草总公司四川省公司重点科技项目(SCYC202003)。

摘　　要：萜类化合物在植物生长发育、胁迫响应以及抵御病原菌侵染过程中发挥重要作用,克隆烟草倍半萜合酶基因并研究其功能可为烟草萜烯合酶的功能鉴定及抗病育种提供理论依据。本研究从普通烟草红花大金元中克隆获得了一个倍半萜合酶基因NtTPS21,通过生物信息学分析、表达模式分析、酶学性质分析以及NtTPS21代谢产物的体外抑菌试验等方法,鉴定NtTPS21的生物学功能。结果表明,NtTPS21开放阅读框为1671bp,编码556个氨基酸,具有典型的萜类合酶催化活性位点,与拟南芥倍半萜合酶AtTPS21具有较高同源性。NtTPS21定位于核质中,能够被烟草青枯病菌感染诱导,且在青枯病抗病品种“岩烟97”中的表达水平显著高于感病品种“长脖黄”。NtTPS21能够以法尼基焦磷酸(FPP)为底物,催化合成倍半萜类化合物β-石竹烯,其代谢产物对烟草青枯病菌的生长具有显著抑制效果,以上结果说明NtTPS21基因及其代谢产物可能与烟草青枯病抗性相关。Terpenoids play an important role in plant growth, development, stress response and resistance to pathogen infection.Cloning the sesquiterpene synthase genes of tobacco and studying their function can provide theoretical basis for functional identification and disease resistance breeding of tobacco. In this study, a sesquiterpene synthase gene NtTPS21 was cloned from common tobacco variety Honghuadajinyuan, and its function was identified by bioinformatic analysis, expression pattern analysis,enzyme property analysis and bacteriostatic test in vitro of NtTPS21 fermentation products. The results showed that the open reading frame of NtTPS21 was 1671 bp which encoded 556 amino acids. NtTPS21 had a typical terpene synthase catalytic active site and high homology with Arabidopsis thaliana sesquiterpene synthase AtTPS21. NtTPS21 was localized in the cytoplasm and could be induced by Granville wilt of tobacco. The expression level of NtTPS21 in the resistant variety “Yanyan 97” was significantly higher than that in the susceptible variety “Changbohuang”. NtTPS21 can catalyze the synthesis of sesquiterpene β-caryophyllene, which has a significant inhibitory effect on the growth of granville wilt. These results indicate that NtTPS21 and its metabolites may be related to the resistence to Ralstonia solanacearum.

关 键 词：烟草 NtTPS21 倍半萜合酶 β-石竹烯 青枯病抗性 

分 类 号：S572.01[农业科学—烟草工业]