烟草中异源表达番茄HQT基因对多酚物质合成的影响  

SlHQT Regulates the Synthesis of Caffeoylquinic Acids and Flavonols in Tobacco

作　　者：丁香玉 王鹏伟 储昭辉[1] 赵翔宇[2] 丁新华[1] 李洋 DING Xiangyu;WANG Pengwei;CHU Zhaohui;ZHAO Xiangyu;DING Xinhua;LI Yang(State Key Laboratory of Crop Biology,College of Plant Protection,Shandong Agricultural University,Tai'an,Shandong 271018,China;College of Life Science,Shandong Agricultural University,Tai'an,Shandong 271018,China)

机构地区：[1]山东农业大学植物保护学院,作物生物学国家重点实验室,山东泰安271018 [2]山东农业大学生命科学学院,山东泰安271018

出　　处：《中国烟草科学》2021年第2期77-83,共7页Chinese Tobacco Science

基　　金：国家自然科学基金青年基金(31801867);山东省重点研发计划(2019JZZY020608)。

摘　　要：羟基肉桂酰辅酶A:奎尼酸羟基肉桂酰转移酶(Hydroxycinnamoyl-CoA:quinate hydroxycinnamoyl transferase,HQT)是催化茄科植物咖啡酰奎尼酸生物合成最后一步的关键酶。为培育富含多酚物质的烟草,提高烟草品质,通过克隆番茄HQT基因,构建P35S::SlHQT超量表达载体,在野生型三生烟草(SS)中异源表达,利用高效液相色谱技术(HPLC)及液质联用(LC-MS/MS)技术对转基因烟草叶片中一咖啡酰奎尼酸及黄酮醇含量进行检测,探究SlHQT基因对三生烟草一咖啡酰奎尼酸及黄酮醇物质合成的影响。结果显示,转SlHQT烟草叶片中一咖啡酰奎尼酸总含量最高可提高14.98倍,芦丁和山奈酚芸香糖苷含量最高可分别提高11.47和9.32倍,且生长表型与野生型烟草没有明显差异;转SlHQT基因烟草叶片中有4种(1-CQA,3-CQA,4-CQA,5-CQA)一咖啡酰奎尼酸,野生型烟草叶片中只检测到3种(3-CQA,4-CQA,5-CQA)。SlHQT基因在烟草中的异源表达促进了1-CQA的生物合成且提高了一咖啡酰奎尼酸总含量,同时增加了黄酮醇的生物合成。Hydroxycinnamoyl-CoA:quinate hydroxycinnamoyl transferase(HQT),is the key enzyme catalyzing the final step in biosynthesis of caffeoylquinic acids.In order to promote the accumulation of tobacco polyphenols,we constructed the vetor P35S::SlHQT which was heterogeneously expressed in Nicotiana tabacum var.Samsun(SS).High performance liquid chromatography(HPLC)and liquid chromatography-mass spectrometry(LC-MS/MS)analysis were used to detect the content of polyphenols in tobacco leaves.The results showed that the concentrations of caffeoylquinic acids were increased by 14.98-fold;rutin and kaempferol rutinoside were increased by 11.47-and 9.32-fold respectively.And the transgenic tobacco exhibited no visible differences compared with wild-type tobacco.There were 4 types of CQAs in the transgenic lines:1-CQA,3-CQA,4-CQA,and 5-CQA(CGA)while only 3 types of CQAs were detected in the wild type tobacco:3-CQA,4-CQA,and 5-CQA(CGA).These results suggested that SlHQT promoted the biosynthesis of 1-CQA and positively regulated the synthesis of flavonols as well as caffeoylquinic acids in tobacco.

关 键 词：烟草 番茄HQT基因 一咖啡酰奎尼酸 1-CQA 黄酮醇 

分 类 号：S572[农业科学—烟草工业]