生姜6-姜酚生物合成相关基因的克隆与表达分析  

Cloning and Expression Analysis of 6-gingerol Biosynthesis Related Genes from Zingiber officinale

作　　者：唐建民[1] 齐力旺[2] 郎美容 张文林 兰建彬[1] 李洪雷 李哲馨 TANG Jian-Min;QI Li-Wang;LANG Mei-Rong;ZHANG Wen-Lin;LAN Jian-Bin;LI Hong-Lei;LI Zhe-Xin(Chongqing Key Laboratory of Economic Plant Biotechnology/Chongqing Special Plant Industry Collaborative InnovationCenter/College of Landscape Architecture and Life Science/Institute of Special Plants,Chongqing University of Arts and Sciences,Chongqing 402160,China;State Key Laboratory of Tree Genetics and Breeding,Research Institute of Forestry,Chinese Academy of Forestry,Beijing 100091,China)

机构地区：[1]重庆文理学院园林与生命科学学院/特色植物研究院/重庆市特色植物产业协同创新中心/重庆市经济植物生物技术重点实验室,重庆402160 [2]中国林业科学研究院林业研究所细胞生物学实验室,北京100091

出　　处：《农业生物技术学报》2021年第3期461-471,共11页Journal of Agricultural Biotechnology

基　　金：重庆市自然科学基金(cstc2019jcyj-msxmX0697);重庆市教育委员会科学技术研究项目(KJQN201901302)。

摘　　要：姜(Zingiber officinale)是药食兼用的高附加值特色经济植物。姜辣素是生姜的主要呈味物质和药用核心成分,6-姜酚则是姜辣素的主要成分。本研究以川渝特色姜品种竹根姜的根状茎为材料,采用液相色谱质谱联用(liquid chromatograph-mass spectrometer, LC-MS)技术检测姜辣素各主要成分的含量;克隆了6-姜酚生物合成途径苯丙氨酸氨裂解酶(phenylalanine ammonia lyase, PAL)、肉桂酸在肉桂酸-4-羟化酶(cinnamate 4-hydroxylase, C4H)、4-香豆酸辅酶A连接酶(4-coumarate:CoA ligase, 4CL)以及咖啡酰辅酶A-O-甲基转移酶(caffeoyl-CoA-O-methyltransferase, CCoAOMT/CCOMT) 4个酶基因的cDNA序列(GenBank No. MN887496, MN887498, MN887497, MN887499);利用qRT-PCR技术检测4个基因在竹根姜不同发育阶段的表达量;并对各基因的表达量与6-姜酚增量进行相关性分析。结果表明,6-姜酚在竹根姜播种后1个月材料中极速积累,并在后期发育阶段少量增加;PAL、C4H、4CL及CCoAOMT基因均在竹根姜播种后1~2个月高表达,与6-姜酚合成存在相关规律;相关性分析结果显示,在PAL、C4H、4CL、CCoAOMT等4个酶基因中,CCoAOMT的表达与6-姜酚合成存在显著正相关性(P<0.05),说明6-姜酚的合成存在阶段特异性,CCoAOMT基因与6-姜酚合成存在正向调控关系。该研究结果将为后期研究6-姜酚合成相关酶基因的功能提供重要基因资源。Ginger(Zingiber officinale) has high economic value as medicinal and food resources. 6-gingerol is high important bioactive component in the pharmacological action of ginger. In the present study, a local ginger cultivar of Chongqing was used as the material. Liquid chromatograph-mass spectrometer(LC-MS)technology was used to detect the main components of pungent principle. Four cDNA sequences of 6-gingerol biosynthetic relative enzyme genes including phenylalanine ammonia lyase(PAL), cinnamate 4-hydroxylase(C4 H), 4-coumarate: CoA ligase(4 CL) and caffeoyl-CoA-o-methyltransferase(CCoAOMT/CCOMT)(GenBank No. MN887496, MN887498, MN887497 and MN887499) were cloned and q RT-PCR was used to detect the expression of the genes in ginger rhizome at different developmental stages. Correlation analysis was performed between the expression of each gene and the 6-gingerol increase. The results showed that 6-gingerol accumulated very quickly in the 1-month ginger material after sowing, and increased slightly in the subsequent stages. PAL, C4 H, 4 CL and CCoAOMT genes were all highly expressed in 1~2 months ginger after sowing, suggesting a correlation between each gene and 6-gingerol synthesis. Correlation analysis showed that the expression of CCoAOMT rather than the others was significantly positively correlated with 6-gingerol synthesis(P<0.05). These results indicated that the biosynthesis of 6-gingerol existed stage specificity, and there was a positive regulatory relationship between CCoAOMT gene and 6-gingerol biosynthesis. The results of this study will lay the foundation for the further study of the function of 6-gingerol synthesis-related enzyme genes in Z. officinale.

关 键 词：生姜 基因克隆 6-姜酚 表达分析 

分 类 号：S632.5[农业科学—蔬菜学]