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2025年8月19日 星期二
农业生物技术学报  2021, Vol. 29 Issue (3): 461-471    DOI: 10.3969/j.issn.1674-7968.2021.03.006
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
生姜6-姜酚生物合成相关基因的克隆与表达分析
唐建民1, 齐力旺2, 郎美容1, 张文林1, 兰建彬1, 李洪雷1, 李哲馨1,*
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
Cloning and Expression Analysis of 6-gingerol Biosynthesis Related Genes from Zingiber officinale
TANG Jian-Min1, QI Li-Wang2, LANG Mei-Rong1, ZHANG Wen-Lin1, LAN Jian-Bin1, LI Hong-Lei1, LI Zhe-Xin1,*
1 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;
2 State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China
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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;PALC4H4CLCCoAOMT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PALC4H4CLCCoAOMT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。
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唐建民
齐力旺
郎美容
张文林
兰建彬
李洪雷
李哲馨
关键词 生姜基因克隆6-姜酚表达分析    
Abstract: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 (C4H), 4-coumarate:CoA ligase (4CL) and caffeoyl-CoA-o-methyltransferase (CCoAOMT/CCOMT) (GenBank No. MN887496, MN887498, MN887497 and MN887499) were cloned and qRT-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, C4H, 4CL 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.
Key wordsZingiber officinale    Gene cloning    6-gingerol    Expression analysis
收稿日期: 2020-05-16      出版日期: 2021-03-01
ZTFLH:  S632.5  
基金资助: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)
通讯作者: *lizhexin_8903@163.com   
引用本文:   
唐建民, 齐力旺, 郎美容, 张文林, 兰建彬, 李洪雷, 李哲馨. 生姜6-姜酚生物合成相关基因的克隆与表达分析[J]. 农业生物技术学报, 2021, 29(3): 461-471.
TANG Jian-Min, QI Li-Wang, LANG Mei-Rong, ZHANG Wen-Lin, LAN Jian-Bin, LI Hong-Lei, LI Zhe-Xin. Cloning and Expression Analysis of 6-gingerol Biosynthesis Related Genes from Zingiber officinale. 农业生物技术学报, 2021, 29(3): 461-471.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.03.006     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I3/461
 
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