基于茉莉酸甲酯诱导金铁锁ptMYC2转录因子调控的初步探究  

Preliminary Study on Regulation of Psammosilene Tunicoides ptMYC2 Transcription Factor Induced by Methyl Jasmonate

作　　者：孟文俊 董金材 杨文婷 李俊楠 李映涛 张爱丽 MENG Wenjun;DONG Jincai;YANG Wenting;LI Junnan;LI Yingtao;ZHANG Aili(Yunnan Medical Health College,Kunming 650106,Yunnan,China;Chenggong Hospital of Kunming Yan′an Hospital,Kunming 650599,Yunnan,China;Yunnan Key Laboratory of Dai and Yi Medicines,Yunnan University of Chinese Medicine,Kunming 650500,Yunnan,China)

机构地区：[1]云南医药健康职业学院,云南昆明650106 [2]昆明市延安医院呈贡医院,云南昆明650599 [3]云南中医药大学云南省傣医药与彝医药重点实验室,云南昆明650500

出　　处：《辽宁中医杂志》2024年第10期128-132,I0001,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81960693);云南省科学技术厅-云南中医学院运用基础联合专项项目(202101AZ070001-230);云南省科技厅基础应用项目面上项目(202201AT070218)。

摘　　要：目的研究外源激素茉莉酸甲酯(MeJA)诱导后金铁锁转录因子ptMYC2-1及其靶基因法尼基焦磷酸合酶(FPS)、鲨烯环氧酶(SE),β-香树素合酶(β-AS)等基因表达量之间的联系,揭示金铁锁三萜皂苷代谢途径中转录因子的调控机制。方法培育遗传背景单一的组培苗,喷施茉莉酸甲酯(100μmol/L)进行诱导实验,分别在0,3,6,12,24 h采集新鲜材料,以β-Actin为内参基因,运用实时荧光定量PCR对转录因子ptMYC2-1及次生代谢途径上的靶基因FPS、SE-1、SE-2、β-AS的表达水平进行检测,并通过R语言和Prism5分析基因表达量的P值和R值。结果喷施MeJA后,转录因子和其靶基因与未进行茉莉酸甲酯处理时相比均有显著性变化。转录因子与靶基因的相关性分析显示pt MYC2-1的表达量与靶基因FPS、SE-1、SE-2、β-AS的表达量均有显著的正相关性。结论实验初步探究了金铁锁转录因子及其靶基因对MeJA诱导的响应模式,为后期研究金铁锁转录因子对靶基因的调控机制奠定了基础。实验材料可能存在基因差异,因此本实验的实验数据不能直接证明转录因子与靶基因的调控关系,还需进一步设计实验进行研究。Objective To study the regulation mechanism of transcription factors in the metabolic pathway of triterpenoid saponins of Psammosilene tunicoides by studying the relationship between the expression of ptMYC2-1 and its target gene farnesyl pyrophosphate synthase(FPS),squalene epoxidase(SE)andβ-amyrin synthase(β-AS)induced by methyl jasmonate(MeJA).Methods The asepsis seedlings were bred with the same genetic background and methyl jasmonate(100μmol/L)was sprayed to induce asepsis seedlings,and fresh materials were collected at 0,3,6,12,24 h,respectively.Withβ-Actin as the internal reference gene,the expression levels of the transcription factor ptMYC2-1 and its predictive target gene(eg:FPS,SE-1,SE-2 andβ-AS)were detected by real-time quantitative PCR,then the P-value and R-value were analyzed by R programming language and Prism5.Results After spraying jasmonate,there were significant changes in expression level of transcription factors and its target genes compared with those without inducing by methyl jasmonate.After correlation analysis between transcription factors and its target genes,ptMYC2-1 was significantly positively correlated with its target genes such as FPS,SE-1,SE-2 andβ-AS.Conclusion The response pattern of transcription factor and its target gene to MeJA induction was preliminarily explored.This study provided the foundation for analysis the regulation of secondary metabolic of Psammosilene tunicoides by ptMYC2 transcription factors.There may be genetic differences in the experimental materials,so the experimental data of this experiment can not directly prove the regulatory relationship between transcription factors and target genes,and further design experiments are needed.

关 键 词：茉莉酸甲酯 转录因子 调控 次生代谢途径 

分 类 号：R284[医药卫生—中药学]