Tis108对天麻GA含量及其失活关键酶GeCYP714A1基因表达的影响  

Influence of Tis108 on GA content and expression of key enzyme GeCYP714A1 involved in GA deactivation of Gastrodia elata

作　　者：张雨轩 樊鑫 张敏[2] 陈虞超[3] 赵玉洋 焦红红 张岗[1] 袁媛 ZHANG Yu-xuan;FAN Xin;ZHANG Min;CHEN Yu-chao;ZHAO Yu-yang;JIAO Hong-hong;ZHANG Gang;YUAN Yuan(College of Pharmacy,Shaanxi University of Chinese Medicine,Xianyang 712000,China;State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Agricultural Biotechnology Center,Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan 750002,China;Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]陕西中医药大学药学院,陕西咸阳712000 [2]中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室,北京100700 [3]宁夏农林科学院农业生物技术研究中心,宁夏银川750002 [4]中国中医科学院医学实验中心,北京100700

出　　处：《中国中药杂志》2024年第15期4007-4014,共8页China Journal of Chinese Materia Medica

基　　金：中央本级重大增减支项目(2060302-2201-21);国家杰出青年科学基金项目(82325049);中国中医科学院科技创新工程项目(CI2023D001,CI2023E002-04)。

摘　　要：为探究独脚金内酯抑制剂Tis108对天麻生长的影响,该研究采用10μmol·L^(-1)的Tis108溶液处理白麻块茎,对块茎的内源激素赤霉素(GA)含量进行测定,通过RT-PCR技术克隆GA失活关键酶GeCYP714A1基因,借助ExPASy、SWISS-MODEL、MEGA等软件对该基因进行生物信息学分析,并测定其在天麻不同组织部位的表达水平。结果显示,Tis108处理后天麻块茎的GA含量显著升高,GA失活关键酶GeCYP714A1的转录水平显著降低。GeCYP714A1基因的编码区全长1173 bp,编码390个氨基酸,蛋白的相对分子质量为44.85 kDa,理论等电点为9.83,不稳定系数为49.20,脂肪系数为89.03,总平均亲水指数平均值为-0.235,属于碱性亲水性不稳定蛋白;且GeCYP714A1蛋白定位于线粒体,无信号肽,不具有跨膜结构。系统进化树分析显示,GeCYP714A1与铁皮石斛DcCYP714C2(PKU78454.1)蛋白亲缘关系最近,序列一致性达67.25%。利用qRT-PCR技术分析天麻GeCYP714A1基因的表达模式,结果显示在天麻块茎中GeCYP714A1转录水平最高,其次是茎和花序。该研究表明Tis108可抑制天麻块茎中GA失活酶GeCYP714A1的转录水平,提高GA的积累量,影响天麻块茎生长,为进一步揭示独脚金内酯调控天麻GA信号和块茎发育奠定基础。To investigate the influence of the strigolactone inhibitor Tis108 on the growth of Gastrodia elata,this study treated G.elata tuber with Tis108 solution of 10μmol·L^(-1)and measured the content of endogenous hormone gibberellin(GA)in the tuber.By using reverse transcription-polymerase chain reaction(RT-PCR)technology,the key enzyme GeCYP714A1 gene involved in GA deactivation was cloned.Bioinformatics analysis on the GeCYP714A1 gene was carried out by using ExPASy,SWISS-MODEL,MEGA,etc.,and its expression levels in different parts of G.elata were determined.The results showed that after Tis108 treatment,GA content in G.elata tuber was significantly increased,and the transcription level of the GeCYP714A1 gene was significantly decreased.The full length of the coding region of the GeCYP714A1 gene is 1173 bp,encoding 390 amino acids.The protein has a molecular weight of 44.85 kDa,a theoretical isoelectric point of 9.83,an instability index of 49.20,an aliphatic index of 89.03,and a grand average of hydropathicity of-0.235,classifying it as an unstable,basic,hydrophilic protein,and the GeCYP714A1 protein was localized in the mitochondria,lacking a signal peptide and a transmembrane structure.Phylogenetic tree analysis revealed that GeCYP714A1 was most closely related to the DcCYP714C2(PKU78454.1)protein from Dendrobium candidum,with a sequence identity of 67.25%.The qRT-PCR analysis of the expression patterns of the GeCYP714A1 gene indicated that GeCYP714A1 had the highest transcription level in G.elata tuber,followed by stem and inflorescence.The study represented that Tis108 inhibited the transcription level of GeCYP714A1 involved in GA deactivation in G.elata tuber,thereby increasing the accumulation of GA and affecting the growth of G.elata tuber.These results provided a basis for further studies of strigolactone regulation of GA signal and tuber development in G.elata.

关 键 词：天麻 Tis108 赤霉素(GA) CYP714A1基因 表达谱分析 

分 类 号：S567.239[农业科学—中草药栽培]