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
张雨轩 1樊鑫 2张敏 3陈虞超 4赵玉洋 3焦红红 1张岗 1袁媛5
作者信息
- 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;宁夏农林科学院 农业生物技术研究中心,宁夏银川 750002
- 3. 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
- 4. 宁夏农林科学院 农业生物技术研究中心,宁夏银川 750002
- 5. 陕西中医药大学 药学院,陕西咸阳 712000;中国中医科学院 医学实验中心,北京 100700
- 折叠
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10 μmol·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.GeCYP714A 1基因的编码区全长1 173 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.
Abstract
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 10 μmol·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 1 173 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Key words
Gastrodia elata/Tis108/gibberellin(GA)/CYP714A1 gene/expression profiles analysis引用本文复制引用
基金项目
中央本级重大增减支项目(2060302-2201-21)
国家杰出青年科学基金(82325049)
中国中医科学院科技创新工程项目(CI2023D001)
中国中医科学院科技创新工程项目(CI2023E002-04)
出版年
2024
NETL
NSTL
万方数据