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.
Zhongguo Zhong Yao Za Zhi. 2024 Aug;49(15):4007-4014. doi: 10.19540/j.cnki.cjcmm.20240517.101.
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.
为研究独脚金内酯抑制剂Tis108对天麻生长的影响，本研究用10μmol·L⁻¹的Tis108溶液处理天麻块茎，并测定块茎中内源激素赤霉素（GA）的含量。利用逆转录-聚合酶链反应（RT-PCR）技术，克隆了参与GA失活的关键酶GeCYP714A1基因。运用ExPASy、SWISS-MODEL、MEGA等软件对GeCYP714A1基因进行生物信息学分析，并测定其在天麻不同部位的表达水平。结果表明，Tis108处理后，天麻块茎中GA含量显著增加，GeCYP714A1基因的转录水平显著降低。GeCYP714A1基因编码区全长1173bp，编码390个氨基酸。该蛋白分子量为44.85kDa，理论等电点为9.83，不稳定指数为49.20，脂肪族指数为89.03，总平均亲水性为-0.235，属于不稳定、碱性、亲水性蛋白，且GeCYP714A1蛋白定位于线粒体，无信号肽和跨膜结构。系统进化树分析显示，GeCYP714A1与铁皮石斛的DcCYP714C2（PKU78454.1）蛋白亲缘关系最近，序列同一性为67.25%。GeCYP714A1基因表达模式的qRT-PCR分析表明，GeCYP714A1在天麻块茎中的转录水平最高，其次是茎和花序。该研究表明，Tis108抑制了天麻块茎中参与GA失活的GeCYP714A1的转录水平，从而增加了GA的积累并影响了天麻块茎的生长。这些结果为进一步研究独脚金内酯对天麻GA信号和块茎发育的调控提供了依据。
