木薯<i>MeERF1.2</i>基因克隆及表达分析

曾坚; 吴伟姗; 谢彩虹; 沈梓欣; 叶晓雪; 颜彦; 李冰; 陈志晟; 彭红元; 胡伟; 曾力旺

doi:10.19303/j.issn.1008-0384.2023.04.003

木薯MeERF1.2基因克隆及表达分析

Cloning and Expression of MeERF1.2 in Cassava

摘要

摘要:
目的乙烯响应因子（Ethylene response factor，ERF）是乙烯信号转导通路的重要成员，克隆并分析其在木薯块根采后生理性变质（Post-harvest physiological deterioration，PPD）过程中的表达情况，能为进一步研究乙烯信号在木薯PPD过程中的功能提供参考。
方法以木薯栽培品种华南8号（SC8）为材料，采用RT-PCR技术克隆木薯MeERF1.2基因，对其进行相关生物信息学分析，如遗传进化关系、结构域、蛋白质结构预测、理化性质等。对MeERF1.2基因在细胞中的亚细胞定位进行确认，并用qRT-PCR技术分析MeERF1.2基因在木薯块根PPD过程中的表达水平。
结果克隆得到的MeERF1.2基因全长为660 bp，编码的氨基酸残基数为219，分子量和等电点分别为25.04 kD和5.61，含有AP2家族结构域，和橡胶HbERF1B-like的亲缘关系最近，序列相似性达到88.74%。MeERF1.2基因定位于细胞核。和对照0 h相比，MeERF1.2基因的表达量在木薯块根的采后过程中表现为显著上升趋势，即MeERF1.2基因的表达受到PPD过程的诱导。
结论克隆得到的MeERF1.2基因包含ERF基因家族的保守结构域，属于ERF基因家族；MeERF1.2基因的表达在采后过程中受到诱导，可能参与了木薯块根的PPD过程，为后续进一步分析乙烯信号转导通路在PPD过程中的作用奠定了基础。

Abstract:
Objective Critical factors in the ethylene signal transduction pathway involving the post-harvest physiological deterioration (PPD) of cassava were investigated.
Method One of the possible ethylene response factors (ERFs) in Manihot esculenta cv.SC8, MeERF1.2, was cloned using RT-PCR to analyze the expressions during PPD. Physicochemical properties, conserved domain, genetic evolutionary relationship, and protein structure of the gene were studied with bioinformatics tools. Distribution of MeERF1.2 in the plant cells was verified by online software and subcellular localization.
Result MeERF1.2 had an ORF of 660 bp encoded 188 amino acids with a molecular weight of 25.04 kD and a pI of 5.61. The protein contained an AP2 domain showing a high sequence similarity of 88.74% with the HbERF1B-like gene. It was localized in the nucleus and significantly upregulated during PPD from 0 to 6, 12, and 48 h displaying an apparent induction trend in the process.
Conclusion MeERF1.2 was one of the ERFs induced in cassava tubers during PPD and played a vital role in the signal transduction pathway of the plant.

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