牵牛胁迫应答基因PnLOG2的克隆及功能验证
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引用本文:侯胜男,金晓霞,陈 超,于丽杰.牵牛胁迫应答基因PnLOG2的克隆及功能验证[J].西北植物学报,2019,39(6):982~990
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作者单位
侯胜男,金晓霞,陈 超,于丽杰* (哈尔滨师范大学 生命科学与技术学院, 哈尔滨 150025) 
基金项目:黑龙江省自然科学基金面上项目(C2017039);
中文摘要:RING型E3泛素连接酶在植物应答非生物胁迫过程中发挥着重要功能。该研究从圆叶牵牛中克隆出RING型E3泛素连接酶基因PnLOG2,该基因序列号为XM_019321049.1。利用ORF Finder预测PnLOG2基因编码开放阅读框长度为912 bp (51~992 bp),编码313个氨基酸,蛋白分子质量34.38 kD,理论等电点为5.14。系统发育分析表明,PnLOG2基因与番茄亲缘关系最近。组织特异性分析表明,PnLOG2基因在牵牛不同组织均有表达,在老茎和新叶中表达量较高。qRT PCR分析结果表明,PnLOG2基因在圆叶牵牛根和叶中受干旱、盐碱胁迫诱导显著上调表达。通过异源表达PnLOG2基因于酵母细胞中,发现干旱、盐碱胁迫下PnLOG2基因提高了重组酵母的耐盐和耐旱能力,但降低了对碱的耐受性。该研究初步阐明了PnLOG2基因在干旱、盐碱胁迫下的功能,为进一步研究RING型E3泛素连接酶在非生物胁迫中的机理提供了理论依据。
中文关键词:干旱      功能验证  泛素连接酶
 
Cloning and Functional Analysis of the PnLOG2 Gene in Response to Stress in Pharbitis purpurea
Abstract:RING type E3 ubiquitin ligase play an important role in plant response to abiotic stress. In this study, we isolated a RING type E3 ubiquitin ligase gene PnLOG2 from Pharbitis purpurea. The gene ID was XM_019321049.1. The length of open reading frame was predicted to be 912 bp (51-992 bp)using ORF Finder. PnLOG2 encoded a protein of 303 residues with a predicted molecular weight of 34.38 kD and an isoelectric point of 5.14. Phylogenetic analysis showed that PnLOG2 gene was clustered with tomato E3 ubiquitin ligase gene. Tissue specific analysis showed that the expression levels of PnLOG2 in different tissues was detected. And the PnLOG2 was highly expressed in the old stem and new leaves. The qRT PCR data showed that the expression levels of PnLOG2 were significantly inducted by drought, salt and alkali stresses in the roots and leaves of P. purpurea. Further, ectopic expression of PnLOG2 in yeast cells enhanced the tolerance to salt and drought stresses, but decreased the tolerance to alkali stress. Therefore, we identified the function of PnLOG2 gene in response to drought, salt and alkali stresses, and provided a theoretical basis for further study RING type E3 ubiquitin in abiotic stress.
keywords:drought  salt  alkali  functional verification  PnLOG2
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