龙葵E2泛素结合酶基因SorUBC克隆及表达特性分析  

Clone and expression pattern analysis of E2 ubiquitin-binding enzyme gene SorUBC in Solanum nigrum L.

作　　者：蔡佳文[1] 金晓霞[1] 于丽杰[1] 崔柏杨 魏迅[1] 董延龙[2] 

机构地区：[1]哈尔滨师范大学生命科学与技术学院,植物生物学黑龙江省高校重点实验室,哈尔滨150025 [2]黑龙江省农业科学院园艺分院,哈尔滨150069

出　　处：《东北农业大学学报》2016年第11期26-36,45,共12页Journal of Northeast Agricultural University

基　　金：黑龙江省教育厅面上项目(12531204,12531178);“植物生物学”黑龙江省高校重点实验室开放课题(ZK201205);哈尔滨师范大学青年学术骨干资助计划项目(KGB201218)

摘　　要：对龙葵SorUBC基因克隆并分析其在多种非生物胁迫处理下表达特性，为E2泛素结合酶在植物逆境胁迫中应用提供理论依据。试验采用电子克隆结合RT—PCR方法从龙葵中克隆得到泛素E2结合酶UBC基因编码区序列，命名为SorUBC，登陆GenBank，编号：KU233684。SorUBC基因编码区全长888bp，编码295个氨基酸。进化树分析显示，该基因编码氨基酸序列与马铃薯、番茄同源性最高，为95％，其次是烟草，为91％。该蛋白在第266—284氨基酸位具有1个跨膜螺旋区域，泛素化位点预测表明其含有8个泛素化位点。利用荧光定量PCR技术分析SorUBC基因在龙葵各器官表达特征和不同非生物胁迫处理下（干旱、盐、碱、高温和低温）根部和叶片表达特性，同时测定龙葵叶片生理响应。结果表明，SorUBC基因表达具有组织差异性，在叶片和果实中表达量较高。非生物胁迫处理（干旱、盐、碱、高温和低温）后根部和叶片基因表达情况显示该基因存在早期（3或5h）应答上调表达，且根部与叶片基因表达量和变化趋势差异显著，根部表达量变化比叶片显著，推测SorUBC基因在龙葵早期响应干旱、盐、碱、高温和低温非生物胁迫中起调控作用。生理特征数据表明，在干旱、盐和高温胁迫中龙葵叶片MDA含量变化差异不显著，POD和SOD活性总体呈先降后升趋势，说明其在胁迫后期（9h）对活性氧造成的损伤起缓解作用。In this research SorUBC gene from Solanum nigrum L. was cloned and the expression pattern under different abiotic stresses were detected. This would help us to predict the characteristics of E2 ubiquitin binding enzyme and provide theoretical basis for the application of E2 ubiquitin binding enzyme in plant stress. The research cloned E2 ubiquitin binding enzyme UBC gene full-length coding region sequence by electronic cloning and RT-PCR method from Solanum nigrum L., named SorUBC,GenBank accession number： KU233684. The coding region sequence was 888 bp, encoded 295 amino acids. Phylogenetic tree analysis showed that SorUBC amino acid sequence was homologous highest with potato and tomato, was 95%, followed by tobacco which was 91%. The protein was predicted to have one transmembrane helical regions in the 266-284 amino acid site, and the ubiquitin-based site prediction showed that it contained eight ubiquitin-based sites. The expression characteristics of SorUBC in various organs and in roots and leaves with abiotic stress treatments （drought, salt, alkali, high temperature and low temperature） of Solarium nigrum L. were detected using quantitative PCR analysis, meanwhile physiological response of leaves in Solarium nigrum L. were determined. The results showed that the expression of SorUBC gene were different in tissues, higher in leaves and fruits. Detection of gene expression in roots and leaves after abiotic stress treatments（drought, salt, alkali, high temperature and low temperature） showed that there was an early （3 or 5 h） response up regulation expression of SorUBC gene, the expression levels and the trends of the gene in roots and leaves had significant difference, the expression in roots were more obvious than in leaves, suggesting that SorUBC gene might play a regulatory role in early stage response to different abiotic stress such as drought, salt, alkali, high temperature and low temperature stress in Solanum nigrum L. Physiological characteristic data showed that no significant chang

关 键 词：龙葵 SorUBC 克隆 表达分析 非生物胁迫 生理特性 

分 类 号：Q785[生物学—分子生物学]