水稻OsMBF1c基因的克隆及表达分析

Cloning and expression analysis of OsMBF1c gene in rice

关键词:  水稻, OsMBF1c, 基因克隆, 表达分析, 镉

Key words:  rice, OsMBF1c, gene cloning, expression analysis, cadmium

摘要:

多蛋白桥联因子1(multi protein bridging factor 1, MBF1)在植物应对逆境胁迫中起着重要的作用,而对于水稻中MBF1是否参与重金属胁迫响应机制目前尚未见相关报道。为了揭示水稻MBF1家族与重金属胁迫的相关性及其潜在作用机制,该研究利用PCR技术克隆水稻OsMBF1c基因的全长编码序列,通过生物信息学对基因功能进行分析和预测,并通过实时荧光定量PCR(RT-qPCR)分析其在镉(Cd)胁迫下的表达特征。结果表明:(1)OsMBF1c的全长编码序列为468 bp,共编码155个氨基酸,相对分子量为16.154 kDa。(2)OsMBF1c与大麦TdMBF1a.1亲缘关系最近,具有光、厌氧等环境因子诱导相关的顺式调节元件。(3)重金属Cd可诱导OsMBF1c表达且在时间上和组织中的表达水平具有特异性,100 μmol·L^-1 Cd 处理1 h 后,地上部分OsMBF1c表达量明显上调,为对照组的7倍; 100 μmol·L^-1 Cd 胁迫处理6 h后,根部OsMBF1c表达量上调为对照组的3倍。该研究结果进一步完善了非生物胁迫下MBF1家族的生物学功能研究。

Abstract:

Multi protein bridging factor 1(MBF1)plays an important role in plant stress resistance. However, there is no report about the specific functional mechanism of MBF1 in rice under heavy metal stress. The purpose of this study was to shed light on the correlation and potential mechanism between MBF1 family and heavy metal stress in rice. In this article, the full length coding sequence of OsMBF1c was cloned by PCR, the function of OsMBF1c was predicted by bioinformatics analysis, and the expression characteristics of OsMBF1c under Cd treatment was analyzed by RT-qPCR. The results were as follows:(1)The full length of OsMBF1c was 468 bp, which encoded 155 amino acids with the relative molecular weight of 16.154 kDa.(2)OsMBF1c was closely related to TdMBF1a.1, and cis-acting elements analysis showed that OsMBF1c was regulated by environmental factors such as light and anaerobic.(3)Gene expression analysis indicated that OsMBF1c was induced by Cd, and the expression level of OsMBF1c varied with different time or different tissues. After treated with 100 μmol·L^-1 Cd, the expression level of OsMBF1c in shoots at 1 h was remarkably up-regulated, which was seven times that of the control group, and the expression level in roots at 6 h was up-regulated to three times that of the control group. In conclusion, this study further refines the biological functions of MBF1 family under abiotic stress.