拟南芥At5g66070基因在ABA处理下的初步研究

Primary Analysis of Arabidopsis thaliana At5g66070 Gene in ABA Treatment

摘要:

摘要：ABA是植物体内非常重要的一种激素，通过诱导某些重要基因的表达从而调节植物响应逆境胁迫。本实验室前期研究中发现一个RING结构的E3连接酶AT5G66070，通过生物信息学分析发现其受ABA诱导，因此我们对其在ABA处理条件下的相关生理功能进行了分析。结果表明，10μMABA处理下该基因的表达量明显升高。亚细胞定位发现AT5G66070定位在细胞核。为进一步研究其功能，通过DNA及RNA水平鉴定了该基因的T-DNA插入的突变体，然后经根瘤农杆菌介导法进行遗传转化，筛选获得了过表达转基因株系。表型分析发现经ABA处理后，突变体相对于野生型而言根长较短，表明突变体对ABA更为敏感。气孔关闭实验发现10μM ABA能诱导突变体气孔关闭，而野生型和过表达无显著影响。初步研究表明At5g66070在拟南芥的ABA胁迫响应中起负调控作用。 关键词：At5g66070;非生物胁迫;拟南芥；ABA信号通路;

Abstract:

Abstract: The phytohormone abscisic acid is a key endogenous messenger in plants’ response to abiotic stress by regulating some crucial gene expression. Founded in previous research in this lab, a RING structure of E3 ligase At5g66070, through bioinformatics analysis found could be induced by ABA.So we analyse its related physiological function under the condition of ABA treatment .Gene At5g66070 was studied before in this laboratory. The results showed that the gene expression was significantly increased after 10μM ABA treatment. The cellular localization of At5g66070 was predominantly found in the nuclear,indicating At5g66070 is a nuclear protein. No full-length At5g66070 transcript was detected in the mutant by DNA and RT-PCR analysis, suggesting that At5g66070 is a loss-of-function mutant.Germinated evenly-grown seedlings were then transferred to MS plates supplemented with or without ABA.At5g66070 mutant showed shorter primary roots compared with the overexpression lines under ABA treatment.It means the mutant is more sensitive than wide type and overpression lines. The 10μM ABA treatment induced stomata closure in mutant, but had no effect on wild type and overexpression lines. Using RT-PCR to analyze the related genes in Arabidopsis ABA singnaling pathway.Our results indicate that At5g66070 is a negative regulator of ABA signaling.