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- 2017
拟南芥RING finger结构域ABRv1基因的功能初步研究
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Abstract:
拟南芥ABscisic acid responsive RING-v protein 1基因(ABRv1)编码一个含有RING finger结构域的E3连接酶。为了分析ABRv1的基因功能,我们首先构建了高表达载体pBI121- ABRv1,经农杆菌转化,花序浸染后得到T0代转基因种子,并经卡那霉素板筛选得到转基因苗,再经过两代的筛选获取了纯合的高表达株系ABRv1。通过DNA及RNA水平鉴定了该基因的T ?CDNA插入突变体abrv1。对突变体、高表达转基因株系及野生型进行ABA诱导处理,检测其萌发率、气孔关闭情况进行,发现ABA处理后突变体的萌发率降低,过表达株系萌发率升高;突变体株系气孔几乎未关闭,过表达株系气孔关闭非常明显。以上结果表明ABRv1基因在拟南芥的ABA信号应答响应中可能起负调控作用,为深入分析该基因的功能打下基础。
ABscisic acid responsive RING-v protein 1(ABRv1) in Arabidopsis thaliana encodes a E3 ligase with a RING finger structure domain. In an effort to analyze the function of ABRv1 in the plant, we performed abscisic acid (ABA) stress experiments on the wild-type, the ARBv1-overexpressing lines and the T-DNA insertion mutants abrv1. The overexpressing lines were acquired through transfection by Agrobacterium with ABRv1 embedded plasmids (pBI121- ABRv1). The mutants obtained were identified by DNA and RNA analysis. With ABA treatment, the mutants presented a lower germination rate than the wild-type while the overexpressing lines showed a higher rate. Little stomatal closure was observed in the the mutants whereas obvious closure was discovered in the overexpresssing lines. The wild-type showed a closure status in between. Therefore, our phenotypic analysis revealed that ABRv1 might participate in the plant response to ABA as a negative regulator