拟南芥AtRaGAP基因功能分析.pdf

摘 要 在植物生长发育的过程中，受到各种因素的刺激，形成了复杂而多样的信号系统， 最终引起相应的生理生化变化和形态反应。ABA 在植物的生长发育过程以及植物抗逆 过程中起重要的作用。已知某些小G 蛋白在 ABA 的信号转导过程中起作用，参与植物 逆境应答、细胞骨架的运动、细胞扩大、根毛发育以及细胞极性生长的信号转导。 RabGAPs 通过催化Rab 水解GTP 使Rab 蛋白在活化形式与非活化形式之间转换，从而 行使分子开关的功能。RabGAPs 大多数含有保守的TBC 结构域，参与胞吞胞吐和极性 运输等过程，但其在植物信号转导过程中的作用还不是很清楚。 我们以拟南芥GAP基因的T-DNA插入缺失纯合突变体为材料，研究了该基因的功 能。表型分析发现，在含有ABA 的MS培养基上，与野生型相比突变体gap在种子萌发和 子叶变绿方面对ABA不敏感。而在失水率方面野生型与突变体没有明显的差异。通过 RT-PCR半定量分析发现，GAP在外源ABA作用下表达量下调，GUS染色得到同样结果。 缺失突变体在用外源ABA处理后体内NCED3表达量下调，CYP707A表达量则上调。GUS 染色显示其在植物根茎叶花中均有表达，气孔中也有微弱表达。 GAP::GFP融合蛋白显 示其定位于细胞质中。因此推测GAP可能在ABA信号通路中起作用。 总之，本研究使人们对GAP的功能有了更进一步的了解，即GAP不仅是Rab 的激活 蛋白，而且可能在ABA 的信号转导过程中起作用。 关键词：拟南芥；GAP；ABA III ABSTRACT Plants could be stimulate by various factors in growth and development process, forming a complex and diverse signaling system. Eventually lead to physiological and biochemical changes and morphological responses. ABA plays an important role in plant growth and development process, and ABA is an important signaling molecule in the process of environmental stresses. Some small G proteins play a role in ABA signal transduction, involved in the movement of the cytoskeleton, cell expansion, root hair development and growth of cell polarity and signal transduction. GTP hydrolysis by Rab protein, which was activated by GAPs, thus GAPs perform the function of the molecular switch through regulating the conversion of Rab between the activated form and non-activated form. GAPs TBC domain containing the majority involved in endocytosis and exocytosis during polar transport, but in plant signal transduction role in the process is not very clear. In this study, we investigated the functions of GA