摘 要 :以水稻(Oryza sativa L. subsp. japonica)品种“中花11”构建的T-DNA插入突变体库为材料,用甲基紫精诱导叶绿体产生活性氧,通过SDS-PAGE电泳分析Rubisco含量相对于野生型的变化,筛选耐氧化胁迫的株系。结果表明,与对照相比,突变体材料各个株系叶片中的Rubisco小亚基的相对含量有较大差异,说明这种筛选Rubisco突变体的方法是可行的,且从中找到了两株Rubisco耐氧化胁迫的株系。
Abstract:Sequencing of the rice genome is nearly complete; therefore, one of the most challenging goals now is to examine the function of its large number of genes. In this article, to get the details about the mechanism how Ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) can keep stable under oxidative stress, we screen the mutant of Rubisco using materials from a T-DNA transposon inserted rice (Oryza sativa L. ssp. japonica cv. Zhonghua 11)transgenic population, by comparing the contents of Rubisco small subunit and large subunit different from these of rice mutants treated with MV (Methyl viologen), a ROS inducer in vivo. We use SDS-PAGE to determine the content of subunits of Rubisco. Compared with the control, Rubisco from the leaves of the mutants had obvious difference in small subunit (SSU)content. We also determined mutants’ and the wild-type’s SPAD of leaves as a relative value of chlorophyll content, to compare the leaf senescence speed under oxidative stress. With this method we got two mutants whose Rubisco showed different tolerance to oxidative stress, demonstrating that this method of screening mutants is viable.