Screening The Mutants Of Rubisco From A T-DNA Inserted Rice Population Under Oxidative Stress

Rice is one of the most important crops in China. It is of great significance to increase largely crop yield per acre on the present basis and discover photosynthetic mechanism by improving photosynthetic efficiency.Sequencing of the rice genome is nearly complete; therefore, one of the most challenging goals now is to examine the functioning 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 screened of rice mutant of Rubisco using materials from a T-DNA transposon inserted rice (Oryza sativa L.subsp.japonica cv.Zhonghua 11) transgenic population. And this will be a guide to manipulate Rubisco and finally increase the yield of rice.Our work compares chlorophyll fluorescence parameters and content of Rubisco of leaves treated treatment with MV (Methyl viologen), a ROS inducer in vivo, by different methods to find a better strategy. And we find that treat the leaves through their petioles have better effect. We also consider the concentration effect. We make this method as the method we use to screen mutant from the T-DNA transposon inserted rice pool.The mutants were screened out by the different responses of the content of the Rubisco small subunit and large subunit to the treatment with MV. We used SDS-PAGE as a method to determine the content of subunits of Rubisco. Compared with the control, Rubisco from the mutants had obvious differences in small subunit (SSU) content. We also determined the leaves of mutants' and the wild-type's SPAD which show a relative value of chlorophyll content up to 5 days, to see the profile of the mutants under non-oxidative stress. With these methods we had got two mutants whose Rubisco show different tolerance to oxidative stress and demonstrated that this method of screening mutants is viable.