Construction Of Indica Rice (Minghui 86)T-DNA Insertional Mutant Library

With the completion of rice (Oryza sativa) genome sequencing, large information will be available which may play important roles in rice functional genomics. Obtaining a mutant library by insertion is a powerful strategy for the research of rice functional genomics.In this study, we adopted promoter trap system, pCASGFP-TRAP,that consisted of modified promoterless gfp reporter gene (mgfp5) located next to the right border, to generate a trapping population of indica rice. Currently, 5944 independent transgenic plants were obtained from indica cultivar Minghui 86, which are first T-DNA insertion indica lines. PCR tests and Southern blot analysis showed that most exogenous genes were integrated into rice genome at lower copies. mgfp5 was found to be expressed in some tissue(s) /organ(s) of the part of transgenic plants via fluorescence microscope, while lots of mutants with distinct phenotype variation were found in T₁ generation.These results indicated that the promoter trap system is efficient in indica rice. In addition, flanking sequences at T-DNA insertion site of the part transformants were obtained. Homologous comparison by BLASTn was conducted in GeneBank.