Effects Of Rna Interference Of A Multiple-stress Responsive Gene,OsMsr16,in Rice

To understand mechanisms underlying stress tolerance and discover novel stress-tolerance genes in rice (Oryza sativa L.), Gene Chip Rice Genome Array (Affymetrix) was used to investigate the global genome expression profiles of the indica cultivar Pei’ai64S subjected to cold, drought or heat stresses. A large number of genes were highly up-and down-regulated under the stresses. OsMsr16(Oryza sativa L. Multiple Stresses Responsive Gene16)was one of the genes highly induced in different plant organs at different development stages by different stresses. In order to understand functions it makes in the rice growth and development. The bio-informatics has took to OsMsrl6, and the construction of RNA interference then delivered it into rice, the transgenic lines were identified by Southern Blot and with the RT-PCR we know the Relative expression of OsMsrl6in rice under the RNA interference.The bio-informatics analysis showed that OsMsr16contains five exons and four introns and encodes a protein with715amino acid residues. Gen Bank sequence database searching and multiple sequence alignments showed that OsMsrl6may belong to the PHD-type contain proteins, it may take actions with regulation the express of PHD-type contain proteins. There were11stresses-responsive cis-acting elements were found in the putative promoter region of OsMsr16which means the gene may play a important role in responding stresses.After obtain the complete ORF through amplification by RT-PCR, the interference vector pRNAi-JIT-OsMsrl6was constructed with a specific sequence contain580bp in OsMsrl6and original vector pCAMBIA1300and pRNAi, the interference vector delivered into rice, and a total of25independent transgenic lines were obtained.Positive plants were selected by PCR and then the interference vector’s copy number was defined by Southern blot.The data of Quantitative real-time RT-PCR show that the interference vector interfered the expression of OsMsr16, and the relative expression of OsMsr16was reducing significantly. I2-KI dyeing the transgenic plants’pollen, we found all the pollen were sterile. At the same time some other stresses sensitivity and infertility phenotypes were observe in transgenic plants.In conclusion, we constructed the interference vector pRNAi-JIT-OsMsrl6, Foretasted and analysised its molecular biological characteristics and functions. the transgenic plants’phenotypes show some stresses sensitivity and infertility phenotypes with interfered the expressing of OsMsrl6. The function of OsMsr16was confirm and the result can provided supports for further study.