The Research Of OsPP6C Gene Overexpression And RNA Interference In Rice

With the improvement of production, rice is not only as an important grain cropin China, but also as an economic crop and forage crop. So it has great value ofdevelopment and utilization, and become more and more important to our life. Inrecent years, although the yield and quality of rice are improved, it is still affectedwith a lot of disadvantages. New varieties cultivation is the only way for thedevelopment of agricultural science and technology in our country. The technique ofmolecular biology and plant gene engineering is the most effective way ofaccomplishing this work.Kinases outnumber phosphatases. However, a relatively limited number ofprotein phosphatases can maintain protein phosphorylation homeostasis in a cell.Protein serine/threonine phosphatases (PSPs) can be categorized into three families—phospho protein phosphatases (PPPs),metal-dependent protein phosphatases(PPMs,PP2Cbelongs to PPMs), and aspartate-based phosphatases such as FCP (TFIIF-associatingcomponent of RNA polymerase II CTD phosphatase) and SCP (small CTD phosphatase).Representative members of the PPP family include protein phosphatase1(PP1), PP2A,PP2B (commonly known as PPP3), PP4, PP5, PP6, and PP7.RNA interference is mediated by double-stranded RNA, typically by causing thedestruction of specific mRNA molecules, which block the corresponding geneexpression of the transcription level of gene silencing mechanism. Most basic RNAiconstructs have an inverted repeat interrupted with a spacer sequence. The spacersequence allows stable replication of RNAi plasmids in E. coli, and sequencesprepared from the β-glucuronidase (GUS) gene, the green fluorescent protein (GFP)gene, and several introns are used as a spacer for the RNAi construct. Several reportsshowed that enhanced gene silencing can be established by using an intron spacerrather than by the GUS or GFP spacers.The Arabidopsis thaliana PP6were found play an important role in cell signaltransduction, especially in relation to auxin, abscisic acid and plant aging. However,there was scarce report in other speices. In this research, to analyze the function ofOsPP6, using the RT-PCR technique, the full length of OsPP6C gene was amplifiedfrom the complementary DNA (cDNA) of rice "Zhonghua11" variety, and it wasfused with GUS to form pBI121-OsPP6C-GUS expression vector. At the same time, RNAi vector of OsPP6C gene have been constructed. We have cloned theOsPP6CRNA interference and OsPP6C gene fragment of rice protein phosphatase6catalytic subunit gene, and constructed the PBI121-OsPP6C expressing vector. Mover,constructed the pCambia2300-RNAi-OsPP6C expressing vector of this gene that hasan inverted repeat interrupted with a intron spacer sequence. As a result, plantover-expression and RNAi inhibition vectors for OsPP6C were constructedsuccessfully. This paper will lay a solid foundation for further studying functions ofOsPP6C gene in rice.