Analysis Of OsPRP-Like-a Interaction Protein With OsHsp90

As the global environment is becoming worse and worse,some abiotic stresses such as drought,cold,heat shock and salination influence the growth and development of crops so badly and it will reduce the production and quality of grain.Therefore,to find news gene related with abiotic stresses and to apply them to screen and improve resistance variety become a hot research topic.In recent years,with the development of molecular biology theory and technique,the research on adversity proteins have made a great progress,including isolation of many genes encoding adversity proteins and finding out many proteins interacted with known resistance genes.We screened many genes related with abiotic stresses and mainly discovered a rice hot shock OsHsp90 before by using Fluorescent Differential Display(FDD) and Macroarry.Then we also found many proteins bound with OsHsp90 by using yeast two-hybrid system.Especially,we discovered a OsPRP-like gene correlated with stress.Thus,we expect to research into the physiological mechanism of these proteins deeply,and to understand their expression and regulation mechanism in stresses.Finally,put them into practice of crop product to increase crop yield.Based on the Former's research in our lab,we take OsPRP-like as the study object. The main results showed as follows:1.Using OsHsp90 as a bait protein,we isolated 5 cDNA of interaction protein from the cDNA library which constructed by rice under cold and drought condition by yeast two-hybrid system.After sequenced and blast,we found that as follow:(1) two genes encoding a same protein is rice proline rich protein(OsPRP) linked with development of cell wall;(2) RBBI3-3 is a protease inhibitor;(3) P450 is interrelated with cytochrome;(4) a unknown gene containing ZIM domain is related with plant transcription factor.2.After transferd the insert library from the AD to DNA-BD vector,we confirmed the reality of interaction between OsPRP and OsHsp90.After inserting the different part of OsPRP to BD vector,we found the most strong interaction site is proline rich region and it interact with the C-teminal of OsHsp90.3.A OsPRP gene family like gene was obtained by RT-PCR,which has 1686 bp length,contains an ORF from 89bp to 1339bp and encodes a 416 amino acids protein(46.5kD).Therefore,we named it OsPRP-like gene.Its main interact site gathers in PEPK repeat region from 54 to 412 AA residues,where we concentrate on the function predict.4.The analysis by Semi-quantitative PCR and real time fluorescence quantitative PCR showed that OsPRP-like is sensitive to cold,heat shock,drought and salination.Compared with the result of OsHsp90,we predict that OsPRP-like may be a precursor for start and expression of OsHsp90,or it may response of adversity damage independently.In addition,OsPRP-like can express specially at the different stage of rice growth.It is likely to participate in regulation of growth and development of crops or transfer the adversity signal.