The Research On The Key DNA Region Of Responsive To SA、ET In Promoter Of OsNPR1

[Objective] NPR1has very important significance in gene function research of plant defence, three homologous genes of NPR1in rice (Oryza sative L.) genome were induced by bacterial leaf blight, rice blast and pathogens benzene defense related material and thiamethoxam diazole, jasmone acid methyl ester and ethylene, and NPR1is salicylic acid (SA) mediated disease resistance response of the key control factor, the study suggests that OsNPR1could control the salicylic acid dependent pathway in rice antagonistic interaction adjustment, this suggests that OsNPRl as a resistance genes can response the hormonal stress response, But OsNPR1for SA and ET induced reactions depend on which a key DNA regions that is still not clear.Therefore,OsNPR1promoter region of research and the role in cis elements were analyzed,is one of the important ways to clarify OsNPRl molecular mechanisms in the defence response of rice. This study used a variety of methods, such as bioinformatics, molecular biology of OsNPR1gene promoter and the study of hormone response.[The method]In this study, to identify genes which is regulated by the hormone in the induction element and related function analysis.To clone gene promoter region, Construct expression vector POsNPR1:GUS, carrier will import of tobacco (Nicotiana tabacum) leaf through a grobacterium tumefaciens mediated,to start the instantaneous expression experiments and study its expression specificity.Using hormone MeJA, ET, SA, ABA, IAA, GA and high salt,low temperature,drought,processing of tobacco seedlings.In order to further determine key DNA gene promoter region area, to delete the sequence of the promoter region, then promoter will be missing pieces with GUS reporter gene fusion, to test the GUS activity. By studying the characteristics of the promoter expression, contribute to more in-depth understanding of the rice disease resistance molecular mechanism of gene expression in response to different stimuli.[Results]1、Amplification from the rice genome of OsNPR1promoter region, to further build with GUS reporter gene expression vector of promoter p1381Z, get the expression vector POsNPR1:GUS. With hormone MeJA, ET, SA, ABA, IAA, GA, high salt, low temperature and drought treatment, GUS staining results showed that the fusion gene expression in the tobacco by SA and ET induced.2、In order to validate the SA and ET response element are located, based on the length OsNPRl promoter sequence, to build a series of5’promoter fragment delete carrier. Mutations of the promoter and the construction of GUS fusion gene by SA and ET induced in transient expression, the results show that presumably the area between-700to-300sequence is the key to the OsNPRl response SA and ET signal area, in combination with PLACE of software prediction, this sequence has a WBOXATNPR1(TTGAC,-485to-485,-501to-496) is one of the important reply response SA components, at the same time T-box (ACTTTG,-648to-643) is the key response ET response element.[The conclusion and significance] By constructing a different promoter fragment of plant expression vector POsNPR1:GUS, SA and ET key DNA response area is identified, thereby for the separation and identification of the gene promoter region which might be responsible for answering different adversity or study the theoretical base of Cis-acting elements, is one way to clarify the molecular mechanisms of inducible defense responses in plants.