Cloning And Primary Functional Analysis Of Rice Blast Resistance Related Gene OsMP1

Rice is one of three major food crops in the world,and nearly half of the world’s population is fed on rice.Rice blast which caused by Magnaprothe oryzae,is one of the most serious rice diseases.Each year,the product of rice worldwide fell by 10%to 30%due to rice blast.Therefore,research on rice blast resistance related genes and their molecular mechanisms involved in resistance,and applied to disease resistance breeding,improving rice blast resistance is important ro the prevention and treatment of the disease.OsMPl is selected through gene chip analysis after Heikezijing infected by M.oryzae and iTRAQ proteomic analysis,which induced by the rice blast significantly.Though RT-PCR,we cloned OsMP1 from Heikezijing,and then we studied the related bioinformatics characteristics,gene expression profiles,Southern blot identify and conducted in rice protoplasts to show the subcellular localization of OsMP1.The mainly results are as follows:1.OsMP1 coding 373 amino acids,which containing 10 transmembrane domain,belongs to a membrane protein.It has single copy in Heikezijing.The result showed that OsMPl can improve the rice resistance to rice blast mabey related to its localization on the plasma membrane.Tissue expression analysis showed similar expression pattern of OsMPl between Heikezijing and Suyunuo.OsMPl constitutively expressed in various rice tissues,with higher expression in the immature panicle,lower expression in stem,leave and leaf shesth,and with slightly higher expression in Heikezijing than in Suyunuo.The expression analysis in resistance varirty Heikezijing and suspective variety Suyunuo induced expression showed,OsMP1 was induced by rice blast significantly.The expression of OsMPl showed a rapid increase in both Heikezijing and Suyunuo at 4 hpi with rice blast Hoku 1,then decreased,with greater induction of OsMPl expression in Heikezijing than that in Suyunuo.Through rice protoplast expression system it was found that the subcellular localization of OsMP1::GFP fusion protein was mainly located in the plasma membrane.2.Screening the interacted proteins of OsMP1 through yeast two-hybrid experiment,OsMP1 was used as "bait" protein,two interacting proteins were found in rice membrance cDNA library,the one named MP-P1 has a ubiquitin domain.It is located in vesicle,which is associated with ubiquitin protein;The other one named MP-P2 has a SNARE_assoc domain which located in the golgi.It’s a Qa-SNARE which is a SNARE associated golgi protein.It was supposed that rice OsMPl possibly played a role in the process of rice blast resistance to penetration through interacting with other proteins.