The Cytology And Proteome Studies On The Resistance Of Wheat Carrying Pm40 Against Blumeria Graminis F.sp.tritici

Wheat powdery mildew caused by Blumeria graminis f.sp.tritici(Bgt),is the main disease affecting the wheat yield seriously in the world.Recently,using the powdery mildew-resistant wheat variety is the most economic,effective and safety method to control the wheat powdery mildew.Wheat powdery mildew resistance gene Pm40 was derived from Elytrigia intermedium,located in 7BS chromosome of wheat.Pm40 plays a potential role in Chinese wheat breeding programs in the post-Pm21 era.The study of the interaction mechanism between wheat and Bgt is helpful to reveal the resistance mechanism of wheat against Bgt and these studies could provide the new though to breed resistant wheat.In the study,the resistant wheat L658 with Pm40 and the susceptible wheat L958 without Pm40were used as materials and wheat L658 and wheat L958 were sister line.The roles of H₂O₂,callose,(1,4)-?-xylan and cellulose were studied using the staining methods of DAB(3,3-diaminobezidin),aniline,LM10-FITC and CBM3a-His-FITC/AMCA.The proteomic differential expression of the leaves affected by Bgt in 24,48,72h was studied using the i TRAQ technique in wheat L658 and wheat L958.The m RNA expression level of genes encoding differentially accumulated proteins in the wheat leaves affected by Bgt were studied by the q RT-PCR technique.The protein expression profile of the Bgt's conidia was provided using the shotgun technique.The main results are listed below:1.The percentage of the papilla in the wheat L658 is two times as much as that in the wheat L958 affected by Bgt at 48 and 72 hpi respectively.The percentage of invasion site producing H₂O₂in the wheat L658 is significantly higher than that in the wheat L958.Papilla and H₂O₂produced by wheat make the contribution to the resistance of the wheat L658.Using the microscope and fluorescence staining technique,the study reveals that the chemistry components of the wheat papilla include callose,(1,4)-?-xylan and cellulose.2.There are 3152 proteins identified in the wheat L658 and L958 affected by Bgt at0,24,48 and 72 hpi using i TRAQ technique.The GO function analysis reveals the upregulated multiple or period of the 27 up-regulated proteins of immune system process in wheat L658 is higher or earlier than in the wheat L958.The KEGG pathway analysis reveals alpha-linolenic acid metabolism,biosynthesis of secondary metabolites,biosynthesis of unsaturated fatty acids,glutathione metabolism,glycolysis/gluconeogenesis,phagosome,and phenylpropanoid biosynthesis play important roles for the resistance of wheat L658.Butanoate metabolism and photosynthesis play important roles for the susceptibility of wheat L958.3.The expression of PR1,12-OPR,PAL is up regulated in wheat L658 and L958,but the upregulated multiple is significantly higher in L658 than in the L958.Salicylic acid,jasmonic acid and phenylpropane pathway play important roles in the wheat resistance response mediated by Pm40.The expression of genes editing identified proteins by i TRAQ in the m RNA level is conformity or inconformity with protein level.The reason of the inconformity may be the post-transcriptional control.Glutathione S-transferase,glucan glucosidases,chitinase,14-3-3 protein,and calreticulin play important roles for the resistance of wheat L658.4.There are 1654 proteins identified in Bgt's conidia using shotgun technique,providing a protein expression profile for Bgt's conidia,including 5 BECs.The GO function analysis reveals Bgt's conidia has 56 proteins with the response to stimulus.The KEGG pathway analysis reveals Bgt's conidia includes alpha-linolenic acid metabolism,biosynthesis of unsaturated fatty acids,glutathione metabolism,glycolysis/gluconeogenesis,phagosome,and butanoate metabolism and the following pathways play key roles in signal transduction of Bgt,including MAPK signaling pathway,c AMP signaling pathway,calcium signaling pathway.