Differential Proteome Between Resistant And Susceptible Varieties To Rice Stripe Tenuivirus

Rice stripe disease is a serious threat for main rice producing areas in China,which is caused by Rice stripe tenuivirus?RSV?.The game of virus invasion,resulting in host defense,large number of host's proteins were differetinal expressed,such as the protein recruited by the virus for its life activities and the protein produced by plant for resistant virus invasion.Therefore,the study of differential expressed proteins under virus stress can easily describe the mechanism such as the disease resistance as well as the formation of symptoms in infectious rice plant.In which study,the effect of RSV infection on the proteome of high resistance and high susceptibility rice varieties were analyzed by using iTRAQ.As the result shows that 496 up regulated proteins and 541 down regulated proteins were identified in resistant line?Zhendao 88?after RSV infection;509 up regulated proteins and 544 down regulated proteins were identified in the susceptible line?Wuyujing 3?after RSV infection.207 up regulated proteins and 213 down regulated proteins were commonly expressed in both resistant and susceptible line.By comparing the fold change?log₂?of the up regulated proteins between two lines,25 induced resistance-related proteins were identified.With the help of gene ontology,these induced resistance-related proteins were described involved in metabolism of reactive oxygen species,biosynthesis of secondary metabolites,programmed cell death,post-translational modification and cell wall structure enhancement.After Gene Ontology and KEGG annotation of 213 down regulated proteins,rate of the chloroplast proteins was found be the most affected by the virus infection.Among them,9 proteins were found associated with the biosynthesis of chlorophyll,such as Geranylgeranyl diphosphate reductase,Protochlorophyllide reductase B,Magnesium-chelatase subunit D?CHLD?,Oxygen-dependent coproporphyrinogen-III oxidase,Uroporphyrinogen decarboxylase 2 and Glutamate-1-semialdehyde2,1-aminomutase;9 proteins were found involved in photosynthesis,such as Ribulose bisphosphate carboxylase small chain,Cytochrome f?Cytf?,Chloroplast photosystem I reaction center subunit II-like protein?PSI-D?,NAD?P?H-quinone oxidoreductase subunit 4L and NAD?P?H-quinone oxidoreductase subunit J.Down-regulation of these proteins may lead to the inhibition of biosynthesis of chlorophyll and photosynthesis,leading to the destruction of the chloroplast structure and casues plant's symptoms of yellowing and chlorosis.In order to verify the differetinail of transcriptional level of differentially expressed proteins by RT-qPCR.Sixdisease-resistantproteins,suchasTubulinalpha-1chain,Dolichyl-diphosphooligosaccharide-protein glycosyltransferase subunit STT3B,Glutaredoxin-C6,Os02g0586400 protein,Superoxide dismutase[Cu-Zn]1,Probable V-type proton ATPase subunit H,involved in disease resistance were selected from the up-regulated proteins;four proteins,including Photosystem I P700 chlorophyll a apoprotein A1,Cytochrome b6-f complex iron-sulfur subunit,Protochlorophyllide reductase B,Magnesium-chelatase subunit D?CHLD?,Probable V-type proton ATPase subunit H,involved in chlorophyll biosynthesis were selected from the down-regulated proteins.The results shows that the 10 proteins were consistent with the results of iTRAQ,indicating that the results of iTRAQ were accurate and reliable.In order to further exploring the mechanism of the formation of rice stripe disease,the virus components interacting with the magnesium ion chelatase D subunit?CHLD?were screened by yeast two-hybrid system.The results showed that RSV specific protein?SP?able to interact with CHLD.Then,the overexpression vector of CHLD was constructed and inherited the rice,and eventually obtained 33overexpressed rice plants of CHLD.Which provided valuable experimental material for further explore the role of CHLD in symptom formation.In addition,the pathogen of pepper virus disease occurred in Guiyang Guizhou was identified by deep sequencing technology.The result showed that the serious viral disease was caused by co-infection of Cucumber mosaic virus?CMV?,Potato virus Y?PVY?,Broad bean wilt virus 2?BBWV2?,Chilli veinal mottle virus?ChiVMV?and Bell pepper endornavirus?BPEV?.At the same time,two RT-PCR detection systems of four pepper viruses,BBWV2,ChiVMV,CMV,PVY and five watermelon virus,Zucchini yellow mosaic virus?ZYMV?,Watermelon mosaic virus?WMV?,Melon aphid-borne yellows virus?MABYV?,Cucumber green mottle mosaic virus?CGMMV?,Citrullus lanatus cryptic virus?CiLCV?,were established for the early diagnosis and effective prevention and control of viral disease.