The Expression,Identification And Mechanism Of Chinese Wheat Mosaic Virus-induced Long Non-coding RNA

Wheat is currently the most widely distributed and the largest cultivated food crop in the world.Once severe wheat virus disease occurs,it will cause serious wheat yield loss.Wheat virus disease transmitted by Polymyxa graminis L.is a worldwide disease.Wheat viruses transmitted by Polymyxa graminis L.are mainly Chinese wheat mosaic virus(CWMV)and wheat yellow mosaic virus(WYMV)in China.CWMV is a single-stranded sense RNA virus(ss RNA).Studies have found that CWMV infection can induce a large number of long non-coding RNAs(lnc RNAs)to be differentially expressed.These lnc RNAs involved in the regulation of host resistance pathway in the viral infection process,the virus involved in the pathogenic process,as well as the mechanism of action of the above involved,are worthy of the in-depth study.In this paper,the model plant-N.benthamiana is the research object.After CWMV infection,the lnc RNA expression characteristics of plants were systematically analyzed,a network map of lnc RNA regulating plant related gene expression was constructed,and Nblnc81288 was used as a mic RNA precursor to regulate virus infection mechanism.The main research content are as follows:(1)Transcriptome sequencing was performed on samples infected with CWMV using high-throughput sequencing technology.4044 differentially expressed m RNAs and 616 differentially expressed Lnc RNAs were identified.There were 1642 genes with significant differential expression,of which 902 genes were significantly up-regulated and 740 genes were significantly down-regulated;375 lnc RNAs were significantly differentially expressed,of which 120 were significantly up-regulated and 256 were down-regulated.Through GO and KEGG enrichment analysis,it was found that a large number of potential target genes that differentially express lnc RNA are widely involved in various signal pathways in plants.These results indicate that lnc RNA may participate in the plant-virus interaction process through affecting various signaling pathways in plants.(2)Identification and cloning of differentially expressed lnc RNAs using RT-PCR technology to obtain 17 lnc RNAs.Spatiotemporal expression analysis of nine lnc RNAs screened by q RT-PCR and Northern-blotting techniques showed that the expression of lnc RNA in plants was tissue-specific.Further analysis revealed that systemic silencing of Nblnc61988,Nblnc13750 and Nblnc91573 promoted CWMV infection,while Nblnc81288 promoted CWMV infection.It showed that different lnc RNAs play different roles in the process of virus inflection.(3)It was proved that Nblnc81288 is the precursor of Nta-mi RNA6151 c.Infection of CWMV can induce the expression of Nblnc81288,so that Nta-mi RNA6151 c accumulates in plants.Further analysis found that Nta-mi RNA6151 c can regulate the expression of the target gene protein-tyrosine-phosphatase IBR5-like(IBR5)at the transcription level,thereby interfering with the SA-mediated disease resistance pathway of the host and promoting CWMV infection.This may be a new mechanism of CWMV pathogenicity,however,specific mechanism of action will need further research.