| Potato virus Y(PVY)is a virus that severely damages solanaceous crops.Revealing the interaction between PVY and the host will provide a theoretical basis for studying the pathogenic mechanism and effective control of PVY.In the previous work,this research group isolated a strain that had passivation effect on PVY from tobacco rhizosphere soil in Shanxi province,and identified it as Serratia marcescen-S3 by 16 S r RNA and Biolog Micro Plate.Nb Hsc70-2 protein was screened by combined proteomics and ubiquitinomics analysis,and it is speculated that the ubiquitination of this protein may play an important role in the mechanism of S3 antiviral-induced resistance.This study hopes to pass a variety of organisms Chemical and molecular biological methods have further revealed the mechanism of the ubiquitination modification of host Nb Hsc70-2 protein in the resistance of Serratia marcescens S3.The main results are as follows:(1)Serratia marcescens S3 is a biologically active strain that inhibits PVY up to 100%.The target response time of S3 to prevent PVY was selected using S3 strain treated with Nicotiana benthamiana.Through a combination of non-standard quantitative technology,high-performance liquid chromatography classification technology,ubiquitinated peptide enrichment technology,and mass spectrometry-based quantitative proteomics technology,a combination of leading-edge technologies for quantitative proteome and The ubiquitinated non-standard quantification omics was studied,and the combined analysis of the proteomics and ubiquitination groups revealed that the protein level of Nb Hsc70-2 was significantly reduced after S3 treatment,whereas the ubiquitination level of this protein was significantly increased.It is suggested that the ubiquitination of Nb Hsc70-2 plays an important role in the mechanism of S3 induced resistance.Western blot technology was used to verify the authenticity of the omics data,and the results showed that the overall ubiquitination level of N.benthamiana was increased after treatment with S3,and the Nb Hsc70-2 protein level in the host was decreased.This is consistent with the results of omics analysis,indicating that the omics The data are credible,and S3 treated plants inhibited Nb Hsc70-2 protein expression in the host.(2)The effects of PVY infection on Nb Hsc70-2 were analyzed by q RT-PCR,Western blot and other techniques.The results showed that PVY infection caused up-regulation of Nb Hsc70-2 m RNA and protein levels in the host.Further use of virus-induced gene silencing(VIGS),transient overexpression and other methods proved that Nb Hsc70-2 plays an important role in plant growth and development,and Nb Hsc70-2 plays an important role in PVY infection and replication effect.(3)Utilizing the ubiquitination inhibitor MG-132 to inhibit the ubiquitination modification in the host,and exploring the role of ubiquitination in the mechanism of S3 antiviral disease,the results show that S3 induces resistance by inhibiting the Nb Hsc70-2 protein Expression to achieve antiviral effects.Inhibition of host ubiquitination modification level inhibits Nb Hsc70-2-mediated S3 antiviral resistance,indicating that the ubiquitination modification of Nb Hsc70-2 may play an important role in the mechanism of S3 antiviral resistance.Based on the above results,it is concluded that after PVY infects N.benthamiana by recruiting host factor Nb Hsc70-2 to promote its own replication and infection,S3 treatment can inhibit the expression of Nb Hsc70-2 in this process and induce resistance in S3 The down-regulation of Nb Hsc70-2 in the mechanism may be due to the increased ubiquitination level of Nb Hsc70-2 after S3 treatment,and the plants acquire antiviral resistance. |
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