Analysis of the Effect of HSVd Targeting ds-sRNA on miRNA Expression in HSVd-Infected Tomato
Exogenous ds-sRNA sequences consistent with the viroid genome can silence the viroid genome in a short time,and miRNAs are widely involved in gene expression regulation,playing an important role in this biological process.In order to explore the core miRNAs involved in the silencing of viroid genomes by exogenous ds-sRNA,ds-sRNA targeting different regions of the genome were applied exogenously to tomato plants infected with hop stun virus(HSVd),and the whole genome titer of the viroid was measured at different periods after application.It was determined that the application of ds-sRNA targeting the central conserved region of HSVd for 36 h had the best silencing efficiency for HSVd.Based on optimal processing,miRNA-seq,degradome sequencing,and quantitative real-time PCR(qRT-PCR)validation of key miRNAs were performed.Compared with the control,significant changes in the expression levels of sly-MIR10528-p3,which is involved in regulating plant resistance,and PTC-MiR162a_R+1,which is involved in regulating gene silencing,were detected by miRNA-seq.The expression trend of the target miRNA detected by qRT-PCR was consistent with the sequencing data.In the degradome sequencing,the expression trends of sly-MIR10528-p3 and PTC-MiR162a_R+1 were consistent with the miRNA-seq results.In addition,significant changes in the expression levels of sly-miR162,which participates in regulating virus defense,vvi-miR3630-3pL_2R-1ss21AT,which regulates abiotic stress resistance,and bna-MIR169c-p3_2ss12GC17TG,which participates in regulating gene silencing,were detected.This test also obtained 52 significantly differentially expressed new miRNAs.This study can provide a new theoretical basis and technical support for tomato disease resistance breeding and molecular biocide development.
万方数据
维普
