RNAi silencing of rice black-streaked dwarf virus P10 and two insect vector genes to reduce virus transmission protects rice plants against RBSDV

Abstract

Rice black-streaked dwarf virus (RBSDV) is a virus transmitted by Laodelphax striatellus (L. striatellus), which poses a serious threat to rice production. P10, a major outer capsid protein of RBSDV, which plays a key role in viral infection. In this study, two L. striatellus transmission-related proteins, novel cuticular protein (NCuP) and receptor for activated protein kinase C (RACK), were identified as interacting with P10. Results demonstrated that virus-induced NCuP and RACK were silenced using double-stranded RNA (dsRNA) to reduce the quantity of virus in L. striatellus. The detection results after inoculation of RBSDV by L. striatellus showed that rice plants sprayed with dsP10 +NCuP and dsP10+RACK and transgenic rice plants expressing both had higher disease resistance than those expressing dsP10, dsNCuP and dsRACK alone. Therefore, simultaneously interfering with both the expression of viral protein and the proteins that interact with them from insect vectors could enhance the control of viral transmission.

Silencing suppressors of rice black‐streaked dwarf virus and rice stripe virus hijack the 26S proteasome of Laodelphax striatellus to facilitate virus accumulation and transmission

“…1(D)). To test the specificity of interactions, the receptor for activated protein kinase C (RACK) of L. striatellus 41 was used. Immunoprecipitation with anti‐GFP antibodies resulted in enrichment of P9‐1–GFP.…”

“…[52][53][54][55] Recent studies have shown that transgenic plants can be engineered to express hairpin dsRNAs targeting genes from insects to increase their resistance to herbivorous insects. 41,56,57 In this work, we used the same strategy to obtain transgenic lines of L. striatellus RPN8 RNAi to investigate its role in viral infection. Although the present results are reliable, the currently available research strategies are relatively simple and inefficient, mostly involving unilateral interference with the expression of host or vector genes to reveal the function of target genes.…”

“…Given that dsRNA is easily expressed in plants and insects, this technique can be applied to the study of functional genomes of plants and insects to silence the expression of specific endogenous genes and genes of invading pathogens 52–55 . Recent studies have shown that transgenic plants can be engineered to express hairpin dsRNAs targeting genes from insects to increase their resistance to herbivorous insects 41,56,57 . In this work, we used the same strategy to obtain transgenic lines of L. striatellus RPN8 RNAi to investigate its role in viral infection.…”

“…Laodelphax striatellus transmission assay were performed as previously described 41 . Two‐week‐old WT and transgenic rice seedlings were placed in single glass jars of 10.5 cm in diameter and 16 cm in height, each with 4 cm of nutrient soil at the bottom.…”

RNA Interference-Based Pesticides and Antiviral Agents: Microbial Overproduction Systems for Double-Stranded RNA for Applications in Agriculture and Aquaculture

“…In addition, although it was suggested that lacY mutation allows uniform entry of IPTG into all cells in the population of the culture and produces a homologous induction level of gene expression by the inducer, the involvement of the lacY mutation in improving dsRNA productivity in the strain is also not certain. However, there have been some reports on successful dsRNA production using M-JM109lacY as the host strain [38][39][40][41][42]. Recently, Ma et al [43] attempted to construct a production system that is more efficient than the conventional system for dsRNA production, L4440-HT115(DE3).…”