关键词:
antibacterial
Nephotettix cincticeps
ovary
peptidoglycan recognition protein (PGRP)
proline-rich protein (prp)
rice dwarf virus (RDV)
symbiotic bacterium
transovarial transmission
摘要:
【Objective】Rice dwarf virus (RDV) causes rice dwarf disease, which is a significant threat to rice production in southern China. This study builds on prior research and employs molecular biology techniques to investigate the role of PGRP6 in regulating the vertical transmission of RDV from symbiotic bacteria within the vector insect Nephotettix cincticeps, thereby establishing a foundation for biological control strategies against rice viral diseases.【Method】Yeast two-hybrid technology was used to screen peptidoglycan recognition proteins (PGRPs) that interact with three proteins (prp, Nasuia porin, and P8) related to transovarial transmission, and the interaction was further confirmed by GST pull-down assay. Real-time fluorescence quantitative PCR technology was employed to assess the differential expression of related genes in nonviruliferous and viruliferous insects, as well as the changes in transcription levels of symbiotic bacteria following interference with PGRP6 and prp. Western blot experiments further confirmed the impact of RNAi treatment on symbiotic bacterial membrane proteins in insects. Following the reduction of PGRP6 expression by RNAi, variations were observed in the survival rate of N. cincticeps, the virus infection rate of the offspring, and the viral nucleic acid level of the infected offspring. Fluorescence in situ hybridization (FISH) and fluorescent immunolabeling techniques were employed to visualize the distribution of prp, PGRP6, Nasuia, and RDV. The functionality of the PGRP6 protein was assessed through inhibition zone assays and experiments on peptidoglycan degradation.【Result】PGRP6 interacted with prp, Nasuia porin, and P8. The expression levels of PGRP6 and prp were significantly elevated in RDV-infected N. cincticeps. Suppression of PGRP6 expression could facilitate the proliferation of symbiotic bacteria Nasuia and Sulcia, while inhibition of prp reduced the proliferation of symbiotic bacteria, acting oppositely to PGRP6. When bot