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Publishing Language: Chinese

PGRP6 Negatively Regulates Symbiotic Bacteria to Prevent the Transovarial Transmission of RDV in Nephotettix cincticeps

YuanYuan XU, DongSheng JIA, Yu BIN, TaiYun WEI()
College of Plant Protection/Vector-Borne Virus Research Center, Fujian Agriculture and Forestry University/Fujian Province Key Laboratory of Plant Virology, Fuzhou 350002
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Abstract

【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 both were interfered simultaneously, there was no significant effect on the proliferation of symbiotic bacteria. Treatment with dsPGRP6 significantly decreased the survival rate of N. cincticeps and enhanced the transmission of RDV to their offspring, resulting in a substantially higher viral load in the progeny compared to the control group. Immunofluorescence analysis revealed that PGRP6 co-localized with prp, Nasuia, and RDV within ovarian tissue, where prp and Nasuia exhibited a wrapping relationship. Ultimately, PGPR6 was identified as possessing antibacterial properties that inhibited the proliferation of symbiotic bacteria.

【Conclusion】

PGRP6 maintains the homeostasis of ovarian tissue in N. cincticeps by inhibiting the proliferation of symbiotic bacteria, which functionally antagonizes prp. This mechanism not only ensures the vitality of N. cincticeps but also regulates the transovarial transmission of RDV associated with bacterial symbionts.

Keywords

rice dwarf virus (RDV)Nephotettix cincticepspeptidoglycan recognition protein (PGRP)proline-rich protein (prp)symbiotic bacteriumovarytransovarial transmissionantibacterial

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Scientia Agricultura Sinica
Volume 58 Issue 5,
March 2025
Pages 907-917
DOI: 10.3864/j.issn.0578-1752.2025.05.007
Cite this article:
XU Y, JIA D, BIN Y, et al. PGRP6 Negatively Regulates Symbiotic Bacteria to Prevent the Transovarial Transmission of RDV in Nephotettix cincticeps. Scientia Agricultura Sinica, 2025, 58(5): 907-917. https://doi.org/10.3864/j.issn.0578-1752.2025.05.007
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