An aspartic protease 47 causes quantitative recessive resistance to rice black-streaked dwarf virus disease and southern rice black-streaked dwarf virus disease.

Rice black-streaked dwarf virus disease (RBSDVD) and southern rice black-streaked dwarf virus disease (SRBSDVD) are the most destructive viral diseases in rice. Progress is limited in breeding due to lack of resistance resource and inadequate knowledge on the underlying functional gene. Using genome-wide association study (GWAS), linkage disequilibrium (LD) decay analyses, RNA-sequencing, and genome editing, we identified a highly RBSDVD-resistant variety and its first functional gene. A highly RBSDVD-resistant variety W44 was identified through extensive evaluation of a diverse international rice panel. Seventeen quantitative trait loci (QTLs) were identified among which qRBSDV6-1 had the largest phenotypic effect. It was finely mapped to a 0.8-1.2 Mb region on chromosome 6, with 62 annotated genes. Analysis of the candidate genes underlying qRBSDV6-1 showed high expression of aspartic proteinase 47 (OsAP47) in a susceptible variety, W122, and a low resistance variety, W44. OsAP47 overexpressing lines exhibited significantly reduced resistance, while the knockout mutants exhibited significantly reduced SRBSDVD and RBSDVD severity. Furthermore, the resistant allele Hap1 of OsAP47 is almost exclusive to Indica, but rare in Japonica. Results suggest that OsAP47 knockout by editing is effective for improving RBSDVD and SRBSDVD resistance. This study provides genetic information for breeding resistant cultivars.

Sublethal concentrations of pymetrozine reduce Sogatella furcifera transmission of Southern rice black-streaked dwarf virus.

BACKGROUND: The transmission of plant viruses is closely associated with the specific probing behaviors of the vectors. Pymetrozine is a pyridine azomethine insecticide that interferes with nervous regulation of feeding behavior of piercing-sucking insects. This study aimed to evaluate the potential of sublethal concentrations of pymetrozine in reducing the transmission of Southern rice black-streaked dwarf virus (SRBSDV) by the planthopper Sogatella furcifera. RESULTS: Laboratory assays showed that both acquisition and inoculation rates of SRBSDV decreased significantly in the planthoppers feeding on plants treated with lethal concentrations 10% and 50% (LC10 and LC50 ) pymetrozine compared with the insects feeding on the control plants, for which significant effects of pymetrozine concentration and time post-treatment were detected. Honeydew excretion of the planthoppers showed significant reduction with increasing concentration of the insecticide but no significant association with time post-treatment. Electrical penetration graph recordings revealed that total durations of each waveform in both acquisition and inoculation were significantly affected by pymetrozine treatment, with total durations of non-probing (NP), penetration initiation (N1), and extracellular activity (N3) elongated whereas those of salivation (N2) and phloem-related activities (N4-a and N4-b) shortened. Additionally, both acquisition and inoculation rates were significantly lower at 168 h than at 6 h post-treatment. CONCLUSION: The results indicate that sublethal concentrations of pymetrozine reduce SRBSDV transmission, which is associated with reduction in feeding and alteration in probing behaviors characterized by the prolonged non-probing, penetration initiation and extracellular activity and shortened salivation and phloem-related activities. © 2023 Society of Chemical Industry.

Rice varietal resistance to the vector Sogatella furcifera hinders transmission of Southern rice black-streaked dwarf virus.

BACKGROUND: The Southern rice black-streaked dwarf virus (SRBSDV) transmitted by Sogatella furcifera constitutes a threat to sustainable rice production. However, most rice varieties are highly vulnerable to SRBSDV, whereas the occurrence of the viral disease varies significantly under field conditions. This study aimed to evaluate the potential of rice varietal resistance to S. furcifera in reducing SRBSDV transmission. RESULTS: Among the five rice varieties, Zhongzheyou8 and Deyou108 exhibited high resistance to S. furcifera, Baixiangnuo33 was susceptible, and TN1 and Diantun502 were highly susceptible. The S. furcifera generally showed non-preference for and low feeding on the Zhongzheyou8 and Deyou108 plants, which may explain the resistance of these varieties to S. furcifera. Transmission of SRBSDV by S. furcifera was significantly impaired on the resistant varieties, both inoculation and acquisition rates were much lower on Zhongzheyou8 than on TN1. The short durations of S. furcifera salivation and phloem-related activities and the low S. furcifera feeding amount may explain the reduced SRBSDV inoculation and acquisition rates associated with Zhongzheyou8. Spearman's rank correlation revealed a significant negative correlation between S. furcifera resistance and SRBSDV transmission among the tested varieties. CONCLUSION: The results indicate that rice varietal resistance to the vector S. furcifera hinders SRBSDV transmission, which is largely associated with the host plant selection and feeding behaviors of the vector. The current findings shed light on the management of the SRBSDV viral disease through incorporation of S. furcifera resistant rice varieties in the management protocol. © 2024 Society of Chemical Industry.

A systematic review of southern rice black-streaked dwarf virus in the age of omics.

Southern rice black-streaked dwarf virus (SRBSDV) is one of the most damaging rice viruses. The virus decreases rice quality and yield, and poses a serious threat to food security. From this perspective, this review performed a survey of published studies in recent years to understand the current status of SRBSDV and white-backed planthopper (WBPH, Sogatella furcifera) transmission processes in rice. Recent studies have shown that the interactions between viral virulence proteins and rice susceptibility factors shape the transmission of SRBSDV. Moreover, the transmission of SRBSDV is influenced by the interactions between viral virulence proteins and S. furcifera susceptibility factors. This review focused on the molecular mechanisms of key genes or proteins associated with SRBSDV infection in rice via the S. furcifera vector, and the host defense response mechanisms against viral infection. A sustainable control strategy using RNAi was summarized to address this pest. Finally, we also present a model for screening anti-SRBSDV inhibitors using viral proteins as targets. © 2023 Society of Chemical Industry.

Resistance Evaluation of Dominant Varieties against Southern Rice Black-Streaked Dwarf Virus in Southern China.

Southern rice black-streaked dwarf virus (SRBSDV), a Fijivirus in the Reoviridae family, is transmitted by the white-backed planthopper (Sogatella furcifera, WBPH), a long-distance migratory insect, and presents a serious threat to rice production in Asia. It was first discovered in China's Guangdong Province in 2001 and has been endemic in the south of China and north of Vietnam for two decades, with serious outbreaks in 2009, 2010, and 2017. In this study, we evaluated the resistance of 10 dominant rice varieties from southern China, where the virus overwinters and accumulates as a source of early spring reinfection, against this virus by artificial inoculation. The results showed that in all tested varieties there was no immune resistance, but there were differences in the infection rate, with incidence rates from 21% to 90.7%, and in symptom severity, with plant weight loss from 66.71% to 91.20% and height loss from 34.1% to 65.06%. Additionally, and valuably, the virus titer and the insect vector virus acquisition potency from diseased plants were significantly different among the varieties: an over sixfold difference was determined between resistant and susceptible varieties, and there was a positive correlation between virus accumulation and insect vector virus acquisition. The results can provide a basis for the selection of rice varieties in southern China to reduce the damage of SRBSDV in this area and to minimize the reinfection source and epidemics of the virus in other rice-growing areas.

Southern rice black-streaked dwarf virus hijacks SNARE complex of its insect vector for its effective transmission to rice.

Vesicular trafficking is an important dynamic process that facilitates intracellular transport of biological macromolecules and their release into the extracellular environment. However, little is known about whether or how plant viruses utilize intracellular vesicles to their advantage. Here, we report that southern rice black-streaked dwarf virus (SRBSDV) enters intracellular vesicles in epithelial cells of its insect vector by engaging VAMP7 and Vti1a proteins in the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex. The major outer capsid protein P10 of SRBSDV was shown to interact with VAMP7 and Vti1a of the white-backed planthopper and promote the fusion of vesicles into a large vesicle, which finally fused with the plasma membrane to release virions from midgut epithelial cells. Downregulation of the expression of either VAMP7 or Vti1a did not affect viral entry and accumulation in the gut, but significantly reduced viral accumulation in the haemolymph. It also did not affect virus acquisition, but significantly reduced the virus transmission efficiency to rice. Our data reveal a critical mechanism by which a plant reovirus hijacks the vesicle transport system to overcome the midgut escape barrier in vector insects and provide new insights into the role of the SNARE complex in viral transmission and the potential for developing novel strategies of viral disease control.

Silicon amendment to rice plants reduces the transmission of southern rice black-streaked dwarf virus by Sogatella furcifera.

BACKGROUND: Plant viruses are transmitted mainly by piercing-sucking herbivores, and viral disease management relies on chemical control of vectors. Southern rice black-streaked dwarf virus (SRBSDV) is transmitted by the white-backed planthopper (WBPH), Sogatella furcifera. This study aimed to evaluate the potential of silicon (Si) amendment for reducing SRBSDV transmission. RESULTS: The settling and ovipositional preferences of WBPH females decreased significantly by 14.6-43.7% for plants treated with either 0.16 g or 0.32 g SiO2  kg-1 soil during SRBSDV acquisition and by 26.2-28.3% for plants treated with 0.32 g SiO2  kg-1 soil during SRBSDV inoculation, compared with controls. Adding either 0.16 or 0.32 g SiO2  kg-1 soil significantly reduced SRBSDV inoculation rate by 31.3% and 45.3%, respectively, and acquisition rate by 25.5% and 66.0%, respectively. Silicification was intensified more in plants treated with 0.32 g SiO2  kg-1 soil than in controls. The nonprobing (np) duration increased, and the phloem sap ingestion (N4-b) duration decreased significantly in the WBPHs feeding on high-rate-Si-supplemented plants compared with control plants during both inoculation and acquisition access. CONCLUSION: This study showed that Si amendment to rice plants decreased the WBPH settling and ovipositional preference and the SRBSDV acquisition and inoculation rates, thereby reducing SRBSDV transmission. The intensified plant silicification and the altered WBPH feeding behaviors (i.e. prolonged np and shortened N4-b) may explain the reduced SRBSDV transmission in Si-amended plants. © 2021 Society of Chemical Industry.

Impact of Two Reoviruses and Their Coinfection on the Rice RNAi System and vsiRNA Production.

Both Southern rice black-streaked dwarf virus (SRBSDV) and Rice ragged stunt virus (RRSV) belong to the family Reoviridae, and synergistic infection of these two viruses commonly occurs in the field. This study revealed that both SRBSDV and RRSV affect the RNA interference (RNAi) pathway and form different virus-derived interfering RNA (vsiRNA) profiles in rice. Co-infection of rice by SRBSDV and RRSV up-regulated the expression of rice DICER-like (DCL) proteins but down-regulated the expression of rice RNA-dependent RNA polymerases (RDRs), and the accumulation of vsiRNAs of either RBSDV or RRSV was decreased compared with that in singly infected plants. The majority of SRBSDV vsiRNAs were 21 nt or 22 nt in length, whether plants were singly infected with SRBSDV or co-infected with RRSV. On the other hand, the majority of RRSV vsiRNAs were 20 nt, 21 nt, or 22 nt in length, among which those 20 nt in length accounted for the largest proportion; co-infection with SRBSDV further increased the proportion of 20 nt vsiRNAs and decreased the proportion of 21 nt vsiRNAs. Co-infection had no effects on the strand favoritism and hot spots of the vsiRNAs, but changed the bias of the 5' terminal nucleotide significantly. This study provides a reference for further study on the pathogenesis and synergistic mechanism of SRBSDV and RRSV.

Metabolomic Changes in Sogatella furcifera under Southern rice black-streaked dwarf virus Infection and Temperature Stress.

Southern rice black-streaked dwarf virus (SRBSDV) is a devastating newly emerged rice reovirus in Eastern and Southeastern Asia transmitted by a long-distance migratory pest, the white-backed planthopper (WBPH). We previously showed that SRBSDV infection decreased the cold tolerance but improved the heat tolerance of its vector, WBPH. Comparative metabolomic analysis was used to explore the potential mechanisms underlying these changes in temperature stress response. Fourth-generation WBPH nymphs were treated with SRBSDV and/or extreme temperature stress and were analyzed using gas chromatography-time of flight-mass spectrometry. A total of 605 distinguishable peaks were identified and 165, 207, and 202 differentially accumulated metabolites were identified in WBPH after virus infection, cold, or heat stress, respectively. The nucleic acids and fatty acids were the major categories of metabolites regulated by SRBSDV infection, whereas temperature stress regulated tricarboxylic acid cycle compounds, sugars, and polyols. For the WBPH samples infected with SRBSDV and subjected to temperature stress, amino acids, sugars, and polyols were the most significant regulated metabolites. The metabolomics study suggests that SRBSDV may influence the extreme temperature tolerance of WBPH by regulating the accumulation of amino acids, sugars, and polyols in the insect body.

Proteomic analysis of interaction between P7-1 of Southern rice black-streaked dwarf virus and the insect vector reveals diverse insect proteins involved in successful transmission.

Southern rice black-streaked dwarf virus (SRBSDV), transmitted by the white-backed planthopper (Sogatella furcifera) in a persistent-propagative manner, has caused serious yield losses in Asia. Here in a yeast two-hybrid system, protein interactions between SRBSDV P7-1 as a bait protein and a cDNA library of S. furcifera as prey protein were assessed. Of 153 proteins identified as putative interactors, 24 were selected for further analysis. Of the 24 proteins, 18 were further confirmed in a chemiluminescent coimmunoprecipitation (Co-IP) assay as true positive interactors with different strengths of interactions. Six potential candidate proteins (neuroglian, myosin light chain 2 [MLC2], polyubiquitin, E3 ubiquitin ligase, ribophorin ii, and profilin) were analyzed for gene expression in five organs by qRT-PCR; mRNA levels were highest in the gut for neuroglian, MLC2, polyubiquitin and profilin, in the salivary glands for ribophorin ii, and in the haemolymph for E3 ubiquitin ligase. A virus-host protein interaction network was constructed using SRBSDV P7-1 and 18 prey positive protein homologs of Drosophila melanogaster. Our findings suggest that these proteins are involved in the complex host reaction to infection by SRBSDV and provide new insights into the molecular basis of transmission. BIOLOGICAL SIGNIFICANCE: Southern rice black-streaked dwarf virus (SRBSDV), transmitted by S. furcifera in a persistent-propagative manner, is a new found virus and a tentative member of the genus Fijivirus in the family Reoviridae. It was widely noted by plant virologist, government officials and the farmers in Asia in recent years because of its epidemic outbreak and causing serious yield losses after 2009. However, the molecular mechanism by which SRBSDV successfully infects and replicates in both plant and insect hosts remains unclear, and much less is known about how the virus spreads from initially infected cells to adjacent cells in the insect vector. In the present study, we examined protein interactions between SRBSDV P7-1 as the bait and cDNA library of WBPH as the prey by using yeast two-hybrid system, 153 proteins were identified as putative interactors and 24 putative proteins were selected for chemiluminescent coimmunoprecipitation (Co-IP) assay, and then constructed a viral protein-host protein interaction network with homologs of D. melanogaster. Six WBPH proteins were confirmed as potential P7-1 partners that take part in a pivotal role for viral movement in insect vector. These findings will greatly facilitate the understanding of the transmission mechanisms of SRBSDV by its insect vector. This is the first to study the molecular interaction between SRBSDV and its insect vector.