Abstract
Rice dwarf virus (RDV) replicates in and is transmitted by a leafhopper vector in a persistent-propagative manner. Previous cytopathologic and genetic data revealed that tubular structures, constructed by the nonstructural viral protein Pns10, contain viral particles and are directly involved in the intercellular spread of RDV among cultured leafhopper cells. Here, we demonstrated that RDV exploited these virus-containing tubules to move along actin-based microvilli of the epithelial cells and muscle fibers of visceral muscle tissues in the alimentary canal, facilitating the spread of virus in the body of its insect vector leafhoppers. In cultured leafhopper cells, the knockdown of Pns10 expression due to RNA interference (RNAi) induced by synthesized dsRNA from Pns10 gene strongly inhibited tubule formation and prevented the spread of virus among insect vector cells. RNAi induced after ingestion of dsRNA from Pns10 gene strongly inhibited formation of tubules, preventing intercellular spread and transmission of the virus by the leafhopper. All these results, for the first time, show that a persistent-propagative virus exploits virus-containing tubules composed of a nonstructural viral protein to traffic along actin-based cellular protrusions, facilitating the intercellular spread of the virus in the vector insect. The RNAi strategy and the insect vector cell culture provide useful tools to investigate the molecular mechanisms enabling efficient transmission of persistent-propagative plant viruses by vector insects.
Highlights
Numerous plant viruses that cause serious losses to agricultural production are transmitted by vector insects, classified according to the type of transmission: nonpersistent, semipersistent and persistent [1]
We took advantage of unique biological tools, including insect vector cell culture and RNA interference (RNAi) induced by synthesized dsRNA, to investigate the molecular mechanisms facilitating the efficient spread of Rice dwarf virus (RDV), a persistentpropagative plant virus, among cells and organs of leafhopper vector
Our experimental evidence shows that RDV exploited virus-containing tubules composed of nonstructural viral protein Pns10 to traffic along actinbased cellular machinery, allowing efficient cell-to-cell spread of the virus in leafhopper vector
Summary
Numerous plant viruses that cause serious losses to agricultural production are transmitted by vector insects, classified according to the type of transmission: nonpersistent, semipersistent and persistent [1]. Detailed analyses of viral propagation in the vector insects in persistent-propagative transmission would help disclose a mechanism that may lead to new strategies to control the transmission of the viruses by vector insects. Plant viruses such as tospoviruses, tenuiviruses, plant rhabdoviruses and plant reoviruses, are transmitted by their respective insect vectors in a persistent-propagative manner, and they are designated as persistent-propagative plant viruses [1,2,3,4]. Acquiring a better understanding of the intercellular spread of plant viruses in insects would lead to better strategies to disrupt the efficient transmission of plant viruses by insect vectors
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