title: Nonstructural protein Pns4 of rice dwarf virus is essential for viral infection in its insect vector.
abstract: BACKGROUND
  Rice dwarf virus (RDV), a plant reovirus, is mainly transmitted by the green rice leafhopper, Nephotettix cincticeps, in a persistent-propagative manner. Plant reoviruses are thought to replicate and assemble within cytoplasmic structures called viroplasms. Nonstructural protein Pns4 of RDV, a phosphoprotein, is localized around the viroplasm matrix and forms minitubules in insect vector cells. However, the functional role of Pns4 minitubules during viral infection in insect vector is still unknown yet.
  
  
  METHODS
  RNA interference (RNAi) system targeting Pns4 gene of RDV was conducted. Double-stranded RNA (dsRNA) specific for Pns4 gene was synthesized in vitro, and introduced into cultured leafhopper cells by transfection or into insect body by microinjection. The effects of the knockdown of Pns4 expression due to RNAi induced by synthesized dsRNA from Pns4 gene on viral replication and spread in cultured cells and insect vector were analyzed using immunofluorescence, western blotting or RT-PCR assays.
  
  
  RESULTS
  In cultured leafhopper cells, the knockdown of Pns4 expression due to RNAi induced by synthesized dsRNA from Pns4 gene strongly inhibited the formation of minitubules, preventing the accumulation of viroplasms and efficient viral infection in insect vector cells. RNAi induced by microinjection of dsRNA from Pns4 gene significantly reduced the viruliferous rate of N. cincticeps. Furthermore, it also strongly inhibited the formation of minitubules and viroplasms, preventing efficient viral spread from the initially infected site in the filter chamber of intact insect vector.
  
  
  CONCLUSIONS
  Pns4 of RDV is essential for viral infection and replication in insect vector. It may directly participate in the functional role of viroplasm for viral replication and assembly of progeny virions during viral infection in leafhopper vector.
authors.affiliation: Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, PR China. chenqian_07@126.com.
authors.forename: Qian
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, PR China. 18659167408@163.com.
authors.forename: Linghua
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, PR China. chy9948@163.com.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Chen
authors.affiliation: Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, PR China. xielh@126.com.
authors.forename: Lianhui
authors.initials: L
authors.lastname: Xie
authors.affiliation: Fujian Province Key Laboratory of Plant Virology, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, PR China. weitaiyun@fafu.edu.cn.
authors.forename: Taiyun
authors.initials: T
authors.lastname: Wei
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques; D006430:Hemiptera / Q000821:virology*; D007303:Insect Vectors / Q000821:virology*; D012087:Reoviridae / Q000235:genetics / Q000502:physiology*; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D017361:Viral Nonstructural Proteins