title: Inoculation of Nicotiana tabacum with recombinant potato virus X induces RNA interference in the solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae).
abstract: OBJECTIVE
  The chitin synthase 1 (CHS1) gene in Phenacoccus solenopsis (PsCHS1) was evaluated as a potential target of RNA interference (RNAi) by using Potato virus X (PVX) as a vector (recombinant PVX) for expressing RNAi triggering elements in Nicotiana tabacum L.
  
  
  RESULTS
  RT-PCR analysis confirmed the expression of PsCHS1 in N. tabacum inoculated with recombinant-PVX-PsCHS1 (treated). RT- and multiplex-PCR further showed a reduction in mRNA levels of the target gene in mealybugs feeding on treated plants. Mortality in parent adults and emerging nymphs (21 and 29%) exposed to the treated plants was significantly higher (P < 0.05) than those exposed to uninoculated (-ve control) or inoculated with non-recombinant PVX (PVX-control). The number of surviving adults and the combined number of adults and nymphs (47 and 60%) was significantly (P < 0.05) lower on the treated plants than the -ve (76%) or PVX (74%) control. The visual observations verified the physical deformities in mealybugs exposed to the treated plants.
  
  
  CONCLUSIONS
  chitin synthase 1 is a potential RNAi target in P. solenopsis and the recombinant PVX can be used as a tool to evaluate candidate RNAi triggering elements in plants.
authors.affiliation: National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan.|Department of Biotechnology, University of Sargodha, Sargodha, Pakistan.
authors.forename: Arif Muhammad
authors.initials: AM
authors.lastname: Khan
authors.affiliation: National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan. mashfaq@uoguelph.ca.|Biodiversity Institute of Ontario, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada. mashfaq@uoguelph.ca.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Ashfaq
authors.affiliation: College of Agriculture, Bahauddin Zakariya University, Bahadur Campus Layyah, Multan, Pakistan.
authors.forename: Azhar Abbas
authors.initials: AA
authors.lastname: Khan
authors.affiliation: National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan.
authors.forename: Akhtar
authors.initials: A
authors.lastname: Rasool
authors.affiliation: School of Life Sciences, Beijing Institute of Technology, Beijing, People's Republic of China.
authors.forename: Javed
authors.initials: J
authors.lastname: Iqbal
authors.affiliation: National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan.
authors.forename: Shahid
authors.initials: S
authors.lastname: Mansoor
mesh_terms: D000818:Animals; D002688:Chitinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006430:Hemiptera / Q000201:enzymology* / Q000502:physiology*; D060885:Multiplex Polymerase Chain Reaction; D009758:Nymph / Q000502:physiology; D010944:Plants; D017863:Potexvirus / Q000235:genetics / Q000254:growth & development*; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016019:Survival Analysis; D014026:Nicotiana / Q000469:parasitology* / Q000821:virology*
chemical_list: D002688:Chitinases
keywords: Mealy bug; Phenacoccus solenopsis; Potato virus X; RNA interference; Recombinant virus; Solenopsis mealybug