title: Cysteine protease enhances plant-mediated bollworm RNA interference.
abstract: Oral ingestion of plant-expressed double stranded RNA (dsRNA) triggers target gene suppression in insect. An important step of this process is the transmission of dsRNA from plant to midgut cells. Insect peritrophic matrix (PM) presents a barrier that prevents large molecules from entering midgut cells. Here, we show that uptake of plant cysteine proteases, such as GhCP1 from cotton (Gossypium hirsutum) and AtCP2 from Arabidopsis, by cotton bollworm (Helicoverpa armigera) larvae resulted in attenuating the PM. When GhCP1 or AtCP2 pre-fed larvae were transferred to gossypol-containing diet, the bollworm accumulated higher content of gossypol in midgut. Larvae previously ingested GhCP1 or AtCP2 were more susceptible to infection by Dendrolimus punctatus cytoplasmic polyhedrosis virus (DpCPV), a dsRNA virus. Furthermore, the pre-fed larvae exhibited enhanced RNAi effects after ingestion of the dsRNA-expressing plant. The bollworm P450 gene CYP6AE14 is involved in the larval tolerance to gossypol; cotton plants producing dsRNA of CYP6AE14 (dsCYP6AE14) were more resistant to bollworm feeding (Mao et al. in Transgenic Res 20:665-673, 2011). We found that cotton plants harboring both 35S:dsCYP6AE14 and 35S:GhCP1 were better protected from bollworm than either of the single-transgene lines. Our results demonstrate that plant cysteine proteases, which have the activity of increasing PM permeability, can be used to improve the plant-mediated RNAi against herbivorous insects.
authors: National Key Laboratory of Plant Molecular Genetics, National Plant Gene Research Center, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, People's Republic of China.
authors: Ying-Bo
authors: YB
authors: Mao
authors: Xue-Yi
authors: XY
authors: Xue
authors: Xiao-Yuan
authors: XY
authors: Tao
authors: Chang-Qing
authors: CQ
authors: Yang
authors: Ling-Jian
authors: LJ
authors: Wang
authors: Xiao-Ya
authors: XY
authors: Chen
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D002463:Cell Membrane Permeability; D057056:Cysteine Proteases / Q000235:genetics / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D003368:Gossypium / Q000201:enzymology* / Q000235:genetics / Q000821:virology; D006072:Gossypol / Q000378:metabolism / Q000494:pharmacology; D060434:Herbivory; D007814:Larva / Q000502:physiology / Q000821:virology; D009036:Moths / Q000502:physiology* / Q000821:virology; D018515:Plant Leaves / Q000201:enzymology / Q000502:physiology / Q000821:virology; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics / Q000821:virology; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D012087:Reoviridae / Q000235:genetics / Q000472:pathogenicity
chemical_list: D012330:RNA, Double-Stranded; D018749:RNA, Plant; D012367:RNA, Viral; D003577:Cytochrome P-450 Enzyme System; D057056:Cysteine Proteases; D006072:Gossypol