Putative protein digestion in a sap-sucking homopteran plant pest (rice brown plant hopper; Nilaparvata lugens: Delphacidae)--identification of trypsin-like and cathepsin B-like proteases.
Sap-sucking phytophagous insect species of the order Hemiptera have been assumed not to carry out digestive proteolysis, but instead to rely on free amino acids in the phloem and xylem saps for their nutritional requirements. Extracts prepared from isolated guts of rice brown planthopper (Nilaparvata lugens), a homopteran crop pest, were shown to contain protease activity, with hydrolysis of both protein and synthetic peptide substrates being observed. Assays with specific inhibitors suggested that a trypsin-like serine protease was responsible for most of hydrolytic activity against synthetic substrates. A cDNA library was prepared from RNA extracted from N. lugens gut tissue, and screened for protease-encoding sequences. cDNAs for a cathepsin B-like protease and a trypsin-like protease were isolated and fully characterised; the latter exhibits a novel C-terminal region and an unusual activation mechanism, and represents a small gene family. Soya bean Kunitz trypsin inhibitor (SKTI) is an effective inhibitor of protein hydrolysis by N. lugens gut extracts in vitro, explaining why transgenic rice plants expressing this protein are partially resistant to the insect (Mol. Breed. 5 (1999) 1). It is suggested that digestive proteolysis may be widespread in sap-sucking homoptera, and can make a significant contribution to nutrition.
Foissac
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X
Crop Protection Group, Department of Biological Sciences, University of Durham, South Road, Durham DH1 3LE, UK.
Edwards
M G
MG
Du
J P
JP
Gatehouse
A M R
AM
Gatehouse
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002401:Cathepsin B / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary; D004064:Digestive System; D006430:Hemiptera / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D014020:Tissue Extracts; D014360:Trypsin Inhibitor, Kunitz Soybean / Q000494:pharmacology; D014361:Trypsin Inhibitors; D000516:alpha-Amylases / Q000037:antagonists & inhibitors
D018076:DNA, Complementary; D010940:Plant Proteins; D014020:Tissue Extracts; D014361:Trypsin Inhibitors; C114880:trypsin-like serine protease; D014360:Trypsin Inhibitor, Kunitz Soybean; D000516:alpha-Amylases; D012697:Serine Endopeptidases; D002401:Cathepsin B