Characterization of midgut trypsinogen-like cDNA and enzymatic activity in Plutella xylostella parasitized by Cotesia vestalis or Diadegma semiclausum.
Protein digestion in insects is a result of the action of a complex of proteinases present in the midgut. In this report we describe the cloning and sequencing of a trypsin cDNA from larvae of the lepidopteran herbivore Plutella xylostella. We investigated the expression of this gene and enzymatic activity of its translation product with N-a-benzoyl-l-arginine p-nitroanilide (BApNA) as substrate in P. xylostella larvae that were either unparasitized or parasitized by Cotesia vestalis or Diadegma semiclausum parasitoids. The full cDNA sequence consisted of an open reading frame (ORF) encoding 273 amino acid residues including 23 residues of a signal peptide, and the predicted mature trypsinogen-like enzyme had a molecular mass of 26.5 kDa. The amino acid sequence of this trypsinogen-like enzyme protein and phylogenetic relationship with other published trypsin enzyme proteins suggested that it may be a new proteinase in the trypsin protein family. Parasitism of D. semiclausum did not significantly change the mRNA transcript level or BApNAase activity in host larvae. By contrast, parasitization by C. vestalis induced higher transcript levels coupled with a higher level of BApNAase activity. The BApNAase activity in the midgut of nonparasitized or parasitized P. xylostella larvae increased to a maximum level at pH 12, and the parasitism by both C. vestalis and D. semiclausum increased sensitivity of the enzyme to pH values ranging from 2 to 9.5. These parasitoid-induced changes may represent host manipulation by the developing parasitoid larva.
Shi
Min
M
Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
Huang
Fang
F
Chen
Ya-Feng
YF
Meng
Xiang-Feng
XF
Chen
Xue-Xin
XX
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism*; D004064:Digestive System / Q000201:enzymology*; D005786:Gene Expression Regulation / Q000502:physiology; D006790:Host-Parasite Interactions; D006863:Hydrogen-Ion Concentration; D007814:Larva / Q000378:metabolism / Q000469:parasitology; D008969:Molecular Sequence Data; D009036:Moths / Q000469:parasitology*; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism; D014362:Trypsinogen / Q000378:metabolism*; D014863:Wasps / Q000502:physiology*
D018076:DNA, Complementary; D012333:RNA, Messenger; D014362:Trypsinogen; D014357:Trypsin