Characterization of major midgut proteinase cDNAs from Helicoverpa armigera larvae and changes in gene expression in response to four proteinase inhibitors in the diet

A Helicoverpa armigera larval midgut cDNA library from larvae raised on an artificial, protein-rich, inhibitor-free diet contained very large numbers of serine proteinase positive clones. DNA sequencing of six random positive cDNAs and 12 PCR derived products identified trypsin genes classifiable into three families, and chymotrypsin and elastase genes classifiable into a single family each. Genomic blots established that the most highly expressed of the trypsin families contained about 18 genes, and that the chymotrypsin and elastase families contained about 14 and 2 genes respectively. The levels of mRNA corresponding to the highly expressed trypsin and chymotrypsin families were determined following chronic ingestion of four proteinase inhibitors. Compared to insects on an inhibitor-free diet, chymotrypsin mRNA was increased by all inhibitors, and trypsin mRNA levels decreased. This occurred independent of whether the inhibitor was solely a trypsin inhibitor (aprotinin), an inhibitor of both trypsin and chymotrypsin (proteinase inhibitor II, soybean trypsin inhibitor) or predominantly a chymotrypsin inhibitor (proteinase inhibitor I). Changing the protein level of the diet did not affect trypsin mRNA levels, but chymotrypsin mRNA levels decreased with increasing dietary protein.