Identification of allatostatins in the brown-winged green bug Plautia stali

Juvenile hormone (JH) biosynthesis is inhibited under short-day conditions in the brown-winged green bug Plautia stali. We investigated allatostatic molecules in the brain of P. stali. Methanol brain extracts strongly inhibited JH biosynthesis. The allatostatic activities of the brain extracts were heat stable but gently suppressed by trypsin treatment, indicating that the allatostatic molecules were peptides. Grybi-MIP1, found in Gryllus bimaculatus as an allatostatic molecule, inhibited JH biosynthesis in P. stali. In contrast, peptides such as Dippu-AST2, 8, and 9, found in Diploptera punctata, did not affect JH biosynthesis in P. stali. We found a cDNA sequence encoding a peptide precursor of myoinhibitory peptides (MIPs), which we named Plast-MIP. Three synthetic peptides, AWKDLSKAW-NH₂ (Plast-MIP1), GWSDLQSAGW-NH₂ (Plast-MIP5), and AADWGSFRGSW-NH₂ (Plast-MIP8), deduced from the precursor sequence, showed clear inhibition of JH biosynthesis in P. stali. Analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and tandem mass spectrometry showed that Plast-MIP8 resides in the brain. Expression of the Plast-MIP mRNA precursor was detected in the brain of insects reared under short- and long-day conditions. These results suggest that Plast-MIP is an allatostatic molecule and that MIPs are synthesized irrespective of photoperiod. To our knowledge, this is the first study to identify Plast-MIP as a functional allatostatin in hemipteran insects.