Neuropeptide and neurohormone precursors in the pea aphid, Acyrthosiphon pisum.

Aphids respond to environmental changes by developing alternative phenotypes with differing reproductive modes. Parthenogenetic reproduction occurs in spring and summer, whereas decreasing day lengths in autumn provoke the production of sexual forms. Changing environmental signals are relayed by brain neuroendocrine signals to the ovarioles. We combined bioinformatic analyses with brain peptidomics and cDNA analyses to establish a catalogue of pea aphid neuropeptides and neurohormones. 42 genes encoding neuropeptides and neurohormones were identified, of which several were supported by expressed sequence tags and/or peptide mass analyses. Interesting features of the pea aphid peptidome are the absence of genes coding for corazonin, vasopressin and sulfakinin and the presence of 10 different genes coding insulin related peptides, one of which appears to be very abundantly expressed.

Innate immunity: eggs of Manduca sexta are able to respond to parasitism by Trichogramma evanescens.

So far, it was unknown whether immune responses of insect eggs are inducible or suppressed by parasitism. We investigated whether transcription of immune related genes in eggs of Manduca sexta changed in response to parasitism by Trichogramma evanescens. First, using DDRT-PCR, several cDNA elements known to represent immune related M. sexta genes inducible by bacterial challenge were isolated from eggs. In addition, two novel cDNAs were found: (a) immulectin-V (IML-V) suggested to be involved in recognition of foreign bodies, and (b) a new like-moricin protein with possible antimicrobial effects (L-Mor). Quantitative real time RT-PCR analyses revealed enhanced transcription in parasitized eggs compared to unparasitized ones for IML-V, prophenoloxidase (ProPO), prophenoloxidase activating protease I (PAP I), and proparalytic peptide (ProPP). No significant differences between parasitized and unparasitized eggs were detected for sequences encoding the antimicrobial peptides L-Mor, leureptin Leu, and attacin II Att II. Transcript levels of other antibacterial peptides were suppressed after parasitization for 3d (cecropin 6, Cec 6) and 2d (gloverin, Glov). While nearly 100% of the Manduca eggs contained Trichogramma specimens 1d after exposure to parasitoids, only 64% of the host eggs harbored parasitoid larvae 4d after parasitization. Our data demonstrate that the immune system of Manduca eggs shows differentiated responses to parasitization and suggest that insect eggs can defend against parasitization.

Molecular characterisation of cDNAs from the fall armyworm Spodoptera frugiperda encoding Manduca sexta allatotropin and allatostatin preprohormone peptides.

Allatotropin (AT) is a 13-residue amidated neuropeptide, first isolated from pharate adult heads of the tobacco hornworm, Manduca sexta (Manse-AT), which strongly stimulates the biosynthesis of juvenile hormones (JH) in the corpora allata (CA) of adult moths. In Spodoptera frugiperda, a cDNA that encodes 134 amino acids, including an AT peptide, has been cloned. The S. frugiperda allatotropin mature peptide (Spofr-AT) [GFKNVEMMTARGFa] is identical to that isolated from M. sexta. The basic organization of the Spofr-AT precursor is similar to that of Agrius convolvuli, M. sexta, Pseudaletia unipuncta, and Bombyx mori with 83-93% amino acid sequence identity. The Spofr-AT gene is expressed in at least three mRNA isoforms with 134, 171 and 200 amino acids, differing from each other by alternative splicing. All allatostatins (AS) have an inhibitory action on the JH biosynthesis in the CA. A cDNA that encodes 125 amino acid residues including one copy of the Manse-AS peptide has been cloned from S. frugiperda (Spofr-AS; QVRFRQCYFNPISCF). The basic organization of the Spofr-AS precursor is similar to that of P. unipuncta with 85% amino acid sequence identity. Using one step RT-PCR for semi-quantification of the gene expression, we showed that the three mRNAs of the Spofr-AT gene and the Spofr-AS gene are expressed in brains of last instar larvae, prepupae, pupae, and adults of both sexes of S. frugiperda with variable intensity.