1a.Objectives (from AD-416):
The objectives of this project are:. 1)produce a transcriptome of codling moth heads;. 2)identify and characterize neuropeptides expressed by codling moth; and. 3)Clone and characterize neuropeptides and receptors from spotted winged Drosophila.
1b.Approach (from AD-416):
1) Messenger RNA will be extracted from heads of codling moth larvae, pupae and adults, and then converted to complementary DNA (cDNA).
2) The sequences of cDNAs representing head mRNA transcripts will be determined using 454 sequencing technology.
3) The sequences of assembled head cDNAs will be used to identify those encoding neuropeptides, peptide hormones, and other potential protein targets for codling moth control.
4) cDNAs from SWD that encode neuropeptides and receptors involved in regulation of feeding and reproduction will be cloned and characterized.
The endocrine system (hormones, neuropeptide and peptide hormones, and their receptors) controls almost every physiological function in the insect including reproduction, regulation of feeding, and development. The goal of this project is to identify neuropeptides and their receptors in both the codling moth, a major pest of apples, and a newly emerging pest insect of tree fruit, spotted-wing Drosophila. Our approach for identification of neuropeptides and receptors in codling moth is through generation of a transcriptome where we will sequence gene transcripts expressed in the brain (the source of neuropeptide production). We have collected thousands of codling moth heads (containing brains), extracted messenger RNA (the protein coding form of RNA) and converted the RNA to a more stable cDNA form. The cDNA has been sent to collaborators at Washington State University (Pullman, WA) to obtain DNA sequence information. For spotted-wing Drosophila, a different approach was used to identify neuropeptides and receptors involved in regulation of feeding and reproduction. Using genome sequences encoding neuropeptides and receptors available from other species of Drosophila; we cloned gene transcripts expressed in spotted-wing Drosophila which should encode five neuropeptides and two receptors that are thought to be involved in the regulation of feeding and one peptide hormone and receptor thought to be involved in controlling female receptiveness during mating and reproduction. The work reported here addresses objectives 1 and 2 in the parent project plan.