Analyzing the evolution of insect TMED gene and the expression pattern of silkworm TMED gene / 生物工程学报

Transmembrane emp24 domain (TMED) gene is closely related to immune response, signal transduction, growth and disease development in mammals. However, only the Drosophila TMED gene has been reported on insects. We identified the TMED family genes of silkworm, Tribolium castaneum, tobacco moth and Italian bee from their genomes, and found that the TMED family gene composition patterns of one α-class, one β-class, one δ-class and several γ-classes arose in the common ancestor of pre-divergent Hymenoptera insects, while the composition of Drosophila TMED family members has evolved in a unique pattern. Insect TMED family γ-class genes have evolved rapidly, diverging into three separate subclasses, TMED6-like, TMED5-like and TMED3-like. The TMED5-like gene was lost in Hymenoptera, duplicated in the ancestors of Lepidoptera and duplicated in Drosophila. Insect TMED protein not only has typical structural characteristics of TMED, but also has obvious signal peptide. There are seven TMED genes in silkworm, distributed in six chromosomes. One of seven is single exon and others are multi-exons. The complete open reading frame (ORF) sequences of seven TMED genes of silkworm were cloned from larval tissues and registered in GenBank database. BmTMED1, BmTMED2 and BmTMED6 were expressed in all stages and tissues of the silkworm, and all genes were expressed in the 4th and 5th instar and silk gland of the silkworm. The present study revealed the composition pattern of TMED family members, their γ class differentiation and their evolutionary history, providing a basis for further studies on TMED genes in silkworm and other insects.

Tissue localization and partial characterization of pheromone biosynthesis activating neuropeptide in Achaea janata.

Female sex pheromone production in certain moth species have been shown to be regulated by a cephalic endocrine peptidic factor: pheromone biosynthesis activating neuropeptide (PBAN), having 33 amino acid residues. Antisera against synthetic Heliothis zea-PBAN were developed. Using these polyclonals, immunoreactivity was mapped in the nervous system of Achaea janata. Three distinct groups of immunopositive secretory neurons were identified in the suboesophageal ganglion; and immunoreactivity was observed in the corpora cardiaca, thoracic and in the abdominal ganglia. From about 6000 brain sub-oesophageal ganglion complexes, the neuropeptide was isolated; and purified sequentially by Sep-pak and reversed phase high performance liquid chromatographic methods. Identity of purified PBAN fraction was confirmed with polyclonal antibody by immunoblotting. Molecular mass of the isolated peptide was determined by matrix-assisted laser desorption/ionization mass spectrometry, and was found to be 3900 Da, same as that of known H. zea-PBAN. Radiochemical bioassay confirmed the pheromonotropic effect of the isolated neuropeptide in this insect.

In vivo enhancement of nucleopolyhedrovirus of oriental Armyworm, Mythimna separata using spindles from Helicoverpa armigera entomopoxvirus.

When the third instar larvae of M. separata were exposed to eight varying concentrations of polyhedral occlusion bodies (POB's) of nucleopolyhedrovirus of M. separata (MsNPV) ranging from 2.6 x 10(-1) to 2.6 x 10(8) POB's/ml, the percent mortality and incubation period ranged from 16-100% and 14 to 9 days respectively. On the other hand when the same third instar larvae of M. separata were exposed to only five varying concentration of POB's of MsNPV ranging from 2.6 x 10(2) to 2.6 x 10(6), POB's/ml along with a constant dose of entomopox viral spindles from Helicoverpa armigera, the per cent mortality ranged from 63 to 100% with reduction in incubation period from 7 to 4 days respectively. The enhancement index (log10) of the virus was 2.76 or reduction of more than 500 times in LC50. The ability and the mechanism of the spindles from H. armigera entomopoxvirus to enhance the infectivity of MsNPV has been discussed.