Genome-wide identification and expression analysis of the mating-responsive genes in the male accessory glands of Spodoptera litura (Lepidoptera: Noctuidae).

BACKGROUND: Mating elicits significant changes in gene expression and leads to subsequent physiological and behavioural modifications in insects. The reproductive success of both sexes is contributed immensely by the male accessory gland (MAG) proteins that are transferred along with sperms to the female reproductive tract during mating where they facilitate several processes that modify the post-mating behaviour. The mating-responsive genes in the MAGs have been identified and reported in many insects but have not been well-characterized in the important agricultural pest Spodoptera litura. Here, we present RNA sequencing analysis to identify mating-responsive genes from the accessory glands of virgin males and males interrupted during mating. RESULTS: Overall, 91,744 unigenes were generated after clustering the assembled transcript sequences of both samples, while the total number of transcripts annotated was 48,708 based on sequence homology against the non-redundant (NR) database. Comparative transcriptomics analysis revealed 16,969 genes that were differentially expressed between the two groups, including 9814 up-regulated and 7155 down-regulated genes. Among the top 80 genes that were selected for heat map analysis, several prominent genes including odorant binding protein, cytochrome P450, heat shock proteins, juvenile hormone binding protein, carboxypeptidases and serine protease were differentially expressed. CONCLUSIONS: The identified genes are known or predicted to promote several processes that modify the female post-mating behaviour. Future studies with the individual MAG protein or in combination will be required to recognize the precise mechanisms by which these proteins alter female physiology and reproductive behaviour. Thus, our study provides essential data to address fundamental questions about reproduction within and among insects and also paves way for further exploration of the functions of these proteins in female insects.

Heterologous expression of a substance which inhibits receptivity and calling in Helicoverpa armigera (Hübner).

The accessory glands of male moths secrete several proteins, which are known to affect post-mating behaviour in females such as calling, reduction in receptivity, rate of egg maturation and laying, sperm maintenance and release and formation of mating plug. Helicoverpa armigera (Hübner) is a polyphagous pest of numerous crops and it is widely distributed on the Indian subcontinent where it causes severe economic losses. In the present study, receptivity- and calling-inhibiting substance (RCIS), a peptide secreted from the accessory glands of male H. armigera, was sequenced, cloned and expressed in a prokaryote, Escherichia coli. RCIS is a peptide comprising 58 amino acids and had a theoretical molecular weight of 6.03 kDa. It showed 64% similarity with pheromonostatic peptide 1, identified in Helicoverpa zea (Kingan et al., 1995) but differed regarding deletion of four and one amino acids at positions 14-17 and 44, respectively, and insertion of one and five amino acids at position 38 and the terminal position of RCIS, respectively. H. armigera females injected with recombinant RCIS showed reduced receptivity and calling behaviour (in 70-80% of the treated individuals), and mating frequencies decreased by 80%. Recombinant RCIS may be employed to artificially induce non-receptivity in virgin females in order to prevent reproduction.