Abstract
Female moths employ their own pheromone blends as a communicational medium in mating behavior. The biosynthesis and release of sex pheromone in female moths are regulated by pheromone biosynthesis activating neuropeptide (PBAN) and the corresponding action of PBAN has been well elucidated in Bombyx mori. However, very little is known about the molecular mechanism regarding the biosynthesis of sex pheromone precursor. In this study, quantitative proteomics was utilized to comprehensively elucidate the expression dynamics of pheromone glands (PGs) during development. Proteomic analysis revealed a serial of differentially expressed sex pheromone biosynthesis-associated proteins at the different time points of B. mori development. Most interestingly B. mori glycerol-3-phosphate O-acyltransferase (BmGPAT) was found to be expressed during the key periods of sex pheromone biosynthesis. RNAi knockdown of BmGPAT confirmed the important function of this protein in the biosynthesis of sex pheromone precursor, triacylglcerol (TAG), and subsequently PBAN-induced production of sex pheromone, bombykol. Behavioral analysis showed that RNAi knockdown of GPAT significantly impaired the ability of females to attract males. Our findings indicate that GPAT acts to regulate the biosynthesis of sex pheromone precursor, TAG, thus influencing PBAN-induced sex pheromone production and subsequent mating behavior.
Highlights
Female moths employ their own pheromone blends as a communicational medium in mating behavior
The biosynthesis and release of sex pheromone in female moths are regulated by pheromone biosynthesis activating neuropeptide (PBAN) and the corresponding action of PBAN has been well elucidated in Bombyx mori
Lepidopteran sex pheromones are de novo synthesized in the pheromone glands (PGs) from acetyl-CoA through fatty acid biosynthesis followed by desaturation, chainshortening, fatty acyl reduction and carbonyl carbon modification[1]
Summary
A series of sex pheromone biosynthesis-associated proteins were highly expressed in 0 h and 72 h PG samples, which are crucial for the biosynthesis and release of sex pheromones (Table 1) These proteins include fatty acid transport protein, acyl CoA desaturase, acyl-CoA binding protein, fatty-acyl reductase, perilipin, Orai[1] alternative splice form A, calcineurin A and acyl carrier protein. Further TAG content analysis manifested that the female PGs injected with GPAT dsRNA synthesized fewer TAGs compared with the control females injected with EGFP dsRNA (Fig. 5D). These results suggested that GPAT significantly affected the biosynthesis of TAGs and LDs. In addition, GPAT-F1 dsRNA had stronger inhibitory effect than GPAT-F2 dsRNA. The results showed that the GPAT knockdown significantly reduced female’s ability to attract males when tested against EGFP control (GPAT-F15EGFP 5 28572; GPAT-F25EGFP 5 33567) (Fig. 6B), similar results were found in the reciprocal experiments of Y-tube left and right portions (data not shown)
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