Zhang Songdou, Liu Xiaoming, Zhu Bin, Yin Xinming, Du Mengfang, Song Qisheng, An Shiheng
State Key Laboratory of Wheat and Maize Crop Science (College of Plant Protection), Henan Agricultural University, Zhengzhou, Henan, P.R. China.
Divisions of Plant Sciences, University of Missouri, Columbia, Missouri, United States of America.
PLoS One. 2014 Oct 20;9(10):e111003. doi: 10.1371/journal.pone.0111003. eCollection 2014.
Mating decreases female receptivity and terminates sex pheromone production in moths. Although significant progress has been made in elucidating the mating-regulated inactivation of pheromone biosynthesis-activating neuropeptide (PBAN) secretion, little is known about the mating induced gene expression profiles in pheromone glands (PGs). In this study, the associated genes involved in Bombyx mori mating were identified through digital gene expression (DGE) profiling and subsequent RNA interference (RNAi) to elucidate the molecular mechanisms underlying the mating-regulated gene expression in PGs.
Eight DGE libraries were constructed from the PGs of mated and virgin females: 1 h mating (M1)/virgin (V1) PGs, 3 h mating (M3)/virgin (V3) PGs, 24 h mating (M24)/virgin (V24) PGs and 48 h mating (M48)/virgin (V48) PGs (M48 and V48). These libraries were used to investigate the gene expression profiles affected by mating. DGE profiling revealed a series of genes showing differential expression in each set of mated and virgin female samples, including immune-associated genes, sex pheromone synthesis-associated genes, juvenile hormone (JH) signal-associated genes, etc. Most interestingly, JH signal was found to be activated by mating. Application of the JH mimics, methoprene to the newly-emerged virgin females leaded to the significant reduction of sex pheromone production. RNAi-mediated knockdown of putative JH receptor gene, Methoprene tolerant 1 (Met1), in female pupa resulted in a significant decrease in sex pheromone production in mature females, suggesting the importance of JH in sex pheromone synthesis.
A series of differentially expressed genes in PGs in response to mating was identified. This study improves our understanding of the role of JH signaling on the mating-elicited termination of sex pheromone production.
交配会降低雌蛾的接受性并终止其性信息素的产生。尽管在阐明交配调节的信息素生物合成激活神经肽(PBAN)分泌失活方面已取得重大进展,但对于交配诱导的性信息素腺(PG)基因表达谱却知之甚少。在本研究中,通过数字基因表达(DGE)分析及随后的RNA干扰(RNAi)鉴定了家蚕交配相关基因,以阐明PG中交配调节基因表达的分子机制。
从已交配和未交配雌蛾的PG构建了8个DGE文库:交配1小时(M1)/未交配(V1)的PG、交配3小时(M3)/未交配(V3)的PG、交配24小时(M24)/未交配(V24)的PG以及交配48小时(M48)/未交配(V48)的PG(M48和V48)。这些文库用于研究受交配影响的基因表达谱。DGE分析揭示了一系列在每组已交配和未交配雌蛾样本中差异表达的基因,包括免疫相关基因、性信息素合成相关基因、保幼激素(JH)信号相关基因等。最有趣的是,发现JH信号因交配而被激活。将JH类似物烯虫酯应用于新羽化的未交配雌蛾会导致性信息素产生显著减少。RNAi介导的雌蛹中假定的JH受体基因甲氧普烯耐受蛋白1(Met1)敲低导致成熟雌蛾中性信息素产生显著减少,表明JH在性信息素合成中具有重要作用。
鉴定了PG中一系列响应交配的差异表达基因。本研究增进了我们对JH信号在交配引发的性信息素产生终止中的作用的理解。