Targeted disruption of genes in the Bombyx mori sex pheromone biosynthetic pathway
Targeted disruption of genes in the Bombyx mori sex pheromone biosynthetic pathway
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DOI：
10.1073/pnas.0511270103
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发表时间：
2006-03-21
影响因子：
11.1
通讯作者：
Matsumoto, S
中科院分区：
文献类型：
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作者：
Ohnishi, A;Hull, JJ;Matsumoto, S
鳞翅目昆虫性信息素的生物合成途径需要多个基因产物的协同作用。一些信息素腺（PG）特异性基因已被克隆，在最近几年，而在体外表征表明功能一致的信息素生产中的作用，有没有明确的示范在体内。使用RNA干扰介导的功能丧失方法，我们将对应于感兴趣基因的dsRNA注入新形成的家蚕蛹[即，PG脂肪酰基还原酶（pgFAR）、B. mori IPG Z11/ Delta 10，12 clesatase（Bmpgdesat 1）、PG酰基辅酶A结合蛋白（pgACBP）、中肠ACBP和信息素生物合成激活神经肽受体（PBANR）]，以评估它们在信息素生成过程中的特定作用。在所有情况下，引入的dsRNA诱导性信息素产生的剂量依赖性减少，转录水平相应降低。未观察到对蛹发育或成虫羽化的影响。破坏PBANR基因导致脂肪酶活性的损失，释放信息素前体，而敲除的pgACBP基因防止日常积累和波动的三酰甘油的功能作为细胞存款的信息素前体。两者合计，我们的研究结果提供了明确的证据表明，pgACBP，Bmpgdesat 1，pgFAR，PBANR基因产物是必不可少的信息素生成过程中，并展示了这种方法的权力解剖的分子相互作用，包括生物合成途径。
The sex pheromone biosynthetic pathways of lepidopterans require the concerted actions of multiple gene products. A number of pheromone gland (PG)-specific genes have been cloned in recent years and, whereas in vitro characterizations have indicated functions consistent with roles in pheromone production, there have been no clear demonstrations in vivo. Using an RNA interference-mediated loss-of-function approach, we injected newly formed Bombyx mori pupae with dsRNAs corresponding to genes of interest [i.e., PG fatty acyl reductase (pgFAR), B. mori IPG Z11/ Delta 10,12 clesaturase (Bmpgdesat1), PG acyl-CoA-binding protein (pgACBP), midgut ACBP, and pheromone biosynthesis activating neuropeptide receptor (PBANR)] to assess their specific roles during pheromonogenesis. In all cases, the introduced dsRNAs induced a dose-dependent reduction in sex pheromone production with the corresponding decrease in transcript levels. No effects on pupal development or adult emergence were observed. Disrupting the PBANR gene resulted in a loss of the lipase activity that liberates pheromone precursors, whereas knockout of the pgACBP gene prevented the daily accumulation and fluctuation of the triacylglycerols that function as the cellular deposits for the pheromone precursors. Taken together, our results provide unequivocal evidence that the pgACBP, Bmpgdesat1, pgFAR, and PBANR gene products are essential during pheromonogenesis and demonstrate the power of this methodology for dissecting the molecular interactions that comprise biosynthetic pathways.