Department of Plant Sciences and Plant Pathology, Montana State University, Bozeman, Montana, USA.
PLoS One. 2010 Jan 13;5(1):e8685. doi: 10.1371/journal.pone.0008685.
The European corn borer (ECB), Ostrinia nubilalis (Hubner), exists as two separate sex pheromone races. ECB(Z) females produce a 97ratio3 blend of Z11- and E11-tetradecenyl acetate whereas ECB(E) females produce an opposite 1ratio99 ratio of the Z and E isomers. Males of each race respond specifically to their conspecific female's blend. A closely related species, the Asian corn borer (ACB), O. furnacalis, uses a 3ratio2 blend of Z12- and E12-tetradecenyl acetate, and is believed to have evolved from an ECB-like ancestor. To further knowledge of the molecular mechanisms of pheromone detection and its evolution among closely related species we identified and characterized sex pheromone receptors from ECB(Z).
Homology-dependent (degenerate PCR primers designed to conserved amino acid motifs) and homology-independent (pyrophosphate sequencing of antennal cDNA) approaches were used to identify candidate sex pheromone transcripts. Expression in male and female antennae was assayed by quantitative real-time PCR. Two-electrode voltage clamp electrophysiology was used to functionally characterize candidate receptors expressed in Xenopus oocytes.
We characterized five sex pheromone receptors, OnOrs1 and 3-6. Their transcripts were 14-100 times more abundant in male compared to female antennae. OnOr6 was highly selective for Z11-tetradecenyl acetate (EC(50) = 0.86+/-0.27 microM) and was at least three orders of magnitude less responsive to E11-tetradecenyl acetate. Surprisingly, OnOr1, 3 and 5 responded to all four pheromones tested (Z11- and E11-tetradecenyl acetate, and Z12- and E12-tetradecenyl acetate) and to Z9-tetradecenyl acetate, a behavioral antagonist. OnOr1 was selective for E12-tetradecenyl acetate based on an efficacy that was at least 5-fold greater compared to the other four components. This combination of specifically- and broadly-responsive pheromone receptors corresponds to published results of sensory neuron activity in vivo. Receptors broadly-responsive to a class of pheromone components may provide a mechanism for variation in the male moth response that enables population level shifts in pheromone blend use.
欧洲玉米螟(ECB),Ostrinia nubilalis(Hubner),存在两种独立的性信息素种族。ECB(Z)雌性产生 Z11-和 E11-十四碳烯基乙酸酯的 97:3 混合物,而 ECB(E)雌性则产生 Z 和 E 异构体的 1:99 混合物。每个种族的雄性都专门对其同种雌性的混合物做出反应。一个密切相关的物种,亚洲玉米螟(ACB),O. furnacalis,使用 Z12-和 E12-十四碳烯基乙酸酯的 3:2 混合物,据信它是从 ECB 样祖先进化而来的。为了进一步了解在密切相关的物种中信息素检测及其进化的分子机制,我们从 ECB(Z)中鉴定并表征了性信息素受体。
同源依赖性(设计用于保守氨基酸基序的简并 PCR 引物)和非同源依赖性(触角 cDNA 的焦磷酸测序)方法用于鉴定候选性信息素转录本。通过定量实时 PCR 测定雄性和雌性触角中的表达。二电极电压钳电生理学用于功能表征在非洲爪蟾卵母细胞中表达的候选受体。
我们描述了五个性信息素受体,OnOrs1 和 3-6。它们的转录物在雄性触角中比在雌性触角中丰富 14-100 倍。OnOr6 对 Z11-十四碳烯基乙酸酯(EC(50)=0.86+/-0.27 microM)具有高度选择性,对 E11-十四碳烯基乙酸酯的反应至少低三个数量级。令人惊讶的是,OnOr1、3 和 5 对所有四种测试的信息素(Z11-和 E11-十四碳烯基乙酸酯以及 Z12-和 E12-十四碳烯基乙酸酯)以及行为拮抗剂 Z9-十四碳烯基乙酸酯有反应。OnOr1 对 E12-十四碳烯基乙酸酯具有选择性,其功效至少比其他四种成分高 5 倍。这种特异性和广泛反应性信息素受体的组合与体内感觉神经元活动的已发表结果相对应。对一类信息素成分广泛反应的受体可能为雄蛾反应的变化提供了一种机制,使信息素混合物的使用在种群水平上发生转移。