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首页> 美国卫生研究院文献>Molecules >Identification and Characterization of trans-Isopentenyl Diphosphate Synthases Involved in Herbivory-Induced Volatile Terpene Formation in Populus trichocarpa
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Identification and Characterization of trans-Isopentenyl Diphosphate Synthases Involved in Herbivory-Induced Volatile Terpene Formation in Populus trichocarpa

机译：涉草诱导的毛果杨挥发性萜形成中反式异戊烯基二磷酸合酶的鉴定与表征

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In response to insect herbivory, poplar releases a blend of volatiles that plays important roles in plant defense. Although the volatile bouquet is highly complex and comprises several classes of compounds, it is dominated by mono- and sesquiterpenes. The most common precursors for mono- and sesquiterpenes, geranyl diphosphate (GPP) and (E,E)-farnesyl diphosphate (FPP), respectively, are in general produced by homodimeric or heterodimeric trans-isopentenyl diphosphate synthases (trans-IDSs) that belong to the family of prenyltransferases. To understand the molecular basis of herbivory-induced terpene formation in poplar, we investigated the trans-IDS gene family in the western balsam poplar Populus trichocarpa. Sequence comparisons suggested that this species possesses a single FPP synthase gene (PtFPPS1) and four genes encoding two large subunits (PtGPPS1.LSU and PtGPPS2.LSU) and two small subunits (PtGPPS.SSU1 and PtGPPS.SSU2) of GPP synthases. Transcript accumulation of PtGPPS1.LSU and PtGPPS.SSU1 was significantly upregulated upon leaf herbivory, while the expression of PtFPPS1, PtGPPS2.LSU, and PtGPPS.SSU2 was not influenced by the herbivore treatment. Heterologous expression and biochemical characterization of recombinant PtFPPS1, PtGPPS1.LSU, and PtGPPS2.LSU confirmed their respective IDS activities. Recombinant PtGPPS.SSU1 and PtGPPS.SSU2, however, had no enzymatic activity on their own, but PtGPPS.SSU1 enhanced the GPP synthase activities of PtGPPS1.LSU and PtGPPS2.LSU in vitro. Altogether, our data suggest that PtGPPS1.LSU and PtGPPS2.LSU in combination with PtGPPS.SSU1 may provide the substrate for herbivory-induced monoterpene formation in P. trichocarpa. The sole FPP synthase PtFPPS1 likely produces FPP for both primary and specialized metabolism in this plant species.
机译：响应昆虫的食草性，白杨释放出挥发物混合物，在植物防御中起重要作用。尽管挥发性花束非常复杂并包含几类化合物，但它主要由单萜和倍半萜组成。单萜和倍半萜的最常见前体，分别是香叶基二磷酸酯（GPP）和（E，E）-法呢基二磷酸酯（FPP），通常由同二聚或异二聚反式异戊烯基二磷酸合酶（trans-IDS）生产异戊二烯基转移酶家族。为了了解草食动物诱导的杨树中萜形成的分子基础，我们研究了苦瓜西部毛杨Populus trichocarpa中的trans-IDS基因家族。序列比较表明，该物种具有单个FPP合酶基因（PtFPPS1）和四个编码GPP合成酶的大亚基（PtGPPS1.LSU和PtGPPS2.LSU）和两个小亚基（PtGPPS.SSU1和PtGPPS.SSU2）的基因。食草动物显着上调了PtGPPS1.LSU和PtGPPS.SSU1的转录本积累，而食草动物处理则不影响PtFPPS1，PtGPPS2.LSU和PtGPPS.SSU2的表达。重组PtFPPS1，PtGPPS1.LSU和PtGPPS2.LSU的异源表达和生化特征证实了它们各自的IDS活性。重组PtGPPS.SSU1和PtGPPS.SSU2本身没有酶活性，但是PtGPPS.SSU1增强了PtGPPS1.LSU和PtGPPS2.LSU的GPP合酶活性。总而言之，我们的数据表明，PtGPPS1.LSU和PtGPPS2.LSU与PtGPPS.SSU1的结合可能为毛果单生象中草食性单萜的形成提供了底物。唯一的FPP合酶PtFPPS1可能为该植物物种的主要代谢和专门代谢产生FPP。

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