Department of Entomology and Center for Chemical Ecology, Pennsylvania State University, University Park, Pennsylvania, United States of America.
PLoS One. 2012;7(7):e41947. doi: 10.1371/journal.pone.0041947. Epub 2012 Jul 25.
The oral secretions of herbivores are important recognition cues that can be used by plants to mediate induced defenses. In this study, a degradation of adenosine-5'-triphosphate (ATP) in tomato leaves was detected after treatment with Helicoverpa zea saliva. Correspondingly, a high level of ATPase activity in saliva was detected and three ATP hydrolyzing enzymes: apyrase, ATP synthase and ATPase 13A1 were identified in salivary glands. To determine the functions of these proteins in mediating defenses, they were cloned from H. zea and expressed in Escherichia coli. By applying the purified expressed apyrase, ATP synthase or ATPase 13A1 to wounded tomato leaves, it was determined that these ATP hydrolyzing enzymes suppressed the defensive genes regulated by the jasmonic acid and ethylene pathways in tomato plant. Suppression of glandular trichome production was also observed after treatment. Blood-feeding arthropods employ 5'-nucleotidase family of apyrases to circumvent host responses and the H. zea apyrase, is also a member of this family. The comparatively high degree of sequence similarity of the H. zea salivary apyrase with mosquito apyrases suggests a broader evolutionary role for salivary apyrases than previously envisioned.
食草动物的口腔分泌物是植物介导诱导防御的重要识别线索。在这项研究中,番茄叶片在处理烟草夜蛾唾液后检测到腺苷-5'-三磷酸(ATP)的降解。相应地,在唾液中检测到高水平的 ATP 酶活性,并在唾液腺中鉴定出三种 ATP 水解酶:apyrase、ATP 合酶和 ATPase 13A1。为了确定这些蛋白质在介导防御中的功能,它们从 H. zea 中克隆并在大肠杆菌中表达。通过将纯化的表达 apyrase、ATP 合酶或 ATPase 13A1 应用于受伤的番茄叶片,确定这些 ATP 水解酶抑制了番茄植物中茉莉酸和乙烯途径调节的防御基因。处理后还观察到腺体毛状体产生的抑制。吸血节肢动物利用 5'-核苷酸酶家族的 apyrase 来规避宿主反应,而 H. zea 的 apyrase 也是该家族的一员。与蚊子 apyrase 相比,H. zea 唾液中的 apyrase 具有相对较高的序列相似性,这表明唾液中的 apyrase 具有比以前想象的更广泛的进化作用。