An Ethylene-Protected Achilles' Heel of Etiolated Seedlings for Arthropod Deterrence.
An Ethylene-Protected Achilles' Heel of Etiolated Seedlings for Arthropod Deterrence.
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DOI：
10.3389/fpls.2016.01246
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发表时间：
2016
影响因子：
5.6
通讯作者：
Reinbothe C
中科院分区：
文献类型：
--
作者：
Boex-Fontvieille E;Rustgi S;von Wettstein D;Pollmann S;Reinbothe S;Reinbothe C
Kunitz蛋白酶抑制剂家族存在于拟南芥中，其中一个成员（由At 1g 72290编码）在植物发育过程中发挥高度特异性的作用。拟南芥Kunitz蛋白酶抑制剂1（Kunitz-PI;1），我们在这里称之为这种蛋白质，作为半胱氨酸PI起作用。活性测量显示，尽管存在保守的Kunitz基序，但细菌表达的Kunitz-PI;1不能抑制丝氨酸蛋白酶如胰蛋白酶和胰凝乳蛋白酶，但非常有效地抑制响应于降解的半胱氨酸蛋白酶21。蛋白质印迹和细胞定位研究，使用单特异性抗体召回Kunitz-PI;1蛋白在花，年轻的角果和黄化幼苗的表达。在黑暗中生长的幼苗，最大Kunitz-PI;1启动子活性检测在下胚轴的顶端钩区和顶端部分。免疫定位证实Kunitz-PI;1在这些器官和组织中表达。在黑暗和变绿过程中，转录因子No transmitting tract（NTT）和HECATE 1（HEC 1）参与调节Kunitz-PI;1的表达，其中NTT起负作用，HEC 1起正作用。实验室饲养实验等足类甲壳类动物，如Porcellio scaber（木虱）和Armadillidium vulgare（pillbug）指出，顶端钩乙烯保护的阿基里斯之踵黄化苗。由于外源应用的乙烯前体1-氨基环丙烷-1-羧酸（ACC）和机械应力（创伤）强烈上调HEC 1依赖Kunitz-PI;1基因的表达，我们的研究结果确定了一个新的电路控制食草动物威慑黄化植物Kunitz-PI;1参与。
A small family of Kunitz protease inhibitors exists in Arabidopsis thaliana, a member of which (encoded by At1g72290) accomplishes highly specific roles during plant development. Arabidopsis Kunitz-protease inhibitor 1 (Kunitz-PI;1), as we dubbed this protein here, is operative as cysteine PI. Activity measurements revealed that despite the presence of the conserved Kunitz-motif the bacterially expressed Kunitz-PI;1 was unable to inhibit serine proteases such as trypsin and chymotrypsin, but very efficiently inhibited the cysteine protease RESPONSIVE TO DESICCATION 21. Western blotting and cytolocalization studies using mono-specific antibodies recalled Kunitz-PI;1 protein expression in flowers, young siliques and etiolated seedlings. In dark-grown seedlings, maximum Kunitz-PI;1 promoter activity was detected in the apical hook region and apical parts of the hypocotyls. Immunolocalization confirmed Kunitz-PI;1 expression in these organs and tissues. No transmitting tract (NTT) and HECATE 1 (HEC1), two transcription factors previously implicated in the formation of the female reproductive tract in flowers of Arabidopsis, were identified to regulate Kunitz-PI;1 expression in the dark and during greening, with NTT acting negatively and HEC1 acting positively. Laboratory feeding experiments with isopod crustaceans such as Porcellio scaber (woodlouse) and Armadillidium vulgare (pillbug) pinpointed the apical hook as ethylene-protected Achilles’ heel of etiolated seedlings. Because exogenous application of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) and mechanical stress (wounding) strongly up-regulated HEC1-dependent Kunitz-PI;1 gene expression, our results identify a new circuit controlling herbivore deterrence of etiolated plants in which Kunitz-PI;1 is involved.
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