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西番莲(黄果西番莲)叶片中的胰蛋白酶抑制剂:对茉莉酸甲酯、机械损伤和食草作用的累积响应

Trypsin inhibitors in passion fruit (Passiflora f. edulis flavicarpa) leaves: accumulation in response to methyl jasmonate, mechanical wounding, and herbivory.

作者信息

Botelho-Júnior Sylvio, Siqueira-Júnior César L, Jardim Bruno C, Machado Olga L T, Neves-Ferreira Ana G C, Perales Jonas, Jacinto Tânia

机构信息

Laboratorio de Biotecnologia, Universidade Estadual do Norte Fluminense, 28013-600 Campos dos Goytacazes, RJ, Brazil.

出版信息

J Agric Food Chem. 2008 Oct 22;56(20):9404-9. doi: 10.1021/jf8013266. Epub 2008 Sep 17.

Abstract

This work investigates the effect of methyl jasmonte (MeJa), mechanical wounding, and herbivory caused by larval feeding of a specialist insect ( Agraulis vanillae vanillae) upon trypsin inhibitory activity in passion fruit leaves. Despite the fact that all treatments caused accumulation of trypsin inhibitors (TIs), higher levels were observed in MeJa treated leaves when plants were assayed 24 and 48 h after stimulus. Concerning both mechanically injured plants and attacked ones, a systemic induction was observed. Partially purified inhibitors from MeJa exposed plants were further characterized by X-ray film contact print technique and N-terminal sequence. Such analysis indicated that the TIs identified belong to the Kunitz family. Moreover, the partially purified inhibitors strongly inhibited trypsin-like digestive enzymes from sugar cane stalk borer ( Diatraea saccharalis) in vitro. Our results further support the protective function of wound-inducible trypsin inhibitors and their potential as tools to improve important crop species against insect predation through genetic engineering.

摘要

本研究调查了茉莉酸甲酯(MeJa)、机械损伤以及由专食性昆虫(香草凤蝶幼虫)取食造成的植食作用对西番莲叶片中胰蛋白酶抑制活性的影响。尽管所有处理均导致胰蛋白酶抑制剂(TIs)积累,但在刺激后24小时和48小时对植株进行检测时,发现经MeJa处理的叶片中胰蛋白酶抑制剂水平更高。对于机械损伤植株和受攻击植株,均观察到了系统性诱导现象。通过X射线胶片接触印片技术和N端序列对从经MeJa处理的植株中部分纯化得到的抑制剂进行了进一步表征。此类分析表明,所鉴定出的胰蛋白酶抑制剂属于库尼茨家族。此外,部分纯化的抑制剂在体外能强烈抑制甘蔗螟虫(蔗扁蛾)的类胰蛋白酶消化酶。我们的结果进一步支持了伤口诱导型胰蛋白酶抑制剂的保护功能及其作为通过基因工程改良重要作物品种以抵御昆虫捕食的工具的潜力。

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