Angelini Riccardo, Tisi Alessandra, Rea Giuseppina, Chen Martha M, Botta Maurizio, Federico Rodolfo, Cona Alessandra
Dipartimento di Biologia, Università Roma Tre, Viale G Marconi 446, 00146, Rome, Italy.
Plant Physiol. 2008 Jan;146(1):162-77. doi: 10.1104/pp.107.108902. Epub 2007 Nov 9.
Hydrogen peroxide (H(2)O(2)) is involved in plant defense responses that follow mechanical damage, such as those that occur during herbivore or insect attacks, as well as pathogen attack. H(2)O(2) accumulation is induced during wound healing processes as well as by treatment with the wound signal jasmonic acid. Plant polyamine oxidases (PAOs) are H(2)O(2) producing enzymes supposedly involved in cell wall differentiation processes and defense responses. Maize (Zea mays) PAO (ZmPAO) is a developmentally regulated flavoprotein abundant in primary and secondary cell walls of several tissues. In this study, we investigated the effect of wounding on ZmPAO gene expression in the outer tissues of the maize mesocotyl and provide evidence that ZmPAO enzyme activity, protein, and mRNA levels increased in response to wounding as well as jasmonic acid treatment. Histochemically detected ZmPAO activity especially intensified in the epidermis and in the wound periderm, suggesting a tissue-specific involvement of ZmPAO in wound healing. The role played by ZmPAO-derived H(2)O(2) production in peroxidase-mediated wall stiffening events was further investigated by exploiting the in vivo use of N-prenylagmatine (G3), a selective and powerful ZmPAO inhibitor, representing a reliable diagnostic tool in discriminating ZmPAO-mediated H(2)O(2) production from that generated by peroxidase, oxalate oxidase, or by NADPH oxidase activity. Here, we demonstrate that G3 inhibits wound-induced H(2)O(2) production and strongly reduces lignin and suberin polyphenolic domain deposition along the wound, while it is ineffective in inhibiting the deposition of suberin aliphatic domain. Moreover, ZmPAO ectopic expression in the cell wall of transgenic tobacco (Nicotiana tabacum) plants strongly enhanced lignosuberization along the wound periderm, providing evidence for a causal relationship between PAO and peroxidase-mediated events during wound healing.
过氧化氢(H₂O₂)参与植物遭受机械损伤后的防御反应,例如食草动物或昆虫攻击以及病原体侵袭时发生的损伤。在伤口愈合过程中以及用伤口信号茉莉酸处理时,会诱导H₂O₂积累。植物多胺氧化酶(PAO)是产生H₂O₂的酶,可能参与细胞壁分化过程和防御反应。玉米(Zea mays)PAO(ZmPAO)是一种受发育调控的黄素蛋白,在多个组织的初生和次生细胞壁中含量丰富。在本研究中,我们调查了创伤对玉米中胚轴外层组织中ZmPAO基因表达的影响,并提供证据表明ZmPAO酶活性、蛋白质和mRNA水平在创伤以及茉莉酸处理后会增加。组织化学检测到的ZmPAO活性在表皮和伤口周皮中尤其增强,表明ZmPAO在伤口愈合中具有组织特异性作用。通过体内使用N-异戊烯基胍丁胺(G3)进一步研究了ZmPAO衍生的H₂O₂产生在过氧化物酶介导的细胞壁硬化事件中所起的作用,G3是一种选择性且强效的ZmPAO抑制剂,是区分ZmPAO介导的H₂O₂产生与过氧化物酶、草酸氧化酶或NADPH氧化酶活性产生的H₂O₂的可靠诊断工具。在此,我们证明G3抑制创伤诱导的H₂O₂产生,并强烈减少伤口处木质素和木栓质多酚结构域的沉积,而对抑制木栓质脂肪族结构域的沉积无效。此外,ZmPAO在转基因烟草(Nicotiana tabacum)植物细胞壁中的异位表达强烈增强了伤口周皮处的木质栓质化,为PAO与创伤愈合过程中过氧化物酶介导的事件之间的因果关系提供了证据。