Moscetti Ilaria, Faoro Franco, Moro Stefano, Sabbadin Davide, Sella Luca, Favaron Francesco, D'Ovidio Renato
Dipartimento di Scienze e Tecnologie per l'Agricoltura, le Foreste, la Natura e l'Energia (DAFNE), Università della Tuscia, Via S. Camillo de Lellis snc, 01100, Viterbo, Italy.
Dipartimento di Scienze Agrarie e Ambientali, Università degli Studi di Milano, Via Celoria 2, 20133, Milano, Italy.
Mol Plant Pathol. 2015 Aug;16(6):583-92. doi: 10.1111/mpp.12215. Epub 2014 Dec 15.
The xylanase inhibitor TAXI-III has been proven to delay Fusarium head blight (FHB) symptoms caused by Fusarium graminearum in transgenic durum wheat plants. To elucidate the molecular mechanism underlying the capacity of the TAXI-III transgenic plants to limit FHB symptoms, we treated wheat tissues with the xylanase FGSG_03624, hitherto shown to induce cell death and hydrogen peroxide accumulation. Experiments performed on lemmas of flowering wheat spikes and wheat cell suspension cultures demonstrated that pre-incubation of xylanase FGSG_03624 with TAXI-III significantly decreased cell death. Most interestingly, a reduced cell death relative to control non-transgenic plants was also obtained by treating, with the same xylanase, lemmas of TAXI-III transgenic plants. Molecular modelling studies predicted an interaction between the TAXI-III residue H395 and residues E122 and E214 belonging to the active site of xylanase FGSG_03624. These results provide, for the first time, clear indications in vitro and in planta that a xylanase inhibitor can prevent the necrotic activity of a xylanase, and suggest that the reduced FHB symptoms on transgenic TAXI-III plants may be a result not only of the direct inhibition of xylanase activity secreted by the pathogen, but also of the capacity of TAXI-III to avoid host cell death.
木聚糖酶抑制剂TAXI-III已被证明可延缓转基因硬粒小麦植株中由禾谷镰刀菌引起的赤霉病(FHB)症状。为了阐明TAXI-III转基因植物限制FHB症状能力背后的分子机制,我们用木聚糖酶FGSG_03624处理小麦组织,该木聚糖酶此前已被证明可诱导细胞死亡和过氧化氢积累。在开花小麦穗的颖片和小麦细胞悬浮培养物上进行的实验表明,木聚糖酶FGSG_03624与TAXI-III预孵育可显著减少细胞死亡。最有趣的是,用相同的木聚糖酶处理TAXI-III转基因植物的颖片,相对于对照非转基因植物,细胞死亡也减少了。分子建模研究预测TAXI-III的H395残基与属于木聚糖酶FGSG_03624活性位点的E122和E214残基之间存在相互作用。这些结果首次在体外和体内明确表明,木聚糖酶抑制剂可以阻止木聚糖酶的坏死活性,并表明转基因TAXI-III植物上FHB症状减轻可能不仅是由于直接抑制病原体分泌的木聚糖酶活性,还由于TAXI-III避免宿主细胞死亡的能力。