Tundo Silvio, Moscetti Ilaria, Faoro Franco, Lafond Mickaël, Giardina Thierry, Favaron Francesco, Sella Luca, 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.
Plant Sci. 2015 Nov;240:161-9. doi: 10.1016/j.plantsci.2015.09.002. Epub 2015 Sep 7.
To shed light on the role of Xylanase Inhibitors (XIs) during Fusarium graminearum infection, we first demonstrated that three out of four F. graminearum xylanases, in addition to their xylan degrading activity, have also the capacity to cause host cell death both in cell suspensions and wheat spike tissue. Subsequently, we demonstrated that TAXI-III and XIP-I prevented both the enzyme and host cell death activities of F. graminearum xylanases. In particular, we showed that the enzymatic inhibition by TAXI-III and XIP-I was competitive and only FGSG_11487 escaped inhibition. The finding that TAXI-III and XIP-I prevented cell death activity of heat inactivated xylanases and that XIP-I precluded the cell death activity of FGSG_11487 - even if XIP-I does not inhibit its enzyme activity - suggests that the catalytic and the cell death activities are separated features of these xylanases. Finally, the efficacy of TAXI-III or XIP-I to prevent host cell death caused by xylanases was confirmed in transgenic plants expressing separately these inhibitors, suggesting that the XIs could limit F. graminearum infection via direct inhibition of xylanase activity and/or by preventing host cell death.
为了阐明木聚糖酶抑制剂(XIs)在禾谷镰刀菌感染过程中的作用,我们首先证明,在四种禾谷镰刀菌木聚糖酶中,有三种除了具有木聚糖降解活性外,在细胞悬浮液和小麦穗组织中还具有导致宿主细胞死亡的能力。随后,我们证明TAXI-III和XIP-I既能阻止禾谷镰刀菌木聚糖酶的酶活性,又能阻止其宿主细胞死亡活性。特别是,我们发现TAXI-III和XIP-I的酶抑制作用具有竞争性,只有FGSG_11487不受抑制。TAXI-III和XIP-I能阻止热灭活木聚糖酶的细胞死亡活性,且XIP-I能阻止FGSG_11487的细胞死亡活性(即使XIP-I不抑制其酶活性),这一发现表明这些木聚糖酶的催化活性和细胞死亡活性是分开的特性。最后,在分别表达这些抑制剂的转基因植物中证实了TAXI-III或XIP-I预防木聚糖酶引起的宿主细胞死亡的功效,这表明木聚糖酶抑制剂可以通过直接抑制木聚糖酶活性和/或防止宿主细胞死亡来限制禾谷镰刀菌的感染。