Institut für Biologie und Biotechnologie der Pflanzen, Westfälische Wilhelms-Universität Münster, Schlossplatz 8, 48143 Münster, Germany.
Sci Rep. 2016 Nov 30;6:38018. doi: 10.1038/srep38018.
To successfully survive in plants, endophytes need strategies to avoid being detected by the plant immune system, as the cell walls of endophytes contain easily detectible chitin. It is possible that endophytes "hide" this chitin from the plant immune system by modifying it, or oligomers derived from it, using chitin deacetylases (CDA). To explore this hypothesis, we identified and expressed a CDA from Pestalotiopsis sp. (PesCDA), an endophytic fungus, in E. coli and characterized this enzyme and its chitosan oligomer products. We found that when PesCDA modifies chitin oligomers, the products are partially deacetylated chitosan oligomers with a specific acetylation pattern: GlcNAc-GlcNAc-(GlcN)-GlcNAc (n ≥ 1). Then, in a bioactivity assay where suspension-cultured rice cells were incubated with the PesCDA products (processed chitin hexamers), we found that, unlike the substrate hexamers, chitosan oligomer products no longer elicited the plant immune system. Thus, this endophytic enzyme can prevent the endophyte from being recognized by the plant immune system; this might represent a more general hypothesis for how certain fungi are able to live in or on their hosts.
为了在植物中成功生存,内生菌需要避免被植物免疫系统检测到的策略,因为内生菌的细胞壁含有容易检测到的几丁质。内生菌可能通过使用几丁质脱乙酰酶(CDA)对其进行修饰,或者对其寡聚物进行修饰,从而“隐藏”来自植物免疫系统的这种几丁质。为了探索这一假设,我们从内生真菌 Pestalotiopsis sp.(PesCDA)中鉴定并表达了一种 CDA,并在大肠杆菌中对该酶及其壳寡糖产物进行了表征。我们发现,当 PesCDA 修饰几丁质寡糖时,产物是部分脱乙酰化的壳寡糖,具有特定的乙酰化模式:GlcNAc-GlcNAc-(GlcN)-GlcNAc(n≥1)。然后,在一个生物活性测定中,悬浮培养的水稻细胞与 PesCDA 产物(处理过的壳六聚体)孵育,我们发现,与底物六聚体不同,壳寡糖产物不再引发植物免疫系统。因此,这种内生酶可以防止内生菌被植物免疫系统识别;这可能代表了某些真菌能够在其宿主中生存或寄生的更普遍的假设。