Puccinia triticina (P. triticina) is one of the most devastating fungal pathogens of wheat which causes significant annual yield loss to the crop. Understanding the gene regulatory mechanism of the biotrophic pathogen is one of the important aspects of host-pathogen interaction studies. Dicer-like genes are considered as important mediators of RNAi-based gene regulation. In this study, we report the presence of three Dicer-like genes (Pt-DCL1, Pt-DCL2, Pt-DCL3) in P. triticina genome identified through computational and biological analyses. Quantitative real-time PCR studies revealed an increase in the expression of these genes in germinating spore stages. Heterologous expression combined with mass spectrometry analysis of Pt-DCL2 confirmed the presence of a canonical Dicer-like gene in P. triticina. Phylogenetic analysis of the Pt-DCLs with the Dicer-like proteins from other organisms showed a distinct cluster of rust pathogens from the order Pucciniales. The results indicated a species-specific duplication of Dicer-like genes within the wheat rust pathogens. This study, for the first time, reports the presence of Dicer-dependent RNAi pathway in P. triticina that may play a role in gene regulatory mechanism of the pathogen during its development. Our study serves as a vital source of information for further RNAi-based molecular studies for better understanding and management of the wheat leaf rust disease.

中文翻译：

---

小麦叶片锈病病原体（小麦锈病）中切丁酶样基因的鉴定与表征。

小麦锈菌（Puccinia triticina）（P. triticina）是小麦中最具毁灭性的真菌病原体之一，每年对作物造成重大的产量损失。了解生物营养性病原体的基因调控机制是宿主-病原体相互作用研究的重要方面之一。切酶样基因被认为是基于RNAi的基因调控的重要介体。在这项研究中，我们报告的存在3 Dicer酶样基因（铂DCL1，铂DCL2，铂DCL3中）小麦叶锈菌通过计算和生物学分析确定的基因组。实时定量PCR研究表明，这些基因在萌芽孢子阶段的表达增加。异源表达结合质谱分析Pt-DCL2证实了小麦中存在典型的Dicer样基因。用来自其他生物的Dicer样蛋白对Pt-DCL进行系统进化分析，结果显示了来自锈菌的锈病病原体明显不同。结果表明，在小麦锈病病原体中，Dicer样基因的物种特异性重复。这项研究首次报道了小麦中的Dicer依赖性RNAi途径的存在。可能在病原体的发展过程中对其基因调控机制起作用。我们的研究为进一步基于RNAi的分子研究提供了重要信息，以更好地了解和管理小麦叶锈病。