Colletotrichum lindemuthianum, the causal agent of anthracnose, is responsible for significant damage in the common bean (Phaseolus vulgaris L.). Unraveling the genetic mechanisms involved in the plant/pathogen interaction is a powerful approach for devising efficient methods to control this disease. In the present study, we employed the Restriction Enzyme-Mediated Integration (REMI) methodology to identify the gene slnCl1, encoding a histidine kinase protein, as involved in pathogenicity. The mutant strain, MutCl1, generated by REMI, showed an insertion in the slnCl1 gene, deficiency of the production and melanization of appressoria, as well as the absence of pathogenicity on bean leaves when compared with the wild-type strain. The slnCl1 gene encodes a histidine kinase class IV called SlnCl1 showing identity of 97% and 83% with histidine kinases from Colletotrichum orbiculare and Colletotrichum gloesporioides, respectively. RNA interference was used for silencing the histidine kinase gene and confirm slnCl1 as a pathogenicity factor. Furthermore, we identified four major genes involved in the RNA interference-mediated gene silencing in Colletotrichum spp. and demonstrated the functionality of this process in C. lindemuthianum. Silencing of the EGFP reporter gene and slnCl1 were demonstrated using qPCR. This work reports for the first time the isolation and characterization of a HK in C. lindemuthianum and the occurrence of gene silencing mediated by RNA interference in this organism, demonstrating its potential use in the functional characterization of pathogenicity genes.

炭疽病的病原体炭疽菌（Colletotrichum lindemuthianum）对普通豆（菜豆）造成了重大损害。揭示涉及植物/病原体相互作用的遗传机制是设计控制此病的有效方法的有力方法。在本研究中，我们采用了限制性酶介导的整合（REMI）方法来鉴定与致病性有关的编码组氨酸激酶蛋白的基因slnCl1。与野生型菌株相比，由REMI产生的突变菌株MutCl1在slnCl1基因中插入，缺乏压感的产生和黑色素化，并且在豆叶上没有致病性。这slnCl1基因编码一个称为SlnCl1的IV类组氨酸激酶，显示与球形炭疽菌和球形炭疽菌的组氨酸激酶分别具有97％和83％的同一性。RNA干扰被用于沉默组氨酸激酶基因，并确认slnCl1是致病因子。此外，我们确定了四个在炭疽菌中干扰RNA介导的基因沉默的主要基因。并演示了该过程在C. lindemuthianum中的功能。在沉默EGFP报告基因和slnCl1使用qPCR进行了验证。这项工作首次报道了在菩提树中HK的分离和鉴定，以及该生物中由RNA干扰介导的基因沉默的发生，证明了其在致病性基因功能鉴定中的潜在用途。