Santi Lucélia, Coutinho-Rodrigues Caio J B, Berger Markus, Klein Lisete A S, De Souza Eduardo M, Rosa Rafael L, Guimarães Jorge A, Yates John R, Perinotto Wendell M S, Bittencourt Vânia R E P, Beys-da-Silva Walter O
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av Ipiranga, 2752, Porto Alegre, RS, 90610-000, Brazil.
Centro de Pesquisa Experimental, Hospital de Clínicas de Porto Alegre, R. Ramiro Barcelos, 2350, Porto Alegre, RS, 90035-903, Brazil.
Folia Microbiol (Praha). 2019 May;64(3):361-372. doi: 10.1007/s12223-018-0659-3. Epub 2018 Oct 25.
Beauveria bassiana is widely studied as an alternative to chemical acaricides in controlling the cattle tick Rhipicephalus microplus. Although its biocontrol efficiency has been proved in laboratory and field scales, there is a need to a better understanding of host interaction process at molecular level related to biocontrol activity. In this work, applying a proteomic technique multidimensional protein identification technology (MudPIT), the differential secretome of B. bassiana induced by the host R. microplus cuticle was evaluated. The use of the host cuticle in a culture medium, mimicking an infection condition, is an established experimental model that triggers the secretion of inducible enzymes. From a total of 236 proteins, 50 proteins were identified exclusively in infection condition, assigned to different aspects of infection like host adhesion, cuticle penetration and fungal defense, and stress. Other 32 proteins were considered up- or down-regulated. In order to get a meaningful global view of the secretome, several bioinformatic analyses were performed. Regarding molecular function classification, the highest number of proteins in the differential secretome was assigned in to hydrolase activity, enzyme class of all cuticle-degrading enzymes like lipases and proteases. These activities were also further validated through enzymatic assays. The results presented here reveal dozens of specific proteins and different processes potentially implicated in cattle tick infection improving the understanding of molecular basis of biocontrol of B. bassiana against R. microplus.
球孢白僵菌作为化学杀螨剂的替代品，在控制微小牛蜱方面得到了广泛研究。尽管其生物防治效率已在实验室和田间规模得到证实，但仍需要在分子水平上更好地理解与生物防治活性相关的宿主相互作用过程。在这项工作中，应用蛋白质组学技术多维蛋白质鉴定技术（MudPIT），评估了由宿主微小牛蜱表皮诱导的球孢白僵菌差异分泌蛋白组。在培养基中使用宿主表皮，模拟感染条件，是一种既定的实验模型，可触发诱导酶的分泌。在总共236种蛋白质中，有50种蛋白质仅在感染条件下被鉴定出来，它们与感染的不同方面相关，如宿主黏附、表皮穿透、真菌防御和应激。另外32种蛋白质被认为是上调或下调的。为了对分泌蛋白组有一个有意义的全局认识，进行了多项生物信息学分析。关于分子功能分类，差异分泌蛋白组中数量最多的蛋白质被归类为水解酶活性，这是所有表皮降解酶如脂肪酶和蛋白酶的酶类。这些活性也通过酶活性测定进一步得到验证。此处呈现的结果揭示了数十种与微小牛蜱感染潜在相关的特定蛋白质和不同过程，增进了对球孢白僵菌对微小牛蜱生物防治分子基础的理解。
