| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1065-1 | ||||
Resumo:The genus Bacillus is home to several species of multifunctional bacteria of agronomic importance and of interest for the research of new bioproducts. In this sense, phenotyping and genotyping studies are important to identify intra and interspecific variability of species of agronomic interest. This study aimed to identify genetic variability among Bacillus spp. using ERIC-PCR primers. 27 Bacillus spp. from the Collection of Phytopathogenic and Multifunctional Microorganisms of Embrapa Rice and Beans. So that the isolates could be submitted to the rep-PCR genomic fingerprint analysis using the pair of ERIC primers (Enterobacterial Repetitive Intergenic Consensus), DNA extraction was initially performed and later PCR (Polymerase Chain Reaction). Amplification was carried out in a thermocycler under the following conditions: an initial cycle at 94°C for 5 minutes, followed by 40 cycles of 1 minute at 94°C, 1 minute at 50°C and 2 minutes at 72°C, and a cycle final 7 minutes at 72°C. After amplification, PCR products were separated by electrophoresis and visualized under UV light on a transilluminator. The 1 Kb molecular weight marker (Ludwig Biotec) was used to determine the size of the generated fragments. The analysis was performed by constructing a binary matrix, recording 1 for presence and 0 for absence of bands. Using the R software version 4.3.1, a matrix of genetic distances was calculated using the complement of the Jaccard similarity index. Through the generated matrix, the dendrogram was produced using the hierarchical grouping algorithm UPGMA (Unweighted Pair Group Method with Arithmetic Mean) and the consistency of the nodes was evaluated through 10.000 bootstraps. Cluster analysis based on generated rep-PCR profiles allowed the differentiation of isolates and the observation of genotypic diversity among the isolates in this study. The ERIC primer generated 18 different fingerprint profiles, the number of bands per isolate ranged from 0 to 8, and polymorphic bands of varying intensity (50 to 3000 base pairs) were observed. The dendrogram allowed the grouping of isolates into two main groups and within them, there was the formation of subgroups. In conclusion, this study identified genetic divergence of 27 Bacillus spp. by genomic fingerprint analysis. In addition, this work emphasizes the importance of this type of molecular characterization, since genetic variability indicates differences between isolates. Palavras-chave: agriculture, beneficial bacteria, fingerprint, rep-PCR Agęncia de fomento:Ministério da Agricultura e Pecuária - MAPA - Brazil | |||||