Molecular phylogenetics and antimicrobial potentials of selected endophytic fungi associated with Curcuma longa Phylogénétique moléculaire et potentiels antimicrobiens d'une sélection de champignons endophytes associés à Curcuma longa
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Abstract
ENGLISH
Background: Curcuma longa, popularly known as turmeric, is a rhizomatous plant belonging to the Zingiberaceae family. This study is necessitated sequel to the dearth of information on endophytic fungi associated with this medicinal plant.
Objectives: This study aimed at isolating endophytic fungi from C. Longa and analyzing their antimicrobial potential.
Methods: The fungal endophytes were isolated from the surface-sterilized rhizomes on potato dextrose agar (PDA) and identified through morphological, microscopical, and internal transcribed spacer (ITS) sequencing. The base calling and consensus sequence was generated using the Geneious software version 9.0.5. The sequence was blasted and submitted to the NCBI database for accession numbers. The Phylogenetic analysis was done using the Geneious software upon MUSCLE alignment. The antimicrobial potential was challenged with some Gram positives and ESKAPE organisms by agar plug and Kirby-Baurer techniques.
Results: A total number of ten (10) fungal endophytes were isolated from thirty sliced samples explored and the preliminary antimicrobial assay revealed that only two isolates, CLR1 and CLR9 could produce antimicrobial secondary metabolites. The BLAST results revealed that they are closely related to Paecilomyces dactylethromorphus CBS 251.55 (NR_149330.1) and Aspergillus ochraceopetaliformis NRRL 4752 (NR_135390.1) at percentage identity of 99.65 % and 99.43 % respectively. Phylogenetic analysis confirmed the relationship between the isolates and known species. Ethyl acetate extract showed zones of growth inhibitions between
10±0.2 mm and 28±1.2 mm in diameter against tested organisms. Their crude extract showed MIC of 1.57 mg/ml to 20 mg/ml and MBC from 2.1 mg/ml to 20.8 mg/ml.
Conclusion: These findings suggest that Curcuma longa hosts fungal endophytes with antimicrobial potential, warranting further biotechnological exploration.
FRENCH
Contexte: Curcuma longa, communément appelé curcuma, est une plante rhizomateuse appartenant à la famille des Zingibéracées. Cette étude est rendue nécessaire par le manque d'informations sur les champignons endophytes associés à cette plante médicinale.
Objectifs: Cette étude vise à isoler les champignons endophytes de C. Longa et à analyser leur potentiel antimicrobien.
Méthodes: Les endophytes fongiques ont été isolés des rhizomes stérilisés en surface sur gélose dextrose de pomme de terre (PDA) et identifiés par séquençage morphologique, microscopique et par séquençage de l'espaceur interne transcrit (ITS). La séquence d'appel de base et la séquence consensus ont été générées à l'aide du logiciel Geneious version 9.0.5. La séquence a été blastée et soumise à la base de données NCBI pour les numéros d'accès. L'analyse phylogénétique a été réalisée à l'aide du logiciel Geneious après alignement MUSCLE. Le potentiel antimicrobien a été testé avec certains organismes Gram positifs et ESKAPE par des techniques de
bouchon d'agar et de Kirby-Baurer.
Résultats: Un nombre total de dix (10) endophytes fongiques ont été isolés à partir de trente échantillons tranchés explorés et l'essai antimicrobien préliminaire a révélé que seuls deux isolats, CLR1 et CLR9, pouvaient produire des métabolites secondaires antimicrobiens. Les résultats du test BLAST ont révélé qu'ils sont étroitement liés à Paecilomyces dactylethromorphe CBS 251,55 (NR_149330.1) et Aspergillus ochraceopetaliformis NRRL 4752 (NR_135390.1) avec des pourcentages d'identité de 99,65% et 99,43% respectivement. L'analyse phylogénétique a confirmé la relation entre les isolats et les espèces connues. L'extrait d'acétate d'éthyle a montré des zones d'inhibition de croissance entre 10±0,2 mm et 28±1,2 mm de diamètre contre les organismes testés. Leur extrait
brut a montré une CMI de 1,57 mg/ml à 20 mg/ml et une CMB de 2,1 mg/ml à 20,8 mg/ml.
Conclusion: Ces résultats suggèrent que Curcuma longa héberge des endophytes fongiques dotés d'un potentiel antimicrobien, ce qui justifie une exploration biotechnologique plus approfondie.
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