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Antifungal performance of extracellular chitinases and culture supernatants of Streptomyces galilaeus CFFSUR-B12 against Mycosphaerella fijiensis Morelet

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Abstract

The tropical and mycoparasite strain Streptomyces galilaeus CFFSUR-B12 was evaluated as an antagonist of Mycosphaerella fijiensis Morelet, causal agent of the Black Sigatoka Disease (BSD) of banana. On zymograms of CFFSUR-B12 culture supernatants, we detected four chitinases of approximately 32 kDa (Chi32), 20 kDa (Chi20), and two with masses well over 170 kDa (ChiU) that showed little migration during denaturing electrophoresis at different concentrations of polyacrylamide. The thymol-sulphuric acid assay showed that the ChiU were glycosylated chitinases. Moreover, matrix assisted laser desorption ionization time-of-flight MS analysis revealed that the ChiU are the same protein and identical to a family 18 chitinase from Streptomyces sp. S4 (gi|498328075). Chi32 was similar to an extracellular protein from Streptomyces albus J1074 (gi|478687481) and Chi20 was non-significantly similar to chitinases from five different strains of Streptomyces (P > 0.05). Subsequently, Chi32 and Chi20 were partially purified by anion exchange and hydrophobic interaction chromatography and tested against M. fijiensis. Chitinases failed to inhibit ascospore germination, but inhibited up to 35 and 62 % of germ tube elongation and mycelial growth, respectively. We found that crude culture supernatant and living cells of S. galilaeus CFFSUR-B12 were the most effective in inhibiting M. fijiensis and are potential biocontrol agents of BSD.

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Acknowledgments

Benjamin Moreno especially thanks CONACYT for the Ph.D. scholarship provided (202979). Thanks also to Alejandra Arteaga, Katy Ortiz, Victor Hernández and Edgardo Madrid for their comments, technical support and advice on this research.

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  1. El Colegio de la Frontera Sur (ECOSUR), Carr. Antiguo Aeropuerto km. 2.5, Apartado Postal 36, 30700, Tapachula, Chiapas, Mexico

    Benjamín Moreno Castillo, Karina Guillén Navarro, Francisco Holguín Meléndez & Graciela Huerta Palacios

  2. Centro de Ciencias Genómicas, Universidad Autónoma de México (UNAM), Campus Morelos, Av. Universidad s/n Col. Chamilpa, 62210, Cuernavaca, Morelos, Mexico

    Michael F. Dunn, Magdalena Hernández Ortiz & Sergio Encarnación Guevara

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  1. Benjamín Moreno Castillo
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Castillo, B.M., Dunn, M.F., Navarro, K.G. et al. Antifungal performance of extracellular chitinases and culture supernatants of Streptomyces galilaeus CFFSUR-B12 against Mycosphaerella fijiensis Morelet. World J Microbiol Biotechnol 32, 44 (2016). https://doi.org/10.1007/s11274-015-1993-0

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