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Extracellular Chitinases of Fluorescent Pseudomonads Antifungal to Fusarium oxysporum f. sp. dianthi Causing Carnation Wilt

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Abstract

Vascular wilt of carnation caused by Fusarium oxysporum f. sp. dianthi (Prill. & Delacr.) W. C. Synder & H.N. Hans inflicts substantial yield and quality loss to the crop. Mycolytic enzymes such as chitinases are antifungal and contribute significantly to the antagonistic activity of fluorescent pseudomonads belonging to plant-growth-promoting rhizobacteria. Fluorescent pseudomonads antagonistic to the vascular wilt pathogen were studied for their ability to grow and produce chitinases on different substrates. Bacterial cells grown on chitin-containing media showed enhanced growth and enzyme production with increased anti-fungal activity against the pathogen. Furthermore, the cell-free bacterial culture filtrate from chitin-containing media also significantly inhibited the mycelial growth. Both the strains and their cell-free culture filtrate from chitin-amended media showed the formation of lytic zones on chitin agar, indicating chitinolytic ability. Extracellular proteins of highly antagonistic bacterial strain were isolated from cell-free extracts of media amended with chitin and fungal cell wall. These cell-free conditioned media contained one to seven polypeptides. Western blot analysis revealed two isoforms of chitinase with molecular masses of 43 and 18.5 kDa. Further plate assay for mycelial growth inhibition showed the 43-kDa protein to be antifungal. The foregoing studies clearly established the significance of chitinases in the antagonism of fluorescent pseudomonads, showing avenues for possible exploitation in carnation wilt management.

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Acknowledgments

We thank the Director, IHBT (CSIR), Palampur for support and encouragement during the course of this investigation. A generous gift of tobacco chitinase antiserum from Prof. Michael Legrand and Pienette Geoffroy, Centre National De La Recherche Scientifique, France is gratefully acknowledged. This work was supported by the Council of Scientific and Industrial Research, Government of India (30-(2050)/SMM 02/2003 dated 19.1.2004) through a co-ordinated network program.

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  1. Floriculture Division, Institute of Himalayan Bioresource Technology, Palampur, 176061, Himachal Pradesh, India

    Naosekpam Singh Ajit, Rajni Verma & V. Shanmugam

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  1. Naosekpam Singh Ajit
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  2. Rajni Verma
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  3. V. Shanmugam
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Correspondence to V. Shanmugam.

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Ajit, N.S., Verma, R. & Shanmugam, V. Extracellular Chitinases of Fluorescent Pseudomonads Antifungal to Fusarium oxysporum f. sp. dianthi Causing Carnation Wilt. Curr Microbiol 52, 310–316 (2006). https://doi.org/10.1007/s00284-005-4589-3

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