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Identification of culturable and originally non-culturable endophytic bacteria isolated from shoot tip cultures of banana cv. Grand Naine

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Abstract

In this article we describe the identification of endophytic bacteria belonging to three groups isolated from shoot tip cultures of banana cv. Grand Naine in a recent study (Thomas et al. 2008) based on partial 16S rRNA gene sequence homology analysis. The first group included banana stocks that displayed obvious colony growth on MS based tissue culture medium during the first in vitro passage. The second group constituted stocks that were tissue index-negative for cultivable bacteria initially but turned index-positive after a few to several (4–8) in vitro passages while the third group formed one sub-stock that turned index-positive after about 18 passages. The organisms belonged to about 20 different genera comprising of α, β, γ-proteobacteria, Gram-positive firmicutes and actinobacteria. Visibly expressing easily cultured organisms during the first in vitro passage included Enterobacter, Klebsiella, Ochrobactrum, Pantoea, Staphylococcus and Bacillus spp. Organisms of second group that were not detected or non-culturable originally constituted Brevundimonas, Methylobacterium, Alcaligenes, Ralstonia, Pseudomonas, Corynebacterium, Microbacterium, Staphylococcus, Oceanobacillus and Bacillus spp. while the third group that turned cultivable after extended in vitro culturing included mostly non-filamentous actinobacteria (Brachybacterium, Brevibacterium, Kocuria and Tetrasphaera spp.). The identification results suggested that the endophytes of second and third groups were not strictly obligate or fastidious microbes but those surviving in viable but-non-culturable (VBNC) state and displaying gradual activation to cultivable form during continuous tissue culturing. Several of the organisms isolated are known as beneficial ones in agriculture while some organisms have possible implications in human health. The use of tissue cultures for isolating uncommon endophytes is discussed.

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Abbreviations

BIM:

Bacteriological indexing medium

NA:

Nutrient agar

OC:

Original culture

TCM:

Tissue culture medium

TSA:

Trypticasein soy agar

VA:

Viss et al. (1991) agar

VBNC:

Viable but non-culturable

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Acknowledgments

This study was supported by a grant from the Department of Biotechnology, Govt. of India under the project ‘Identification of covert endophytic microbes in plant tissue cultures and their management and control’. Contributions from D. P. Prakash and V. Bidari during their brief association with the project and laboratory assistance by N. Shivrudriah and B. Hanumanthraju are acknowledged. This publication bears IIHR Contribution No. 10/2007.

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Author notes

  1. Ganiga K. Swarna

    Present address: Department of Biotechnology, Dr. S. G. Reddy College, Domlur, Bangalore, 560071, India

  2. Pulak K. Roy

    Present address: Faculty Centre of Integrated Rural and Tribal Development, Ramakrishna Mission Vivekananda University, Morabadi, Ranchi, 834008, Jharkhand, India

Authors and Affiliations

  1. Division of Biotechnology, Indian Institute of Horticultural Research, Hessarghatta Lake, Bangalore, 560089, India

    Pious Thomas, Ganiga K. Swarna & Pulak K. Roy

  2. AICRP on Tropical Fruit Crops, Indian Institute of Horticultural Research, Hessarghatta Lake, Bangalore, 560089, India

    Prakash Patil

Authors
  1. Pious Thomas
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  2. Ganiga K. Swarna
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Corresponding author

Correspondence to Pious Thomas.

Additional information

Supply of live bacterial cultures or genetic material for research purpose is subject to their revival from glycerol stocks (as some of the organisms showed poor tolerance) and the requestor obtaining written permission from the Director General, Indian Council of Agricultural Research, New Delhi-110001.

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Thomas, P., Swarna, G.K., Roy, P.K. et al. Identification of culturable and originally non-culturable endophytic bacteria isolated from shoot tip cultures of banana cv. Grand Naine. Plant Cell Tiss Organ Cult 93, 55–63 (2008). https://doi.org/10.1007/s11240-008-9341-9

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