Document Detail

Isolation of Bacillus pumilus from in vitro grapes as a long-term alcohol-surviving and rhizogenesis inducing covert endophyte.
MedLine Citation:
PMID:  15186448     Owner:  NLM     Status:  MEDLINE    
AIMS: To isolate and characterize the alcohol-surviving covert bacterium associated with grape tissue culture. METHODS AND RESULTS: Single colony was isolated by plating the spent rectified spirit used during grape culturing and the organism was identified as Bacillus pumilus using partial 16S rDNA sequence data. Spotting tests (1 microl) using 3-day-old broth culture having a spore content of 20-30% showed similar bacterial survival in 25-90% (v/v) aqueous ethanol for 14 days. Survival in 90% ethanol and 90% rectified spirit appeared affected thereafter with no colony growth from 1 microl samples after 4 months. Plating the samples at this stage gave similar CFU ml(-1) for 25, 50 and 70% ethanol, a significant reduction in 80% ethanol and very few colonies in 90% ethanol and rectified spirit. B. pumilus-inoculated grape microcuttings showed substantial endophytic colonization of original cuttings (7.4 x 10(6) CFU g(-1)) followed by the sprout (5.9 x 10(5)) and roots (2.0 x 10(4)). The bacterium although a poor root colonizer, induced early rooting and more roots in vitro. Inoculation at ex vitro planting resulted in significantly more roots, root weight and shoot growth. CONCLUSIONS: Bacillus pumilus could remain as a covert endophyte in grape tissue cultures and survive in aqueous ethanol for extended periods. 90% ethanol was the most effective bactericidal concentration. The bacterium showed endophytic colonization and root and shoot growth promotion. SIGNIFICANCE AND IMPACT OF THE STUDY: The revelation that general recommendation of 70-80% ethanol may not be the most effective bactericidal concentration for all bacteria, elucidation of the possibility of covert bacterial survival in vitro plant cultures and isolation of a potential plant growth promoting endophyte in grape.
P Thomas
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of applied microbiology     Volume:  97     ISSN:  1364-5072     ISO Abbreviation:  J. Appl. Microbiol.     Publication Date:  2004  
Date Detail:
Created Date:  2004-06-09     Completed Date:  2004-10-19     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  9706280     Medline TA:  J Appl Microbiol     Country:  England    
Other Details:
Languages:  eng     Pagination:  114-23     Citation Subset:  IM    
Division of Biotechnology, Indian Institute of Horticultural Research, Hessarghatta Lake, Bangalore, India.
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MeSH Terms
Bacillus / growth & development,  isolation & purification*
Base Sequence
Culture Techniques
DNA, Bacterial / analysis
Ethanol / pharmacology
Food Contamination*
Molecular Sequence Data
Plant Shoots
Spores, Bacterial
Vitis / growth & development,  microbiology*
Reg. No./Substance:
0/DNA, Bacterial; 64-17-5/Ethanol

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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