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Studies on Endophytic Colonization Ability of Two Upland Rice Endophytes, Rhizobium sp. and Burkholderia sp., Using Green Fluorescent Protein Reporter

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Abstract

Colonization ability of the two endophytic bacteria, isolated from surface sterilized roots of upland cultivated rice viz., Rhizobium sp. and Burkholderia sp., was compared after genetically tagging them with a constitutively expressing green fluorescent protein gene (gfp/gusA). Confocal laser scanning microscopy (CLSM) of gnotobiotically grown seedlings of Narendradhan 97, inoculated with gfp/gusA-tagged endophytes, revealed that both Rhizobium sp. and Burkholderia sp. colonized the intercellular spaces in the root cortex when inoculated separately. Colonization by gfp/gusA-tagged Rhizobium sp. was severely inhibited when co-inoculated with an equal number (106 cfu ml−1) of wild type Burkholderia sp. Burkholderia sp. was a more aggressive endophytic colonizer of rice than Rhizobium sp. The potential of using gfp/gusA reporter and CLSM as tools in evaluating competitive ability of colonization among endophytes is demonstrated in this study.

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Acknowledgments

Authors are grateful to Prof S.C. Lakhotia, DST-National facility for Multiphoton Confocal Laser Scanning Microscopy for permission to use the facility and for valuable suggestions. We also thank Prof Humberto J.O. Ramos (Curitiba, PR, Brazil) for providing the plasmid, pHRGFPGUS. MKS is thankful to UGC research fellowship and Department of Science and Technology, Government of India for the financial support.

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Authors and Affiliations

  1. Microbial Genetics Laboratory, Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, 221005, India

    Manoj Kumar Singh & Ramesh Kumar Singh

  2. Department of Mycology and Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, 221005, India

    Chanda Kushwaha

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  1. Manoj Kumar Singh
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  2. Chanda Kushwaha
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  3. Ramesh Kumar Singh
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Correspondence to Manoj Kumar Singh.

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Singh, M.K., Kushwaha, C. & Singh, R.K. Studies on Endophytic Colonization Ability of Two Upland Rice Endophytes, Rhizobium sp. and Burkholderia sp., Using Green Fluorescent Protein Reporter. Curr Microbiol 59, 240–243 (2009). https://doi.org/10.1007/s00284-009-9419-6

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  • DOI: https://doi.org/10.1007/s00284-009-9419-6

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