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Deciphering the diversity of culturable thermotolerant bacteria from Manikaran hot springs

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Abstract

The aim of this study was to analyze and characterize the diversity of culturable thermotolerant bacteria in Manikaran hot springs. A total of 235 isolates were obtained employing different media, and screened for temperature tolerance (40 °C–70 °C). A set of 85 isolates tolerant to 45 °C or above were placed in 42 phylogenetic clusters after amplified ribosomal DNA restriction analysis (16S rRNA-ARDRA). Sequencing of the 16S rRNA gene of 42 representative isolates followed by BLAST search revealed that the majority of isolates belonged to Firmicutes, followed by equal representation of Actinobacteria and Proteobacteria. Screening of representative isolates (42 ARDRA phylotypes) for amylase activity revealed that 26 % of the isolates were positive, while 45 % exhibited protease activity, among which one amylase and six protease producers were tolerant up to 70 °C. BIOLOG-based identification of 13 isolates exhibiting temperature tolerance up to 70 °C, using carbon utilization patterns and sensitivity to chemicals, revealed a high degree of correlation with identification based on 16S rRNA gene sequencing for all isolates, except one (M48). These promising isolates showing a range of useful metabolic attributes demand to be explored further for industrial and agricultural applications.

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Acknowledgments

The first author is thankful to Department of Science and Technology, Government of India for providing fellowship for his PhD program and Post Graduate School, IARI. All the authors are thankful to the Division of Microbiology, IARI, New Delhi for providing the necessary facilities for undertaking this study. The financial support from National Agricultural Innovation Project (Indian Council of Agricultural Research) under the project “Diversity analysis of Bacillus and other predominant genera in extreme environments and its utilization in Agriculture” is gratefully acknowledged. The authors also sincerely thank the technical assistance of Central Instrumentation Facility, Delhi University for BIOLOG analysis.

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  1. Division of Microbiology, Indian Agricultural Research Institute (IARI), New Delhi, 110012, India

    Murugan Kumar, Ajar Nath Yadav, Rameshwar Tiwari, Radha Prasanna & Anil Kumar Saxena

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  1. Murugan Kumar
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  2. Ajar Nath Yadav
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  3. Rameshwar Tiwari
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Correspondence to Anil Kumar Saxena.

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Kumar, M., Yadav, A.N., Tiwari, R. et al. Deciphering the diversity of culturable thermotolerant bacteria from Manikaran hot springs. Ann Microbiol 64, 741–751 (2014). https://doi.org/10.1007/s13213-013-0709-7

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