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Elimination and molecular identification of endophytic bacterial contaminants during in vitro propagation of Bambusa balcooa

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Abstract

Bambusa balcooa is an economically important, multipurpose bamboo species, decidedly used in construction industry. Availability of natural bamboo is depleting very rapidly due to accelerated deforestation and its unrestrained use. The large number and timely supply of saplings are the need of the hour for the restoration of bamboo stands. Micropropagation, being the potent alternative for season independent rapid regeneration, is restricted in bamboo because of endophytic contamination. An in vitro attempt has been taken to overcome the endophytic contamination by using broad spectrum antibiotics as surface sterilant as well as a media component. Ampicillin sodium salt (5 mg/ml for 30 min) as a surface sterilant was found as the best treatment for high bud breaking (80%) coupled with high branching and low contamination (20%) but it was found ineffective to control the contamination during multiplication stage. Then, two endophytes were isolated and minimum inhibitory concentration was determined through antibiotic susceptibility test for successful eradication at multiplication stage. Finally, contamination free cultures were obtained when streptocycline (100 μg/ml) and gentamicin sulphate (75 μg/ml) were added into the medium. The two isolated endophytes, BB1 and BB2, were identified through 16S rDNA techniques and NCBI-BLAST algorithm with 99% sequence similarity with those of Janibacter sp. (KX423734) and Serratia marcescens strain (KX423735). To our knowledge, this is the first report for B. balcooa where antibiotics were used as surface sterilant as well as medium component, to control endophytic bacterial contaminants, followed by their identification.

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References

  • Agnihotri RK, Nandi SK (2009) In vitro shoot cut: a high-frequency multiplication and rooting method in bamboo Dendrocalamus hamiltonii. Biotechnology 8(2):259–263

    Article  Google Scholar 

  • Ali H, Nirmala C, Sharma M (2009) Control of in vitro contamination in bamboos. Plant Cell Biotechnol Mol Biol 10(3&4):119–124

    Google Scholar 

  • Anand M, Brar J, Sood A (2013) In vitro propagation of an edible bamboo Bambusa bambos and assessment of clonal fidelity through molecular markers. J Med Bioeng 2(4):257–261

    CAS  Google Scholar 

  • Andrews MJ (2001) Determination of minimum inhibitory concentrations. J Antimicrob Chemother 48(1):5–16

    Article  CAS  Google Scholar 

  • Attree SM, Sheffield E (1986) An evaluation of Ficoll density gradient centrifugation as a method for eliminating microbial contamination and purifying plant protoplasts. Plant Cell Rep 5(4):288–291

    Article  CAS  Google Scholar 

  • Brar J, Anand M, Sood A (2013) In vitro seed germination of economically important edible bamboo Dendrocalamus membranaceus Munro. Indian J Exp Biol 51(1):88–96

    CAS  Google Scholar 

  • Eed AM, Reddy SA, Reddy KM, Silva JAT, Reddy PV, Beghum H, Venkatsubbaiah PY (2010) Effect of antibiotics and fungicides on the in vitro production of Citrus limonia Osbeck nodal segment and shoot tip explants. Asian Australas J Plant Sci Biotechnol 4(1):66–70

    Google Scholar 

  • Fang JY, Hsu YR (2012) Molecular identification and antibiotic control of endophytic bacterial contaminants from micropropagated Aglaonema cultures. Plant Cell Tiss Org 110(1):53–62

    Article  CAS  Google Scholar 

  • Gaintait S, Pramanik BR, Banerjee M (2016) Optimization of planting materials for large scale plantation of Bambusa balcooa Roxb.: influence of propagation methods. J Saudi Soc Agric Sci. doi:10.1016/j.jssas.2015.11.008

    Google Scholar 

  • Ghosh JS, Chaudhuri S, Dey N, Pal A (2013) Functional characterization of a serine-threonine protein kinase from Bambusa balcooa that implicates in cellulose overproduction and superior quality fiber formation. BMC Plant Biol 13(1):128. doi:10.1186/1471-2229-13-128

    Article  Google Scholar 

  • Goyal AK, Pradhan S, Basistha BC, Sen A (2015) Micropropagation and assessment of genetic fidelity of Dendrocalamus strictus (Roxb.) nees using RAPD and ISSR markers. 3 Biotech 5(4):473–482

    Article  Google Scholar 

  • Hugenholtz P (2002) Exploring prokaryotic diversity in the genomic era. Genome Biol 3(2):1. doi:10.1186/gb-2002-3-2-reviews0003

    Article  Google Scholar 

  • Kulkarni AA, Kelkar SM, Watve MG, Krishnamurthy KV (2007) Characterization and control of endophytic bacterial contaminants in in vitro cultures of Piper spp., Taxus baccata subsp. wallichiana, and Withania somnifera. Can J Microbiol 53(1):63–74. doi:10.1139/w06-106

    Article  CAS  Google Scholar 

  • Leifert C, Cassells AC (2001) Microbial hazards in plant tissue and cell cultures. In Vitro Cell Dev Plant 37(2):133–138

    Article  Google Scholar 

  • Leifert C, Ritchie JY, Waites WM (1991) Contaminants of plant-tissue and cell cultures. World J Microbiol Biotechnol 7(4):452–469

    Article  CAS  Google Scholar 

  • Luna C, Collavino M, Mroginski L, Sansberro P (2008) Identification and control of bacterial contaminants from Ilex dumosa nodal segments culture in a temporal immersion bioreactor system using 16S rDNA analysis. Plant Cell Tiss Org 95(1):13–19

    Article  CAS  Google Scholar 

  • Mehta R, Sharma V, Sood A, Sharma M, Sharma RK (2011) Induction of somatic embryogenesis and analysis of genetic fidelity of in vitro-derived plantlets of Bambusa nutans Wall., using AFLP markers. Eur J For Res 130(5):729–736

    Article  Google Scholar 

  • Mishra Y, Patel PK, Yadav S, Shirin F, Ansari SA (2008) A micropropagation system for cloning of Bambusa tulda Roxb. Sci Hortic Amst 115(3):315–318

    Article  CAS  Google Scholar 

  • Misra P, Gupta N, Toppo DD, Pandey V, Mishra MK, Tuli R (2010) Establishment of long-term proliferating shoot cultures of elite Jatropha curcas L. by controlling endophytic bacterial contamination. Plant Cell Tiss Org 100(2):189–197

    Article  Google Scholar 

  • Moore E, Arnscheidt A, Kruger A, Strompl C, Mau M (2004) Simplified protocols for the preparation of genomic DNA from bacterial cultures. In: Molecular microbial ecology manual, 2nd edn. Kluwer Academic Publishers, Dordrecht, pp 3–18

  • Msogoya T, Kanyagha H, Mutigitu J, Kulebelwa M, Mamiro D (2012) Identification and management of microbial contaminants of banana in vitro cultures. J Appl Biosci 55:3987–3994

    Google Scholar 

  • Mudoi KD, Borthakur M (2009) In vitro micropropagation of Bambusa balcooa Roxb. through nodal explants from field-grown culms and scope for upscaling. Curr Sci India 96(7):962–966

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497

    Article  CAS  Google Scholar 

  • Nadha HK, Salwan R, Kasana RC, Anand M, Sood A (2012) Identification and elimination of bacterial contamination during in vitro propagation of Guadua angustifolia Kunth. Pharmacogn Mag 8(30):93–97

    Article  Google Scholar 

  • Nair DN, Padmavathy S (2014) Impact of endophytic microorganisms on plants, environment, and humans. Sci World J. doi:10.1155/2014/250693

    Google Scholar 

  • Negi D, Saxena S (2011) Micropropagation of Bambusa balcooa Roxb. through axillary shoot proliferation. In Vitro Cell Dev Plant 47(5):604–610

    Article  CAS  Google Scholar 

  • Niedz RP (1998) Using isothiazolone biocides to control microbial and fungal contaminants in plant tissue cultures. HortTechnology 8(4):598–601

    Google Scholar 

  • Niedz RP, Bausher MG (2002) Control of in vitro contamination of explants from greenhouse-and field-grown trees. In Vitro Cell Dev Plant 38(5):468–471

    Article  Google Scholar 

  • Patel B, Gami B, Patel N, Bariya V (2015) One step pre-hardening micropropagation of Bambusa balcooa Roxb. J Phytol 7:1–9

    Article  CAS  Google Scholar 

  • Pollock K, Barfield DG, Shields R (1983) The toxicity of antibiotics to plant cell cultures. Plant Cell Rep 2(1):36–39

    CAS  Google Scholar 

  • Reed BM, Tanprasert P (1995) Detection and control of bacterial contaminants of plant tissue cultures. A review of recent literature. Plant Tiss Cult Biotechnol 1(3):137–142

    Google Scholar 

  • Reed BM, Mentzer J, Tanprasert P, Yu X (1998) Internal bacterial contamination of micropropogated hazelnut: identification and antibiotic treatment. Plant Cell Tiss Org 52(1):67–70

    Article  CAS  Google Scholar 

  • Sarangthem K, Singh TN (2003) Biosynthesis of succulent bamboo shoots of Bambusa balcooa into phytosterols and its biotransformation into ADD. Acta Bot Sin 45(1):114–117

    Google Scholar 

  • Sharma P, Sarma KP (2011) In vitro propagation of Bambusa balcooa for a better environment. In: International conference on advances in biotechnology and pharmaceutical sciences (ICABPS’2011), Bangkok, pp 248–252

  • Sharma P, Sarma KP (2014) In vitro propagation of Bambusa nutan in commercial scale in Assam. J Environ Res Dev 9(2):348–355

    Google Scholar 

  • Sivabalan S, Ramkumar P, Satheesh PM, Ahmed SM (2014) Detection of albino in micropropagated shoots of Bambusa balcooa Roxb, using PCR based techniques. Int J Eng Res Technol 3(3):596–605

    Google Scholar 

  • Tewari DN (1992) A monograph on bamboo. International Book Distributors, Dehradun

    Google Scholar 

  • Thakur R, Sood A (2006) An efficient method for explant sterilization for reduced contamination. Plant Cell Tiss Org 84(3):369–371

    Article  Google Scholar 

  • Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173(2):697–703

    Article  CAS  Google Scholar 

  • Wilson ZA, Power JB (1989) Elimination of systemic contamination in explant and protoplast cultures of rubber (Hevea brasiliensis Muell. Arg.). Plant Cell Rep 7(8):622–625

    CAS  Google Scholar 

  • Woese CR, Kandler O, Wheelis ML (1990) Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya. Proc Natl Acad Sci USA 87(12):4576–4579. doi:10.1073/pnas.87.12.4576

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are highly indebted to DST, Govt. of West Bengal, India for providing a fund to this project.

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Authors and Affiliations

  1. IRDM Faculty Centre, Ramakrishna Mission Vivekananda University, Ramakrishna Mission Ashrama, Narendrapur, Kolkata, 700103, India

    Syandan Sinha Ray, Shibasis Mukherjee & Gautam Chatterjee

  2. Department of Agricultural Biotechnology, Faculty of Agriculture, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia, West Bengal, India

    Md. Nasim Ali

  3. West Bengal State Council of Science and Technology, Vigyan Chetana Bhavan, DD-26/B, Salt Lake, Kolkata, 700064, India

    Maitreyi Banerjee

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  1. Syandan Sinha Ray
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  2. Md. Nasim Ali
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  3. Shibasis Mukherjee
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  4. Gautam Chatterjee
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  5. Maitreyi Banerjee
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Correspondence to Md. Nasim Ali.

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Ray, S.S., Ali, M.N., Mukherjee, S. et al. Elimination and molecular identification of endophytic bacterial contaminants during in vitro propagation of Bambusa balcooa . World J Microbiol Biotechnol 33, 31 (2017). https://doi.org/10.1007/s11274-016-2196-z

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  • DOI: https://doi.org/10.1007/s11274-016-2196-z

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