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Non-Destructive Assessment of Growth Performance of Embryogenic Suspension Culture of Pinus kesiya (Royle ex. Gord.) in Shake-Flask and Self-Designed Bubble Bioreactor and Successful Regeneration of Plantlets from the Culture Systems

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Abstract

Embryogenic suspensions of Pinus kesiya (Khasi pine) derived from embryogenic cultures obtained from immature zygotic embryos (intact megagametophytes) of Pk-04 genotype were grown in shake-flasks and self-designed bubble bioreactors. Conifer somatic embryogenesis could find its utility for rapid mass propagation if grown as suspension cultures. To minimize the destruction of embryogenic cultures during study, a few growth characteristics viz., sedimented cell volume and number of stage-I (early stage) and -III (cotyledonary) embryo formation in suspensions were determined over a 21-day period. Compared to shake-flasks, proliferation in bioreactors resulted in increased biomass. The suspension culture showed a mixture of freely suspended as well as aggregates of stage-I (early stage) embryos. Maturation of somatic embryos was done on gelled medium and in submerged culture where gelled solid medium was covered with a layer of liquid medium. The number of stage-III (cotyledonary) embryos formed in bubble bioreactor were significantly higher than shake-flask cultures with both solid medium and submerged cultures. The cotyledonary embryos germinated successfully producing plantlets on germination medium.

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Acknowledgments

Laboratory facilities provided by the Department of Botany and other logistics by the Department of Basic Sciences and Social Sciences, North-Eastern Hill University, Shillong, Meghalaya, India are gratefully acknowledged.

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Authors and Affiliations

  1. Biotechnology Division, Department of Basic Sciences & Social Sciences, School of Technology, North-Eastern Hill University, Shillong, 793022, Meghalaya, India

    Hiranjit Choudhury

  2. Division of Plant Biotechnology, Department of Botany, School of Life Sciences, North-Eastern Hill University, Shillong, 793022, Meghalaya, India

    Pramod Tandon

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  1. Hiranjit Choudhury
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  2. Pramod Tandon
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Correspondence to Hiranjit Choudhury.

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Choudhury, H., Tandon, P. Non-Destructive Assessment of Growth Performance of Embryogenic Suspension Culture of Pinus kesiya (Royle ex. Gord.) in Shake-Flask and Self-Designed Bubble Bioreactor and Successful Regeneration of Plantlets from the Culture Systems. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 84, 771–777 (2014). https://doi.org/10.1007/s40011-013-0253-z

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  • DOI: https://doi.org/10.1007/s40011-013-0253-z

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