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In vitro propagation of Dendrobium aphyllum (Orchidaceae)—seed germination to flowering

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Abstract

This communication describes asymbiotic seed germination, protocorm development, micropropagation and flowering in in vitro and hardened seedlings of Dendrobium aphyllum (Roxb.) C.E.C. Fischer. Effects of four culture media viz., Murashige and Skoog (MS); Phytamax (Sigma Chemical Co. USA; PM); Mitra et al. (M) and Knudson ‘C’ (KC), 6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D), peptone and activated charcoal were studied on seed germination and protocorm development. Maximum germination (97 %) was recorded in PM basal medium. Peptone (2.0 gl−1) remarkably enhanced germination percentage (100 %), vigorous growth, high survival and subsequent development of protocorms, while in activated charcoal the response was not encouraging. BAP improved germination percentage, however, 2,4-D showed noticeably low seed germination. The morphogenetic response of protocorms and nodal segments of in vitro raised seedlings varied depending on type of explants and concentrations and combinations of plant growth regulators used. Stout root system was induced in 1/2PM + 0.5 mgl−1 IAA. Approximately 10 % of the in vitro raised plants (4–5 cm) with 3–4 leaves flowered in vitro irrespective of flowering season. The well-rooted plants showed 80 % survival under green house conditions and flowering was noticed after 5–6 months in 10 % of hardened plants.

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Abbreviations

BAP:

6-Benzylaminopurine

IAA:

Indole-3-acetic acid

2,4-D:

2,4-dichlorophenoxy acetic acid

NAA:

α-Naphthaleneacetic acid

PLBs:

Protocorm like bodies

MS:

Murashige and Skoog medium

PM:

Phytamax medium (Sigma Chemical Co. USA)

M:

Mitra et al. medium

KC:

Knudson ‘C’ medium

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Acknowledgements

Authors thank the Director, CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, India for providing laboratory facilities. DBT-TWAS for providing a Postgraduate Fellowship to MM Hossain and financial assistance from Council of Scientific and Industrial Research is gratefully acknowledged.

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  1. Mohammad Musharof Hossain

    Present address: Department of Botany, University of Chittagong, Chittagong, 4331, Bangladesh

Authors and Affiliations

  1. Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur, 176 061, Himachal Pradesh, India

    Madhu Sharma

  2. Department of Botany, Panjab University, Chandigarh, 160 014, India

    Promila Pathak

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  1. Mohammad Musharof Hossain
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Correspondence to Madhu Sharma.

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Hossain, M.M., Sharma, M. & Pathak, P. In vitro propagation of Dendrobium aphyllum (Orchidaceae)—seed germination to flowering. J. Plant Biochem. Biotechnol. 22, 157–167 (2013). https://doi.org/10.1007/s13562-012-0124-3

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