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Induction of somatic embryogenesis and analysis of genetic fidelity of in vitro-derived plantlets of Bambusa nutans Wall., using AFLP markers

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Abstract

Bambusa nutans Wall., is an evergreen, perennial, and multipurpose bamboo having strong culms, which are largely used for construction, scaffolding, craft purposes, pulp, and paper industry. Multiple shoots from nodal segments (3–4 cm) of young branches of mature culms were established in Murashige and Skoog (1962) (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BAP) (1.0–6.0 mg l−1) or in combination with α-naphthaleneacetic acid (NAA) (0.5–1.0 mg l−1) or kinetin (Kn) (1.0–2.0 mg l−1). February–March and December were found to be the best seasons for culture establishments. Maximum shoots were achieved on MS medium fortified with BAP (2.0 mg l−1). Embryogenic callus (slightly greenish compact, globular, and slow growing) was initiated from the base of severed sprouted buds in 2–3 subsequent subcultures on MS medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) (5.0 mg l−1) under dark incubations. Maturation and germination of well-organized somatic embryos was achieved on MS medium containing BAP and 2,4-D (1.0 mg l−1 each) with 20.0 mg l−1 ascorbic acid. Full-strength MS medium supplemented with 2% glucose favored further development of proliferated somatic embryos into plantlets. Genetic variations of field-established B. nutans plants regenerated through tissue cultures were assessed by amplified fragment length polymorphism (AFLP) analysis using 6 primer combinations. Four hundred and seven scorable fragments were amplified, of which 402 (98.8%) have recorded conservation at various morphogenetic stages leading to plantlets regeneration, therefore, revealed a high level of genetic stability.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxy acetic acid

AFLP:

Amplified fragment length polymorphism

BAP:

6-Benzylaminopurine

Kn:

Kinetin

MS:

Murashige & Skoog (1962) medium

NAA:

α-Naphthaleneacetic acid

PGR:

Plant growth regulator

masl:

Meter above sea level

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Acknowledgments

We are thankful to Director, IHBT for providing necessary facilities. Financial assistance from Department of Biotechnology, Government of India, New Delhi, is gratefully acknowledged. This is IHBT publication 0698.

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Authors and Affiliations

  1. Biotechnology Division, Institute of Himalayan Bioresource Technology, Council of Scientific & Industrial Research, Palampur, 176061, Himachal Pradesh, India

    Rupali Mehta, Vikas Sharma, Anil Sood, Madhu Sharma & Ram Kumar Sharma

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  1. Rupali Mehta
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  2. Vikas Sharma
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  3. Anil Sood
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  4. Madhu Sharma
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  5. Ram Kumar Sharma
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Correspondence to Anil Sood or Ram Kumar Sharma.

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Communicated by R. Matyssek.

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Mehta, R., Sharma, V., Sood, A. et al. Induction of somatic embryogenesis and analysis of genetic fidelity of in vitro-derived plantlets of Bambusa nutans Wall., using AFLP markers. Eur J Forest Res 130, 729–736 (2011). https://doi.org/10.1007/s10342-010-0462-4

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  • DOI: https://doi.org/10.1007/s10342-010-0462-4

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