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In vitro regeneration of Aristolochia tagala and production of artificial seeds

  • Original Papers
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Biologia Plantarum

Abstract

Protocols for in vitro plant multiplication from somatic tissues and production of artificial seeds through encapsulation of nodes were developed for Aristolochia tagala Cham., a rare and valuable medicinal plant, as a measure of conservation and as a prerequisite for genetic transformation procedure. A maximum number of adventitious shoots were regenerated from leaf-derived callus on Murashige and Skoog (MS) medium containing 6-benzylaminopurine (BAP; 2 μM), α-naphthaleneacetic acid (NAA; 0.5 μM), and phloroglucinol (PG; 10μM). Nodes collected from in vitro established shoot cultures were encapsulated in 3 % (m/v) sodium alginate and 1 % (m/v) calcium chloride. Multiple shoots were successfully regenerated from the encapsulated nodes cultured on MS medium supplemented with 3 μM BAP and 0.5 μM kinetin (KIN). Regenerated shoots from callus and artificial seeds were successfully rooted and acclimated to greenhouse conditions. Since roots of A. tagala are primarily used in traditional medicine, a protocol for regenerating roots directly from the leaf derived callus was also developed. Maximum root length was obtained when the callus was cultured in MS medium supplemented with KIN (1 μM), indole acetic acid (IAA; 0.5 μM), NAA (0.1 μM), and PG (10 μM). Biochemical parameters were studied in calli grown with and without PG in the medium to establish a correlation between these parameters and shoot morphogenesis. An increment of antioxidant enzymes (peroxidase and catalase) and metabolites (sugars and proteins), and a decrease in the amount of polyphenol oxidase was observed in the calli which were grown in the presence of PG.

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Abbreviations

AC:

activated charcoal

BAP:

6-benzylaminopurine

2,4-D:

2,4-dichlorophenoxyacetic acid

IAA:

indole acetic acid

IBA:

indolebutyric acid

KIN:

kinetin

MS:

Murashige and Skoog

NAA:

α-naphthaleneacetic acid

PG:

phloroglucinol

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Authors and Affiliations

  1. Department of Biotechnology, Vinayaka Missions University, Aarupadai Veedu Institute of Technology, Kancheepuram, 603104, Tamil Nadu, India

    M. Remya

  2. Department of Botany, Bharathiar University, Coimbatore, 641046, Tamil Nadu, India

    V. Narmatha Bai & V. N. Mutharaian

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  1. M. Remya
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  2. V. Narmatha Bai
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  3. V. N. Mutharaian
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Correspondence to M. Remya.

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Acknowledgements: The authors express their sincere thanks to Dr. R. Gopalan, Botanical Survey of India, Coimbatore, Tamil Nadu and Dr. Venkata Subramaniam, Institute Of Forest Genetics and Tree Breeding, Coimbatore, Tamil Nadu, for their valuable assistance in collection and identification of the plant.

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Remya, M., Narmatha Bai, V. & Mutharaian, V.N. In vitro regeneration of Aristolochia tagala and production of artificial seeds. Biol Plant 57, 210–218 (2013). https://doi.org/10.1007/s10535-012-0280-2

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