In Vitro Propagation, Encapsulation, and Genetic Fidelity Analysis of Terminalia arjuna: a Cardioprotective Medicinal Tree

• Published in: Applied biochemistry and biotechnology 2014-07, Vol.173 (6), p.1481-1494
• Main Authors: Gupta, Amit K, Harish, Rai, Manoj K, Phulwaria, Mahendra, Agarwal, Tanvi, Shekhawat, N. S
• Format: Article
• Language: English
• Subjects: Acetic acid; agar; Agricultural biotechnology; artificial seeds; Biochemistry; Biological and medical sciences; Biotechnology; calcium chloride; Cardiotonic Agents - isolation & purification; Cardiovascular disease; Chemistry; Chemistry and Materials Science; DNA Fingerprinting; DNA, Plant - genetics; encapsulation; explants; Fundamental and applied biological sciences. Psychology; Genetic diversity; genetic variation; genotype; germplasm exchange; greenhouses; Growth regulators; Herbal medicine; in vitro regeneration; In Vitro Techniques; indole butyric acid; mature plants; medicinal plants; micropropagation; Naphthalene; naphthaleneacetic acid; Plant growth; Plant Roots - growth & development; Plant Shoots - growth & development; plantlets; Plants, Medicinal - genetics; Plants, Medicinal - growth & development; Plants, Medicinal - physiology; Propagation; random amplified polymorphic DNA technique; Regeneration; rooting; Seeds; Shoots; sodium alginate; temperature; Terminalia - genetics; Terminalia - growth & development; Terminalia - physiology; Terminalia arjuna; Trees
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-014-0920-4

The present study described an improved and reproducible in vitro regeneration system for Terminalia arjuna using nodal segment explants obtained from a mature plant. Shoot tips excised from in vitro proliferated shoots were encapsulated in 3 % sodium alginate and 100 mM CaCl₂[Symbol: see text]2H₂O for the development of synthetic seeds which may be applicable in short-term storage and germplasm exchange of elite genotype. Shoot multiplication was significantly influenced by a number of factors, namely types and concentrations of plant growth regulators, medium composition, repeated transfer of mother explants, subculturing of in vitro regenerated shoot clumps, agar concentrations, and temperature. Maximum numbers of shoots (16.50 ± 3.67) were observed on modified Murashige and Skoog (MMS) medium containing 0.5 mg l⁻¹ of benzylaminopurine (BAP) and 0.1 mg l⁻¹ of naphthalene acetic acid (NAA). To shortening the regeneration pathway, rooting of micropropagated shoots under in vitro condition was excluded and an experiment on ex vitro rooting was conducted and it was observed that the highest percentage of shoots rooted ex vitro when treated with indole-3-butyric acid (IBA, 250 mg l⁻¹) + 2-naphthoxy acetic acid (NOA, 250 mg l⁻¹) for 5 min. The well-developed ex vitro rooted shoots were acclimatized successfully in soilrite under greenhouse conditions with 80 % survival of plants. Randomly amplified polymorphic DNA (RAPD) analysis confirmed that all the regenerated plants were genetically identical to the mother plant, suggesting the absence of detectable genetic variation in the regenerated plantlets. To the best of our knowledge, this is the first report on synthetic seed production as well as ex vitro rooting and genetic fidelity assessment of micropropagated shoots of T. arjuna.