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Induction of somatic embryogenesis in endangered butterfly ginger Hedychium coronarium J. Koenig.

Indian journal of experimental biology (2013-08-31)
Manju Verma, Y K Bansal
ABSTRACT

An efficient protocol has been developed for regeneration of complete plants through somatic embryogenesis in H. coronarium. Creamish white, pale yellow and brown calli were obtained on MS medium supplemented with different concentrations of auxins [2, 4-Dichlorophenoxy acetic acid (2, 4-D), Indole-3 acetic acid (IAA) and 1-Naphthylacetic acid (NAA)] after 4 weeks. Creamy white calli developed on 0.5 mg L(-1) 2, 4-D turned embryogenic when subcultured on basal medium and produced small globular somatic embryos after 6 weeks. Further growth of somatic embryos required their transfer to medium containing 6-benzylaminopurine (BAP) or kinetin (KN). BAP was more effective than KN in promoting shoot proliferation. Maximum shoot length was obtained with 0.5 mg L(-1) BAP whereas maximum shoot number was obtained with 1.0 mg L(-1) BAP. The plantlets thus formed were successfully hardened, and transferred to sand-soil and farm yard manure (1:1:1) with 95% survival.

MATERIALS
Product Number
Brand
Product Description

Supelco
2,4-D, certified reference material, TraceCERT®
Supelco
6-Benzylaminopurine, PESTANAL®, analytical standard
Supelco
2,4-D, PESTANAL®, analytical standard
Sigma-Aldrich
1-Naphthaleneacetic acid, suitable for plant cell culture, BioReagent, ≥95%, crystalline
Sigma-Aldrich
3-Indoleacetic acid, suitable for plant cell culture, crystalline
Supelco
2,4-D, PESTANAL®, analytical standard
Sigma-Aldrich
6-Benzylaminopurine, suitable for plant cell culture
Sigma-Aldrich
1-Naphthaleneacetic acid, technical grade
Supelco
2,4-D solution, certified reference material, 5000 μg/mL in acetonitrile
Sigma-Aldrich
3-Indoleacetic acid, 98%
Sigma-Aldrich
6-Benzylaminopurine, ReagentPlus®, ≥99.0% (HPLC)
Supelco
3-Indoleacetic acid, PESTANAL®, analytical standard
Supelco
1-Naphthaleneacetic acid, PESTANAL®, analytical standard
Sigma-Aldrich
1-Naphthylacetic acid, 1 mg/mL, BioReagent, suitable for plant cell culture
Sigma-Aldrich
2,4-Dichlorophenoxyacetic acid, ≥95%, crystalline
Sigma-Aldrich
2,4-Dichlorophenoxyacetic acid, 97%