MilliporeSigma
  • Direct and indirect method of plant regeneration from root explants of Caesalpinia bonduc (L.) Roxb.--a threatened medicinal plant of Western Ghats.

Direct and indirect method of plant regeneration from root explants of Caesalpinia bonduc (L.) Roxb.--a threatened medicinal plant of Western Ghats.

Indian journal of experimental biology (2013-08-31)
S R Santosh Kumar, V Krishna, Venkatesh, K Pradeepa, K Girish Kumar, A U Gnanesh
ABSTRACT

An in vitro regeneration protocol has been standardized via direct and indirect methods from excised root explants of C. bonduc, a threatened woody legume used for the treatment of contagious diseases, inflammation, leprosy, antiperiodic, febrifuge, anthelmenthic, urinary disorders, leucorrhoea, piles and to heal wounds. MS medium supplemented with 17.75 micromol BAP and 2.46 micromol IBA, induced a mean of 3.40 +/- 1.07 shoots directly from the surface of excised root explant. Subsequently, the shoots rooted readily on MS half strength medium with out growth regulators. In indirect organogenesis, callogenic frequency was optimized (96.66%) at the concentration of 9.04 micromol 2, 4-D and 0.88 micromol BAP. An average, 15.30 +/- 5.25 shoots were differentiated from the root callus at the concentration of 17.57 micromol BAP and 2.85 micromol IAA. Shoots regenerated through callus were rooted well on MS half strength medium with growth regulators at 2.95 micromol IBA. Rooted plantlets were transferred to the pots containing sterilized soil and were successfully hardened at greenhouse condition for three weeks then exposed to the natural environment. Survival rate was more (95%) in plantlets derived through direct organogenesis than (60%) the plantlets regenerated through root calli.

MATERIALS
Product Number
Brand
Product Description

Supelco
2,4-D, PESTANAL®, analytical standard
Sigma-Aldrich
Kollidon® 25
Povidone, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Adenine, ≥99%
Sigma-Aldrich
Adenine, BioReagent, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Adenine, BioReagent, suitable for cell culture
Supelco
2,4-D, PESTANAL®, analytical standard
Sigma-Aldrich
3-Indoleacetic acid, suitable for plant cell culture, crystalline
Sigma-Aldrich
Povidone, meets USP testing specifications
Supelco
3-Indoleacetic acid, PESTANAL®, analytical standard
Sigma-Aldrich
6-Benzylaminopurine, suitable for plant cell culture
Sigma-Aldrich
6-Benzylaminopurine, ReagentPlus®, ≥99.0% (HPLC)
Sigma-Aldrich
3-Indoleacetic acid, 98%
Supelco
2,4-D solution, certified reference material, 5000 μg/mL in acetonitrile
Supelco
2,4-D, certified reference material, TraceCERT®
Supelco
6-Benzylaminopurine, PESTANAL®, analytical standard
Adenine, European Pharmacopoeia (EP) Reference Standard
Supelco
Povidone, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
2,4-Dichlorophenoxyacetic acid, ≥95%, crystalline
Sigma-Aldrich
Polyvinylpyrrolidone, average mol wt 10,000
Sigma-Aldrich
Polyvinylpyrrolidone, suitable for plant cell culture, average mol wt 10,000
Sigma-Aldrich
Polyvinylpyrrolidone, for molecular biology, nucleic acid hybridization tested, mol wt 360,000
Sigma-Aldrich
Polyvinylpyrrolidone, average mol wt 40,000
Sigma-Aldrich
Polyvinylpyrrolidone, powder, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
Polyvinylpyrrolidone, mol wt (number average molecular weight Mn 360kDa)
Sigma-Aldrich
Polyvinylpyrrolidone, powder, average Mw ~29,000
Sigma-Aldrich
Polyvinylpyrrolidone, powder, average Mw ~55,000
Sigma-Aldrich
Polyvinylpyrrolidone, K 30
Sigma-Aldrich
Polyvinylpyrrolidone solution, K 60, 45% in H2O
Sigma-Aldrich
Polyvinylpyrrolidone, K 90