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In Vitro Regeneration of Okra (abelmoschus Esculentus L Moench.) (2015-2016)

M. R. Kabir
Horticulture Research Centre, BARI, Joydebpur, Gazipur, Bangladesh

S. Ahmed
Horticulture Research Centre, BARI, Joydebpur, Gazipur, Bangladesh

M. A. Y. Akhond
Horticulture Research Centre, BARI, Joydebpur, Gazipur, Bangladesh

Abstract/ Executive Summary:

Seedling-derived cotyledonary nodes and hypocotyl explants of BARI Dherosh-1 were cultured in vitro on MS medium supplemented with varying concentrations of 2,4-D, BAP, TDZ, BAP with NAA, BAP IAA and Zeatin with IAA along with a control. Shooting response (100%) with callus was only observed from cotyledonary nodes on TDZ where hypocotyls produced only callus or callus with roots on different concentrations of plant growth regulators. Considering the shooting response, the cotyledonary nodes of were cultured on various concentrations of TDZ for regeneration. The highest percentage (64.0) with maximum number (6.8) of shoots per explant were observed in 0.044 µM TDZ in 8.4 days. The regenerated shoots were rooted on ½ strength MS, MS supplemented with 2.46 µM IBA and 0.53 µM NAA. The highest percentage (83.3) and minimum days (9.7) required for root induction were recorded in 2.46 µM IBA. The rooted plantlets were transferred to soil and hardened in the plastic pots under green house conditions.

Keywords: Okra, regeneration, Transformation

Location:

Start Date: 00-00-2015

End Date: 00-00-2016

Program Area: Variety and Species

Commodity: Okra

Objectives:

To develop a simple and efficient regeneration protocol of okra aiming at the future genetic transformation.

Methodology:

BARI Dherosh-1 seeds were collected from Horticulture Research Centre, Bangladesh Agricultural Research Institute, Gazipur. Clean and healthy seeds were washed with 100% ethanol and sterilized for 20 minutes in 1% (v/v) sodium hypochlorite containing 3-4 drops Tween20 in a laminar airflow cabinet. Afterward, they were rinsed 4 times with sterile distilled water and transferred to ½ strength MS medium (Murashige and Skoog, 1962). The pH of the medium was adjusted to 5.8 before autoclaving at 121°C, 15 psi for 20 min. Cotyledonary nodes and hypocotyls were cultured on MS medium supplemented with varying concentrations of 2,4-D (2.26, 4.52, 6.78 and 9.04 µM), BAP (2.22, 4.44, 6.66 and 8.88 µM), TDZ (0.022, 0.044, 0.22, 0.45 and 2.25 µM), BAP and NAA in combinations (4.44+0.53, 8.88+0.53 µM), BAP and IAA in combinations (4.44+0.57, 8.88+0.57 µM), Zeatin and IAA in combinations (4.56+0.57, 9.12+0.57 µM) along with a control treatment without any growth regulators. Based on the response of two different types of explants the cotyledonary nodes were cultured on media having five different concentrations of TDZ for further regeneration. Explants were subcultured in every 14 days on the same media and kept under same conditions. When shoots attained a height of 2.0-2.5 cm they were cleaned, excised and transferred to ½ strength of MS, MS supplemented with 2.46 µM IBA and 0.53 µM NAA for root induction. Well-developed rooted shoots were transferred to pots with sterile soil mix and enclosed with polythene bags to maintain high humidity. The plantlets were kept in the greenhouse and watered once or twice a week while keeping covered. After 2 weeks, the bags were removed and the plantlets were transferred to the field for growing to maturity. The experiment was set up in a completely randomized design (CRD) with three independent replicates. The analysis of variance for different parameters was performed and the means were compared by R programme using STAR software at a 5% level of significance.

Source of Information: Annual Research Report 2015-2016, Horticulture Research Centre, BARI, Joydebpur, Gazipur, Bangladesh

Article Link (Online Journal):

Funding Source:

1.

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The development of a highly efficient regeneration protocol based on using cotyledonary nodes as explants for the induction of multiple shoots from 0.044 µM TDZ in okra is described here. The time taken from culture initiation to the establishment of plants in the greenhouse was shorter (about 3 months) as compared with the longer periods reported in previously published regeneration protocols. The important feature of the present protocol is a shortening of regeneration time, as well as the induction of high number of multiple shoots per explant. Besides, this stable and reliable regeneration protocol is expected to be helpful for future research on genetic transformation on okra.

Knowledge Management: Report/Proceedings