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In Vitro Mass Propagation of Plumbago Zeylanica L., Through Direct Organogenesis

A. K. M. S. Hassan
Department of Botany, Savar College, Savar, Dhaka-1340,

F. Haque
Biological Research Division, BCSIR Laboratories, Dhaka-1205

M. A. A. Jahan
Biological Research Division, BCSIR Laboratories, Dhaka-1205

S. K. Roy
Department of Botany, Jahangirnagar University,Savar, Dhaka-1342, Bangladesh

Abstract/ Executive Summary:

An efficient protocol was developed for in vitro mass propagation of an important medicinal shrub, Plumbago zeylanica L., (Plumbaginaceae) through direct organogenesis using shoot tip and nodal explants. Best shoot induction was observed on MS basal medium supplemented with 0.5 mg/l BAP, in which 86.4% of nodal explants responded to produce maximum number (12.4 ± 0.66) of shoots per culture. In vitro raised shoots rooted on half strength MS medium with 0.5 mg/l IAA. For acclimatization and transplantation, the plantlets in the rooted culture tubes were kept in normal room temperature for 7 days before transplanting in pots where plantlets were reared for three weeks. The survival rate of regenerated plantlets was 85%.

Keywords: Plumbago zeylanica, Medicinal plant, Organogenesis, Mass propagation, Acclimatization

Location: BAU, Mymensingh

Start Date:

End Date:

Program Area: Variety and Species

Commodity:

Objectives:

To develop a protocol for in vitro mass propagation of this important medicinal shrub through direct organogenesis using shoot tip and nodal explants.

Methodology:

Plumbago zeylanica L., grown at Medicinal Plants Garden of Bangladesh Agricultural University, Mymensingh was used as a source of explants. Shoot tip and nodal explants with a single axillary bud were used for this purpose. The explants were washed thoroughly under running tap water, pre-soaked in liquid detergent for about 30 min, wiped with cottonand dipped in 70% (v/v) ethanol for 1 min. They were then surface-sterilized with 0.1% (w/v) mercuric chloride for 5 min, followed by five times rinse with sterile distilled water under laminar air flow cabinet. The surface-sterilized explants were sized to 1-1.5 cm length containing a single node with an axillary bud or a shoot tip with an apical bud. The explants were placed vertically on the culture medium. The new shoots induced from the in vitro cultures were further used as an explants for adventitious shoot regeneration. MS (Murashige and Skoog, 1962) basal medium was used for shoot proliferation and adventitious shoot regeneration and half strength MS was used for in vitro root induction. All media were supplemented with 30 g/l sucrose, 7 g/l agar (Difco) and dispensed into 15x150 mm culture tubes and 250 ml conical flasks. The pH of the media was adjusted to 5.8 before autoclaving at 1.9 kg/cm² pressure at 121°C for 20 min. The cultures were incubated for a 16 h photoperiod at 24 ± 2^oC under 1200 lux/m² fluorescent light. Shoot proliferation from shoot tip and nodal explants was obtained in two separate sets of experiments. In the first experiment 0.5-2.0 mg/l BAP and 0.5-2.0 mg/l Kn were incorporated into MS media to select the best cytokinin for the response of shoot induction. In the second set, combination of BAP (0.5-2.0 mg/l) with NAA (0.1-0.5 mg/l) and BAP (0.5-2.0 mg/l) with IAA (0.1-0.5 mg/l) were assessed for shoot multiplication. Number of new shoot proliferation of each culture was recorded after every week of inoculation. For in vitro rooting, individual shoots (3-5 cm) were excised from the proliferated shoot cultures and implanted onto half strength MS media with different concentrations and combinations of NAA, IBA and IAA. The rooted plants were taken out from the culture tubes, washed to remove agar gel adhered to the roots and transplanted to plastic pots with soil and compost (1: 1) for hardening. The plantlets were kept in a polychamber at 80% relative humidity, 32 ± 2^oC temperatures for a 12 h photoperiod under 1500 lux/m² sun light for acclimation. Established plants were transplanted in earthen pots under natural conditions and the survival rate was recorded

Source of Information: Bangladesh J. Sci. Ind. Res. 47(3), 297-302, 2012

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Best shoot induction was observed on MS basal medium supplemented with 0.5 mg/l BAP, in which 86.4% of nodal explants responded to produce maximum number (12.4 ± 0.66) of shoots per culture. In vitro raised shoots rooted on half strength MS medium with 0.5 mg/l IAA. For acclimatization and transplantation, the plantlets in the rooted culture tubes were kept in normal room temperature for 7 days before transplanting in pots where plantlets were reared for three weeks. The survival rate of regenerated plantlets was 85%.

Knowledge Management: Journal