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High Frequency in Vitro Regeneration of Dianthus Caryophyllus L., a Herbaceous Perennial Ornamental Plant

A. K. M. Sayeed Hassan
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh.

J. L. Munshi
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh.

R. Sultana
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh.

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

R. Khatun
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh.

Abstract/ Executive Summary:

High frequency plant regeneration was established from shoot tips and nodal explants of a perennial ornamental plant, Dianthus caryophyllus L. Best shoot induction was observed on MS basal medium supplemented with 0.5 mg/l BAP + 0.1 mg/l NAA, in which 82% of the explants responded to produce maximum number of shoots (38) per culture. In vitro raised healthy shoots were rooted on half strength MS medium with 0.5 mg/l IBA +0.5 mg/l NAA. For acclimatization and transplantation, the plantlets in the rooting culture tubes were kept in normal room temperature for 7 days before transplanting in pots where plant lets were reared for three weeks. The survival rate of regenerated plantlets was 78%.

Keywords: Dianthus caryophyllus, Shoot proliferation, Micropropagation, Acclimatization

Location: BCSIR

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Ornamental Plant(s)

Objectives:

To develop a reliable repeatable and efficient method for rapid disease free micropropagation of Dianthus caryophyllus L.

Methodology:

The experiment was conducted at Biological Research Division in Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka. Healthy and profusely growing vine of Dianthus caryophyllus L.was collected from the nursery of BCSIR Campus, Dhaka and used as source of explants. Shoot tips and nodal explants 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 cotton and 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 121°C for 20 min. The cultures were incubated for a 16 h photoperiod at 24 ± 2oC under a fluorescent light. Shoot induction and proliferation from Shoot tips and nodal explants was obtained in two separate sets of experiments. In the first experiment 0-2.0 mg/l BAP and 0-2.0 mg/l Kn were incorporated into MS media for shoot induction. In the second set, combination of BAP (0-2.0 mg/l) with NAA (0.1-0.5 mg/l) and BAP (0-2.0 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 every week after 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 trans planted to plastic pots with soil and compost (1: 1) for hardening. The plantlets were kept in a polychamber at 80% relative humidity, 32 ± 2oC under a 12 h photoperiod, 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. 46(4), 495-498, 2011

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In vitro raised healthy shoots were rooted on half strength MS medium with 0.5 mg/l IBA +0.5 mg/l NAA. For acclimatization and transplantation, the plantlets in the rooting 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 78%.

Knowledge Management: Journal