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In Vitro Plantlets Regeneration of Rose

Protul Kumar Roy
Plant Biotechnology and Genetic Engineering Division, Institute of Food and Radiation Biology, Atomic Energy Research Establishment (AERE), GPO Box-3787, Dhaka-l000, Bangladesh

A. N. K. Mamun
Plant Biotechnology and Genetic Engineering Division, Institute of Food and Radiation Biology, Atomic Energy Research Establishment (AERE), GPO Box-3787, Dhaka-l000, Bangladesh

Golam Ahmed
Plant Biotechnology and Genetic Engineering Division, Institute of Food and Radiation Biology, Atomic Energy Research Establishment (AERE), GPO Box-3787, Dhaka-l000, Bangladesh

Abstract/ Executive Summary:

Nodal segments and shoot tips were cultured on MS supplemented with different concentrations and combinations of BAP, NAA and zeatin for multiplying axillary shoots. Among two types of explants, the nodal explants produced the highest number of shoots when they were cultured on MS + 1.5 mg/l BAP + 0.25 mg/l zeatin + l00 mg/l CH. The number of shoots per explant in initial cultures varied from five to seven. But after three weeks when they were subcultured in the same medium, the number of shoots increased with an average of 15 per culture. Shoot tip explants also produced multiple shoots, but their performance was not as good as nodal explants. For best rooting, 34 cm long shoots were excised and implanted individually onto the rooting medium containing half strength MS fortified with 1.0 mg/l IBA and 0.5 mg/l IAA. Within four weeks of transfer to the rooting medium, 85% of the microcuttings produced seven - ten roots. The regenerated plantlets were successfully transferred to soil and the percentage of survivability under ex vitro condition was 80.

Keywords: Plantlet regeneration, Nodal segments, Shoot tips, Rose

Location: Institute of Food & Radiation Biology, Atomic Energy Research Establishment, Savar, Dhaka.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Rose

Objectives:

To establish an efficient and reproducible method for rapid and large scale propagation of Rosa spp.

Methodology:

Shoot tips and nodal segments were taken from garden grown rose plants in the Institute of Food & Radiation Biology, Atomic Energy Research Establishment, Savar, Dhaka. These were washed thoroughly with running tap water for one h; thereafter they were kept in a beaker for 30 min in order to remove dirt on the stem surface. Next, these were cut into small pieces and surface sterilized with aqueous solution of 0.1 % HgCl₂ for six min under aseptic condition and rinsed five times with autoclaved distilled water to wash away the traces of HgCl₂ completely. Shoot tips and nodal segments, approximately 2.0 cm in length were cut from sterilized stem for explants. MS basal medium was used for regeneration of plantlets through in vitro culture. Shoot tips and nodal segment explants were cultured on MS supplemented with different concentrations of cytokinin (BAP, Kn, zeatin) and auxin (NAA) singly or in combination for shoot regeneration. Half strength MS supplemented with auxin such as IBA, IAA and NAA were used for root formation. Casein hydrolysate (CH) was used in the media for shoot multiplication. The pH of the media was adjusted to 5.8 before adding agar. All media were gelled with 0.6% agar. The cultures were incubated at 26 ± 1°C under cool white fluorescent light for a daily 16 h photoperiod. Subculturing was done every three weeks. In vitro rooted plantlets were taken out from the test tubes and gently washed to free agar from surface. In vitro rooted plantlets were then transferred to small earthen pots containing a mixture of soil, sand and compost (2 : 1 : 1) and covered with transparent polyethylene bag.

Source of Information: Plant Tissue Cult. 14 (2) : 149-154, 2004 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The best response in shoot regeneration and multiplication was observed, when the culture was initiated during the winter season. Seasonal variation of in vitro shoot induction was also reported in many other plant materials. For further development of the medium and enhanced shoot proliferation, casein hydrolysate (CH) was added (75 - 150 mg/l) to the medium. Addition of l00 mg/l CH to the medium increased the number of shoots (15 and 10 in case of nodal segments and shoot tips, respectively) per culture. Best response was observed when 1.0 mg/l IBA and 0.5 mg/l IAA were added to half strength MS. In this combination, it was observed that 85% shoots rooted within four weeks of culture and each microcutting produced seven - ten roots. 0.2 mg/l IBA and 0.2 mg/l IAA was best combination with half strength MS for root formation in Chrysanthemum morifolium. For hardening and plant establishment under the natural conditions, the well rooted plantlets were transferred to small earthen pots containing soil, sand and compost (2 : 1 : 1). During hardening 80% plantlets survived and those were subsequently transferred to field. The results of the present investigation demonstrate a reproducible and efficient regeneration protocol of in vitro micropropgation of Rosa spp.

Knowledge Management: Journal