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Effects of Plant Growth Regulators on Callus Proliferation, Plantlet Regeneration and Growth of Plantlets Dooritaenopsis Orchid

Israt Chowdhury
Department of Crop Botany, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.

Abu Reza Md. Mahfuzur Rahman
Department of Crop Botany, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.

M. Obaidul Islam
Department of Crop Botany, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.

S. Matsui
Department of Plant Production, Faculty of Agriculture, Gifu University, Gifu-shi, 501-1193, Japan.

Abstract/ Executive Summary:

Supplement of 2, 4-D and BAP into New Phalaenopsis (NP) medium was not effective for the growth of callus in Doritaenopsis orchid. Among the different combinations of BAP and NAA, 5.0 mg l^-1 BAP + 0.1 mg l^-1 NAA highly enhanced PLB formation from calli. For plantlet initiation from PLBs 0.5 mg l^-1 BAP was suitable and higher concentration was inhibitory. Similar effect to that of BAP was found for NAA (0.5 mg l^-1) and BAP + NAA (0.5 mg l^-1 + 0.5 mg l^-1) in plantlet initiation. For in vitro growth of plant lets, 0.5 mg l^-1 of BAP was the best for leaf and shoot growth compared to that of for BAP and NAA. However, for root growth of plant lets 1.0 mg l^-1 BAP + 1.0 mg l^-1 NAA was the most suitable.

Keywords: Orchid,Callus growth, Plantlet regeneration, Doritaenopsis orchid, Plant growth regulator

Location: Department of Crop Botany, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Orchid

Objectives:

To investigate the effects of different plant growth regulators on callus growth, PLB initiation from callus, plantlet regeneration and the subsequent growth of plant lets in Doritaenopsis orchid.

Methodology:

Embryogenic calli of Doritaenopsis which were developed earlier by culturing flower stalk were maintained by sub culturing monthly on NP medium supplemented with 20 g l^-1 of sucrose. At the beginning, 1.0 g of source callus was sub cultured twice on 20 ml of NP medium supplemented with 20 g l^-1 of sucrose in 50 ml flasks at monthly intervals to obtain homogeneous callus. Uniform, friable, translucent, yellowish callus was used as plant material. Two cultures were performed in callus culture, the first culture (Ist 8 weeks) and second culture (2nd 8 weeks) have been indicated as 8 weeks culture and 8 weeks x 2 culture respectively in the subsequent results and discussion. In the first culture, 0.2 g of homogeneous callus was cultured on 20 ml of NP medium supplemented with 2, 4-D + BAP for 8 weeks to evaluate their effects on callus growth. For the second culture, 0.2 g of proliferated callus obtained from the first culture was cultured again on the same fresh medium for another 8 weeks with same treatments. In separate cultures, effects of NAA and BAP and PLB initiation from callus were investigated. PLBs obtained from second culture of callus were cultured on 20 ml NP medium supplemented with BAP and NAA for 10 weeks to investigate their effects on plantlet initiation from PLBs. Larger than 2 mm sized, uniform 8 PLBs were used in culture in each flask. Mini plant lets regenerated from PLBs were culture for 12 weeks on 20 ml NP medium with same supplements used in PLB culture to evaluate their effects on growth of plant lets. The basal medium (BM) was New Phalaenopsis (NP) medium. Sucrose 20 g l^-1 and gelrite 3 g l^-1 were added and the pH was adjusted at 5.6 prior to autoclaving. After dissolving the solidifier, 20 ml of hot medium was dispensed into 100 ml conical flasks. The conical flasks containing the medium were autoclaved at 121oC with 1.16 kg cm-2 of pressure for 20 minutes. After autoclaving the flasks containing the medium were allowed to cool and explants were cultured. Cultures were maintained in a growth room and allowed to grow at 25±1oC under 16 h photo period of 50 μ mol m^-2s^-1 illuminated with fluorescent tubes. After each culture, proliferated callus or PLB were weighed and the number of initiated PLBs was counted. Data were also recorded on different parameters to estimate growth of plant lets.

Source of Information: Biotechnology, 2(3): 214-221, 2003, ISSN 1682-2978.

Article Link (Online Journal): DOI: 10.3923/biotech.2003.214.221, URL: http://scialert.net/abstract/?doi=biotech.2003.214.221

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

It might be concluded that ,supplement of PGRs into BM is not effective for callus growth of Doritaenopsis. For PLB production from callus, 5.0 mg l^-1 BAP + 0.1 mg l^-1 NAA is suitable. Plant lets is effective at 0.5 ml-1 of BAP for the subsequent growth of plant lets in vitro 0.5 mg l^-1 BAP or 1.0 mg l^-1 BAP + 1.0 mg l^-1 NAA to be supplemented into BM regeneration from PLB.

Knowledge Management: Journal