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Large Scale Plant Regeneration in Vitro from Leaf Derived Callus Cultures of Pineapple [ananas Comosus (l.) Merr. Cv. Giant Kew]

M. N. Amin
Professor
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

M. M. Rahman
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

K. W. Rahman
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

R. Ahmed
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

M. S. Hossain
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

M. B. Ahmed
Department of Botany, University of Rajshahi, Rajshshi 6205, Bangladesh.

Abstract/ Executive Summary:

The leaf base explants from the in vitro established shoot cultures were induced to form callus and subsequently to differentiate into shoots on MS medium supplemented with different concentrations and combinations of cytokinins and auxins. The cultured explants produced calli from their cut margins within four weeks of incubation on media supplemented with 0.5-3.0 mg L^-1 2,4-D alone and in combination with 0.5-3.0 mg L^-1 BA. Maximum number of shoot buds with optimum callus growth was observed on MS medium containing 1.0 mg L^-1 BA and 0.1 mg L^-1 NAA after six weeks of culture. Rooting was induced in the in vitro regenerated shoots on 2 MS medium with different concentrations and combinations of NAA, IBA or IAA. Rooting performance was best when the microshoots were rooted on 2 MS medium containing 0.2 mg L^-1 IBA + 0.2 mg L^-1 NAA. The regenerated plant lets were successfully transferred to soil and percentage of their survivability under ex vitro condition was almost ninety.

Keywords: Plant regeneration, In vitro, Callus cultures, Pineapple

Location: Department of Botany, University of Rajshahi, Bangladesh.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Pineapple

Objectives:

To establish a suitable in vitro plant regeneration protocol for pineapple cv. Giant Kew through callus culture from leaf base explant.

Methodology:

The creamy-white leaf base segments were excised aseptically from in vitro grown cultures, originally established from the crown tip explants of field grown mature plant (cf.). Excised leaves were cut into pieces (1-1.5 mm) from basal site and placed on the medium. The explants were initially cultured on MS medium supplemented with various concentrations of either 2, 4-D alone or in combination with BA for callus induction. Four-week old calli were subcultured on MS medium containing different combinations of BA with NAA, IBA or IAA for shoot differentiation. The calli were sub cultured at monthly intervals on same medium for growth, elongation and proliferation of shoots rapidly. The shoots having more than 4 cm length (from base to leaf) were harvested from the proliferating cultures and cultured individually for rooting on ₂ MS (half strength of both major and minor salts) medium supplemented with NAA, IBA, IAA. The pH value of all media was adjusted to 5.7±0.1 before addition of agar and sterilized by autoclaving for 20 minutes at 1.1 Kg/cm^-2pressure at 121°C. For solidifying the medium, 7-8 gm L^-1 agar was used as gelling agent. The tubes or flasks containing explants and proliferating cultures were incubated on culture racks in the growth chamber. The cultures were maintained at 25±2°C under the cool white fluorescent lights for 16 h photo period with a photon flux density of about 60 μ mol.m^-2 s^-1.

Source of Information: International Journal of Botany, 1(2): 128-132, 2005, ISSN 1811-9700.

Article Link (Online Journal): DOI: 10.3923/ijb.2005.128.132; URL: http://scialert.net/abstract/?doi=ijb.2005.128.132

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Three auxins were used at different concentrations and combinations to induce and differentiate adventitious roots. Among the three auxins, IBA and NAA were found to show comparatively better response than IAA for production of roots. Reasonably better rooting was observed on 2 MS medium having combined auxins than single auxin. The best rooting was observed in 2 MS medium supplemented with 0.2 mg L^-1 IBA + 0.2 mg L^-1 NAA where 100 percent of micro shoots produced root and number of roots per micro shoots was 6.50±0.22. The regenerated micro shoots of pineapple produced roots on MS medium supplemented with different types of auxins and a cytokinin, BA. However, there are some other reports on root induction of pineapple micro shoots on half strength of MS medium supplemented with different auxins viz. IBA and NAA. The complete plant lets thus produced from the in vitro regenerated micro shoots were transferred to soil. On an average 90% survival of the regenerated plant lets of pineapple was achieved in the present study.

Knowledge Management: Journal