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Callus Induction and Plant Regeneration from Internodal and Leaf Explants of Four Potato (solanum Tuberosum L.) Cultivars

Farzana Shirin
Plant Breeding and Gene Engineering Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

M. Hossain
Plant Breeding and Gene Engineering Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

M. F. Kabir
Plant Breeding and Gene Engineering Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

M. Roy
Plant Breeding and Gene Engineering Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

S.R. Sarker
Plant Breeding and Gene Engineering Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

Abstract/ Executive Summary:

The present investigation was conducted to develop a protocol for rapid callus induction and plant regeneration of potato. The internodal and leaf explants of four potato (Solanum tuberosum L.) cultivars viz. Diamant, Multa, Atlus and Lalpakri were cultured for callus induction and plant regeneration. The explants were cultured onto MS media supplemented with different concentrations and combinations of 2, 4-D, NAA alone and NAA with BA for callus induction The highest 80% internodal explants of Atlus induced to develop callus in medium containing 3.0 mg L^-1 2, 4-D. Of all cultivars the callusing response of both types of explants was the best in 3.0 mg L^-1 2, 4-D containing MS media. MS medium supplemented with different concentrations and combinations of BA, NAA and KIN were employed for shoot regeneration. MS medium containing 4.0 mg L^-1 KIN + 0.5 mg L ^-1 NAA was the best for maximum shoot regeneration from the internode and leaf derived calli in most of the cultivars. The regenerated shoots were rooted in MS_o medium and successfully transplanted to the field.

Keywords: Callus induction, Plant regeneration, Cultivar, Potato

Location: Plant Breeding and Gene Engineering Laboratory, Department of Botany, University of Rajshahi, Bangladesh

Start Date: 00-12-2004

End Date: 00-11-2005

Program Area: Variety and Species

Commodity: Potato

Objectives:

To establish an effective protocol for rapid plant regeneration from internodes and leaf explants derived callus of potato cultivars cultivated to a great extent in Bangladesh.

Methodology:

The research work was carried out during the period from December, 2004 to November 2005 at the Plant Breeding and Gene Engineering Laboratory, Department of Botany, University of Rajshahi, Bangladesh to evaluate the response of potato genotypes cultivated widely throughout the country. Internode and leaf segments of four in vitro grown potato cultivars viz Diamant, Multa, Atlus and Lalpakri were used for callus induction and plant regeneration. MS semi-solid media with different concentration and combination of auxin (2, 4-D, NAA) and cytokinin (BA, kinetin) were used for this purpose. In each treatment 10 explants were inoculated. Explants were cultured in culture tubes containing MS medium (30 g L^-1 sucrose, 7 g L^-1 agar). The pH of the medium was adjusted to 5.8 by using 1N NaOH. The media were autoclaved at 121°C for 20 min after adjusting the pH. The explants were incubated on callus induction medium at 25±2°C for 3-6 weeks. The calli were transferred to the fresh callus inducing medium about 21 days interval for further proliferation and maintenance. After 40-60 days of incubation in the dark, the callus induction frequency was determined and the well developed calli were selected and sub-cultured on regeneration media. MS medium was supplemented with different concentration and combination of KIN, BA and NAA for shoot regeneration. The cultures were incubated at 25±2°C under white light (2500-3000 lux). After 4-6 weeks differentiation shoot formation was observed. The shoots were excised and transferred to MS_o (without growth regulators) medium for root induction. Percentage of explants induced callus proliferation, callus color, degree of callus formation, percentage of calli regenerated shoots and No. of shoots per callus were recorded to investigate the effect of different treatments and response of different genotypes.

Source of Information: World Journal of Agricultural Sciences 3 (1): 01-06,2007, ISSN 1817-3047

Article Link (Online Journal): http://www.idosi.org/wjas/wjas3(1)/1.pdf

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In this investigation it was observed that auxin alone and in combination with cytokinin can produce callus but 2, 4-D was the most effective for callus induction and proliferation. Rapid shoot regeneration was achieved when the calli were sub-cultured in KIN with NAA supplemented MS medium. The regenerated shoots were excised and individually transferred onto semi solid MS_o rooting medium. Cent percent shoots were induced to develop root within 2 weeks of culture. The plantlets were finally and successfully transferred to the field and grown to maturity. The present study describes an efficient method for in vitro regeneration of potato cultivars which could be considered for large scale multiplication and propagation of this important vegetable crop.

Knowledge Management: Journal