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Callus Culture and Plant Regeneration of Tomato from Leaf Explant

K. Khatun
Department of Biotechnology,Bangladesh Agricultural University, Mymensingh

M.S. Haque
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh

Abstract/ Executive Summary:

The effects of different concentrations and combinations of plant growth regulators (BAP, NAA, and IBA) were studied to develop a protocol for efficient in vitro regeneration of tomato. Regeneration ability of two tomato (BINA tomato 4 and 5) cultivars were investigated via callus culture of leaf as explants. Explants were first cultured on MS medium supplemented with different concentrations and combinations of NAA and BAP for callus induction. Among the combinations used BAP at 3.0 mgl^-1and NAA at 0.2 mgl^-1exhibited best performance of 88.00% callus initiation from leaf. Shoots were induced from callus cultured on a medium supplemented with different concentrations and combinations of both BAP and NAA. Maximum percentages of shoot formation from leaf calli were 91.00 % on MS medium supplemented with BAP at 4.0 mgl^-1and NAA at 2.0 mgl^-1. The regenerated shoots developed roots on MS medium supplemented with different concentrations of IBA (0.0, 0.5, 1.0, 1.5, 2.0 and 2.5 mgl^-1). The best IBA concentration for rooting was 2.0 mgl^-1with a rooting frequency of 97.67%.

Keywords: Callus, Regeneration, NAA, BAP, IBA, Explants tomato

Location: Mymensingh

Start Date: 00-06-2008

End Date: 00-11-2008

Program Area: Variety and Species

Commodity: Tomato

Objectives:

To develop a protocol for efficient in vitro regeneration of tomato

Methodology:

An experiment was carried out in the Biotechnology Laboratory of the Bangladesh Agricultural University, Mymensingh, during the period from June 2008 to November 2008. Two tomato varieties namely BINA tomato 4 and BINA tomato 5 were used as experimental materials. Leaf of two tomato varieties were used as explants for callus inoculation and plant regeneration. The seeds were washed in distilled water for 3-5 minute then sterilized with 70% ethanol for 5 minutes followed by sodium hypochloride for 12 minutes. The explants were then washed five times with sterilized distilled water. Sterilized seeds were germinated on MS medium without hormone. The pH of all of the tissue culture media used in this study was adjusted to 5.8 before autoclaving at 121^oC for 30 minutes. Leaf segments of 2 to 3 mm in length were cut from four-day-old germinated seedlings and were placed on callus induction medium which was same as basal medium except for the addition of 1, 2, 3 and 4, mgl^-1BAP with 0.1, 0.2, 0.3 and 0.4 mgl^-1NAA. Basal medium cultured on MS medium containing with different concentrations and combinations of BAP (0.0, 1.0, 2.0, 3.0, and 4.0 mgl^-1) and NAA (0.0, 0.1, 0.2, 0.3 and 0.4 mgl^-1)was used as basal medium. Calli were initiated after17 days of inoculation and after 4 weeks of initiation, calli were transferred on fresh medium. Subculture was carried out at 21 days interval and finally transferred to MS medium with different concentrations of BAP (0.0, 1.0, 2.0, 3.0, 4.0 and 5.0 mgl^-1) and NAA (0.0, 0.1, 0.2, 0.3 and 0.4 mgl^-1) for shoot regeneration. The regenerated shoots produced roots in MS medium containing (0.0, 0.5, 1.0, 1.5, 2.0, and 2.5 mgl^-1 IBA). All cultures were maintained at 26±1^OC under white fluorescent tubes with a 16h photoperiod.

Source of Information: Bangladesh J. Seed Sci. Technol. 13(1&2): 57–60 (2009),ISSN 1029-8800

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The experiment found that BAP at 3.0 mgl^-1and NAA at 0.2 mgl^-1is best for callus induction whereas BAP at 4.0 mgl^-1and NAA at 2.0 mgl^-1is more appropriate for shoot induction and IBA at 2.0 mgl^-1 resulted highest rooting for in vitro regeneration of tomato from leaf explants.

Knowledge Management: Journal