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In Vitro Callus Initiation and Regeneration of Potato

A. U. HAQUE
Scientific Officer
Tuber Crops Research Centre, BARI, Joydebpur, Gazipur

M. A. SAMAD
Senior Scientific Officer
Bangladesh Institute of Nuclear Agriculture, Mymensingh

T. L. SHAPLA
Assistant Professor
Sher-e-Bangla Agricultural University, Dhaka-1206, Bangladesh

Abstract/ Executive Summary:

The first experiment involving different explants and concentrations of 2,4-D and kinetin showed highly significant differences for length and weight of callus formed except interaction of callus weight. Leaf explant appeared to be best of all for callus length and weight when 1.0 mg/L 2,4-D + 0.25 mg/L kinetin concentration was used. Similarly, different explants versus different concentrations of BAP/GA₃/IAA showed significant differences for shoot length and leaf number per plantlet and also for root length. However, interaction term confirmed node and node/internode explants produced better results in shoot length and number of leaves per plantlet when concentrations 1 .0 mg/L BAP + 0.1 mg/L GA₃and 1.0 mg/L BAP + 0.2 mg/L GA₃, 1.0 mg/L BAP + 0.4 mg/L GA₃, respectively, were used. Similarly, internode explants produced better results for root length after 21 days plantlet -1 when concentration of 1.0 mg/L IAA + 0.25 mg/L GA₃was used. Shoot tip explants also produced better results in root length after 28 days plantlet -1 when concentrations 1.0 mg/L IAA + 0.25 mg/L GA₃ were used.

Keywords: In vitro, Callus initiation, Regeneration, Potato

Location: BARI, Gazipur

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Potato

Objectives:

To develop a callus induction and regeneration protocol cv. Diamant from leaf discs, nodes, internodes and shoot tips.
To identify the suitable explants for large scale utilization in tissue culture technique.

Methodology:

In vitro plant materials of potato (Solanum tuberosum) cv. Diamant, developed using leaf, node, internode, and shoot tip were grown on medium consisting of Murashige and Skoog basal medium (Murashige and Skoog, 1962). For callus formation, fresh MS medium was supplemented 2,4-D (0.0, 1.0, 2.0, 3.0 and 4.0 mg/L) and kinetin was 0.25 mg/L in each case. The solid MS medium was prepared having the pH of 5.8 and was sterilized at 121°C for 20 minutes at 15 psi pressure. Glassware and other instruments were also sterilized in an autoclave at the same temperature and pressure. Aseptic conditions were maintained throughout the experimental period. The sprouts of potato were rapidly washed in 70% alcohol and then immersed in 0.1% HgCl₂ solution for 15 minutes and then washed in sterilized water for several times and they are placed in MS medium. Leaf, node, internode, and shoot tip of one month old microplant (4-5 cm in height) were cut into small pieces (2-5 mm) and placed in MS media containing different concentrations of 2,4-D and kinetin. Culture was kept in a growth room at 25±2 °C having 1.83 m fluorescent tubes and was illuminated 16 h daily with a light intensity of 1500 lux. Callus initiated after 10-14 days of incubation. Four weeks after inoculation, explants were removed aseptically from the Petridish on a sterilized glass plate inside the laminar air flow cabinet and were placed again on freshly prepared sterilized medium containing fixed concentrations of BAP (1.0 mg/L) and GA₃(0.0, 0.05, 0.1, 0.2, 0.4 mg/L) for shoot initiation. The subcultured plants were then incubated at 22±2°C with 16 h photoperiod. After shoot initiation, more light intensity was given for shoot elongation. When these shoots grew about 3-4 cm in length, rescued aseptically, separated from each other and again subcultured with freshly prepared root induction medium supplemented with IAA (1.0 mg/L) and GA₃ (0.0, 0.25, 0.5, 1 .0, 2.0 mg/L) to induced root. The experiment was laid down in the CRD with 5 replications.

Source of Information: Bangladesh J. Agril. Res. 34(3) : 449-456, September 2009, ISSN 0258-7122

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

From the above discussion, it is revealed that the variation of callus development among the different types of explants were not so wide. In the conclusion, the findings of the present study could be useful to develop protocol to identify the appropriate explants of potato with exact concentration. Furthermore, the results could be used to produce large scale production of healthy and disease free planting materials commercially.

Knowledge Management: Journal