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In Vitro Plant Regeneration of Potato (solanum Tuberosum L.) at the Rate of Different Hormonal Concentration

Sayeed Shahriyar
Department of Biotechnology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Soleh Akram
Department of Genetics and Plant Breeding, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Koushik Khan
Department of Biotechnology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Md. Faruk Miya
Department of Biotechnology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Md. Abdur Rauf Sarkar
Department of Genetic Engineering and Biotechnology, Jessore University of Science and Technology, Jessore-7400, Bangladesh

Abstract/ Executive Summary:

One of the goals of the experiment is to standardization of HgCl₂ treatment for explants sterilization. The objectives also include developing a reproducible cost effective protocol for large scale production of Solanum tuberosum of Cardinal variety plantlets from selectively better clones through plant in vitro propagation methods. Selection of growth regulators for proper multiple shoots regeneration, elongation and root induction. To produce genetically uniform plantlets within a short time capable surviving in natural condition raised in in vitro environment. Shoot tip and nodal segment explants from field grown plants were used as experimental materials in this investigation. All explants were cultured on Murashige and Skoog medium supplemented with various plant growth regulators. For surface sterilization of explants, HgCl₂ (0.1%) for 2 minutes was found to be most effective for complete destroying of surface pathogens and getting healthy tissues. Shoot regeneration was observed from both shoot tips and nodal explants for the studied plant. Maximum number of shoot per culture (17) was recorded and it also obtained the highest average length of the shoot (5cm) in Murashige and Skoog medium containing no hormone. On the other hand 6-benzyl amino purine (0.2mg/l) in 3 media showed the highest rate of shoot multiplication (73%) and the highest average length (4cm). In case of Gibberellic acid (0.1mg/l) in Murashige and Skoog media showed its highest rate of shoot regeneration (82%) and the highest average length (4.5cm). From the overall experiment it was observed that shoot tips are more responsive for micro propagation. In root induction Murashige and Skoog medium supplemented with different concentration (0.5, 1, 1.5 and 2mg/l) of indol-3-acetic acid and kinetin. Indol-3-acetic acid and kinetin (1.5+1.5 mg/l) showed its lowest rate of root regeneration (40%) and the average length of the root (1.5 cm). On the contrary Murashige and Skoog medium with no hormone showed the rate of root regeneration (96%) and the highest average length of the root (2.5 cm). The supplemented Murashige and Skoog media with no hormone showed the best performance for root regeneration.

Keywords: Regeneration; Potato; Indole acetic acid (IAA); Kinetin (KIN)

Location: Department of Genetic Engineering and Biotechnology, Jessore University of Science and Technology, Bangladesh.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Potato

Objectives:

To develop a reproducible cost effective protocol for large scale production of Solanum tuberosum of Cardinal variety plantlets from selectively better clones through plant in vitro propagation methods.
Selection of growth regulators for proper multiple shoots regeneration, elongation and root induction.
To produced genetically uniform plantlets.
To obtain a large number of plantlets within a short time. To establish the in vitro raised plantlets in the natural conditions.

Methodology:

The experiment was conducted at the Department of Genetic Engineering and Biotechnology, Jessore University of Science and Technology, Bangladesh. Shoot tips and nodal segments were used for micro propagation of Solanum tuberosum (potato) a cultivar of cardinal. The surface sterilized explants were sized 1.0-1.5 cm in length.HgCl₂(0.1%) was used as surface sterilizing agent at different time duration ranging from one to five minutes then washed thoroughly under running tap water and then treated with two drop of Tween-20 (wetting agent) for 5-6 minutes and then the materials were washed several times with distil water. Then savlon (ACI Ltd. BD) was used as detergent and surfactant. For plants nutrient basal salts were used which contains macro, micro nutrients and vitamins. In the present experiment different culture media with various growth regulators and additive were used for shoot tip and nodal segment culture: (a) Murashige and Skoog medium with different concentrations of 6-benzyl amino purine, Gibberellic acid singly was used for shoot induction .For carbon source 3% sugar was used and the medium was solidified with 6-6.2% agar.(b) Half and full strength of medium with different concentration of indol-3-acetic acid and kinetin were used for root induction. The pH of the medium was adjusted to 5.8 by using 1N NaOH. The media were autoclaved at 121⁰C for 20 min after adjusting the pH. The explants were inoculated on callus induction medium at 25±2⁰ C for 3-6 weeks under white light (2500-3000 lux). The photoperiod was maintained generally 16 hours and 8 hours dark. The culture vessels such as test tube, bottle conical flask, measuring cylinders, glass rods, beakers, pipette pumps, parafilm, cotton plug, rubber bands, filter paper, aluminum foils, forceps, fire box, marker pen, spirit lam, needle, sharp blade, stereomicroscope, electronic balance, autoclave, pH meter, magnet stirrer, laminar airflow machine etc. were used as the present investigation. Data were recorded on the following parameters, different time duration for surface sterilization of explants, days to shoot regeneration, number of shoot per explants, days to root induction, number of root per explants.

Source of Information: Asian J. Med. Biol. Res. 2015, 1 (2), 297-303; doi: 10.3329/ajmbr.v1i2.25625; ISSN 2411-4472

Article Link (Online Journal):

Funding Source:

1.

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The result of this study set a standard of HgCl₂ treatment for explants sterilization of Solanum tuberosum. More importantly the study provides a suitable result in selection of growth regulators for proper multiple shoots regeneration, elongation and root induction in case of producing genetically uniform plantlets. The techniques are becoming popular as an alternative means of vegetative propagation for commercially important plants like potato. The in vitro technique of propagation has a number of advantages over conventional method and it can be widely used for commercial micro propagation.

Knowledge Management: Journal, Online Circulation