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In Vitro Response of Strawberry (fragaria X Ananassa Dutch.) for Callus Induction and Shoot Regeneration

M Rezaul Karim
Department of Botany, University of Rajshahi, Rajshahi 6205, Bangladesh.

M Abdul Aziz
Department of Botany, University of Rajshahi, Rajshahi 6205, Bangladesh.

Uthpal Krishna Roy
Department of Botany, University of Rajshahi, Rajshahi 6205, Bangladesh.

M Aminul Hoque
Department of Agronomy and Agricultural Extension, University of Rajshahi, Rajshahi 6205, Bangladesh.

M Monzur Hossain
Department of Botany, University of Rajshahi, Rajshahi 6205, Bangladesh.

Abstract/ Executive Summary:

Effect of growth regulators on in vitro responses of strawberry for callus induction and shoot regeneration has been investigated. MS (Murashige and Skoog) medium supplemented with 2,4 - diclorophenoxyacetic acid (3.0mg/l) and 6 - Benzyl adenine (0.5mg/l) were found most effective in terms of percentage to callus inductions and degree of callus development. Calli derive d strawberry explants were cultured in shoot formatting media supplemented with different concentrations of plant growth regulators. The combination of BA (1.5 mg/l) and α- Napthalene acetic acid (0.75mg/l) showed highest percentage of shoot induction (55.6 ±0.81) and multiple shoot regeneration. Combination of BA, NAA and KIN (6 - Furfuryl amino purine/kinetin) found to be most efficient in shoot induction (60.7%) and multiplications (16.7±0.48). After successful shoot regeneration, regenerated shoots were cultured for root induction in MS₀. The developed plantlets through in vitro culture technique were acclimated and successfully transferred to in vivo condition for further evaluations.

Keywords: Callus induction, Regeneration, Strawberry

Location: Plant Breeding and Gene Engineering Laboratory of Rajshahi University of Bangladesh.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Strawberry

Objectives:

To develop an efficient protocol for in vitro callus induction and regeneration of strawberry for somaclonal variation.

Methodology:

For callus induction, two or three weeks old immature in vitro leaves were collected from Plant Breeding and Gene Engineering Laboratory of Rajshahi University of Bangladesh. Explants was surface sterilization by using mercuric chloride (HgCl₂ ) as surface sterilizing agent, tween - 80 and savlon (ACI Pharma, Bangladesh) as surfactant cum detergent. Then the leaves were cut into small segments (0.5 - 1.0cm) and notched by scalpel in laminar air flow and were cultured on a semisolid MS medium supplemented with different concentrations (1.0, 1.5, 2.0 and 3.0) of 2,4 - diclorophenoxyacetic acid (2,4 - D) and α - Napthalene acetic acid (NAA) alone and different concentrations and combinations (2.0+0.5, 2.0+1.0, 3. 0+0.5, 3.0+1.0, 4.0+0.5 and 4.0+1.0) of 2,4 - D (2,4 - diclorophenoxyacetic acid)+BA (6 - Benzyl adenine) and α - Napthalene acetic acid (NAA)+6 - Benzyl adenine (BA). Then the cultures were incubated in dark condition around 15 days for callus induction. The responded calli were further sub cultured on MS media containing low concentrations and combinations of BA, NAA and KIN (6 - Furfuryl amino purine/kinetin) for shoot regeneration. The pH of all media was adjusted to 5.7 before addition of agar and sterilized by autoclaving for 20 minutes at 15lb. pressure and at 121ºC temperature. The cultures were maintained at 25 ºC±2ºC (room temperature) under the cool white fluorescent lights for 16 hours photoperiod at 2000 - 3000 lux.

Source of Information: Int. J. Agr. & Agri. R. Vol. 1, No. 1, p. 29-36, 2011, ISSN:2223-7054

Article Link (Online Journal): http://www.innspub.net/wp-content/uploads/2012/01/IJAAR-V1-No-1-p29-36.pdf

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

After successful shoot regeneration, regenerated shoots were cultured for root induction in MS₀. The developed plantlets through in vitro culture technique were acclimated and successfully transferred to in vivo condition for further evaluations. Present observation shows that the high incidence of callus induction and regeneration for somaclonal variations could be used in breeding programme for improvement of strawberry in Bangladesh.

Knowledge Management: Journal