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Micropropagation of Strawberry (fragaria X Ananassa Duch.)

M. N. HASAN
Lecturer
Genetic Engineering and Biotechnology Department, Jessore Science and Technology University, Jessore, Bangladesh

S. NIGAR
Lecturer
Department of Nutrition and Food Technology, Jessore Science and Technology University, Jessore, Bangladesh

M. S. RAHMAN
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna Bangladesh

Abstract/ Executive Summary:

The experiment was conducted from March 02 to September 25, 2010 at Plant Tissue Culture Laboratory of Genetic Engineering and Biotechnology Department, Jessore Science and Technology University, Jessore, Bangladesh to clarify the effective concentration of growth regulators for micropropagation of strawberry. Response of medial additives like adenine sulfate (120 mgL^-1) and coconut water (150 mg^L-1) were also investigated. The concentration of cytokinin-auxin was ranged from 1.0-3.0 mgL^-1 and 0.1-0.3 mgL^-1respectively. Addition of adenine sulfate, gibberellic acid and coconut water was insignificant than combine concentration of cytokinin and auxin regarding usable shoots. But shoot length was the highest (2.4 cm) which is significantly different than that of gibberellic acid and coconut water. Maximum number of nods was also observed in the presence of adenine sulfate. For rooting the shoots were transferred to MS medium with four different combination of IBA (0.1, 0.2, 0.5, 1.0) mgL^-1 and IAA (0.1, 0.2, 0.5, 1.0) mgL^-1 individually. Low concentration of IAA produces maximum number of roots which was 16.5 and significantly different than that IBA or IBA IAA combination. But the highest root length was found in the combined treatment of IBA and IAA (0.1+0.1) mgL^-1.

Keywords: Micropropagation, Strawberry, Adenine sulfate, MS medium

Location: Plant Tissue Culture Laboratory of Genetic Engineering and Biotechnology Department, Jessore Science and Technology University, Jessore

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Strawberry

Objectives:

To find out the optimum concentration of cytokinins (BA Benzyladenine and Adenine) and auxins (IBA and NAA, Alphanaphthalene acetic acid) for high multiplication rate of strawberry, effects of additives (coconut water, adenine) on high frequency shoot proliferation, effects of gibberelic acid (GA₃) on the structure of the strawberry plant and response of different concentration and combination of auxins on root induction.

Methodology:

The experiment was conducted at Plant Tissue Culture Laboratory of Genetic Engineering and Biotechnology Department, Jessore Science and Technology University, Jessore, Bangladesh. The nodal segments of mature strawberry plant were used as explant that’s collected from Sathe Nursery, Jainagar, Ishardi, Pabna, Bangladesh. Sterilization of plant material: At first, explants were washed under running tap water for 20-25 min. Then they were rinsed with sterilized distilled water. Finally, explants were subjected to incision into appropriate sizes (around 1-1.5 cm long), and then transferred to the laminar hood where the processed explants were sterilized in 0.1% HgCl₂ with 2 drops of tween 80 for five minutes followed by three rinsing with sterile distilled water for 4 minutes each. Afterwards explants were resized by scalpels and forceps. Trace of water remaining on the surface of the explants was soaked with sterilized filter paper. Thus surface sterilization was completed and the material was ready for inoculation on appropriate nutrient medium. Media for Micropropagation: MS basal media supplemented with different concentrations and combinations of growth regulator were prepared as a media for Micropropagation. Media for shoot proliferation: For shoot proliferation, nodal segments of mature strawberry plant were taken. Node was cultured on MS basal medium supplemented along with the concentration of cytokinin, auxin, additives and GA₃. After mixing all stock solutions and growth regulators at appropriate volume, 3% sucrose was added. The pH of the medium was adjusted to 5.7-5.8 and then agar (0.7%) was added and dissolved. The media were dispensed in the 40×150 mm glass bottles in a volume of 20-25 ml. Each treatment consisted of 10 glass bottles. The media were sterilized by autoclaving at 121ºC for 20 minutes. Culture media were dispensed in the glass bottles and sterilized as mentioned above. About 3-4 weeks period was required for shoot proliferation. Media for root induction: For rooting, regenerated shoots were sub-cultured in solidified MS medium supplemented along with (Table 2) concentration of IBA or IAA and their combinations. After mixing all stock solutions and growth regulators at appropriate volume, 3% sucrose was added. The pH of the medium was adjusted to 5.7-5.8 and then agar (0.7%) was added and dissolved. The media were dispensed in the 40×150 mm glass bottles in a volume of 20-25 ml. Each treatment consisted of 10 glass bottles. The media were sterilized by autoclaving at 121ºC for 20 minutes. Culture media were dispensed in the glass bottles and sterilized as mentioned above. About 3-4 weeks period was required for root induction. Data collection and Statistical analysis: Weekly Visual observation of culture was made and frequency of culture showing plantlet, shoot and root formation and multiplication was recorded. The data pertaining to shooting and rooting per culture were analyzed by using Duncan’s new multiple range tests.Transfer to the soil: Rooted plantlets were taken out from culture tubes and washed thoroughly with tap water to remove the culture medium from the roots. Washed plantlets were sprayed with fungicide and planted to normal and sterilized soil in polybags. After 7 days the hardened plantlets were planted in soil.

Source of Information: Int. J. Sustain. Crop Prod. 5(4):36-41, (2010)

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In this experiment, as the concentration of growth regulators decreased shoot formation increased. After four to six weeks the well developed in-vitro shoots were sub-cultured on MS medium supplemented with the same concentration of growth regulators. After four to six weeks of sub-culture, these were transferred to the rooting medium. For rooting the shoots were transferred to MS medium with four different concentration of IBA (0.1, 0.2, 0.5, 1.0), and IAA (0.1, 0.2, 0.5, 1.0 mg/L) individually. The combination of IBA and IAA were also used on the concentration of 0.1, 0.2 and 0.5 mg/L. After four weeks the highest numbers of roots were observed in IAA 0.1 mg/L.

Knowledge Management: Journal