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Standardization of Protocol for in Vitro Propagation of Banana (musa Sapientum)

Farhana Hoque
Department of Horticulture, Patuakhali Science and Technology University, Dumki, Patuakhali-8602, Bangladesh

Mahbub Robbani
Department of Horticulture, Patuakhali Science and Technology University, Dumki, Patuakhali-8602, Bangladesh

Md. Fakhrul Hasan
Department of Horticulture, Patuakhali Science and Technology University, Dumki, Patuakhali-8602, Bangladesh

Jahanara Parvin
Department of Horticulture, Patuakhali Science and Technology University, Dumki, Patuakhali-8602, Bangladesh

Abstract/ Executive Summary:

An experiment was conducted at the Plant Biotechnology Laboratory, Department of Horticulture, Patuakhali Science and Technology University during the period from August 2016 to April 2017 to standardize the protocol for in vitro propagation of banana. The experiment was laid out in completely randomized design with four replications. Three to four months aged field grown rhizome attached shoots were used as explants and cultured on MS medium with different concentrations and combinations of BAP (6-benzylamino purine), BAP + KIN (Kinetin) + NAA (Naphthalene Acetic Acid) and IBA (Indole3- Butyric Acid) + IAA (Indole-3- Acetic Acid) to observe their efficacy on single shoot initiation, shoot multiplication and root formation respectively. Minimum number of days required for shoot initiation (9.07) with highest shoot initiation percentage (91.14) and the longest shoot (2.23 cm) was found in MS medium supplemented with 5.0 mg/L BAP. On the other hand, highest shoot multiplication percentage (80.99) with maximum number of shoots per explant (4.47), the highest length of shoots (4.17 cm) and maximum number of leaves (4.04) was observed in MS medium supplemented with 4.0 mg/L BAP + 2.0 mg/L KIN + 2.0 mg/L NAA. In case of root regeneration, the best results on days required for root initiation (9.00), the highest root initiation percentage (85.05), maximum number of roots per plantlet (5.83) and the highest length of roots (4.17 cm) was obtained in MS medium supplemented with1.5 mg/L IBA + 0.5 mg/L IAA. After 5-7 days of hardening in room temperature, established plantlets were ready for planting.

Keywords: Banana, In vitro propagation, Plant growth regulators

Location: Plant Biotechnology Laboratory, Department of Horticulture, Patuakhali Science and Technology University

Start Date: 00-08-2016

End Date: 00-04-2017

Program Area: Crop-Soil-Water Management

Commodity: Banana

Objectives:

The present study was undertaken to evaluate the propagation rate of M. sapientum under the influence of PGRs as well as large scale production of banana.

Methodology:

Young, healthy, disease free, 3–4 old months amritasagar plants were collected from Germplasm Center, Department of Horticulture, PSTU. The rhizome attached shoot of amritasagar was used as plant material in this research work. Outer leaves, roots, dust, other debris and outer tissues were removed until the explant measured about 2.0–3.0 cm in length and 2.0 cm width with the help of sharp knife. Then the explants washed with distilled water until complete removal of all traces. Surface sterilization of explant was done under laminar airflow cabinet with a few drops of savlon, tween 20, 0.1% mercuric chloride and 70% ethyl alcohol respectively. Then the explants rinsed with sterile distilled water three to four times. All the plant growth regulators, stock solutions were prepared before preparing the culture media. The media were adjusted to pH 5.8 with 1 N NaOH or 1 N HCl and autoclaved at 121°C for 45 minutes at 15 psi pressure. The isolated and surface sterilized explants were directly inoculated individually to each of the culture tube containing 25 ml of MS medium supplemented with different concentration and combinations of auxins (IAA, IBA, NAA) and cytokinins (BAP and KIN) to conduct three experiments. Five different concentration of BAP (1.0, 3.0, 5.0, 7.0, 9.0) mg/L was used for single shoot regeneration. After raising several growing points, the in vitro grown shoot-tips were cultured on MS medium supplemented with BAP (1.0, 2.0, 3.0, 4.0, 5.0) mg/L, KIN (2.0 mg/L) and NAA (2.0 mg/L) for shoot multiplication. When the regenerated shoots became 2–3 cm in length with 3–6 well developed leaves, they were rescued aseptically from the culture tubes. After separating from each other, they were again cultured on freshly prepared medium containing different concentrations and combinations of IBA (1.0, 1.5) mg/L and IAA (0.5, 1.0, 1.5) mg/L for root initiation. The cultures were incubated at 25±2°C by an air conditioner with light intensity varied from 2000-3000 lux under 16 hours light and 8 hours dark. When the regenerated plantlets produced 3–5 cm roots with vigorous shoot, they were transferred in soil. Plantlets were planted in poly bags containing garden soil + compost + sand (1:1:1). The experiments were arranged in completely randomized design (CRD) with 4 replications. Each treatment consisted of 10 culture tubes per replication. Data were collected on the effect of different treatments on shoot proliferation and rooting. The significance among the means of treatments was evaluated at 5% level of significance by Duncan’s Multiple Range Test (DMRT, Gomez and Gomez, 1984) by using MSTAT-C statistical package (Steel et al. 1997).

Source of Information: J Bangladesh Agril Univ 16(1): 27–30, 2018 ISSN 1810-3030 (Print) 2408-8684 (Online)

Article Link (Online Journal): doi: 10.3329/jbau.v16i1.36477

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Tissue culture technologies are applicable for the mass propagation of Amritasagar to get true to type plantlets. Amritasagar shows better response in in vitro condition with plant growth regulators. Further comprehensive study is needed to test the performance of tissue culture derived plantlets.

Knowledge Management: Journal