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Effect of Bap and Iba on in Vitro Regeneration of Local Banana Variety of Sabri

Fahima Khatun*
Department of Biotechnology, Sher-e-Bangla Agricultural University, Bangladesh

M. E. Hoque
Department of Biotechnology, Sher-e-Bangla Agricultural University, Bangladesh

Homayra Huq
Department of Biotechnology, Sher-e-Bangla Agricultural University, Bangladesh

Md. Adil
Department of Plant Breeding, Bangladesh Rice Research Institute, Bangladesh.

Kh. Ashraf-Uz-Zaman
Department of Agricultural Chemistry, Sher-e-Bangla Agricultural University, Bangladesh.

Mominul Haque Rabin
Department of Agricultural Chemistry, Sher-e-Bangla Agricultural University, Bangladesh.

Abstract/ Executive Summary:

The experiment was carried out to study the effect of benzyl amino purine (BAP) (1.0, 2.0, 3.0, 4.0 and 5.0 mg/L) and indole-3-butyric acid (IBA) (0.5, 1.0, 1.5, 2.0 and 2.5 mg/L) for in vitroregeneration of sabri variety of Banana using shoot tip explants. The highest response of explants (84%) and maximum number of shoots per explant (3.4) were observed with 5.0 mg/L BAP in sabri. In contrast, due to the combined effect, 5.0 mg/L BAP+2.5 mg/L IBA showed best response (90%). The highest shoot number per explant (3.4) was found with 5.0 mg/L BAP+2.0 mg/L IBA. The maximum number of roots (3.4 and 5.2) was observed in 1.5 mg/L and 3.0 mg/L BAP+1.0 mg/L of IBA. In a controlled environment, the regenerated healthy rooted plantlets were transferred from culture media to soil in plastic pots where 90% plantlets were survived and in open atmosphere, the survival rate was 88.89%. This protocol has the applicability in vitro rapid propagation of Banana.

Keywords: In vitro; Regeneration; BAP; IBA; Survival rate

Location: In Bangladesh

Start Date:

End Date:

Program Area: Resource Development and Management

Commodity: Banana, In vitro

Objectives:

The present study was undertaken with the following objectives: i. To study the individual effect of BAP and IBA on in vitro regeneration of sabri variety of Banana.

ii. To study the combined effect of BAP and IBA on in vitro regeneration of sabri variety of Banana, and iii. To develop an efficient protocol for in vitro rapid propagation of Banana varieties.

Methodology:

2.1 Plant Materials The experiment was conducted at the Biotechnology Laboratory, Department of Biotechnology, Sher-e-Bangla Agricultural University, Bangladesh for in vitro regeneration of sabri variety of Banana. The three months age of sabri variety was used as experimental materials in the present investigation and collected from Agargau nursery, Sher-e-Bangla nagar, Dhaka. 2.2 Explants The healthy, disease-free shoot tips of 2 cm length were used as explants for the study for in vitro regeneration. 2.3 Culture Media MS (Murashige and Skoog) medium supplemented with different phytohormones as per treatments were used as culture medium for shoot induction, shoot multiplication, maintenance, and regeneration of roots. Hormones (BAP and IBA) were added either in combination or separately to the media according to the treatment. Five levels of BAP (1.0, 2.0, 3.0, 4.0 and 5.0 mg/L) and 5 levels of IBA (0.5, 1.0, 1.5, 2.0 and 2.5 mg/L) were used separately and three levels of BAP (3.0, 4.0 and 5.0 mg/L) with 5 levels of IBA (0.5, 1.0, 1.5, 2.0 and 2.5 mg/L) were used as treatment. The pH was adjusted to 5.8 before placing in the microwave oven which was used for melting agar (semi solidifying agent). 2.4 Surface Sterilization Isolated shoot tips were collected from Banana suckers of three months having 4 cm in length were surface sterilized with 70% ethanol for 1 min and 0.2% HgCl2 separately for 12 min, respectively followed by 3-4 times rinse with sterilized distilled water and then outer layer of explants were removed carefully. Further, the explants were sterilized with 0.1% HgCl2 for 5 min then washed for at least 3 times with sterilized distilled water. Another outer layer of the explants were removed carefully. The final size of explants were 1-2 cm which had 6/8 overlapping leaf base enclosing auxiliary bud. 2.5 Culture of Explants The isolated and surface sterilized shoot tips were collected carefully through maintaining aseptic condition inside the laminar airflow cabinet and inoculated to each of the culture tube containing 50 mL of MS medium supplemented with different concentrations of hormones as per treatment. The culture vials were transferred to culture racks and allowed to grow within 25±1°C temperature by an air conditioner and 16 hour photoperiod was maintained along with light intensity of 3000 lux for proper growth and development of culture. To control the blackening of the tissues on the explants which occurred within 6-7 days after inoculation were removed and the explants were transferred to similar fresh medium. It was repeated at the interval of 10 days interval for about one month to minimize further blackening of the tissues. 2.6 Maintenance of Proliferating Shoots For subculturing, the entire samples of in vitro shoot were cut into small pieces so that each piece would contain about one shoot and inoculated into a similar fresh medium. The subculturing was done at the interval of 20-25 days. 2.7 Regeneration of Plants from in vitroProliferated Buds In vitro proliferated micro shoots were separated and each of the micro shoot was placed on culture medium, which was supplemented with a particular concentration of hormone for shoot differentiation. When the shoots grew about 3-5 cm in length with 3-6 well-developed leaves they were cultured on freshly prepared medium containing different combinations of hormonal supplements for root induction. 2.8 Establishment of Plantlets Regenerated plantlets were transplanted to pots (10×15 cm) containing sandy soil and cowdung in 1:1 ratio. An occasional spray of water was done to prevent sudden desiccations and maintain high humidity (98%) around the plantlets. Initially the plantlets were hardened in a controlled environment. Then after 2 weeks, exposed to lower humidity and higher light intensity (3000 lux). Finally, after 20 days plantlets were transferred to natural environment. 2.9 Data Collection and Statistical Analysis of Data Data were recorded at 2, 4 and 6 weeks after inoculation (WAI) on the effect of different treatments on shoot and root proliferation. Data were analyzed using MSTAT-C statistical package where means were compared by Duncan’s Multiple Range Test (DMRT). The experiment was conducted in Completely Randomized Design (CRD) with five replications in the culture room.

Source of Information: Biotechnology Journal International 18(1): 1-10, 2017; Article no.BJI.31592

Article Link (Online Journal): DOI: 10.9734/BJI/2017/31592

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In present study we found that the 5.0 mg/L BAP and 1.5 mg/L IBA concentration performed best in in vitro shoot and root regeneration. In case of combined effect, 5.0 mg/L BAP ± 2.5 mg/L IBA and 5.0 mg/L BAP ± 2.0 mg/L IBA showed the highest results for in vitro regeneration of sabri variety of banana. Therefore, suitable protocol was established which could be used for in vitro rapid propagation of Banana plantlets. In future more research works have been conducted to assess the field performance for the ultimate improvement of Banana under Bangladesh condition.

Knowledge Management: Journal