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Bap and Iba Pulsing for in Vitro Multiplication of Banana Cultivars Through Shoot-tip Culture

M. H. Ferdousa
Dept. of Genetics and Plant Breeding, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

H. Mehrajc
The United Graduate School of Agricultural Sciences, Ehime University, 3-5-7 Tarami, Matsumaya, Ehime 790-8556, Japan

A. F. M. Jamal Uddin
Associate Professor
Dept. of Horticulture, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

T. Taufiqued
Dept. of Horticulture, Sher-e-Bangla Agricultural University, Dhaka-1207, Bangladesh

A. A. Masum Billahb
Dept. of Agriculture Extension, Ministry of Agriculture, Iswardi, Pabna, Bangladesh

Abstract/ Executive Summary:

The study was undertaken with a view to establish a protocol for in vitro plant regeneration from shoot tip explants of banana. Different concentrations of BAP (0.0, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0 and 7.0 mg/l) and IBA (0.0, 1.0, 2.0, 3.0, 4.0 and 5.0 mg/l) was used in MS medium to assess the influence on in vitro shoot regeneration and subsequent root formation of Amritasagar and Sabri banana cultivars. Maximum single shoot formation (50.0% and 30.0%), number of single shoot (3.50 and 2.00 in), longest shoot (2.64 cm and 2.16 cm) were found from 0.5 mg/l BAP while maximum number of roots (3.83 and 2.50) and longest root (3.60 cm and 3.10 cm) was found from 0.3 mg/l IBA in Amritasagar and Sabri respectively. The survival of the plantlets of both cultivars was more than 82% under ex vitro condition. 0.5 mg/l BAP and 0.3 mg/l IBA can be used with MS media for shoot and root formation of Amritasagar and Sabri banana cultivars through shoot tip culture.

Keywords: Banana,Ggrowth regulators, In vitro shoot-tip culture, Plantlets regeneration

Location: Proshika Tissue Culture Centre Trust, Manikgonj, Bangladesh

Start Date: 00-04-2008

End Date: 00-11-2008

Program Area: Variety and Species

Commodity: Banana

Objectives:

To assess the suitable concentration of BAP for in vitro shoot proliferation and root induction of two banana cultivars, namely, cv. Amritasagar and cv. Sabri, for plantlets regeneration.

Methodology:

Laboratory and period of the experiment: The study was carried out at the laboratory of Proshika Tissue Culture Centre Trust, Manikgonj, Bangladesh from the April, 2008 to November, 2008.

Genetic materials: Banana cv. Amritasagar (Musa sapientum, Genotype AAA) and Sabri (Musa sapientum, Genotype AAA) were used in the experiment.
Concentrations of BAP and IBA: Different concentrations of BAP (0.0, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0 and 7.0 mg/l) on shoot multiplication and IBA (0.0, 1.0, 2.0, 3.0, 4.0, and 5.0 mg/l) for rooting of two cultivars of banana was studied.
Culture media for in vitro: Both for shoot regeneration and rooting of multiplied shoots MS medium was used with different vitamins and hormonal supplementation. Hormones were added separately to different media according to the requirements. For the preparation of media, stock solutions were prepared at the beginning and stored at 4±1⁰C temperature. The respective media were prepared from the stock solutions. The culture tubes containing media, beakers, pipettes, measuring cylinder, metal instruments were autoclaved at 15 psi pressure at 121⁰C for 20 minutes. The medium was then cooled at room temperature before use. Laminar Airflow Cabinet was usually sterilized by switching on the UV light of the cabinet for 30 minutes and wiping the working surface with 70% ethyl alcohol for 30 minutes before starting the transfer work.
Sample collection and preparation for in vitro culture: Meristem was collected from developing suckers from field for both cultivars. The suckers were washed thoroughly under running tap water. The roots and outer tissues of the suckers were removed with the help of a sharp knife. A number of outer leaves were removed until the shoot measured about 1.5-2.0 cm in length and 1.0 cm width at the base. The prepared explants were taken into a conical flask and were washed with distilled water containing 1% savlon and 3-4 drops of Tween-80 for 20 minutes to remove dusty substance and followed by successive 3 times washing with distilled water to make the materials free from savlon and Tween-80. Subsequently the materials were transferred to running Laminar Airflow Cabinet.
Surface sterilization: The surface sterilization of explants was carried out in 0.1% HgCl₂ for different periods (10, 12, 14, 16 minutes). Then the materials were washed 3-5 times with distilled water to remove all traces of HgCl₂.
Inoculation of plant materials: The shoot tip explants of about 0.5 cm long with 3-4 leaf primordial were prepared. The individual shoot tip was directly inoculated to each culture vessel and covered with plastic cap. After that the caps were sealed with parafilm.

Transfer to growth chamber: The culture vessels were transferred to growth room and were allowed to grow in controlled environment (25±2⁰C temperature, 16-hour light period and 2000 lux light intensity) for the growth and development of the cultures.
Sub-culturing: For sub-culturing, the entire samples of in vitro shoots were cut into small pieces so that each piece would contain about one shoot. Leaf and blackish or browned basal tissues were removed to expose the meristem. Each piece was inoculated into a similar fresh medium. It was practiced at the interval of every one month. In vitro proliferated micro shoots were separated and each of the micro shoots was placed on culture medium.

Source of Information: Journal of Bioscience and Agriculture Research, Vol. 03, Issue 02: 87-95, 2015

Article Link (Online Journal): http://www.journalbinet.com/uploads/2/1/0/0/21005390/bap_and_iba_pulsing_for_in_vitro_multiplication_of_banana_cultivars_through_shoot-tip_culture.pdf

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Application of 0.5 mg/l BAP and 0.3 mg/l IBA as growth supplements on MS culture media help to perform better for new plantlet regeneration capability through In vitro shoot-tip culture of Banana. BAP concentration 0.5 mg/l and IBA concentration of 0.3 mg/l was found best concentration for shoot proliferation and root elongation in both cultivars of banana. On the other hand, Application of more than 0.5 mg/l BAP and 0.3 mg/l IBA gradually decreases In vitro culture developments for all parameters as observed for shoot proliferation and root elongation. But for the commercial plantlet regeneration, Amritasagar was found better than Sabri cultivar with 0.5 mg/l BAP and 0.3 mg/l IBA for shoot regeneration and root formation respectively.

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