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In Vitro Regeneration Protocol for Anupam and Chini Champa: Two Banana (musa Sapientum) Cultivars of Bangladesh

Rabab Mahdi
Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1209, Bangladesh.

Md. Jahidul Islam
Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1209, Bangladesh.

Md. Ataur Rahman
Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1209, Bangladesh.

Anup Biswas
Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1209, Bangladesh.

F.M. Safiul Azam
Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1209, Bangladesh.

Mohammed Rahmatullah
Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1209, Bangladesh.

Abstract/ Executive Summary:

Anupam and Chini Chmpa are the two most favored banana (Musa sapientum) cultivars in Bangladesh. The fruit, inflorescences and pseudostems are the edible parts and rich in carbohydrate, protein, minerals and vitamins. It’s now a matter for grower to have disease free planting materials in large quantities because of transmission of viral disease through conventional propagation by suckers that reduces banana yield in the country. Our objective of this study was to develop an in vitro method of clonal propagation of these two cultivars. The experimental design was conducted to examine the effect of phytohormones in vitro on these cultivars. For in vitro study, field grown suckers were collected to isolate the meristemic part of the pseudostems and cultured in Murshige and Skoog (MS) medium containing plant growth hormones after surface sterilization by 0.1% HgCl2 containing 6-benzyleaminopurine (BAP), kinetin (Kn), indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) for six weeks. Buds were sprouted when cultured in MS + BAP [4.5 mg/l] for Chini Champa, and when MS was upplemented with BAP [2.0mg/l] then 75% response was obtained with Anupam. Synergistic effect of cytokinin and auxin was found effective in improving shoot proliferation rate for both the cultivars. In Chini Champa, BAP [5.0 mg/l] + IAA [2.5 mg/l] showed 85% frequency with an average of 1.9±0.09 cm and in Anupam shoot length attained up to 2.9±0.11 cm in MS + BAP [5.0 mg/l] + IAA [1.5 mg/l]. Combination of BAP and Kn was found effective only in Anupam. Tallest shoot (3.5±0.09) was observed in MS + Kn [2.0 mg/l] + IAA [0.25 mg/l]. The regenerated micro-shoots were then cultured to induce roots in MS + IBA [1.0 mg/l]. The plantlets were transplanted in to soil followed by hardening and acclimatization and 85% survival rate was obtained after eight weeks. Our developed protocol can play a role for in vitro studies on banana and as well as can produce plantlets for these cultivars.

Keywords: Regeneration, Musa sapientum, Anupam, Chini Champa, AAB Genome

Location: Natore district, Bangladesh

Start Date:

End Date:

Program Area: Resource Development and Management

Commodity: Banana, In vitro

Objectives:

To meet the demand of the farmers and for commercialization of good varieties like Anupam and Chini Champa and increase the total yield, large number of plantlets is needed within a specific time period. Thus in vitro clonal propagation could play a role to reduce this scarcity of plantlets and can serve to develop in vitro techniques on other cultivars of banana.

Methodology:

Explant preparation. Newly sprouted suckers (2- 3 weeks old) were collected from different banana farmlands of Natore district, Bangladesh. The mother plants for each cultivar were carefully inspected to make sure that they were healthy and phenotypically true-to-type and then these suckers were moved to Dhaka. The experiment was conducted at Plant Cell and Tissue Culture Laboratory at the University of Development Alternative. The meristemic part from each pseudostems were trimmed from the sucker and used to prepare explant. Then the shoot tips were decapitated and a block of tissue (1.5 cm × 1.5 cm × 1.5 cm) was excised and washed under running tap water for 15-20 minutes. The explants were soaked in Tween 80 solution for 7 minutes and then washed thrice with distilled water. The tissues were then surface sterilized under laminar air flow by immersion in 0.1% HgCl2 solution for 15 minutes and washed thoroughly four times with distilled autoclaved water to remove any traces of HgCl2. Explants were implanted onto Murashige and Skoog's (MS) medium [15] containing different phytohormones (6-benzylaminopurine (BAP), indole-3-acetic acid (IAA), Kinetin (Kn) and Indole3-butyric acid (IBA) supplemented with 3% sucrose and 0.8% Difco Bacto-agar in test tubes and culture vessels. The pH of the medium was adjusted to 5.8 before autoclaving and the culture condition was maintained at 24±2oC under 12 h cool white fluorescent tube light (4000 lux).

Effect of cytokinins. The excised cormlets were transferred aseptically in MS medium supplemented with different concentrations (1.00, 1.50, 2.00, 2.50, 3.00, 3.50, 4.00, 4.50 and 5.00 mg/l) of BAP for single shoot regeneration. Effect of cytokinins-auxin combinations. The synergistic effect of auxin-cytokinin (BAP + IAA, Kn + IAA) was observed with the sterile explants cultured in MS medium containing different combinations of auxin and cytokinins. Root induction phase. In vitro regenerated shootlets were inoculated in ½ MS medium fortified with IBA (0.1 to 0.8 mg/l) for root induction. Establishment into soil. After root induction, good and healthy plantlets were properly hardened and transplanted in soil composed of humus and garden soil (1:1) into plastic pots and kept in the shade house at a relative humidity of 80-90%.

Data assembly and scrutiny. Weekly growth observations were made up to six weeks after setup of each treatment and experimental data were recorded. The data collected were: a) percentage of response (%); b) shoot length in cm and number of leaves/shoot; c) number of roots/shoot and d) days required for response. The data then were analyzed as mean ± SEM according to Mian and Miyan.

Source of Information: American-Eurasian Journal of Sustainable Agriculture 8(1), January, Pages: 28-33

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

This protocol for clonal propagation can serve as a convenient method for in vitro plantlets production of M. sapientum cv. Chini Champa and M. sapientum cv. Anupam to meet the demand for disease free, healthy plants and ultimately it can enhance the economic benefit of the cultivators. This study brought out the selection of suitable phythormone level and media composition for bud sprouting, shoot proliferation and consequently, root formation of microshoots in vitro for Chini Champa and Anupam.

Knowledge Management: Journal