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Natural Growth Substances Has Effective Role in Callus Culture of Banana (musa Spp.) Cultivar ‘anupam’ (aab Genome, Sapientum Subgroup)

Mst. Rabeya Pervin
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.

Abstract/ Executive Summary:

This experimental design was conducted to establish the effect of organic nutrients for in vitro morphogenesis on callus of Musa sapientum cv Anupam (Family: Musaceae). This cultivar is a well distributed herbaceous flowering plant in Bangladesh and highly favored by the people for its edible parts (fruit, flower and stem) and serves as a regular nutritional supplement in diet for notably carbohydrate, protein, vitamin B, potassium and calcium. The objective of the study was to develop a suitable protocol for callus culture of this species using organic supplements. In vitro generated calluses of this cultivar which were induced from leaf and stem segments were cultured onto Murashige and Skoog (MS) medium containing 2,4-Dichlorophenoxyacetic acid [2,4-D] (3mg/L) + 1-Naphthaleneacetic acid [NAA] (0.5mg/L), and was used as experimental plant resource. In vitro grown calluses were sub-cultured on MS medium containing different concentrations of coconut water (CW), casein hydrolysate (CH), biotin (Bn), peptone (Pn), yeast extract (YE) and activated charcoal (AC). The pH of the medium was adjusted at 5.8. The highest average callus index number (CIN) (2.65) was recorded in MS + CW (15%) after 28 days. Colors of induced calluses were varied with respect to different additives. In the MS medium containing 15% coconut water, 100% were greenish and fragile and where 60% were non-embryonic and 70% showed root formation. 90% root formation was observed when MS medium was enriched with 20% CW. Addition of CH (200mg/l) and Bn (0.5mg/l) in MS medium was also found to be effective in promoting callus growth and rhizogenesis in calluses. These observations revealed that morphogenic responses could play a role for physiological studies in vitro and served to develop a suitable composition of medium for callus culture for this species from natural and organic sources.

Keywords: Callus induction, Organic supplements, Rhizogenesis, Musa sapientum cv. Anupam

Location: Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1209, Bangladesh.

Start Date:

End Date:

Program Area: Resource Development and Management

Commodity: Banana, Callus

Objectives:

The plant is tall and bears bunch with an average weight of 10 kg. A bunch contains 85-120 fingers, fruits are medium-sized, firm in texture, sweet and tasty. The cultivar is widely grown in the north and western areas of Bangladesh. Objectives of our study were to select the suitable media composition for callus induction through using organic additives and activated charcoal.

Methodology:

Explant source and preparation. The experimental material was in vitro generated calluses of Musa sapientum cv. Anupam, which was induced from leaf and stem segments (0.8-1.0cm) cultured onto MS (Murashige and Skoog, 1962) medium containing 2,4-D (3.0 mg/l) and NAA (0.5 mg/l). In vitro grown microcallus were sub-cultured in Murashige-Skoog (MS) medium (Murashige, T. and F. Skoog, 1962) containing different concentrations (see detail in Results section) of organic supplements including biotin (w/v), peptone (w/v ), casein hydrolysate (w/v), yeast extract (w/v), coconut water (v/v), activated charcoal (w/v), sucrose (3%) and agar (7%). Twenty replicates were inoculated for each treatment. The pH of all media was adjusted to 5.8 before autoclaving and media was sterilized by autoclaving for 20 min at 121ºC and 1.05 kg/cm2. Culture of explants. Cultures were maintained at 26 ± 2ºC with a photoperiod of 16/8 h under an illumination of 20 ìmol m-2s -1 provided by cool white fluorescence lamps. Data recording and analysis. Weekly visual observation of culture was made and data of callus growth were recorded as callus index number (CIN); its structure, morphology, color and texture was determined for 4 weeks and tabulated for detailed study. The callus index number refers to the product value of length (cm) and width (cm) of callus which indicates the area (cm2 ) of the callus and is expressed as Mean ± SEM.

Source of Information: American-Eurasian Journal of Sustainable Agriculture, 7(3): 149-154, 2013 ISSN 1995-0748

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

It is concluded that organic supplements and activated charcoal has enormous and diverse effect on cell growth and organogenic response on callus of banana (M. sapientum cv. Anupam). The results of current study can play a role for in vitro research on organogenesis and cellular morphology because we found notable points from our experiment, as outlined below. 1. Callus growth varied with nutrient supplements. 2. Features of callus (color, texture and morphology) depended on culture medium. 3. Rhizogenesis can be achieved from addition of additives. 4. This protocol would be a fundamental step towards artificial seed production, breeding programs and for genetic manipulation. 5. Supplementations of organic growth adjuncts in culture medium is a simple, practical, and beneficial method to improve media used for commercial production.

Knowledge Management: Journal