An Efficient Protocol for In vitro Regeneration and Acclimatization of Banana (Musa spp.) cv. Grand Naine
H. B. Medhane
Department of Plant Biotechnology, K. K. Wagh College of Agricultural Biotechnology, Nashik- 422003, Mahatma Phule Krishi Vidyapeeth, Maharashtra, India.
R. S. Bhadange
Department of Plant Biotechnology, K. K. Wagh College of Agricultural Biotechnology, Nashik- 422003, Mahatma Phule Krishi Vidyapeeth, Maharashtra, India.
S. D. Surbhaiyya *
Department of Plant Biotechnology, K. K. Wagh College of Agricultural Biotechnology, Nashik- 422003, Mahatma Phule Krishi Vidyapeeth, Maharashtra, India.
N. S. Pachpor
Department of Plant Biotechnology, K. K. Wagh College of Agricultural Biotechnology, Nashik- 422003, Mahatma Phule Krishi Vidyapeeth, Maharashtra, India.
R. S. Nandre
Department of Plant Biotechnology, K. K. Wagh College of Agricultural Biotechnology, Nashik- 422003, Mahatma Phule Krishi Vidyapeeth, Maharashtra, India.
*Author to whom correspondence should be addressed.
Abstract
Aim: The study was undertaken with a view to standardize a protocol for in vitro regeneration of Banana cv. Grand naine using shoot tip of sucker as an explant
Methodology: In present study, explants sterilized with different sterilizing agents such as Tween-20 (1%), Bavistin (0.5-1%), Streptomycin sulphate (250 mg/L), Ascorbic acid (150 mg/L) + Citric acid (100 mg/L), HgCl2 (0.1%) and 70% ethanol.Sterilized explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of plant growth hormones for shoot initiation BAP alone (1.5, 2.0, 4.0, 6.0 mg/L) and BAP (3.0 mg/L) in combination with IAA and IBA (2.0 mg/L), elongation BAP (3.0 mg/L) and NAA (1.0, 1.5, 2.0 mg/L) and rooting IAA (1.0, 1.5 mg/L) and IBA (1.0, 1.5 mg/L). Primary and secondaryhardening was done in potting mixture containing autoclaved black soil: vermicompost: cocopeat (1:1:1) and garden black soil, cocopeat and red soil (1:1:1) respectively.
Results: In present investigation 1% Bavistin (fungicide) showed maximum respond to prevent fungal contamination. Highest shoot initiation (100%) was observed on a MS medium fortified with BAP (1.5 mg/L). Maximum shoot length (10.7 cm) was recorded on a MS medium supplemented with BAP (3.0 mg/L) + NAA (2.0 mg/L) + Activated charcoal. Maximum root initiation was observed on half strength MS medium supplemented with IAA (1.5 mg/L). In vitro regenerated plantlets hardened on the mixture of autoclaved black soil: vermicompost: cocopeat (1:1:1). After 14 weeks In vitro plantlets transferred in green house for acclimatization where, 80% survival rate was recorded.
Conclusion: Regeneration protocol was successfully standardized. Therefore, itcan be used for large scale propagation of healthy, disease and virus free planting material and In vitro propagation helps to meet higher demand of healthy planting material within shorter period.
Keywords: Banana, grand naine, in vitro, suckers, activated charcoal