The healthy four week old sword suckers from Musaparadisiaca Monthan cv. Karibale weighing about one kilogramuniformly sized were identified from elite plants of superior qualities such asbunch weight of above 40 kg’s, more than ten number of hands in a bunch, morethan eighteen number of fingers in second hand from top and length of pedunclemore than 20 to 24 inches was selected and cut from the pseudostem 15 cm above thebase level, weighing 500 to 1500 g were collected from the banana fields of aprogressive farmer and used as starting material for micropropagation.
Thesesuckers were transported to the Department of Biotechnology and Bioinformatics, Kuvempu University, Shankaraghatta and arranged on sand bed under shade and watered regularly. Periodically Fungicides/ Bactericides are applied to thesesuckers till they are taken for initiation process. The disinfection of suckerswas carried out thoroughly washed with tap water, soaked in a solution of 0.2%Bavistin and 0.1% Streptocyclin for overnight. One whorl of leaf sheath wasremoved and corm portion trimmed to a width of 1 inch x 1 inch and height about2 inches, dipped in 0.
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001 M Teepol solution for an hour and finally rinsedthree to four time using distilled water to remove residual Teepol. From such trimmed suckers shoot tip measuringabout 2.0 to 3.0 cm containing several sheathing leaf bases and axillary budalong with underlying rhizome tissue were isolated.Surface sterilization:The sucker cubes were transferred to the laminar airflowchamber. Tissue blocks containing shoot-tips and rhizomatous baseswere surface sterilized for six min in 70% ethanol, 0.
2% Mercuric Chloride solution for ten minutes and rinsed three times repeatedly with sterile distilled water forfive minutes. One whorl of leaf sheath was removed, corm portion trimmed andtreated with 70% alcohol for six minutes, 0.1 % mercuric chloride for tenminutes and rinsed three times repeatedly with sterile distilled water forfive minutes.
Further, a whorl of leaf sheath was removed, corm portion wastrimmed, dipped in 1 % sodium hypochlorite for fifteen minutes and rinsed threetimes repeatedly with sterile distilled water for five minutes. Then the cut surface of the sucker tissuewas further trimmed and dipped in antibiotic solution containing Cifotaxime0.1% and Gentamicin 0.05 % for 5 min. Finally, after the antibiotic treatmentthe suckers were treated with Ascorbic acid (100mg/l) for 10 min to avoidblackening of tissues due to phenolic exudation.
