Callus formation and somatic embryogenesis in sugarcane (Saccharum spp L.) using various concentrations of 2, 4-D and RAPD analysis of regenerated plants
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Abstract
The present study was carried out to build up an efficient protocol for various concentrations of 2, 4-D for “Callus formation and somatic embryogenesis in sugarcane (Saccharum spp L.) using various concentrations of 2, 4-D and RAPD analysis of regenerated plants in the laboratory of Biotechnology Nuclear Institute of Agriculture, Tandojam during the year 2015 to 2016. Three sugarcane varieties Bl4, NIA-2010, NIA-2011 with different concentrations of 2, 4-D (0.5.1.0, 2.0, 3.0 and 4.0 mg L-1) were used respectively , while 3.0 mg L-1Kin+IAA+IBA was used for callus proliferation and shoot formations. Four different concentration of IBA were used for rooting purposes 0.5, 1.0, 2.0 and 3.0 mgL-1+20 g Sugar. Embryogenic callus was obtained by culturing young apical meristem. Three sugarcane clones BL4, NIA-2010 and NIA-2 011 were developed in the field area for eight month and sand was used for tissue culture somatic embryonic callus study. Apical meristematic region was used for callus formation and somatic embryogenesis induction on 0.5, 1.0, 2.0, 3.0 and 4.0 mg L-1.2, 4-D actively growing callus was subcultured on kin. IAA, IBA, 3.0 mg L-1. Maximum callus proliferation and number of plantlets shoot length and regeneration growth were observed in plants that 1.0 and 2.0 mg L-12, 4 -D callus was taken from. Maximum chlorophyll mutation frequency was observed in NIA-2010 and BL4 grown on 1.0 mg L-1 2, 4-D. Maximum number of roots were observed in BL4 when 1.0 mg L-1IBA+ 20% sugar was applied; variability was also obtained through callus culture and confirmed through random amplified polymorphic DNA (RAPD) techniques.
Keywords: Embryonic callus induction; Callus proliferation; Chlorophyll mutant regeneration and root formation