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Somatic embryogenesis and callus formation in sugarcane (Saccharum SPP L.) using different concentration of 2, 4-D and RAPD analysis of plants regenerates

Misbah Naz*, Ghulam Sughar, Zahoor Ahmed Soomro, Imtaiz Ahmed, Neghat Seema, Ghulam Shah Nizamani, Saboohi, Muhammad Rashid Nizamani

Department of Biotechnology, Sindh Agriculture University, Tando Jam, Pakistan

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Present research was conduct to build up an efficient protocol for various concentrations of 2, 4-D for ”Callus formation and somatic embryogenesis in sugarcane (saccharin 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 2013- 2014. Three sugarcane varieties Bl4, NIA-2010, NIA-2011 with various concentrations of 2, 4-D (, 2.0, 3.0 and 4.0 mg l-1). While 3.0 mg l-1 Kin+IAA+IBA were used for callus proliferation and shoot formations. Four different concentration of IBA were used for rooting purpose 0.5, 1.0, 2.0 and 3.0 mgl-1+20 g Sugar. Embryonic callus was obtained by culturing young apical merited. Eight-month field grown three sugarcane clones via; BL4, NIA-2010 and NIA-2 011 we use for tissue culture somatic embryonic callus studies. The 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 a number of plantlets shoot length and regeneration growth was observed in those plants that call taken from 1.0 and 2.0 mg l-1.2, 4-D Maximum chlorophyll mutation frequency was recorded in NIA-2010 and BL4 grown on 1.0 mg l-1 2, 4-D. The maximum number of roots was observed in BL4 when 1.0 mg l-1IBA+ 20% sugar was applied, variability obtained through callus culture and also confirmed through random amplified polymorphic DNA (RAPD) techniques.

Callus prolifereation, Chlrophyll mutant regenretion and root formation, Embryonic Callus induction.
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