SOMATIC EMBRYOGENESIS AND PLANTLET REGENERATON VIA EMBRYOGENIC SUSPENSIONS OF GRAPE (VITIS VINIFERA L.)D.K. SHARMA, M.K. TRIPATHI, R. TIWARI, R.P. PATEL, B. S. BAGHEL AND ASHOK AHUJA
A simplified procedure for establishment of embryogenic cell suspension culture from the friable embryogenic callus of Vitis vinifera L. was accomplished by transferring 6-8 weeks-old embryogenic calli achieved from nodal segment and leaf disc explants in liquid media. The asynchronous Embryogenic cultures were swamped with clumps of proliferating embryos of various developmental stages along with modest non-embryogenic tissues. The frequency of embryo proliferation was recorded to determine growth rate of embryogenic tissues under diverse conditions. Modulation of initiation and proliferation of embryogenic cell suspension culture was under regulation of the influence of exogenous plant growth regulators added in culture medium at different concentrations and combinations. Higher relative growth of embryogenic cell suspension cultures was recorded with combination of culture medium MS+2.0mgL-12,4- D+0.5mgL-1Kn+30.0gL-1 sucrose, whereas, cell clumps/embryoids in higher numbers were attained on culture medium (MS+2.0mgL-12,4-D+0.5 mgL-1BA+30.0gL-1 sucrose). For subsequent subculturing, reduced level of 2,4-D (1.0 mgL-1) in combination with 0.5 mgL-1 BAP accelerated somatic embryogenesis. High frequency plantlet regeneration was recorded on regeneration medium supplemented with 0.5 mgL-1, each of BAP, Kn and NAA. In terms of root proliferating ability and root of higher length, rooting medium MS.5IB.5Kn(MS+0.5mgL-1IBA+0.5mgL-1Kn+15.0gL-1 sucrose+7.5gL-1 agar) was found superior among the tested media combinations. However, root formation in higher frequency was attained with media supplemented with 2.0mgL-1 IBA. Among two genotypes. Thompson seedless was found consistently superior to Karnet for all attributes investigated. Plantlets regenerated showed normal development and were established successfully in the field after hardening.
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