Optimizing Culture Condition for High Frequency Regeneration in Vitro Tissue Culture in Daucus Carota L. and Genetic Stability Assessment

Abstract

Carrot is well known for medicinal purposes. For this reason optimization of tissue culture is necessary and first step for this purpose. In the present study, an efficient in vitro direct and indirect regeneration of Daucus carota L. was performed. The root, shoot, leaf and nodal explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentration and combinations of plant growth regulators (PGRs). Direct multiple shoot regeneration was induced on seventeen media culture varied in the type of culture medium and PGRs, the MS medium containing 1 mg/l Benzylaminopurine (BAP) + 2 mg/l NAA (Naphthylacetic Acid) was the best medium for direct shoot regeneration. In indirect regeneration the highest callus induction, embryogenesis and the shooting rate obtained from shoot segments followed by leaf segments cultured on the MS medium containing 2 mg/l NAA, respectively. The highest rooting percentage (95%) was recorded on the MS and ½MS medium both containing 1 mg/l NAA. Suggestion, medium 1 including MS + 2 mg/l NAA + 1 mg/l BAP best medium for direct regeneration, medium 6 including MS + 2 mg/l NAA, the best medium for indirect regeneration. Percentage of adaptation was 45%. Intra shoot variability of the shoot forming capacity indicate in the Nantes was depends on the shoot position. Shoot differentiation from leaf tissue indicate highest shoot- forming capacity was obtained of two nearest explants to apex of 1 mg/l NAA of 2 mg/l. Results of Genetic fidelity with ISSR markers indicate that of 6 regeneration plants, one plants not same with mother plants in genetic stability, but five regeneration plants same with mother plant in genetic stability. This results shown genetic stability in mother plant and regeneration plants is very high. In this research was obtain highest amount of callus induction, embryogenesis and shooting.