Role of Interaction of BA and 2,4 – D in the Growth and Differentiation of the Callus of (Nigella sativa L.) Plants and Their Relation with the Level of Its Effective Compounds

Abstract

This study demonstrated the role of some standard growth regulators in the induction, growth and differentiation of callus of Nigella sativa L.. Benzyl Adenine (BA) and 2,4-Dichlorophenoxyacetic acid (2,4-D) were used with concentrations (10-3, 10-4 , 10-6 , 10-8 , 10-10 ) molar for each separately on one hand and their interference’s. The results show that the best media for the induction and growth of callus are those which contain 2,4-D (10-6)M as the fresh weight of the growing callus reached (3.58), (12.89) gm at 45,75 days respectively. Also best differentiation among shoots was with the media supplying 2,4-D (10-10)M with BA (10-6)M. But, the root growth was not observed before the callus reached the age of 100 days on the media which contain 2,4-D alone. This study explored the content of the callus of black seed plant like the active compounds and their comparison with the seeds contents. This was carried out using high performance liquid chromatographic technique HPLC. This technique proved that the callus content of active compounds almost equal or higher than those in the seeds bearing in mind that callus content differs according to the used growth regulator,it was found that compounds separated according to their Retention time which ranged between 4to 14 mint ,these compounds were closely similar ( according to the separated peaks) those compounds separated from the seed extract (Alcohol soluble phase).