Abstract

EFFECT OF ZnSO4 AND CuSO4 ON REGENERATION AND XANTHOTOXIN CONTENT IN AMMI MAJUS AND AGROBACTERIUM- MEDIATED TRANSFORMATION

*Deepshikha Pande Katare , Amit Chandra Kharakwal and Priyanka Sharma

ABSTRACT

The present study investigates the effects of zinc and copper on morphogenic, biochemical and phytochemical responses of Ammi majus L. and determines the highest tolerance levels of zinc and copper to which tolerant cultures can be raised. Incorporation of zinc sulphate (100-400μM) and copper sulphate (25-100 μM) in Murashige and Skoog medium + (in mg l-1) 0.5 indole-3-acetic acid + 2.0 kinetin + 500 casein hydrolysate + 40 adenine elicited variation in growth and percent regeneration of calli. Zinc sulphate at 200 μM proved to be the best and promoted high percent (15.5±0.22) regeneration, growth and enhanced xanthotoxin production (12.8±0.03) whereas copper tolerance was only up to 25 μM. Variation in xanthotoxin was observed with highest concentration (12.8+ 0.04 mg/g DW) at 200 μM zinc sulphate. A simple technical procedure is described here using Agrobacterium tumefaciens for the efficient transformation method for this important medicinal plant. The procedure begins with inoculation of leaf explants in bacterial suspension followed by co-cultivation. Transformed shoots were selected on the regeneration medium containing antibiotic and the presence of the inserted genes was checked by gus assay.

Keywords: Ammi majus, CuSO4, gus, transformation, xanthotoxin, ZnSO4.