Maintaining the artemisinin content through direct and indirect in vitro regeneration and their assessment of variations with the field grown mother plants of Artemisia annua L.

Abstract

An efficient protocol for maintaining the artemisinin content in tissue culture and high frequency of in vitro direct and indirect regenerations of multiple shoots of high artemisinin yielding genotypes of Artemisia annua has been developed and their comparison with field grown mother plant has been carried out. The eleven elite genotypes (containing more than 1% artemisinin) were tested on seven different modified media formulations. Modified half MS (Murashige and Skoog’s) media containing 100 mg L−1 myo-inositol, 0.5 g L−1 casine hydrolysate, 5 mg L−1 biotin, 2 mL L−1 RT (Revised Tobacco) vitamin stock, 0.5 mg L−1 BAP and 0.01 mg L−1 NAA showed best regeneration while, above modified MS medium containing 0.2 mg L−1 BAP and 0.2 mg L−1 NAA showed maximum shoot multiplication with maintained artemisinin content. Based on the chemical profiles of both the systems, minor difference was observed in their artemisinin content. A large scale culture of these plants maintained the normal growth index along with the artemisinin content and could be a better alternative to maintain the high artemisinin yielding genotypes with their genetic constraint in specific media combinations and also used as base material for further genetic improvement. Graphical abstract: [Figure not available: see fulltext.].