Shoot regeneration of young leaf explants from basil (Ocimum basilicum L.)

An efficient plant regeneration protocol was successfully developed for bas il (Ocimum basilicum L.). Explants from 1 mo. old seedlings yielded the hig hest frequency of 85% regeneration with an average of 5.1 sheets per explan t. The regeneration protocol was performed on three basil varieties (Sweet Dani; methylcinnamate; Green Purple Ruffles). Callus and shoot induction wa s initiated on Murashige and Skoog basal medium supplemented with thidiazur on (16.8 El mu M) for approximately 30 d. Shoot induction and development w ere achieved by refreshing the induction medium after 14 d. The most morpho genetically responsive explants were from the first fully expanded true lea ves of greenhouse-grown basil seedlings. All developing bud tissue demonstr ated temporary anthocyanin expression; however, anthocyanin expression in G reen Purple Ruffles remained stable until maturity. Developing shoots were rooted in the dark on media with thidiazuron removed. Within 20 d, rooted p lantlets were transferred and acclimatized under greenhouse conditions wher e they developed normal morphological characteristics. This is the first re port of a successful in vitro regeneration system for basil through primary callus.