Multiple shoot formation from mature seeds in lentils (Lens culinaris) G

A protocol based on seed culture was developed for efficient in vitro propagation of lentil (Lens culinaris Medik). Benzyladenine (BA), thidiazuron (TDZ), and kinetin all induced multiple shoot formation. In terms of the number of long shoots ( > 2.0 cm) produced per seed, BA and TDZ at optimum concentrations (0.2–0.4 and 0.1 mg/litre, respectively) had similar efficiency, whereas kinetin produced less shoots. Murashige and Skoog (MS) salt composition was better than that of Gamborge (B5) for shoot induction. Increasing calcium (Ca) concentration was necessary to overcome shoot-tip necrosis. For shoot elongation, fresh medium of the same composition of shoot induction medium could be used for stumps from medium with low BA ( < 0.8 mg/litre) or TDZ ( < 0.4 mg/litre). Otherwise, MS medium supplemented with 0.5 mg/ litre gibberellic acid (GA 3 ) or 0.05 mg/litre α-naphthaleneacetic acid (NAA) was appropriate. Multiple shoots were induced from 10 inbred of four species. Best rooting was achieved using MS medium supplemented with 1.5 mg/litre NAA. Using this rooting medium more than 50% shoots from the 10 inbreds rooted. From 80 to 100% plantlets of the 10 inbreds survived after being transferred to potting mixture in glass house. © 2002 Taylor & Francis Group, LLC.