Efficient plant regeneration from multiple shoots formed in the leaf-derived callus of Lavandula vera, using the “open culture system”

Abstract

To develop an efficient procedure for plant regeneration from leaf-derived callus of Lavandula vera DC, the production of multiple shoots and the formation of roots from these shoots were studied. When calli were cultured in a medium with 4.0×10 −7 M N-(2-chloro-4-pyridyl)- N′-phenylurea (CPPU), urea-type cytokinin, multiple shoots were obtained from the greenish surface of the callus efficiently at a rate of 52.2%. For multiple shoot development, the shoots were divided into 7–20 shoot clusters with 2–3 shoots and subcultured every 2 weeks. Two modified culture-systems were employed for subculture; a “closed culture system” and an “open culture system”. In the open-system, shoot clusters grew and elongated vigorously. In the closed-system, most of the clusters showed vitrification and did not elongate. In root induction culture, shoot clusters in the open-system were well rooted and grew vigorously, the highest rate of root formation being 74.0%, in the 1 2 MS medium supplemented with 1.0×10 −6 M indoleacetic acid (IAA). On the other hand, shoot clusters in the closed-system showed vitrification and did not root well. About seven times more regenerants were obtained from the multiple shoots than from normal shoots treated with 4.4×10 −6 M 6-benzylamino purine (BA), a purine-type cytokinin. These results indicate that efficient plant regeneration in L. vera from callus using multiple shoots is possible by means of an open-system.