Somatic Embryogenesis from Leaf Explants of Gladiolus anatolicus (Boiss.) Stapf

Abstract

An in vitro micropropagation method by somatic embryogenesis was developed for Gladiolus anatolicus (Boiss.) Stapf using leaves of in vitro shoots obtained from lateral buds. Lateral buds removed from sterilized fresh corms were placed on Murashige and Skoog (MS) medium supplemented with various concentrations of N6-benzyladenine (BA) for shoot culture establishment. The highest number of shoot per lateral bud explant was on MS medium supplemented with 2 mg L(-1) BA (11.00 +/- 0.38). To induce somatic embryogenesis, leaves of in vitro shoots obtained from lateral buds were used as explant. Calli were obtained from middle and basal region of leaf explant cultured on MS basal medium supplemented with different concentrations of alpha-naphthaleneacetic acid: N6-benzyladenine (NAA:BA) ratio and without growth regulators. The highest rate of callus formation was obtained from basal part of leaves cultured on MS medium containing 5 mg L(-1) NAA in darkness (80 +/- 0.41%). Creamy-white and friable calli produced numerous somatic embryos on MS basal medium supplemented with 0.1 mg L(-1) BA within 4 weeks in light (On avarage 30 structures per callus). Well-developed somatic embryos were germinated on MS medium supplemented with 0.1 mg L(-1) BA and reduced sucrose concentration (20 g L(-1)). On this medium 40% of the somatic embryos developed into plantlets. Cormlet formation was observed on MS basal medium (30 g L(-1) sucrose) containing same concentration of BA.