Regeneration of garlic plants (allium sativum L., CV. “Chonan”) via cell culture in liquid medium

Abstract

Aiming at the genetic improvement of garlic cultivars, a cell suspension protocol was established which includes the induction of friable callus, establishment of cells in liquid medium, plating, regeneration, and bulb formation. Calluses of various textures from compact to friable and from green to yellowish were obtained by culturing explants excised from inner leaves of garlic bulbs on Marashig-Shoog (MS) medium with 2,4 dichlorophenoxy acetic acid (2,4-D), (1.1 mg/liter [5.0 µM]), picloram (1.2 mg/liter [5.0 µM]), and kinetin (2.1 mg/liter [10 µM]). Friable callus occurred on MS-A contained 2,4-D alone (1.0 mg/liter [4.52 µM]) and this callus was used to develop cell suspension cultures, which were maintained in liquid MS-B medium with a 2,4-D/benzyl adenine (BA) (0.5 mg/liter [2.25 µM]: 0.5 mg/liter [2.22 µM]) ratio. High plating efficiency was obtained on MS-C medium with different naphthalene acetic acid/BA combinations. Regeneration occurred after transfer of the caulogenic mass to MS-C medium containing 10 mg/liter (74.02 µM) and 20 mg/liter (148.04 µM) adenine for 60 days, followed by transfer to adenine-free medium. Plantlets transplanted to soil showed normal phenology. Shoots grown on modified MS medium supplemented with indolylbutryic acid (3.0 mg/liter [14.7 µM]) stimulated bulb formation by 30 days in culture.