Plant Regeneration from Uninuclear Microspore Suspension Cultures of Aesculus hippocastanum L.

Abstract

Androgenesis induction in suspension culture of uninuclear microspores of Aesculus hippocastanum L. was achieved in MS liquid medium supplemented with MS mineral solution (Murashige, Skoog, 1962), 5% sucrose, mixture of vitamins (Radojevic, 1991), l-proline (Pro), casein hydrolysate (CH, 200 mgL⁻¹, each), 2,4 — dichlorphenoxyacetic acid (2,4-D) and 6- furfurylaminopurine (KIN, 4.5 µM, each). Formation of androgenic embryos (AE) in different stages of development i.e. globular (GE), heart — shaped (HE) and embryo with cotyledons (AECo) were obtained in liquid and/or solid MS medium 4 weeks after suspension plating. The germination (100%) of AECo was achieved in agar medium supplemented with 400 mgL⁻¹ l-glutamine (Glu), 0.1% activated charcoal (AC) and 0.5% polyethylene glycol (PEG). The best plant conversion rate of AECo (11%) was obtained after the successive transfer from agar M₂ (M₀ + AC 0.5%) medium to solid N medium without hormones.