Makowczyńska J and Andrzejewska-Golec E, Acta Soc. Bo t. Pol., 69: 245-250 (2000)
Summary
Early stages of direct somatic embryogenesis in Plantago asiatica were observed by light microscopy. In calli of Plantago asiatica two phenomena occurred simultaneously: somatic organogenesis and somatic embryogenesis.
Objectives
Induction of somatic embryogenesis in Plantago asiatica L.
Materials
Seeds of Plantago asiatica L. from Botanical Garden Faculty of Science, University of Tokyo
Explant
Seeds
Initial culture
Seeds of Plantago asiatica L. were surface sterilized by immersion in 2.0% sodium hypochlorite for 10 min, then rinsed 3 times in sterile water. Then, they were germinated in aseptic conditions on Murashige and Skoog medium supplemeted with 0.2 mg/l kinetin and 1 mg/l giberellic acid and 0.7% Difco Bacto Agar. The cultures were placed in the dark and after germination they were transferred into light. 4 week-old seedling were used as explants (hypocotyls, roots, cotyledons).
Shoot multiplication
For embryogenic callus initiation, explants were placed on the MS medium with 0.5 or 1 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.2 mg/l 6-benzylaminopurin (BAP). The best result for callus initiation was observed with the root as explant, and somatic embryos; globular stage embryos and polycotyledonary torpedo stage embryos occurred on the root tissue cultured with 1 mg/l 2,4-D and 0.2 mg/l BAP (passage 0). The resulting calli after 6 weeks of passage 0 were transferred onto agar medium MS with 0.5 mg/l 2,4-D and 0.2 mg/l BAP or 0.2 mg/l α-naphthaleneacetic acid (NAA) and 0.2 mg/l BAP or only 0.5 mg/l BAP-6 weeks of passage 1. In calli of various origin histological study showed globular, heart and aberrant torpedo stage embryos and rhizogenesis was observed. Resulting microshoots had well-shaped chloroplasts, stomata and headed hairs typical for Plantaginaceae family. The calli obtained from the first passage were grown on MS medium supplemented with 0.5 mg/l abcisic acid (ABA) for prevention of aberrations in somatic embryogenesis and 1 mg/l BAP-passage 2-5. In passage 5, many microshoots formed on the callus induced from root.