Efficient adventitious shoot regeneration and somatic embryogenesis in pea

Abstract

A rapid and simple method for adventitious shoot regeneration and somatic embryogenesis from immature cotyledon explants of pea (Pisum sativum L.) is described. Cotyledon size and the explant orientation to the medium surface were shown to have a clear effect on shoot regeneration. The highest frequency of shoot regeneration was achieved when the distal end of the greenest cotyledons (7–8 mm in size) were placed in contact with the agar surface. Shoots rooted at a frequency of 80–90% and grew into normal fertile plants. Somatic embryos were induced in cultures of immature cotyledons on modified MS medium containing high levels of α-naphthaleneacetic acid (27–215 μM) and 2,4-dichlorophenoxyacetic acid (23–181 μM). A higher frequency of somatic embryos with a normal morphology were induced using α-naphthaleneacetic acid.