Isolation and culturing of protoplasts in endangered Lachenalia viridiflora

Abstract

Lachenalia viridiflora is a critically endangered geophyte. This study aimed to develop a protocol for protoplast isolation and culture to enhance biotechnological tools for the improvement of this species. Protoplasts were isolated from micropropagated bulbils with leaves. Isolation yielded 1.2 × 10⁵ to 1.7 × 10⁵ protoplasts per gram of tissue, with 60–70 % viability. Protoplast-derived cells underwent divisions that resulted in microcallus formation and later shoot regeneration, demonstrating the potential of an elaborated protocol for successful protoplast-to-plant regeneration.