Plant regeneration from protoplasts of Gentiana straminea Maxim

• Published in: Open life sciences Vol. 11; no. 1; pp. 55 - 60
• Main Authors: Shi, Guomin, Yang, Lina, He, Tao
• Format: Journal Article
• Language: English
• Published: De Gruyter Open 01.01.2016; De Gruyter
• Subjects: 2,4-D; Agar-pool method; callus; cell division; endo-1,4-beta-glucanase; Gentiana straminea; gentiana straminea maxim; liquids; Maxim; Plant regeneration; plantlets; Protoplast culture; protoplasts; somatic embryogenesis; somatic embryos; sorbitol
• ISSN: 2391-5412; 2391-5412
• DOI: 10.1515/biol-2016-0007

A protocol is described for plant regeneration from protoplasts of Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 10 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L N -benzylaminopurine (BA). Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.