Influences of factors affecting the induction of high protoplast yield and callus induction in two different carnations

This study aims to establish an efficient protocol for protoplast isolation, cell division, and callus induction in two carnation cultivars, Chabaud and Giant Chabaud, by examining the influence of different types of plant tissue, enzyme concentrations, incubation times, cotyledon ages, and medium compositions. Our results indicate that protoplast yield varies significantly between different plant tissues, with true leaves offering the highest yield and viability, especially under a 0.1% driselase concentration and a 6-h incubation time. We observed that increasing the driselase concentration to 1.0% significantly reduced protoplast yields in all tissues tested. In addition, the age of the cotyledons notably affected protoplast yield, with younger cotyledons providing higher yields. The Murashige and Skoog medium supplemented with 1 mg/L zeatin and 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) proved to be the most effective for promoting cell division and colony formation from protoplasts derived from cotyledons and true leaves. The study also found that plant growth regulators (PGRs) significantly influence callus proliferation, with differences observed between protoplast sources.