Plant regeneration from seed-derived embryogenic callus and cell suspension cultures of bahiagrass ( Paspalum notatum)

Abstract

We have established a high-frequency plant regeneration system via somatic embryogenesis from seed-derived callus and cell suspension cultures in 6 genotypes of bahiagrass ( Paspalum notatum). Embryogenic callus was initiated from mature seeds on Murashige and Skoog (MS) medium supplemented with 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 3.0% sucrose and 0.3% Gellan Gum in the dark. Culture response was found to be correlated with genotype. ‘Pensacola’ had the best response in embryogenic callus formation, and 74% of the calli regenerated plants. Cell suspension cultures were established from embryogenic callus of ‘Pensacola’ in modified N 6 medium containing 1.0 mg/l 2,4-D. The suspension was composed of compact cell clusters. When smaller clusters (approx. 2.0–3.3 mm in diameter) were transferred to a solid MS medium with 3.0% sucrose but without hormones, plant regeneration was initiated at high frequency (28.6%). Morphological evidence is provided that regeneration of suspension cells occurred via embryogenesis. Regenerated plants were established to soil after culture in water at room temperature for 14 days for acclimatization.