Abstract
To establish a simple and constant suspension culture system for gene manipulation, the leaflets of “C1” sexual bahiagrass line were used for embryogenic callus formation on Murashige and Skoog’s medium (MS) supplemented with 2mg l-1 2, 4-dichlorophenoxyacetic acid (2, 4-D). The obtained embryogenic calli were broken into pieces for suspension culture in N6 liquid medium (Chu et al., 1975) supplemented with 1mg l-1 2, 4-D. After 2 weeks of cultures two kinds of bigger and smaller calli were obtained. Another 2 weeks later, the bigger (≥4mm diameter) became brown color and vacuolated, and the smaller (≤2mm diameter) showed uniform size and dense cytoplasm. The smaller calli were then selected and moved onto the solid medium for regeneration. When the cultures were Continued, shoots and plants were regenerated. The abilities of shoot regeneration from suspension culture could be kept at least for 12 months. This study provides a simple and constant plant regeneration system available for making transgenic apomictic plants based on embryogenic suspension culture derived from leaflets of sexual bahiagrass.
