Study On The Tissue Culture Regeneration System Of Trifolium Hybridum L. And Trifolium Repens L.

Triflium L. is the most important fodder crop plant and turf grass. The tissue culture regeneration system will setαgood base for the application of other technologies and gene transform in the future. The effect of different media, different concentrations combination of phytohormones on embryogenic callus inducing, embryoid differentiation and plant regeneration were studied,αfull regeneration system of clover was achieved. The results of this research are followed:Trifolium hybridum L.,Haifa,Huia and Rivendel of Trifolium repens L. were tested in this research, among which, Callus induction rate of Hypocotyls of Trifolium hybridum L. was 97.6%; Hypocotyls is the most effective explants. Cotyledon of Trifolium hybridum L. and Trifolium repens L. (Rivendel) made difference when cultured on different initiation basal media (contained the same regulators) for callus induction, modified SH was the optimum medium. It was found 2,4-D essential for embryogenic callus induction, but not NAA. The best phytohormones for embryogenic callus induction was 4mg·L^-12,4-D+0.5mg·L^-16-BA , The embryogenic callus induction rate of Trifolium hybridum L. and Trifolium repens L. (Rivendel) reached 43.6% and 34.1% respectively. Light condition has predominant effect on callus generated time and callus texture, but has no effect on callus induction rate.Callus in all combination was heterogeneous. There were EC and NEC. Cell of EC were small and regular and exhibited embryogenic competence. But EC was on the contrary. When EC of Trifolium hybridum L. and Trifolium repens L.(Rivendel) cultured on different differentiation media, MSO and Modified SH were more effective than SH and MS. The best embryoid differentiation for Trifolium hybridum L. was MSO+0.5mg·L^-1NAA+1.0mg·L^-16-BA, for Trifolium repens L. (Rivendel) was MSO+0.5mg·L^-1NAA+2.0mg·L^-16-BA. The presence of AgNO3 stimulated embryogenesis, 1mg·L^-1AgNO3 was the best concentration. Adding 20g·L^-1 Sucrose, The embryoid differentiation from Trifolium hybridum L. and Trifolium repens L. (Rivendel) was up to 31.4% and 24.4%.Many of the embryiods were abnormal. Isolated embryoids did not germinate in...