The European Horse Chestnut Somatic Embryos And Plant Regeneration

It was of great value to propagate Aesculus hippocastanum L rapidly, which was an important landscape tree and with great value for medical treatment, by the means of cell engineering.Somatic embryogenesis systems were investigated by means of vitro culture from young leaf of Aesculus hippocastanum L. The results were showed that embryogenic calli were induced from young leaves, which were cultured on MS medium supplemented with 2,4-D 2mg/L and KT 0.2mg/L. For the proliferation of embryogenic calli, the suitable culture medium was MS+BA 8mg/L +NAA 1mg/L. The development and maturation of somatic embryo could be much improved by using the medium of MS+ZT 2mg/L or BA 5mg/L +IAA 0.2mg/L. For the induction of secondary somatic embryo from integral somatic embryo, the suitable culture medium was MS+KT 0.1mg/L+NAA 0.01 mg/L or MS+ZT 0.1mg/L+NAA 0.01 mg/L, the proliferation frequency is 214%, 256% respectively.The cotyledonary generated from somatic embryos of Aesculus hippocastanum L. can be used to induce secondary somatic embryogenesis or bud differentiation in vitro culture also. The results were showed that somatic embryos can be directly re-induced from cotyledonary, which was cultured on MS medium supplemented with ZTO.5-2 mg/L, BA 1mg/L and NAA 0.2mg/L, and the induction rate was over 80%. For the bud directly differentiation , the suitable culture medium was MS+BA 2mg/L +NAA 0.2mg/L or MS+TDZ0.01mg/L +NAA0.2 mg/L, of which the induction rate was 88% and 100% respectively.An efficient tissue culture system has been developed with the bud of mature seed of Aesculus hippocastanum L. as explants . Buds were induced from 2cm high young plantlet cultured on MS medium supplemented with 0.6 mg/L 6-BA plus 0.1mg/L or 0.4~0.6mg/L ZT plus 0.1 mg/L NAA for 15 d, and the induction rate was 100%, the mean No. of buds was 35.7; The combination of MS+0.2mg/L 6-BA +0.1mg/L NAA + 10mg/L AD was the suitable culture medium for elongation of the buds. The medium of 1/2MS+ 0.4mg/L NAA +0,2mg/L IBA was used for rooting, and the rooting rate was 75%.The origin and development of somatic embryos was observed by histological method. It was found that the somatic embryos of Aesculus hippocastanum L were developed from single cell. Somatic embryos can be produced both from the surface and the inner of the calli. Embryogenic cell has normal shape, dense cytoplasm , clear nucleolus, hearty metabolize; Plantlet regenerations were developed from global embryo, heart-shaped embryo, torpedo-shaped embryo and cotyledon embryo, in which the V-shaped vessel vascular bundles and shoot tip meristem and root tip meristem were observed.Inconclution, for produce Aesculus hippocastanum L in a mass, we should induce secondary somatic embryos with integral somatic embryos, the suitable culture medium is MS+KT 0.1mg/L+NAA 0.01 mg/L or MS+ZT 0.1mg/L+NAA 0.01 mg/L.