A Study On Tissue Culture For Three Wild Species Of Clematis Genus In China

The regeneration system in this paper was established by culturing stems with new born budsfrom three wild species of Clematis genus in China,which provided technical support for therapid promotion of wild.Then the inducing somatic embryogenesis and the cytologycharacteristics were studied on C. huchouensis Tamura, to explore the high frequency plantregeneration and morphogenetic pathway in vitro conditions, and guide the plantlet production.The results were as follows:1．The rapid propagation on three wild species of Clematis genus in China(1)The explants in vitro culture of proliferation basic medium was1/2MS culture medium.Rooting of basic culture medium was MS culture medium.(2)Proliferation: The suitable of axillary buds proliferation medium was1/2MS+0.5mg/L6-BA+0.3mg/L IBA+Sucrose30.0g/L for C. huchouensis Tamura the proliferation coefficientgot4.43; The suitable of axillary buds proliferation medium was1/2MS+0.5mg/L6-BA+0.3mg/L IBA+Sucrose40g/L for C. caerulea, and the proliferation coefficient was4.42; Thesuitable of axillary buds proliferation medium was1/2MS+0.3mg/L6-BA+0.1mg/L IBA+Sucrose20g/L for C. cadmia Buch, and the proliferation coefficient got4.23.(3)Rooting:The rooting suit culture medium was MS+NAA0.05mg/L for C. huchou-ensis Tamura, and the rooting rate was60%; The rooting siut culture medium was MS+0.08mg/L NAA+0.1mg/L IBA for C. caerulea, and the rooting rate was50%; The rooting siutculture medium was MS+0.1mg/L NAA+0.5mg/L IBA for C. cadmia Buch, rooting rate got43.3%.(4)Transplant:Rooting culture transplant was suitable in spring, the matrix to perlite andvermiculite2:1ratio was relatively appropriate,and the survival rate could reach to more than80%.2．Somatic embryogenesis of C. huchouensis Tamura(1)The embryo callus induction medium suitable is1/2MS+0.2mg/L TDZ+0.2mg/LNAA+100mg/L inositol+200mg/L casein hydrolysat+1g/L activated carbon. Additionalsucrose30g/L, AGAR5.8g/L, pH5.8, dark treatment.(2)The callus proliferation of embryo subculture suitable medium is1/2MS+0.05mg/LTDZ+0.05mg/L NAA+100mg/L inositol+200mg/L casein hydrolysate+1g/L activated carbon.subculture after five times the proliferation coefficient be more stable.(3)Embryonic cells are formed better in1/2MS+0.03mg/L ABA+0.05mg/L NAA,somatic embryos induction rate can amount to10.526%.3．Histological study on the origin and development of somatic embryos,it was found thatsomatic embryos were mostly developed from single embryonic cells that possessed the ability ofembryoid occurrence form the surface or near of the callus. somatic cells after spherical embryos,heart-shaped embryos, torpedo embryo and then develop into a complete cotyledons embryos ofplants.