Establishment Of High Efficient Regeneration System And Research On Genetic Transformation Of Lilium Spp.

Lily (Liliu spp.) is an important ornamental flower of perennial bulb plants in the world. It has great value because of its big flowers, various colors and pleasant fragrance and they are widely used as cut flowers, potted flowers and in gardens. Traditional propagation methods can't meet the requirements to lily. Modern molecular breeding has become an important complement and tissue culture and genetic engineering play an significant role in practice and theory.In this thesis, research on plant regeneration with high efficiency from leaf explants and genetic transformation were studied. The resulte are as follows:(1) Plant regeneration system in vitro culture was established from leaf explants of Lilium. longiflorum Thunb.'Snow Queen'. Results showed that the differentiation ability of lamina was stronger than that of petiole. The best medium for shoot multiplication was MS medium incorporated with NAA 0.5 mg/L and 6-BA 2.0 mg/L with the mean multiplication rate of 3.5. The shoots formed roots best on MS medium containing NAA 0.2 mg/L.(2) Plant regeneration system in vitro culture was also established from leaf explants of Oriential Lilium'Tiber'. Results showed that the differentiation ability of petiole was stronger than that of lamina. The best medium for shoot multiplication was MS medium incorporated with NAA 2.0 mg/L and 6-BA 0.1 mg/L. The shoots formed roots best on 1/2MS medium containing NAA 0.2 mg/L.(3) Effects of sucrose concentration on enlargements of bulblets from Lilium. longiflorum Thunb. were studied and results showed that 60 g/L sucrose was perhaps sufficient to support the growth of bulblets.(4) The resistance of lamina and bulbscale chips explants from Lilium. longiflorum Thunb. to Hyg for selecting culture were studied and the optimum concentrations respectively were 1.0 mg/L and 2.0 mg/L.(5) The transient expression of GUS showed that it was feasible to infect lamina explants from Lilium. longiflorum Thunb. through Agrobacterium-meditation. Blue spots were also examined in cut places and middle sites of lamina through Sanitation-aided Agrobacterium transformation.(6) Regeneration plants of Lilium. longiflorum Thunb. resistant to Hyg were obtained by Agrobacterium strain EHA105 with pCAMBIA1304+anti lLACO1. Six plants were positive by PCR analysis.(7) Regeneration plants of Lilium. longiflorum Thunb. resistant to Kan were obtained by Agrobacterium strain EHA105 with pCAMBIA2300 + PTA. Six plants were positive by PCR analysis. This work lays a good foundation for research on genetic transformation of lily by Agrobacterium-meditation and improvements of its traits.