Research On Eucalyptus Regeneration System Establishment And Gene Transformation With Agrobacterium Tumefaciences

Eucalyptus. which originally grows in Australia. is one of the three most rapidly growing trees of the world. It has become the strategic plant species for developing the fast-growing forestry in southern China and the main resources of papermaking industry. Controlling the plant phenylpropanoid in Eucalyptus is essential for reducing the pollution and promoting the producing efficiency. This research focuses on transplanting the anti- 4-coumarate into Eucalyptus to reduce the henylpropanoid. By using the established regeneration and transformation system. one of the transgenic callus had been tested positive through PCR.A primary tissue culture system for callus induction and plant regeneration had been established by using different growth regulators on medium MS. 1/2MS. White and WPM. At the same time. explants sterilization. multiplication and rooting had also been researched. Bud explants first cleaned with eradicator. washed under tap water for 30min. and then sterilized by 70% ethanol for 30sec and 0.5% NaClO for 8min. The explants multiplicities in high rate on MS + IBA 0.25mg/L + 6-BA 0.5mg/L + Suger 30g/L + Agar 6g/L and roots well on 1/2White + IBA0.5 mg/L + Suger 40g + Agra 6g/L. Leaf calluses had been induced on WPM+6-BA 0.1mg/L+NAA 3.0mg/L+Suger 50g/L+Casein 1g/L+ Agra 5g/L with nearly 100% rate and sprouts can be induced on WPM+6-BA 0.3mg/L+ NAA 0.03mg/L+ Suger 50g/L+ Agra 5g/L from these calluses.Research on gene transformation system has found the suitable Kam concentrations for callus. bud and root inducing selection and the time for Agrobacterium tumefaciences infection and incubation. The Kam concentrations for callus. bud and root inducing selection are 30. 60. 80mg/L respectively. Leaves infected with Agrobacterium tumefaciences for 8min and incubated for 2 days gained the highest transformation rate. The transformed leaves were cultured on callus inducing medium and formed Kam...