Studies On Somatic Embryogenesis From Cell Suspension Culture Of Lilium Tenuifolium (Orirntal×Trumpet)

In plant genetic transformation, a good planting system is very important. Transformation through embryogenesis has advantages like high regeneration rate, less chimeras, less mutation. However, in genetic transformation of lily, there are many problems such as scanty materials, slower proliferation, low transformation rate and the existing research is limited on the lilies and the new iron cannon lily. Therefore, this study focused on the factors of generation oflily （Lilium tenuifolium Oriental×tumpe‘Robina’）.The main results were as follows:（1） To study the growing condition of callus inducted by two kinds of hormones, bulbscales，filament， ovary and stem axis of Lilium tenuifolium oriental×tumpe‘tRobina’ wereused as explants, and then inducted callus were performed suspension culture. The resultsshowed that: the optimum medium for callus induction of bulb scales and filament isMS+Picloram1.0mg/L，however for the ovary is MS+Picloram1.5mg·L^-1.The stem oxis hasthe highest rate of callus induction on medium MS+NAA2.2mg·L^-1+TDZ0.1mg·L^-1，to89.20%. During suspension culturing, the suspension cultures were established by thecombination of NAA and TDZ with2～5mm cell clusters, which cost long time comparingwith the suspension cultures established by picloram with1～3mm cell clusters.In threesuspension cultures induced by picloram，the optimum callus was derived filament onproliferation and regeneration. The plantlet inducing rate of suspension cultures,which wasderived stem oxis,was higher than other three suspension culture induced by picloram. VectorpCAMBIA1301was transformed into different suspension culure（initiated from filament andstem oxis）by agrobacterium mediation. The frequency of transient expression of suspensionculure from stem oxi（sinduced by the combination of NAA and TDZ） was higher, comparedwith which from filament.（2） Adventitious buds which differentiated from bulb scales, filaments and stalk ofLilium tenuifoliu oriental×tumpet‘Robina’were used to form embryogenic callus. It producedthe most embrogenic callus when explants were cultured on induction medium whichcontained0.5mg·L^-1NAA. The best result of callus proliferation was made via a alternativeculture method which callus was transferred on solid and liquid medium by turns. The GUSgene contained in vector pCAMBIA1301was introduced into the embryogenic cultures byAgrobacterium-mediated tracient expression and then GUS chemical tissues staining was conducted to observe. The result showed that it was no significant transient expressionefficiency variance among callus of three cultivation methods.（3）Plasmid of pCAMBIA1301was used to study the best condition of transformation.The best infection time was10min. Transgenic rate reached26.8%when embryogenic calluswhich was induced from filments was used as explants.Whereas, stem callus induced on medium with NAA and TDZ gave a diiferent resultwhich the best infection time was8min,and the transgenic efficiency was28.3%.The best infection time of callus induced on medium with NAA was10min. Transgenicrusluts were not different.