In Vitro Germination,Storage And Genetic Transformation Of Hemerocallis Fulva Pollen

Genetic transformation plays an important role in gene function verification and cultivation of fine crops.At present,most genetic transformation methods need to be combined with tissue culture regeneration,which involves a complex and long process;especially many important economic crops are difficult to regenerate.Using pollen as genetic transformation acceptor,the transformed pollen can directly pollinate to produce transgenic seeds,which avoid the obstacles of tissue culture and greatly shorten the period of obtaining transgenic germplasm.The results showed that the optimum medium for pollen germination of Hemerocallis fulva was 10% sucrose,0.05% boric acid,0.3% calcium chloride,0.2% magnesium chloride,0.1% potassium nitrate,0.035% gibberellin and 1% agar,and the optimum culture temperature was 25 ℃.The results showed that the best storage condition of Hemerocallis fulva pollen was drying pollen for 6 hours and then stored at-80℃.The feasibility of electroporation and gene gun transformation of Hemerocallis fulva pollen was evaluated by the method of in vitro germination and trypan blue staining.The results showed that the electroporation rate could be improved by selecting the electric shock conditions that could not only make the pollen of Hemerocallis fulva in large proportion,but also keep the pollen activity;the parameters of gene gun could be adjusted to make the micro particles carrying foreign genes into the pollen by bombardment Transformation is the main concern in our future research.After magnetic transfection of Hemerocallis fulva pollen,p CAMBIA 3301 plasmid with GUS gene was loaded on magnetic nano gene vector.Under the external magnetic field,the vector gene complex was introduced into the pollen and successfully integrated into the Hemerocallis genome for expression.The study of pollen in vitro germination,storage and genetic transformation of Hemerocallis fulva laid a foundation for molecular biology research of Hemerocallis fulva,and promoted the research of gene function of Liliaceae plants and the cultivation of transgenic new varieties.