Influencing factors on somatic embryo induction and proliferation of 'Xilin No.3' walnut
[Objective]To select the best conditions for induction and proliferation of somatic embryo of'Xilin No.3'walnut.[Method]Through different disinfection methods,different explants,different sampling periods,different hormone concentration ratio,different sucrose and glutamine concentration treatment,the induction of embryogenic callus of walnut was studied,and the best induction conditions were selected.At the same time,the effects of different hormone concentrations on the proliferation rate of walnut somatic embryos were compared,and the best breeding medium was selected.[Result]The best disinfection methods for male flowers(anthers),seeds and young embryos were soaked in 70%ethanol for 15 s+10%NaClO for 4 min,70%ethanol for 30 s and 70%ethanol for 5 min,respectively.After 4 days of culture,the survival rate was high and the contamination rate was low.Young embryos were the best material for induction of embryogenic callus,and May 31 was the best sampling time,and the induction rates were(65.00±7.07)%and(85.83±1.17)%,respectively.When the callus was used as the material,the optimal hormone concentration ratio medium was modified DKW medium supplemented with 1.0 mg/L 6-BA,2.0 mg/L KT and 0.01 mg/L IBA,and the induction rate was(37.94±1.33)%.When callus was used as the material,the optimal concentration of sucrose was 30 g/L,and the induction rate was(82.67±0.94)%.Glutamine was not an influential factor in inducing embryogenic callus.The optimal hormone concentration ratio medium for somatic embryo proliferation was modified DKW medium supplemented with 0.5 mg/L 6-BA,1.0 mg/L KT and 0.005 mg/L IBA,and the proliferation rate was 98%.[Conclusion]The selection of explants and sampling time directly affects the result of somatic embryo induction.For the internal materials such as young embryos,only simple and quick disinfection can be done.Hormone and carbon source are the key factors of embryogenic callus induction and occurrence,glutamine is not the influential factor of embryogenic callus induction.During the secondary proliferation of somatic embryo,appropriate addition of hormones can inhibit somatic embryo maturation and increase the proliferation rate.
NETL
NSTL
万方数据
