Maize Embryo Callus Induction And Establishment Of Regenerated Plant System

In maize tissue culture,young embryos are widely used as common exosomes for work such as gene conversion,so it is esserntial to establish an efficient maize young embryo tissue culture system.In this experiment,we used the young embryos of ten self-crossing lines of maize Mo17,b101,4F1A,121-B73,b105,08y44,H35,gns12059,gns12062,and 4144 as exosomes,and designed the method to study the effects of post-pollination time,medium type,2,4-D,TDZ,and 6-BA concentrations on healing tissue induction and healing tissue differentiation of maize young embryos,and the effect of IBA concentration on root system induction of maize young embryos by orthogonal experiments.The results show that.1.After pollination,the time of embryo taking is the main factor affecting the rate of induction of young embryos,except for the self-crossing line GNS12059,the best time of embryo taking is 14 d;the best time of embryo taking for the remaining nine self-crossing lines is the 10th d after pollination;and there is no significant difference in the medium type in the process of induction of wounded tissue;when the concentration of 2,4-D is 1-2 mg/L,the growth of wounded tissue in all self-crossing lines is the best;the addition of TDZ to the medium can significantly improve the rate of induction of wounded tissue in maize self-crossing lines,indicating that TDZ has an obvious promoting effect on the induction of wounded tissue.2.The healing tissue differentiation of young maize embryos is influenced by genotype.Screening of different maize self-crossing lineage differentiation cultures resulted in the optimal combination of b 101,4F1A,b105 and 08y44 healing tissue differentiation as MS+0.0 mg/L 6-BA+0.0 mg/L TDZ;121-B73 and gns12062 healing tissue differentiation as MS+1.0 mg/L 6-BA+1.0 mg/L TDZ;gns12059 healing tissue differentiation as MS+2.0 mg/L 6-BA+0.5 mg/L TDZ;H35 healing tissue differentiation as N6+1.0 mg/L 6-BA+0.5 mg/L TDZ;4144 healing tissue differentiation as NB+0.0 mg/L 6-BA+0.5 mg/L TDZ;Mo17 healing tissue differentiation as NB+2.0 mg/L 6-BA+1.0 mg/L TDZ.3.The highest rooting rates of different maize self-crossing lines were achieved at IB A concentrations of 0.5 mg/L-0.6 mg/L in 1/2 MS of rooting medium.4.Significant differences were observed in the induction of octocotyledons of mature embryos from 10 maize autocross lines under different concentrations of 2,4-D and NAA added to N6.During the induction process,the induction rate of the tip and middle 1 sites was significantly better than that of the middle 2 and the tail,where the tip induction rate reached more than 80%and the middle 1 site induction rate reached more than 90%.