| The stem segment vitro culture and the establishment of leaf regeneration system of test-tube plantlets of columnar apple Runtai No.1 were studied,the effects of different shading,different disinfection time,different stem length and different medium adjunct treatments on the explant contamination rate,browning rate and germination rate were also studied.The adventitious buds with different hormone concentrations were induced from the leaves of sterile materials.At the same time,the effects of different hormone concentrations and culture medium on the strong seedlings and rooting of apple rootstock M9-T337 were studied.The main results were as follows:1.The results of different shading time on the aseptic material of Runtai No.1 showed that the pollution rate and browning rate of explants was decreased with the increase of shading time,while the growth potential of explants was weaker.the best material was selected for 7 days of dark light culture,and the pollution rate was 23.33,the browning rate was 25,the germination rate was 51.67 in this study.The pollution rate was increased with the increase of the number of axillary buds in the treatment of different number of axillary buds,while there was no significant difference between the browning rates of the two axillary buds.The germination rate of two axillary buds was the highest,reaching 50%.For different disinfection time,the pollution rate was decreased with the increase of disinfection time,and the browning rate was increased with the increase of disinfection time.The best disinfection time was 8 minutes,the pollution rate was 33.33,the browning rate was 23.33 and the germination rate was 53.33.There was no significant difference in the contamination rate among the different adjuncts,and the browning rate was the lowest in the medium supplemented with 100mg·L-1 Cef,and there was significant difference in the germination rate.The best medium for the addition was MS+100 mg·L-1 Cef.When the adventitious buds were induced,the induction rate of adventitious buds was increased with the increase of the number of days of induction,and the effect of inducing adventitious buds on 50 days was better,and the most suitable adventitious bud induction was MS+ 4.0 mg·L-1 6-BA+0.5 mg·L-1 NAA.2.The results of different hormone concentration on proliferation and growth of M9-T337 showed that the number of effective proliferating buds was higher,regardless of the number of proliferating buds,when the concentration of 6-BA was 1.0 mg·L-1 and 2.0 mg·L-1.The number of effective proliferating buds was less,and the leaf edge was scorched,when the concentration of 6-BA was 3.0 mg·L-1 and 4.0 mg·L-1.The best culture medium was MS+2.0 mg·L-1 6-BA+0.1 mg·L-1 NAA+0.3 mg·L-1 GA3,and the multiplication coefficient was 3.93.Different concentrations of hormone in seedling medium screening showed that NAA has great effect on the growth seedling.When the concentration of NAA was 0.5 mg·L-1,the plantlets in tissued culture had strong growth,with a few or no yellow leaves,and with the presence of lateral branches.When the concentration of NAA was 1.0 mg·L-1 and the medium without any hormones,the growth of the plantlets in tissued culture was weaker and there were more yellow leaves,no lateral branch.The results showed that the best medium was MS+4.0 mg·L-1 6-BA+0.5 mg·L-1 NAA.3.The results showed that the best rooting medium was 1/2 MS+0.3 mg·L-1 IBA,the total number of rooting was 117,and the average number of rooting was 13,when the plantlets growing to 1.5cm were inoculated into 1/2 MS containing different concentrations of IBA,and the shape of the root was slender and without lateral roots.The rootless seedlings were cultured in 1/2 MS medium for a week in dark and then inoculated into MS medium without hormone.The results showed that the best rooting medium was MS+0.6 mg·L-1 IBA.The total number of rooting was 102,the average number of rooting was 11.33,as well as the shape of roots was short and thick and had more lateral roots.In the study,0.3 mg·L-1 IBA treatment and no IBA treatment were added to the 1/2 MS and MS medium.The results showed that in the medium supplemented with IBA,the root system was short,the root system was longer in the medium without IBA,and the length of the culture medium was not greatly affected. |
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