Induction of Dioscorea alata Callus and Establishment of Its Suspension Cell Culture System
In order to determine the better hormone concentration and suspension cell culture conditions induced by Dioscorea alata callus,the axillary bud stem segments of three genotypes of Dioscorea alata's tissue culture seedlings were used as materials.And a three-factor,three-level orthogonal experiment was used to investigate the effects of different combinations of 6-BA,NAA and PIC on the induction of healing tissues.In order to initially establish a healing tissue suspension cell culture system,single-factor experiments were used to explore the effects of initial suspension cell inoculum,sucrose concentration,shaker speed,casein concentration,and light time on their cell biomass.The experimental results showed that under the combination of 1.0 mg/L 6-BA+1.0 mg/L NAA+0.5 mg/L PIC hormone concentration,the healing induction rate of Da70 was 90％and the healing status was better,and the effects of different hormones on the healing induction rate were as follows:NAA＞PIC＞6-BA.In the sin-gle-factor suspension cell culture experiment,an initial suspension cell inoculum of 10 mL,a sucrose concentra-tion of 10 g/L,a shaker speed of 100 r/min,a casein concentration of 50 mg/L,and 0 h of light facilitated the in-crease in cell biomass.The growth of the suspension cells of the crop showed an overall"S"shape and reached a peak of 0.159 3 g fresh weight,0.042 3 g dry weight and 0.485 0 cell viability OD at the 18th day.The results of this study can provide the basis for the subsequent isolation of protoplasts and the subcellular localization of functional genes.
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