Plant Regeneration and Transformation of the Potato Cultivar'Kexin No.13'
This study aims to explore the potato plant regeneration and transformation technology.The stem segment,leaf,and minituber of'Kexin No.13'were used as explants and inoculated into the media with different plant hormone ratios for callus induction and differentiation.Furthermore,the Agrobacterium tumefaciens-mediated transformation method was employed to establish a genetic transformation and regeneration system for potatoes.The results showed that the stem segment was the ideal explant of'Kexin No.13'.The optimal medium for callus induction was 4.4 g/L MS + 30 g/L sucrose + 8 g/L agar + 2 mg/L 6-BA + 0.5 mg/L 2,4-D + 0.4 mg/L KT,in which the callus induction rate reached above 95%.The optimal differentiation medium was 4.4 g/L MS + 30 g/L sucrose + 8 g/L agar + 2 mg/L 6-BA + 3 mg/L KT + 0.5 mg/L GA3 + 0.1 g/L inositol,in which the emergence rate was 56.25%.The transformation experiment was carried out in a procedure of explant pre-culture for 2 d,soaking in the A.tumefaciens suspension(OD600=0.5)for 7 min,and co-culture for 3 d.Subsequently,50 mg/L kanamycin and 200 mg/L timentin were used for screening of the seedlings carrying the target gene Cas9p.
potato'Kexin No.13'plant regenerationtransformation system
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