Establishment of genetic transformation and regeneration systems for different melon varieties and rapid acquisition of gene edited plants
[Objective]Study on establishment of genetic transformation and regeneration systems for dif-ferent melon varieties and rapid acquisition of gene edited plants.[Methods]The cotyledons of four melon va-rieties,including Bawudun,Laohangua,Qiligan,and Xinmiza No.11(86-1).The sweet melon cotyledons were infected by Agrobacterium tumefaciens mediated method,and a genetic transformation and regeneration system of Laohan melon was established by tissue culture method and then genetically edited plants were ob-tained through PCR detection.[Results]The results showed that Laohan melon was selected as the dominant variety from four melon varieties as the explants.It was pre-cultured on MS+6-BA 1.0 mg/L+NAA 0.05 mg/L medium for 2 days and co-cultured for 3 days,then it was further cultured on MS+6-BA 1.0 mg/L+NAA 0.05 mg/L+hygromycin 5 mg/L+cephalosporin 250 mg/L screened medium for 7 days,after that,small buds were seen growing on MS+6-BA 1.0 mg/L+NAA 0.05 mg/L+hygromycin 5 mg/L+cephalo-sporin 250 mg/L adventitious bud induction medium for 1-2 weeks.The induced buds were placed on both non hygromycin and hygromycin added bud elongation medium for cultivation.It was found that the growth rate of the induced buds on the non hygromycin added bud elongation medium was faster than that on the hygromy-cin added medium,which shortened the growth of the buds to the two leaf seedling stage by 3 weeks.One bud DNA was cut off in a sterile environment and 21 transformed plants were examined by RT-PCR.[Conclu-sion]Gene edited plant buds were successfully selected on a shoot elongation medium with added hygromycin.The use of a shoot elongation medium without added hygromycin can shorten the time for obtaining gene edited plants by 3 weeks.
万方数据
维普
