The selection of Agrobacterium-mediated genetic transformation system for hornworts provides a theoretical basis for the future study of early terrestrial plant evolution.In this study,Agrobacterium AGL1 carrying the plasmid pCAMBIA1305.2 was used to infect the explants of Folioceros fuciformis.With hygro-mycin as a screening marker,the pre-culture time of explants,the OD value and addition amount of the bacte-rial suspension,the concentration of acetosyringone(AS),and the co-culture time were optimized for the es-tablishment of the genetic transformation system.The results showed that the optimum concentration of hy-gromycin was 15 mg/L for the screening of resistant seedlings,and the state of the explants was the best af-ter 4 d of pre-culture.The optimal infection conditions were addition of 80 μL Agrobacterium suspension with OD600=0.8 in the co-culture medium,AS concentration of 100 μmol/L,and co-culture time of 3 d.The transgenic plants were identified by hygromycin screening,PCR,RT-qPCR and beta-glucuronidase(GUS)staining.The findings provide technical support for the subsequent research on the gene functions of horn-worts.
关键词
褐角苔/外植体/农杆菌介导法/遗传转化体系
Key words
Folioceros fuciformis/explants/Agrobacterium-mediated method/genetic transformation system
维普
万方数据