发根农杆菌介导的甘薯遗传转化体系优化.

Genetic transformation system is of great significance for the verification of plant gene function. In order to establish Agrobacterium rhizogenes mediated genetic transformation system in sweet potato(Ipomoea batatas L.), a three‑factor design of 3×3×5 was used to study the effects of infection time, explant type, and A. rhizogenes strains on the induction rate of hairy roots in sweet potato. The appropriate explant type and infection time were determined, and the 5×5 two‑factor design was used to study the effects of five A. rhizomei strains and sweet potato varieties(lines)on the induction rates of sweet potato. The detection of induced hairy roots by PCR confirmed that the rolB gene of the A. rhizogenes Ri plasmid was integrated into the genome of sweet potato cells. The results showed that the infection time, explant type, A. rhizogenes strain, and sweet potato variety all affected the induction rate of hairy roots, and there was an interaction between the factors. Stem segment was the most suitable explants for hair roots, and the optimal infection time was 20 min. MSU440 was the suitable strain for‘Xushu 22’. The suitable strain for‘Taizhong 6’‘YS’were K599. The suitable strain for‘1610’was C58C1. The suitable strain for‘Huishu’was C58C1. Using the A. rhizogenes strain MSU440, stem segments of‘Xushu 22’were used as explants and Agrobacterium infection for 20 min, and the highest induction rate of hairy root was 79.63%. The transgenic efficiency of the hairy root of sweet potato was found to be 38.1% by PCR amplification of the target gene. A genetic transformation system of sweet potato mediated by A. rhizogenes was established, which lays the foundation for further transgenic breeding of sweet potato. [ABSTRACT FROM AUTHOR]