Agrobacterium mediated method was used to transfer the IPT gene into Chrysanthemum grandiflorum'Ziyan'and the optimal genetic transformation system was established.The specific operation was to cut the leaves of the Chrysan-themum grandiflorum'Ziyan'into small leaves of 0.5 cm x0.5 cm under sterile conditions and pre-cultivated them on MS+6-BA 2.0 mg/L+NAA 1.0 mg/L medium for 2 days.Then,infected them in a agrobacterium bacterial solution with an OD600 of 0.5 for 3 minutes and inoculated them onto co-culture medium(same as pre-culture medium)and black cultured at 28 ℃ for 36 hours.Later,induced leaf differentiation and bacterial removal on medium(same as pre-culture medium)with a concentration of 3mg/L of Hyg and 500mg/L of Cephalosporin.After the resistant callus tissue differentia-ted into adventitious buds,cut off about 1.5 cm of adventitious buds and inoculated them into a 1/2 MS+NAA 0.2 mg/L+Hyg 14 mg/L medium for rooting culture.By using this system to transform the leaves of Chrysanthemum grandiflorum'Ziyan',resistant plants with good rooting and growth could be obtained.
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