Study on the Genetic Transformation System of the IPT Gene in Chrysanthemum Grandiflorum'Ziyan'
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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