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用根癌农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓ）共培养法将外源基因导入青花菜（Ｂｒａｓｓｉｃａｏｌｅｒａｃｅａｖａｒ．ｉｔａｌｉｃａ）上海１号的无菌苗子叶及下胚轴原生质体。菌株为ＥＨＡ１０５，携带质粒ｐＭＯＧ４１０，含ＧＵＳ和ＮＰＴⅡ基因。实验得出其子叶芽分化率最高的激素组合为６－ＢＡ５ｍｇ／Ｌ＋ＩＡＡ０．１ｍｇ／Ｌ。原生质体以８～１０℃暗培养５～６ｄ，然后于２５℃光照８ｈ生长的无菌苗下胚轴切片游离出的密度最高。Ｋ８Ｐ培养基激素组合（ｍｇ／Ｌ）以２，４－Ｄ１．０＋ＢＡ０．５＋ＮＡＡ０．１为最好。共培养的最适温度为１８～２０℃。经检测由子叶转化得到了转基因植株；原生质体转化得到了抗性愈伤组织。
The exogenous genes were introduced into the sterile seedling cotyledons and hypocotyl protoplasts of Brassica oleraceavar.italica Shanghai No.1 with Agrobacterium tumefaciens co-culture method. The strain is EHA105, carrying the plasmid pMOG410, containing the GUS and NPTII genes. The result showed that the hormone combination with the highest rate of cotyledon bud differentiation was 6-BA5mg / L + IAA0.1mg / L. Protoplasts were dark-cultured at 8 ~ 10 ℃ for 5 ~ 6d, then the hypocotyl cuttings of sterile seedlings grown at 25 ℃ for 8h had the highest density. K8P medium hormone combination (mg / L) to 2,4-D1.0 + BA0.5 + NAA0.1 is best. Co-culture of the optimum temperature of 18 ~ 20 ℃. Transgenic plants were obtained from the cotyledons tested; protoplasts transformed to obtain resistant callus.