农杆菌介导的玉米叶片遗传转化体系的建立

盛奇明, 徐永盛, 荆风雪, 苏纯洁, 解光宁, 孙晓莎

Establishment of Agrobacterium-Mediated Genetic Transformation System for Maize Leaves

Sheng Qiming, Xu Yongsheng, Jing Fengxue, Su Chunjie, Xie Guangning, Sun Xiaosha

表1 试验所用培养基

Table 1 Media used in the test

培养基Medium 培养基组分Medium composition 种子萌发培养基 Seed germination medium (G) MS+20 g/L蔗糖+5 g/L琼脂+10 mg/L噻苯唑，pH 5.6 侵染液培养基 Infection medium (I) 1.204 g/L MgSO4+5 g/L麦芽糖+20 mg/L乙酰丁香酮+0.5 mg/L羟敌草腈+0.01% (v/v) BREAK-THRU S233，pH 5.6 共培培养基 Co-cultivation medium (C) MS+20 g/L麦芽糖+10 g/L葡萄糖+0.8 mg/L CuSO4+0.7 g/L脯氨酸+0.5 g/L 2-吗啉乙磺酸+2 mg/L 2,4-二氯苯氧乙酸+10 mg/L维生素C+20 mg/L乙酰丁香酮+3.4 mg/L硝酸银+300 mg/L半胱氨酸+8 g/L琼脂，pH 5.6 恢复培养基 Resting medium (Rs) M5605 (Phytotech)+20 g/L蔗糖+0.6 g/L葡萄糖+0.3 g/L酸水解酪蛋白+0.8 mg/L酸水解酪蛋白+1.2 mg/L麦草畏+3.4 mg/L硝酸银+200 mg/L特美汀+0.025 mg/L植物磺肽素+6 g/L 琼脂，pH 5.6 愈伤组织诱导培养1 Calli inducing medium 1 (CIM1) N492 (Phytotech)+20 g/L蔗糖+2.88 g/L脯氨酸+650 mg/L MgCl2+300 mg/L谷氨酰胺+0.1 g/L酸水解酪蛋白+ 2.2 mg/L 2,4-二氯苯氧乙酸+10 mg/L硝酸银+200 mg/L特美汀+0.025 mg/L植物磺肽素+8 g/L琼脂，pH 5.8 愈伤组织诱导培养2 Calli inducing medium 2 (CIM2) N492+20 g/L蔗糖+2.88 g/L脯氨酸+650 mg/L MgCl2+0.1 g/L酸水解酪蛋白+2.2 mg/L 2,4-二氯苯氧乙酸+ 10 mg/L硝酸银+200 mg/L特美汀+0.025 mg/L植物磺肽素+8 g/L琼脂，pH 5.8 分化培养基 Maturation medium (M) MS+30 g/L蔗糖+0.5 g/L 2-吗啉乙磺酸+1.3 mg/L CuSO4+0.1 mg/L噻苯隆+0.5 mg/L玉米素+1 mg/L吲哚乙酸+ 0.1 μmol/L脱落酸+200 mg/L特美汀+3.5 g/L植物凝胶，pH 5.8 生根培养基Rooting medium (R) MS+30 g/L蔗糖+0.5 g/L 2-吗啉乙磺酸+1.3 mg/L CuSO4+200 mg/L特美汀+3.5 g/L植物凝胶，pH 5.8