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以‘Old Home’梨无菌苗叶片为外植体,利用根癌农杆菌介导将专一在韧皮部表达启动子AtSUC2驱动下的GUS基因转入‘Old Home’中。筛选出了‘Old Home’梨叶片高效遗传转化的最佳条件:农杆菌和外植体在液体培养基上共培比在固体培养基上共培转化率提高5倍。PCR分析和Southern杂交鉴定表明GUS基因已整合到转基因植株的基因组中,组织化学检测和Western杂交鉴定证明了GUS基因在转基因植株韧皮部中的表达。
Using ’Old Home’ alum seedlings as explants, Agrobacterium tumefaciens was used to transfer the GUS gene driven by phloem-specific promoter AtSUC2 into ’Old Home’. The optimal conditions for efficient transformation of ’Old Home’ pear leaves were screened out: Agrobacterium and explants were cultured on liquid medium five times more efficiently than those cultured on solid medium. PCR and Southern blotting showed that the GUS gene was integrated into the genome of transgenic plants. The expression of GUS gene in the phloem of transgenic plants was confirmed by histochemical detection and Western hybridization.