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采用‘吉塞拉6号’甜樱桃矮化砧木(Prunus cerasus×P.canescens)离体叶片外植体,在再生培养基附加生长素的条件下通过根癌农杆菌(Agrobacterium tumefaciens)EHA105(p35SGUS intron)介导研究了β-葡萄糖醛酸酶基因(GUS)的瞬时表达、稳定表达和转基因植株再生,证明了培养基中生长素(IBA或NAA)的存在可促进基因转化,转化效率比对照提高2倍以上。将500个叶片外植体与EHA105(p35SGUS intron)株系在含有生长素的培养基中共培养,获得了11个转基因株系。采用PCR分析和South-ern Blotting核酸杂交,确定GUS基因已整合到矮化砧木‘吉塞拉6号’植株的染色体上。组织化学染色确定了GUS基因在植株体内的表达。
Prunus cerasus × P. canescens in vitro leaf explants were cultured with Agrobacterium tumefaciens EHA105 (p35SGUS) under the conditions of regeneration medium supplemented with auxin intron mediated the transient expression of β-glucuronidase gene (GUS), stable expression and regeneration of transgenic plants, proved that the presence of auxin (IBA or NAA) in the medium can promote gene transformation, the transformation efficiency than the control Increase more than 2 times. 500 leaf explants were co-cultured with EHA105 (p35SGUS intron) lines in auxin-containing medium to obtain 11 transgenic lines. PCR analysis and South-ern Blotting nucleic acid hybridization confirmed that the GUS gene was integrated into the chromosome of dwarf rootstock ’Gisela 6’. Histochemical staining confirmed the GUS gene expression in plants.