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以2日龄西瓜(Citrullus lanatus(Thunb.)Mansfeld)无菌苗子叶为外植体,通过与根癌农杆菌(Agrobacterium tumefaciens)进行叶盘共培养建立了西瓜的遗传转化系统。所用根癌农杆菌中含有改建后分别携带嵌合NPTⅡ基因和番茄的ACC合成酶基因及其反义基因的质粒。外植体在MSA培养基(MS盐类、B_5维生素、1.0mg/L BA、0.2mg/L IAA)上预培养3～4d后,与根癌农杆菌共培养4d,随后转移外植体至附加100mg/L卡那霉素、300mg/L头孢菌素的MSA培养基上筛选转化芽。将带芽外植体移入含有100mg/L卡那霉素、300mg/L头孢菌素的伸长培养基(MS+0.2mg/L KT)上进行芽伸长,切取2～3cm高的伸长芽移入生根培养基(1/2MS+0.1mg/L NAA)生根。Southern blot结果证明获得转基因植株,乙烯释放指标表明转入的正义和反义ACC合成酶基因得到不同程度的表达。
Two-day-old sterile seedlings of Citrullus lanatus (Thunb.) Mansfeld were used as explants to establish a genetic transformation system of watermelon by leaf disk co-culture with Agrobacterium tumefaciens. The Agrobacterium tumefaciens used contained the plasmids carrying the ACC synthase gene and the antisense gene of the chimeric NPTII gene and tomato, respectively, after reconstitution. The explants were pre-cultured for 3 to 4 days in MSA medium (MS salts, B_5 vitamins, 1.0 mg / L BA, 0.2 mg / L IAA) and co-cultured with Agrobacterium tumefaciens for 4 days before explants were transferred Transformation shoots were screened on MSA medium supplemented with 100 mg / L kanamycin and 300 mg / L cephalosporins. Bud explants were transferred to elongation medium (MS + 0.2 mg / L KT) containing 100 mg / L kanamycin and 300 mg / L cephalosporins and bud elongation was performed to cut 2 to 3 cm high elongation The shoots were rooted in rooting medium (1 / 2MS + 0.1 mg / L NAA). Southern blot results showed that transgenic plants were obtained. The ethylene release index indicated that the genes of sense and antisense ACC synthase introduced into the plant were expressed to different extents.