枸杞、番茄体细胞原生质体培养体系研究

利用组培技术,以宁杞1号枸杞和农大1号番茄种子的无菌苗叶片、下胚轴为外植体,脱分化培养分别获得其愈伤组织,经继代增殖和酶解处理获得质量较好的原生质体。结果表明,枸杞、番茄无菌苗的下胚轴分别在附加有1.0mg/L2,4-D+1.5mg/LKT和1.0mg/L2,4-D+1.0mg/LKT的MS固体培养基中诱导的愈伤组织(诱导率分别是68.45%、64.33%)无褐化或褐化率最低,产生的愈伤组织适合于原生质体培养且效果最好;将0.5%果胶酶、1%纤维素酶、0.2%离析酶R-10、0.2%半纤维素酶配合使用,在pH5.0~6.0、温度30℃下振荡酶解12~16h,所得枸杞、番茄原生质体均在Km8P液体培养基中纯化培养,原生质体及活力状态均可保持2d左右。

Study on the protoplast culture of somatic embryogenesis of Lycium barbarum. L. and Lycopersicon esculentum Mill.

The sterilized seeds of Ningqi 1of Lycium barbarum L. and Nongda 1of Lycopersicon esculentum Mill were germinated on MS or 1/2MS medium to acquire aseptic seedlings, and then, using the seedlings to induce callus on MS medium supplemented. A stable cell suspension culture system was established with callus of L. barbarum L. and L. esculentum Mill after several rounds of subculture.The results showed that supplement of 1.0 mg/L 2, 4-D and 1.5 mg/L KT for L. barbarum L. and 1.0 mg/L 2, 4-D and 1.0 mg/L KT for L. esculentum Mill in MS solid culture medium are the best condition for inducing callus both of them, and achieved 68.45% and 64.33% induction rate respectively. Treating suspension cell of 0.5% enzyme(Pectinase), 1% cellulase(Cellulase), isolated and R-10 0.2%(Macerozyme R-10), 0.2%, hemicellulase (Hemicellulase) at 30, pH 5.0-6.0 with a 12~16 h shaking resulted in the protoplast actvitity for 2 days.