林 勤, 黄述生, 李 锋,等.枸杞、番茄体细胞原生质体培养体系研究[J].广东农业科学,2015,42(3):24-30 |
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枸杞、番茄体细胞原生质体培养体系研究 |
Study on the protoplast culture of somatic embryogenesis of Lycium barbarum. L. and Lycopersicon esculentum Mill. |
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DOI: |
中文关键词: 枸杞 番茄 愈伤组织 酶解法 原生质体 |
英文关键词: Lycium barbarum. L. Lycopersicon esculentum Mill callus protoplast enzymatic hydrolysis |
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中文摘要: |
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 1.0 mg/L 2,4-D+1.5 mg/L KT 和 1.0 mg/L 2,4-D+1.0 mg/L KT 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 pH 5.0~6.0、温度 30益下振荡酶解 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。 |
英文摘要: |
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. |
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