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Position: Home > Articles > Protoplast preparation and regeneration of Tilletia foetida China Plant Protection 2017,37 (10) 14-18

小麦光腥黑粉菌原生质体的制备与再生

作  者：
沈慧敏;高利;李超;刘太国;刘博;陈万权
单  位：
中国农业科学院植物保护研究所
关键词：
小麦光腥黑粉菌;原生质体;制备;再生
摘  要：
以优化原生质体的制备方法,建立小麦光腥黑粉菌的遗传转化体系为目的,研究了小麦光腥黑粉菌菌株培养时间、细胞壁裂解酶、酶解时间和渗透压稳定剂等对原生质体制备的影响。结果表明,以水琼脂培养基上培养7d的菌丝为初始材料,配置1.5%崩溃酶+1.5%溶壁酶+1.5%蜗牛酶的复合酶液,28℃酶解2 h后原生质体的获得率最高;以1.2 mol/L的氯化钾为渗透压稳定剂,得到的原生质体数量最多,且释放的原生质体能够均匀散乱分布,不聚集成堆;较适宜原生质体再生的培养基为TB3培养基,可产生较多的单菌落。
译  名：
Protoplast preparation and regeneration of Tilletia foetida
作  者：
Shen Huimin;Gao Li;Li Chao;Liu Taiguo;Liu Bo;Chen Wanquan;State Key Laboratory for the Biology of Plant Diseases and Insects Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences;
单  位：
State Key Laboratory for the Biology of Plant Diseases and Insects Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences
关键词：
Tilletia foetida;;protoplast;;preparation;;regeneration
摘  要：
To establish protoplast-mediated genetic transformation system of Tilletia foetida, conditions for protoplast isolation and regeneration of T. foetida were optimized. The results showed that mycelia incubated for 7 days were the most suitable for protoplast release. Mixture solution of 1.5% driselase, 1.5% lyase and 1.5% snailase were the most favorable for protoplast preparation. The most suitable incubation time with enzyme for maximum release of protoplasts was 2 h at 28 ℃. The most effective osmotic stabilizer for the protoplast release was 1.2 mol/L KCl, and protoplast also scattered distribution. For regeneration of the protoplast prepared freshly, TB3 medium was better than PDA medium, which could produce more single colonies.

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