小麦矮腥黑粉菌原生质体的制备与再生  

Protoplast preparation and regeneration of Tilletiacontroversa

作　　者：沈慧敏[1] 李超[1] 高利[1] 刘太国[1] 刘博[1] 陈万权[1] SHEN Huimin;LI Chao;GAO Li;LIU Taiguo;LIU Bo;CHEN Wanquan(State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing　100193,China)

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193

出　　处：《植物保护》2018年第2期140-144,共5页Plant Protection

基　　金：国家自然科学基金(31571965)

摘　　要：本文优化了小麦矮腥黑粉菌原生质体的制备与再生条件。结果表明:小麦矮腥黑粉菌的冬孢子在土壤浸提液固体培养基中萌发后转接入T19液体培养基,培养45d,以菌丝作为酶解初始材料,用1.5%崩溃酶+1.5%溶壁酶+1.5%蜗牛酶复合酶液28℃酶解2.5h原生质体的获得率最高;以1.2mol/L的氯化钾为渗透压稳定剂,所得原生质体数量最多,且释放的原生质体能均匀分散分布,不聚集成堆;小麦矮腥黑粉菌的原生质体在TB3培养基上能长出较多的单菌落。Protoplast is the receptor cells for genetic transformation of fungi.To establish protoplast-mediated genetic transformation system of Tilletia controversa,conditions for the protoplast isolation and regeneration of T.controversa were optimized,including incubation time,various enzymes,digestion time and osmotic stabilizers.The results showed that T.controversa teliospores cultured in soil extract medium for 45 days and then transferred into T19 medium were most suitable for protoplast release.The mixture solution of 1.5%driselase,1.5%lyase and 1.5%snailase was most favorable for protoplast preparation.The suitable incubation time with enzyme for the maximum release of protoplasts was 2.5 h at 28℃.The most effective osmotic stabilizer for the protoplast release was 1.2 mol/L KCl,and protoplast was also well dispersed.For the regeneration of the protoplast,TB3 medium was the best,which can produce more single colony under the same condition.

关 键 词：小麦矮腥黑粉菌 原生质体 制备 再生 

分 类 号：S435.121[农业科学—农业昆虫与害虫防治]