产苦马豆素疯草内生真菌Undifilum oxytropis原生质体的制备和再生  

Preparation and Regeneration of Protoplasts of Swainsonine-producting Endophytic Fungi Undifilum oxytropis from Locoweed

作　　者：张蕾蕾[1,2] 余永涛[1] 何生虎[1] 赵清梅[3,4] 葛松[1] 

机构地区：[1]宁夏大学农学院,宁夏银川750021 [2]韩城市金城办畜牧兽医工作站,陕西韩城715400 [3]北方民族大学生物科学与工程学院,宁夏银川750021 [4]国家民委发酵酿造工程生物技术重点实验室,宁夏银川750021

出　　处：《动物医学进展》2015年第2期53-58,共6页Progress In Veterinary Medicine

基　　金：宁夏自然科学基金项目(NZ1118)

摘　　要：以产苦马豆素内生真菌Undifilum oxytropis NX-FEL001为供试菌株,研究了培养方式、菌龄、酶解时间和酶解温度等,对U.oxytropis原生质体制备和再生的影响。结果表明,U.oxytropis菌丝在质量分数为10g/L纤维素酶+10g/L蜗牛酶+1g/L溶壁酶组成的复合酶溶液,35℃恒温,80r/min振荡孵育3h左右,原生质体释放量最高;原生质体在YCM培养基上,22℃培养9d可以再生形成菌落,培养15d其再生率可达8.5%;原生质体在YCM培养基上再生后其菌落颜色与野生型U.oxytropis不一致,呈现白色;菌丝形态与野生型U.oxytropis相一致,经检测再生后菌丝仍然能够产生苦马豆素。U.oxytropis原生质体的制备,将为进行疯草内生真菌的遗传转化和基因操作研究提供重要工具,为进一步开展疯草内生真菌合成苦马豆素的分子调控机制奠定了基础。The swainsonine-pruducting endophytie fungi, Undifilum oxytropis, was used to develop protoplasts. The effects of some factors on formation and regeneration of protoplasts were investigated, including culture method, mycelium age, digesting time and temperature. The results showed that the highest preparation rate was achieved through mixture solution of 10 g/L cellulase＋10 g/L snailase＋1g/L lysing enzyme, at 35 ℃, 80 r/min for 3 h. The regeneration colonies were achieved when cultured in YCM medium for 9 d at 22 ℃ and the highest regeneration rate （8.5%） was achieved when cultured in YCM medium for 15 d at 22 ℃. The color of regeneration colonies was different from wild strain and the form of regeneration mycelia was the same as wild strain when cultured in YCM medium. The regeneration mycelia could still produce swainsonine. The preparation and regeneration of protoplasts of the swainsonine-producting endophytic fungi, Undifilum oxytropis, would provide important tool for the research of genetic transfor- mation and genetic manipulation and science basis for further elucidating the synthesis of swainsonine in molecular level.

关 键 词：苦马豆素 Undifilum OXYTROPIS 原生质体 制备及再生 

分 类 号：S856.9[农业科学—临床兽医学]