CAJ | 학술논문

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10 g／L纤维素酶＋10 g／L蜗牛酶＋1 g／L溶壁酶组成的复合酶溶液，35℃恒温，80 r／min振荡孵育3 h左右，原生质体释放量最高；原生质体在Y C M 培养基上，22℃培养9 d可以再生形成菌落，培养15 d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基础。
이산고마두소내생진균Undifilum oxytropis NX‐FEL001위공시균주，연구료배양방식、균령、매해시간화매해온도등，대U ．oxytropis원생질체제비화재생적영향。결과표명，U ．oxytropis균사재질량분수위10 g／L섬유소매＋10 g／L와우매＋1 g／L용벽매조성적복합매용액，35℃항온，80 r／min진탕부육3 h좌우，원생질체석방량최고；원생질체재Y C M 배양기상，22℃배양9 d가이재생형성균락，배양15 d기재생솔가체8．5％；원생질체재YCM 배양기상재생후기균락안색여야생형U ．oxytropis불일치，정현백색；균사형태여야생형U ．oxytropis상일치，경검측재생후균사잉연능구산생고마두소。U ．oxytropis원생질체적제비，장위진행풍초내생진균적유전전화화기인조작연구제공중요공구，위진일보개전풍초내생진균합성고마두소적분자조공궤제전정기출。
The swainsonine‐pruducting endophytic fungi ,Undi f ilum oxytropis ,was used to develop pro‐toplasts .The effects of some factors on formation and regeneration of protoplasts were investigated ,inclu‐ding culture method ,mycelium age ,digesting time and temperature .The results showed that the highest preparation rate was achieved when prepared through mixture solution of 10 g/L cellulase+10 g/L snailase+1 g/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 mycelium was the same as wild strain when cultured in YCM medium .The regenera‐tion mycelium could still produce swainsonine . The preparation and regeneration of protoplasts of the swainsonine‐productingendophyticfungi,Undifilumoxytropis,whichwouldprovidingimportanttoolfor the research of genetic transformation and genetic manipulation and science basis for further to elucidate the synthesis of swainsonine in molecular level .