双亲灭活原生质体融合技术在草菇耐低温菌株选育上的应用  

Application of Inactivated Parental Strain Protoplasts Fusion Technology in Selection of Volvariella volvacea with Higher Antifreeze Capacity

作　　者：陈建中[1,2] 祝子坪[2] 吴潇[2] 何建华[2] 蒋玮[2] 王金斌[2] 唐雪明[2] 

机构地区：[1]上海海洋大学水产与生命科学学院,上海201306 [2]上海市农业科学院生物技术研究所,上海市农业遗传育种重点实验室,上海201106

出　　处：《中国农业科技导报》2013年第5期166-172,共7页Journal of Agricultural Science and Technology

基　　金：上海市科技兴农重点攻关项目[沪农科攻字(2011)第1-2号]资助

摘　　要：为了选育耐低温草菇菌株,以草菇V23、V3552为亲本,采用酶解法制备原生质体,用紫外诱变、化学诱变两种方法对草菇菌株进行诱变,筛选到耐低温的突变株;然后利用紫外灭活(20 W,30 cm,110 s)和热灭活(50℃3 min)的双亲灭活标记法对突变株进行化学融合,结果表明在400 g/L的PEG6000、pH 8.0、融合时间30 min和融合温度32℃的条件下融合率最高,达到0.517%,共获得200个融合子。经过0℃低温筛选,最终获得15株草菇耐低温菌株,菌丝在0℃的耐冻能力提高了4.5倍。经出菇实验证明其子实体与出发菌株相比具有明显的耐低温性,液化现象明显推迟,说明该方法筛选出的菌株具有进一步应用开发的价值。The protoplasts of VolvarieUa volvacea V23 and V3552 were prepared by enzymatic method, and then were taken mutagenesis by UV and chemical 2 methods. A fusant displaying higher antifreeze capacity, as compared to that of its parents, was obtained by PEG-induced fusion between UV-inactivated protoplasts of V3552 EMS and heat-inactivated protoplasts of V23 UV through low temperature （ 0℃ ） selection. The best fusion condition is gained at 400g/LPEG6000, pH 8.0, for 30min fusion time, at 32℃ fusion temperature. The fusion rate could be as high as 0. 517%. And 200 fusants were obtained. Through screening with 0℃ low temperature, 15 VolvarieUa volvacea cold tolerant bacterial strains were obtained. The cold tolerant ability of hypha at 0℃ was improved by 4. 5 times. The results proved that the fruit displayed higher antifreeze capacity, as compared with bacterial strains; and the liquefaction phenomenon was obviously postponed. This indicated that strains screened by this method were of further development value.

关 键 词：草菇 原生质体 紫外诱变 化学诱变 双亲灭活 PEG融合 

分 类 号：S646.13[农业科学—蔬菜学]