都雯玥陆锡宏李雪虎周 翔梁剑平辛志君

1（中国科学院近代物理研究所 兰州 730000）2（兰州理工大学生命科学与工程学院 兰州 730000）



碳离子束辐照选育截短侧耳素菌原生质体

都雯玥1,2陆锡宏1李雪虎1周 翔1梁剑平1,2辛志君1

1（中国科学院近代物理研究所 兰州 730000）
2（兰州理工大学生命科学与工程学院 兰州 730000）

摘要使用12C6+离子束辐照Clitopilus pinsitus 原生质体至不同吸收剂量，运用琼脂柱预筛和96孔板固体发酵选育截短侧耳素高产变异株。结果显示，最佳12C6+离子束吸收剂量为1.5 Gy，在该吸收剂量下正变异率为37.58％。选育出C.pin15I5E和C.pin15II6B两株正变异株，其产量较出发菌株分别提高16.17％和15.47％。表明重离子束辐照原生质体是行之有效的工业微生物诱变育种方法。

关键词碳离子束，辐照，Clitopilus pinsitus原生质体，高产变异株，截短侧耳素

Supported by National High Technology Research and Development Program of China (863Program) (2011AA10A214) and Gansu Local Pharmaceutical Industry Development Fund (Y361010SJ0)

Frist author: DU Wenyue, female, was born in June 1988 and graduated from Gansu Agricultural University in 2011. Now she is a master candidate of Lanzhou University of Technology, majoring in microbial mutation breeding

Received 10 September 2015, accepted 10 November 2015

Breeding pleuromutilin protoplasts strains by carbon ion beam irradiation

DU Wenyue1,2LU Xihong1LI Xuehu1ZHOU Xiang1LIANG Jianping1,2XIN Zhijun1

1(Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000, China)
2(Life Science and Engineering College, Lanzhou University of Technology, Lanzhou 730000, China)

ABSTRACT Clitopilus pinsitus protoplast was irradiated by12C6+ion beam at varied absorbed doses for mutagenesis, and high-yielding pleuromutilin mutant strains were screened using high-throughput method and solid fermentation. The results showed that the optimum12C6+ion beam absorbed dose is 1.5 Gy, and under this condition the positive mutation rate is 37.58％. By agar column prescreening and 96-plate solid fermentation, the mutant strains, C.pin15I5E and C.pin15II6B, were selected. The yields of the strain increased by 16.17％ and 15.47％, respectively, compared with that of the original strain. The research suggests that applying heavy ion irradiation to protoplast is a possible method to breed mutant strains.

KEYWORDS Carbon ion beam, Irradiation, Clitopilus pinsitus protoplast, High-yielding mutant strain, Pleuromutilin

CLC Q691.5, TL99

截短侧耳素是斜盖菇属(Clitopilus)真菌产生的一种二萜抗生素，对支原体和一些革兰氏阳性菌有较强抗菌活性，其衍生物如泰妙菌素具有生物利用率高、毒性低和残留少的优势，作为饲料添加剂应用于家畜家禽养殖[1-3]。目前，国内截短侧耳素生产企业少，发酵技术不成熟，菌株截短侧耳素产量低，发酵周期长[2,4]，通过诱变选育截短侧耳素高产菌株具有可观的经济效益。

重离子束辐照是一种新型的诱变手段，具有突变频率高、突变谱广、能够打破基因连锁、促进基因重组等优点[5-7]。离子注入细胞，可以引起DNA损伤，激发细胞修复机制，最终形成碱基缺失和颠换，DNA片段缺失和插入等突变[8-10]。陈宇等[11]使用N+离子束辐照红霉素产生菌，经过筛选得到高产突变菌，其摇瓶发酵证明产量较原始菌株提高20％以上。……