作者：杨小帆, 宋丽菊, 齐盼盼, 吴道艳, 闵霞, 李彩云, 彭静珊, 赵建

作者单位：四川大学生命科学学院资源微生物及微生物生物技术重点实验室, 四川 成都 610064

关键词：有柄石韦;内生细菌;抑菌活性;GC-MS检测;代谢产物

摘要：

为探究Pyrrosia petiolosa内生细菌的抑菌活性，采用植物组织分离和微生物传统培养法从其根、茎、叶共分离检测到内生细菌14株。通过滤纸片抑菌法筛选两株有显著抑菌活性的内生细菌G7、Y12，并借助GC-MS技术对这两株内生细菌进行抑菌活性成分检测与分析。研究表明：两株内生细菌经16S rDNA测序比对分别鉴定为Lysinibacillus sphaericus C3-41和Bacillus amyloliquefaciens FZB42。生长曲线测定结果显示菌株G7、Y12分别在16 h和12 h后进入稳定期。GC-MS检测鉴定到两株内生细菌发酵上清抑菌活性物质主要是有机酸类和2，5-二酮哌嗪衍生物。该研究可为挖掘P. petiolosa内生细菌药物活性、筛选抗菌药物新来源提供参考。

Screening of endophytic bacteria in Pyrrosia petiolosa and analysis of its antimicrobial active ingredients

YANG Xiaofan, SONG Liju, QI Panpan, WU Daoyan, MIN Xia, LI Caiyun, PENG Jingshan, ZHAO Jian

Key Laboratory of Microbiological Resource and Technology, College of Life Sciences, Sichuan University, Chengdu 610064, China

Abstract: In order to study antimicrobial activity of endophytic bacteria in Pyrrosia petiolosa, plant tissue separation and traditional microbial culture methods were employed, by which fourteen bacteria strains were isolated from the roots, stems and leaves of P. petiolosa. Endophytic bacteria G7 and Y12 were screened out due to their significant antimicrobial activity through filter paper bacteriostatic method. The antimicrobial active ingredients were detected and analyzed by GC-MS technology. Strains G7 and Y12 were identified as Lysinibacillus sphaericus C3-41 and Bacillus amyloliquefaciens FZB42 respectively according to the 16S rDNA sequence analysis. Growth curves of strains G7 and Y12 showed that the stationary phase begins after 16h and 12h, respectively. The antimicrobial active materials mainly consisted of organic acids and 2, 5-piperazinedione derivative through GC-MS detection. The study laid an research foundation for discovering the pharmaceutical activity of endophytic bacteria in P. petiolosa and screening a new source of antimicrobial agents.

Keywords: P. petiolosa;endophytic bacteria;antimicrobial activity;GC-MS analysis;metabolites

2016, 42(7): 47-52  收稿日期: 2016-2-3;收到修改稿日期: 2016-3-10

基金项目: 国家自然科学基金（31270175）；教育部新世纪人才和川大优秀青年学者项目（NCET-13-0397，2013SCU04B14）

作者简介: 杨小帆(1991-),女,湖北安陆市人,硕士研究生,专业方向为资源微生物应用。

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