生物技术通报 ›› 2017, Vol. 33 ›› Issue (2): 137-142.doi: 10.13560/j.cnki.biotech.bull.1985.2017.02.020

• 研究报告 • 上一篇    下一篇

豨莶草内生抗感染细菌的分离鉴定及生长特性

陈少红, 赵朋超, 陈明钊, 李洛宜, 李悦, 贾靖靖   

  1. 河南科技大学医学技术与工程学院,洛阳 471023
  • 收稿日期:2016-06-14 出版日期:2017-02-26 发布日期:2017-02-08
  • 作者简介:陈少红,女,本科,研究方向:微生物学;E- mail:2351538594@qq.com
  • 基金资助:
    河南省高等学校重点科研项目(15A180038),河南科技大学博士科研启动基金项目(4022/13480021),国家级大学生创新创业训练计划项目(201510464048),河南科技大学大学生科研训练计划项目(2015125)

Species Identification and Growth Characteristics of Endophytic Anti-infective Bacteria from Herba siegesbeckiae

CHEN Shao-hong, ZHAO Peng-chao, CHEN Ming-zhao, LI Luo-yi, LI Yue, JIA Jing-jing   

  1. College of Medical Technology and Engineering,Henan University of Science and Technology,Luoyang 471023
  • Received:2016-06-14 Published:2017-02-26 Online:2017-02-08

摘要: 豨莶草有多种药用价值,旨在从豨莶草果实中分离和筛选出高拮抗活性的内生细菌。采用了形态学观察、生理生化试验、16S rRNA 序列比对、系统发育树分析以及OD600值、药敏试验和超滤等方法对该抗感染细菌C-5-1进行了相关生物学特性评价。结果显示,C-5-1为侧孢短芽孢杆菌(Brevibacillus laterosporus),对多种病原细菌均敏感,尤其是对铜绿假单胞菌;低pH和高NaCl盐度环境中耐受性有限,在pH≤5.0或盐度≥15%时其生长受到了完全的抑制甚至是死亡,但在pH范围7.0-11.0和盐度范围0-5%下均能生长良好,说明该菌株为耐碱,但不耐酸和盐;筛选出了对菌株C-5-1高度敏感而对5种指示病原菌耐药的抗生素,即氨苄西林;C-5-1发酵上清液中存在着两种抗菌物质,且它们的相对分子质量均小于2 000。因此豨莶草有着优良微生物资源,可以成为开发抗感染细菌药物的重要资源。

关键词: 豨莶草, 侧孢短芽孢杆菌, 氨苄西林, 耐碱, 超滤

Abstract: Herba siegesbeckiae has many healing values,and our aim is to isolate and screen highly active antagonistic endophytes from its fruits. The biological characteristics of anti-infective bacterium C-5-1 were determined by antagonistic experiment,morphological observation,physiological and biochemical tests,sequence alignment of 16S rRNA gene,analysis of phylogenetic tree,measuring OD600,drug sensitive tests,and ultra-filtration centrifugation. The results showed that the C-5-1 was identified as Brevibacillus laterosporus and was sensitive to various pathogenic bacteria,especially Pseudomonas aeruginosa. The strain C-5-1 had limited tolerance to low pH and highly saline environments,i.e.,inhibited completely or even died in pH≤5 or more than 15% NaCl,but it grew well in pH≥7 and 0-5% NaCl,suggesting that C-5-1 had good alkali resistance,but not resistant to acid and salt. Screening experiment revealed that the antibiotic ampicillin was highly sensitive to C-5-1,but resistant to 5 pathogenic bacteria. Fermentation supernatant of C-5-1 contained two antibacterial substances and relative molecular mass of both was less than 2 000. Conclusively,H. siegesbeckiae having excellent microbial resources will be a key resource for the development of anti-bacteria drugs.

Key words: Herba siegesbeckiae, Brevibacillus laterosporus, ampicillin, alkali-resistant, ultra-filtration centrifugation