• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

啟良重楼内生真菌的分离鉴定及抑菌活性

Isolation, Identification, and Antibacterial Activity of Endophytic Fungi from Paris qiliangiana

  • 摘要:
      目的  从药用植物啟良重楼中分离鉴定内生真菌,研究其对常见病原细菌的抑菌活性,并分析其次级代谢产物,为利用传统药用植物内生菌开发新型抗生素药物提供新的思路。
      方法  采用平板分离法分离纯化啟良重楼内生真菌,提取其基因组DNA,PCR扩增ITS序列并测序,鉴定内生真菌的种类;采用滤纸片法研究其次级代谢产物对12种供试菌的抑菌活性;采用GC-MS技术分析次级代谢产物的种类。
      结果  从啟良重楼中分离了2株内生真菌,分别鉴定为Setophoma caverna CLZ1和Psilocybe sp.CLZ2;S. caverna CLZ1对肺炎克雷伯菌(Klebsiella pneumonia ATCC 13883)、藤黄微球菌(Micrococcus luteus ATCC 4698)、苏云金芽孢杆菌(Bacillus thuringiensis ATCC 10792)、多耐铜绿假单胞菌(Pseudomonas aeruginosa ATCC 9027)和耐甲氧西林金黄色葡萄球菌(Staphylococcus aureus ATCC 43300,MRSA)等5种供试菌有中度敏感抑制作用,其最小抑菌浓度(MIC)为0.78~3.13 mg·mL−1Psilocybe sp.CLZ2对MRSA有中度敏感抑制作用,其MIC值为3.13 mg·mL−1。运用GC-MS从S. caverna CLZ1提取物中鉴定出36个化合物,包括吡啶(0.91%)、烟酰胺(0.61%)、邻苯二甲酸二丁酯(1.42%)、油酸甲酯(2.28%)和硬脂酸甲酯(0.83%)等5个抑菌活性化合物;从Psilocybe sp. CLZ2提取物中鉴定出44个化合物,包括吡啶(0.106%)和苯乙醇(0.073%)等2个抑菌活性化合物。
      结论  从啟良重楼根茎中分离出内生真菌S. caverna CLZ1和Psilocybe sp.CLZ2,发现S. caverna CLZ1的次级代谢产物具有广抑菌谱,并从中鉴定出5个可能具有抑菌活性的次生代谢产物。

     

    Abstract:
      Objective   Endophytes from the medicinal Paris qiliangiana plants were isolated to identify effective antipathogenic metabolites secreted.The in vitro activity of their secondary metabolites might lead to the development of new antibiotics for disease control on plants.
      Method   Endophytic fungi of P. qiliangiana plants were isolated in a disc diffusion method and identified by ITS gene sequencing. The secondary metabolites from each isolate were obtained by filter paper separation and tested for antibacterial activity against 12 pathogens. Active components were analyzed by GC-MS.
      Results   Setophoma caverna CLZ1 and Psilocybe sp. CLZ2 were the two functional endophytic fungi identified. The former showed a broad spectrum against the tested gram-negative and gram-positive bacteria including Klebsiella pneumonia ATCC 13883, Micrococcus luteus ATCC 4698, Bacillus thuringiensis ATCC 10792, Pseudomonas aeruginosa ATCC 9027, and Staphylococcus aureus ATCC 43300. Their intermediate degree of bacteriostatic activities on the pathogens had the MICs ranging from 0.78 to 3.13 mg·mL−1. The latter exhibited a similar degree of inhibition against S. aureus at an MIC of 3.13 mg·mL−1. The GC–MS analysis identified 36 chemicals in the S. caverna CLZ1 culture filtrate that included pyridine, nicotinamide, dibutyl phthalate, 9-octadecenoic acid (Z)-, and methyl ester methyl stearate. In the Psilocybe sp. CLZ2 filtrate, 44 compounds, including antibiotic pyridine and phenylethanols, were found.
      Conclusion   The endophytic fungi, S. caverna CLZ1 and Psilocybe sp. CLZ2, isolated from the roots of the new variety of P. qiliangiana displayed significant antibacterial property.

     

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