橘绿木霉GF-11的哌珀霉素鉴定及活性研究

Identification and bioactivity detection of peptaibols from Trichoderma citrinoviride GF-11

[目的]为将橘绿木霉GF-11应用于生防实践,研究了该菌的哌珀霉素及其生物活性。[方法]使用浸提法提取该菌的哌珀霉素,运用TLC及LC-MS/MS鉴定哌珀霉素,并采用稀释平板法和牛津杯法测定抑菌效果,应用染色法、电导率测定、核酸和蛋白质含量测定、DNA降解等指标初步分析了该粗提物对链格孢菌的抑菌机制。[结果]TLC和LC-MS/MS结果显示该粗提物有5种新的哌珀霉素;粗提物对链格孢菌、细极链格孢菌、高氏白粉菌的抑菌率分别为61.2%、58.2%、35.7%,对链格孢菌孢子萌发的抑制率为85%;对大肠杆菌等细菌抑制效果较弱。粗提物可改变链格孢菌丝形态、破坏细胞膜结构,致使胞内电解质及大分子物质外泄,降解基因组DNA。[结论]橘绿木霉GF-11粗提物中鉴定出5种新的哌珀霉素;该粗提物对真菌抑菌效果较好,对细菌抑菌效果较差,并可通过破坏病原菌细胞膜结构达到抑菌效果。

[Aims]This study investigated the biological activity of the peptaibols from Trichoderma citrinoviride GF-11 for the biocontrol application.[Methods]The extracted peptaibols were identified using TLC and LC-MS/MS.Antibacterial effects of the peptaibols was tested by Oxford cup method and plate dilution method,the fungicidal mechanism of the peptaibols against Alternaria alternata was analyzed by staining method,measurement of nucleic acid and protein content,detection of the electrical conductivity and DNA degradation.[Results]The extract having 5 new peptaibols exhibited inhibiting effects against A.alternate,A.tenuissima and Golovinomyces cichoracearum,with the inhibitory rates of 61.2%(inhibitory rates of spore germination is 85%),58.2%and 35.7%,respectively,while the inhibiting effects of the extract against Escherichia coli and other bacteria are weak.The results of the antimicrobial tests showed that the extract could change the mycelium morphology,break the cell membrane,cause the leakage of electrolyte and macromolecular,degrade genomic DNA.[Conclusions]5 New peptaibols were identified from the extract of T.citrinoviride GF-11.The extract has good activity against the tested fungi but weak activity against the tested bacteria,and the fungicidal activity can be achieved by destroying the membrane structure of pathogenic fungi.