基于非靶向代谢组学解析猬木霉挥发性物质对香梨采后链格孢的抑制作用

Non-targeted Metabolomics Analysis of the Inhibitory effect of Trichoderma erinaceum Volatile Substances on Alternaria alternata Isolated from postharvest Korla Fragrant pear

本研究探讨猬木霉M6-5挥发性物质对香梨采后黑斑病菌链格孢(Alternaria alternata)代谢物质、呼吸和能量代谢的影响。基于非靶向代谢组学技术通过液相色谱-质谱联用仪测定链格孢受M6-5挥发性物质处理后体内代谢物质的变化;测定M6-5挥发性物质熏蒸处理后0、3、5、7d的链格孢呼吸速率,计算呼吸抑制率;采用液相色谱仪测定不同培养时间处理组和对照组链格孢ATP、ADP、AMP和能荷的影响。结果表明:M6-5挥发性物质处理后链格孢菌丝体上调的差异代谢物有6-戊基-2H-吡喃-2-酮、4-乙基苯甲醛、3,4-二甲基苯甲酸、4-苯基丁酸、2-甲氧基间苯二酚等,下调的差异代谢物有6-氨基烟酰胺、11-酮基乙酰胆甾醇、3-羟基康力龙、茉莉酸、苯甲酰芽子碱等。利用京都基因与基因组百科全书通路分析发现差异显著的通路为次生代谢物的生物合成。在培养时间结束(7d)时,M6-5挥发性物质对链格孢的呼吸抑制率为(49.64±2.04)%;与对照组相比,M6-5挥发性物质处理显著降低了链格孢ATP、ADP、AMP的含量和能荷水平,说明M6-5挥发性物质处理使链格孢胞内维持在更低的能荷水平,能量代谢失衡,呼吸速率受抑制,进而影响了代谢物质的变化。

In this study,the effects of Trichoderma erinaceum M6-5 volatile components on metabolites,respiration and energy metabolism of Alternaria alternata from postharvest Korla fragrant pear were investigated.Non-targeted metabolomics based on liquid chromatography-mass spectrometry(LC-MS)was used to determine the changes of metabolites in A.alternata after treatment with M6-5 volatile substances.At 0,3,5 and 7 days after fumigation with M6-5 volatile substances,the respiratory rate of A.alternata was measured,and the respiration inhibition rate were calculated.The levels of ATP,ADP,AMP and energy charge in the M6-5-treated and control groups as a function of culture time were determined by liquid chromatography(LC).The results showed that among the identified differential metabolites,6-amyl-2H-pyrano-2-one(6pp),4-ethylbenzaldehyde,3,4-dimethylbenzoic acid,4-phenylbutyric acid and 2-methoxyresorcinol were up-regulated,while 6-aminonicotinamide,11-ketoacetyl cholesterol,3-hydroxyconilone,jasmonic acid and benzoylsphenine were down-regulated by M6-5 volatile substances.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis showed that the significantly differential pathway was involved in the biosynthesis of secondary metabolites.At the end of the 7 day incubation period,the respiration inhibition rate of A.alternata by M6-5 volatile substances was(49.64±2.04)%.Compared with the control group,treatment with M6-5 volatile substances significantly reduced the levels of ATP,ADP,AMP and energy charge,indicating that it can keep the energy charge at lower levels,thereby resulting in imbalanced energy metabolism and inhibited respiratory rate and affecting metabolite changes.