Metabolome of nematicidal fungus Esteya vermicola in carbon and nitrogen culture
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摘要:
目的 比较伊氏杀线虫真菌Esteya vermicola(EV菌)在碳、氮营养源培养下的代谢差异，并找到重要代谢物或信号分子。 方法 选取培养真菌的碳培养基[主要为马铃薯葡萄糖肉汤培养基(PDB)]和培养细菌的氮培养基(主要为酵母粉)，将EV菌在2种培养基上25 ℃条件下培养7 d，收获菌丝体并提取代谢产物。采用非靶标的高效液相色谱-质谱联用技术(HPLC-MS)，在阴、阳离子模式下对代谢物组分进行分析和鉴定，并分析差异显著代谢物的代谢通路。 结果 共得到498种代谢物，阴、阳离子模式分别有176和362种，其中2种模式共同含有40种。差异显著的代谢物共有444种，占总数的89.2%，其中阴、阳离子模式分别有162和310种，有28种为2种模式共有。主成分和偏最小二乘判别分析均可使碳、氮培养条件下的代谢物聚为不同的簇并显著分离。氮培养条件下，磷酸胍基乙酸酯和对甲酚硫酸盐是大量产生且特有的代谢物；重要代谢物尿囊素、光色素、吲哚和海藻糖产量显著上调。通路分析将显著上调和下调的代谢物分别富集到氨基酸和糖类代谢相关的代谢通路。 结论 EV菌在碳培养和氮培养条件下呈现明显的代谢差异，代谢通路主要涉及糖类和氨基酸代谢。重要代谢物可为EV菌的高效培养和应用提供基础。图3表2参31 Abstract:Objective The objective is to compare the metabolic differences of the nematode-killing fungus Esteya vermicola (EV) cultured in carbon and nitrogen nutrient sources and to identify key metabolites or signal molecules. Method The carbon medium (mainly composed of PDB) for culturing fungi and the nitrogen medium (mainly composed of yeast powder) for culturing bacteria were selected. EV bacteria were cultured on two kinds of culture media at 25 ℃ for 7 days. The mycelia were harvested and the metabolites were extracted. Non-target high performance liquid chromatography-mass spectrometry(HPLC-MS) was used to analyze and identify metabolite components in both positive and negative ion modes. The metabolic pathways of metabolites with significant differences were analyzed. Result A total of 498 metabolites were identified, including 176 negative and 362 positive ion modes and 40 metabolites in both modes. There were 444 metabolites with significant differences, accounting for 89.2% of the total, among which 162 were negative and 310 were positive, and 28 were common to the two modes. Both principal component analysis and partial least square discriminant analysis could cluster the metabolites into different clusters and separate them significantly in carbon and nitrogen culture. In nitrogen culture, guanidine phosphate acetate and p-cresol sulfate were abundant and unique metabolites, and the yield of allantoin, photopigment, indole, and trehalose were significantly up-regulated. Pathway analysis enriched the significantly up-regulated and down-regulated metabolites into the metabolic pathways related to amino acid and carbohydrate metabolism, respectively. Conclusion EV bacteria showed significant metabolic differences in carbon and nitrogen culture. The metabolic pathway mainly involves carbohydrate and amino acid metabolism. The important metabolites will provide a theoretical basis for efficient culture and application of EV. [Ch, 3 fig. 2 tab. 31 ref.] -
Key words:
- biocontrol fungi /
- Esteya vermicola /
- metabonomics /
- LC-MS
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图 1 碳、氮培养条件下EV菌代谢物的PCA和PLS-DA分析
Figure 1 PCA and PLS-DA of EV metabolites under carbon and nitrogen culture conditions
图 2 碳、氮培养条件下EV菌的差异代谢物
Figure 2 Differential metabolites of EV bacteria under carbon and nitrogen culture conditions
图 3 差异代谢产物富集到的显著代谢通路
Figure 3 Significant metabolic pathways enriched by differential metabolites
表 1 部分差异化合物
Table 1. Part of significantly different metabolites
化合物名称 质荷比 保留时间/min 二级数据库得分 变化倍数 P KEGG编号 磷酸胍基乙酸酯 198.03 9.86 62 12.46 1.13E−13 C03166 对甲酚硫酸盐 187.01 6.17 53 12.41 8.88E−15 邻氨基苯甲酸酯 136.04 2.00 50 9.93 1.67E−08 C00108 尿酸 190.05 1.29 40 8.26 6.26E−07 C01717 尼古丁 163.12 4.68 44 5.51 1.14E−06 C16150 4-羟基-2-喹啉羧酸 188.03 5.76 30 5.48 9.62E−08 C01717 烟酸 124.04 3.50 41 5.17 1.71E−07 C00253 尿囊素 159.05 5.46 50 5.09 2.53E−04 C01551 吲哚丙烯酸 188.07 7.98 98 4.20 4.68E−08 2-吡咯烷酮 86.06 3.79 41 4.10 2.57E−05 C11118 咪唑乙酸 127.05 9.62 69 3.54 1.56E−07 C02835 组胺 112.09 5.74 45 3.34 2.79E−08 C00388 谷胱甘肽 306.08 1.02 58 3.26 5.97E−04 C00051 光色素 243.09 1.58 70 3.00 2.79E−04 C01727 肌肽 227.11 12.66 55 2.40 3.30E−05 C00386 甜菜碱醛 102.09 6.73 68 2.25 9.68E−03 C00576 吲哚 118.07 8.32 57 2.12 4.40E−03 C00463 苹果酸 133.01 0.90 43 1.90 2.86E−05 C00711 甜菜碱 118.09 8.30 51 1.88 2.79E−08 C00719 胍基乙酸d 118.06 8.26 38 1.87 1.19E−03 C00581 葫芦巴碱 138.06 9.05 38 1.59 1.71E−07 C01004 海藻糖 341.11 1.04 78 1.34 6.16E−05 C01083 左旋肉碱 162.11 9.31 72 1.34 2.97E−06 C00318 邻乙酰左旋肉碱 204.12 8.54 73 1.32 1.69E−06 C02571 茶碱 181.07 2.24 55 −1.64 6.07E−07 C07130 二乙醇胺 106.09 8.16 45 −1.73 1.42E−04 C06772 去甲肾上腺素 170.08 5.83 57 −2.40 1.52E−05 C00547 阿魏酸盐 193.05 5.36 32 −3.58 2.87E−05 C01494 核糖醇 151.06 1.01 46 −4.08 3.13E−08 C00474 甲基咪唑乙酸 141.07 9.52 41 −4.51 2.90E−08 C05828 赤藓糖醇 121.05 1.04 75 −5.20 2.80E−05 C00503 
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表 2 碳、氮培养条件下EV菌差异代谢物富集到的KEGG通路
Table 2. Enriched KEGG pathways by differential metabolites of EV under carbon and nitrogen culture conditions
代谢通路编号 代谢通路名称 匹配情况 P 影响大小 上调 sce00970 氨酰基tRNA的生物合成 18/46 4.38E−04 0.11 sce00330 精氨酸和脯氨酸代谢 10/25 7.53E−03 0.33 sce00220 精氨酸生物合成 8/18 8.05E−03 0.60 sce00430 牛磺酸和低牛磺酸代谢 4/7 2.28E−02 1.00 sce00250 丙氨酸、天冬氨酸和谷氨酸代谢 8/22 3.06E−02 0.81 下调 sce00520 氨基糖和核苷酸糖代谢 8/24 3.41E−04 0.48 sce00052 半乳糖代谢 5/17 8.98E−03 0.82 sce00040 戊糖和葡萄糖醛酸的相互转化 4/12 1.22E−02 0.27 sce00500 淀粉和蔗糖代谢 4/15 2.79E−02 0.35 说明：匹配情况为匹配化合物个数/通路化合物总数
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