Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (8): 162-175.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0273
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LIU Chuan-he1(), HE Han1, HE Xiu-gu2, LIU Kai1, SHAO Xue-hua1, LAI Duo1, KUANG Shi-zi1, XIAO Wei-qiang1
Received:
2021-03-10
Online:
2021-08-26
Published:
2021-09-10
LIU Chuan-he, HE Han, HE Xiu-gu, LIU Kai, SHAO Xue-hua, LAI Duo, KUANG Shi-zi, XIAO Wei-qiang. Analysis of Differential Metabolites and Bacterial Community Structure in Soils of a Pineapple Orchard in Different Continuous-cropping Years[J]. Biotechnology Bulletin, 2021, 37(8): 162-175.
项目Item | 参数Parameter |
---|---|
进样量Sample volume | 1 μL |
分流模式Front inlet mode | 不分流模式Splitless mode |
隔垫吹扫流速 Front inlet septum purge flow | 3 mL/min |
载气Carrier gas | 氦Helium |
色谱柱Column | DB-5MS(30 m×250 μm×0.25 μm) |
柱流速Column flow | 1 mL/min |
柱箱升温程序Oven temperature ramp | 50℃保持1 min,以10℃/min的速度升至 310℃,并保持8 min 50℃ Stay at 50℃ for 1 min,raise to 310℃ at a rate of 10℃/min,Stay at 50℃ for 8 min |
前进样口温度 Front injection temperature | 280℃ |
传输线温度 Transfer line temperature | 280℃ |
离子源温度 Ion source temperature | 250℃ |
电离电压Electron energy | -70 eV |
质量范围Mass range | m/z:50-500 |
扫描速率Acquisition rate | 12.5光谱/秒Spectra per second |
溶剂延迟Solvent delay | 6.35 min |
Table 1 Instrument parameters
项目Item | 参数Parameter |
---|---|
进样量Sample volume | 1 μL |
分流模式Front inlet mode | 不分流模式Splitless mode |
隔垫吹扫流速 Front inlet septum purge flow | 3 mL/min |
载气Carrier gas | 氦Helium |
色谱柱Column | DB-5MS(30 m×250 μm×0.25 μm) |
柱流速Column flow | 1 mL/min |
柱箱升温程序Oven temperature ramp | 50℃保持1 min,以10℃/min的速度升至 310℃,并保持8 min 50℃ Stay at 50℃ for 1 min,raise to 310℃ at a rate of 10℃/min,Stay at 50℃ for 8 min |
前进样口温度 Front injection temperature | 280℃ |
传输线温度 Transfer line temperature | 280℃ |
离子源温度 Ion source temperature | 250℃ |
电离电压Electron energy | -70 eV |
质量范围Mass range | m/z:50-500 |
扫描速率Acquisition rate | 12.5光谱/秒Spectra per second |
溶剂延迟Solvent delay | 6.35 min |
Fig.2 Volcano plot of differential metabolites A and B are the comparison groups of T0-T5 and T0-T15,respectively. Each point from the volcano plot refers to a metabolite,the abscissa refers to the multiple change of each sample in the group(take the logarithm based on 2),and the ordinate refers to the P-value of the t test(take the logarithm based on 10),the size of the scatter points refers to the VIP value of the OPLS-DA model. The green point refers to the down-regulated differential metabolites,the red point refers to up-regulated the differential metabolites,and the black point refers to the differential metabolites with no significant difference
代谢物序号 Metabolite ID | 代谢物名称 Metabolite name | 分类Classification | 表达差异倍数 log2Fold Change | VIP值 VIP value | ||||
---|---|---|---|---|---|---|---|---|
T0-T5 | T0-T15 | T0-T5 | T0-T15 | |||||
meta_395 | (2R,3S)-2-羟基-3-异丙基丁二酸 (2R,3S)-2-hydroxy-3-isopro pylbutanedioic acid | 有机酸Organic acid | 1.924 | 2.099 | 1.504 | 1.685 | ||
meta_423 | 4-羟基苯甲酸4-Hydroxybenzoic acid | 有机酸Organic acid | 1.538 | 2.191 | 1.524 | 1.635 | ||
meta_465 | 4-羟基-3-甲氧基苯甲酸 4-hydroxy-3-methoxybenzoic acid | 有机酸Organic acid | 1.569 | 1.879 | 1.496 | 1.665 | ||
meta_484 | 3,4-二羟基苯甲酸3,4-dihydroxybenzoic acid | 有机酸Organic acid | 1.740 | 1.746 | 1.430 | 1.642 | ||
meta_304 | N-乙酰基-β-丙氨酸N-Acetyl-beta-alanine | 氨基酸衍生物 Amino acid derivatives | 2.211 | 3.186 | 1.491 | 1.726 | ||
meta_489 | N-氨基甲酰谷氨酸N-Carbamylglutamate | 氨基酸衍生物 Amino acid derivatives | 2.327 | 3.440 | 1.506 | 1.686 | ||
meta_504 | 葡萄糖Glucose | 糖类Carbohydrates | 4.878 | 1.580 | 2.840 | 1.634 | ||
meta_706 | 槐糖Sophorose | 糖类Carbohydrates | 4.201 | -3.892 | 1.471 | 1.112 | ||
meta_789 | 松三糖Melezitose | 糖类Carbohydrates | 1.292 | -1.386 | 1.368 | 1.156 | ||
meta_783 | (3β,5α,6β)-胆固醇3,5,6-三醇 (3β,5α,6β)-Cholestane-3,5,6-triol | 醇类Alcohols | 3.071 | 1.568 | 1.428 | 1.469 | ||
meta_513 | 山梨糖醇Sorbitol | 醇类Alcohols | 2.244 | 1.901 | 1.493 | 1.607 |
Table 2 Differential metabolites among the comparison groups
代谢物序号 Metabolite ID | 代谢物名称 Metabolite name | 分类Classification | 表达差异倍数 log2Fold Change | VIP值 VIP value | ||||
---|---|---|---|---|---|---|---|---|
T0-T5 | T0-T15 | T0-T5 | T0-T15 | |||||
meta_395 | (2R,3S)-2-羟基-3-异丙基丁二酸 (2R,3S)-2-hydroxy-3-isopro pylbutanedioic acid | 有机酸Organic acid | 1.924 | 2.099 | 1.504 | 1.685 | ||
meta_423 | 4-羟基苯甲酸4-Hydroxybenzoic acid | 有机酸Organic acid | 1.538 | 2.191 | 1.524 | 1.635 | ||
meta_465 | 4-羟基-3-甲氧基苯甲酸 4-hydroxy-3-methoxybenzoic acid | 有机酸Organic acid | 1.569 | 1.879 | 1.496 | 1.665 | ||
meta_484 | 3,4-二羟基苯甲酸3,4-dihydroxybenzoic acid | 有机酸Organic acid | 1.740 | 1.746 | 1.430 | 1.642 | ||
meta_304 | N-乙酰基-β-丙氨酸N-Acetyl-beta-alanine | 氨基酸衍生物 Amino acid derivatives | 2.211 | 3.186 | 1.491 | 1.726 | ||
meta_489 | N-氨基甲酰谷氨酸N-Carbamylglutamate | 氨基酸衍生物 Amino acid derivatives | 2.327 | 3.440 | 1.506 | 1.686 | ||
meta_504 | 葡萄糖Glucose | 糖类Carbohydrates | 4.878 | 1.580 | 2.840 | 1.634 | ||
meta_706 | 槐糖Sophorose | 糖类Carbohydrates | 4.201 | -3.892 | 1.471 | 1.112 | ||
meta_789 | 松三糖Melezitose | 糖类Carbohydrates | 1.292 | -1.386 | 1.368 | 1.156 | ||
meta_783 | (3β,5α,6β)-胆固醇3,5,6-三醇 (3β,5α,6β)-Cholestane-3,5,6-triol | 醇类Alcohols | 3.071 | 1.568 | 1.428 | 1.469 | ||
meta_513 | 山梨糖醇Sorbitol | 醇类Alcohols | 2.244 | 1.901 | 1.493 | 1.607 |
代谢通路ID Metabolic pathway ID | 代谢通路名称 Pathway name | -lgp | 代谢物序号 Metabolite ID | |
---|---|---|---|---|
T0-T5 | T0-T15 | |||
ko00051 | 果糖和甘露糖代谢 Fructose and mannose metabolism | 5.5963 | 4.6088 | 山梨糖醇meta_513 Sorbitol meta_513 |
ko00100 | 类固醇生物合成 Steroid biosynthesis | 2.3240 | 7.2183 | 胆固醇meta_762 Cholesterol meta_762 |
ko00940 | 苯丙烷生物合成 Phenylpropanoid biosynthesis | 3.3804 | 3.2558 | 4-羟基肉桂酸meta_517 4-Hydroxycinnamic acid meta_517 |
ko00130 | 泛醌和其他萜类醌的生物合成 Ubiquinone and other terpenoid-quinone biosynthesis | 3.3901 | 3.8455 | 4-羟基苯甲酸meta_423;4-羟基肉桂酸meta_517 4-Hydroxybenzoic acid meta_423;4-Hydroxycinnamic acid meta_517 |
ko00350 | 酪氨酸代谢 Tyrosine metabolism | 3.3733 | 1.6328 | 4-羟基肉桂酸meta_517 4-Hydroxycinnamic acid meta_517 |
ko00360 | 苯丙氨酸代谢 Phenylalanine metabolism | 2.7754 | 2.7181 | 4-羟基苯甲酸meta_423;4-羟基肉桂酸meta_517 4-Hydroxybenzoic acid meta_423;4-Hydroxycinnamic acid meta_517 |
ko00400 | 苯丙氨酸,酪氨酸和色氨酸的生物合成 Phenylalanine,tyrosine and tryptophan biosynthesis | 3.4807 | 1.9702 | 3,4-二羟基苯甲酸meta_484 3,4-Dihydroxybenzoic acid meta_484 |
ko00591 | 亚油酸代谢 Linoleic acid metabolism | 2.3376 | 4.1084 | 亚油酸meta_594 Linoleic acid meta_594 |
ko00790 | 叶酸生物合成 Folate biosynthesis | 6.7045 | 3.6467 | 4-羟基苯甲酸meta_423 4-Hydroxybenzoic acid meta_423 |
ko00950 | 异喹啉生物碱的生物合成 Isoquinoline alkaloid biosynthesis | 3.0892 | 4.2224 | 4-羟基肉桂酸meta_517 4-Hydroxycinnamic acid meta_517 |
ko01040 | 不饱和脂肪酸的生物合成 Biosynthesis of unsaturated fatty acids | 2.7263 | 2.9874 | 亚油酸meta_594;山嵛酸meta_681;木质酸meta_714 Linoleic acid meta_594. Behenic acid meta_681.Lignoceric acid meta_714 |
Table 3 Metabolic pathway analysis of the differential metabolites
代谢通路ID Metabolic pathway ID | 代谢通路名称 Pathway name | -lgp | 代谢物序号 Metabolite ID | |
---|---|---|---|---|
T0-T5 | T0-T15 | |||
ko00051 | 果糖和甘露糖代谢 Fructose and mannose metabolism | 5.5963 | 4.6088 | 山梨糖醇meta_513 Sorbitol meta_513 |
ko00100 | 类固醇生物合成 Steroid biosynthesis | 2.3240 | 7.2183 | 胆固醇meta_762 Cholesterol meta_762 |
ko00940 | 苯丙烷生物合成 Phenylpropanoid biosynthesis | 3.3804 | 3.2558 | 4-羟基肉桂酸meta_517 4-Hydroxycinnamic acid meta_517 |
ko00130 | 泛醌和其他萜类醌的生物合成 Ubiquinone and other terpenoid-quinone biosynthesis | 3.3901 | 3.8455 | 4-羟基苯甲酸meta_423;4-羟基肉桂酸meta_517 4-Hydroxybenzoic acid meta_423;4-Hydroxycinnamic acid meta_517 |
ko00350 | 酪氨酸代谢 Tyrosine metabolism | 3.3733 | 1.6328 | 4-羟基肉桂酸meta_517 4-Hydroxycinnamic acid meta_517 |
ko00360 | 苯丙氨酸代谢 Phenylalanine metabolism | 2.7754 | 2.7181 | 4-羟基苯甲酸meta_423;4-羟基肉桂酸meta_517 4-Hydroxybenzoic acid meta_423;4-Hydroxycinnamic acid meta_517 |
ko00400 | 苯丙氨酸,酪氨酸和色氨酸的生物合成 Phenylalanine,tyrosine and tryptophan biosynthesis | 3.4807 | 1.9702 | 3,4-二羟基苯甲酸meta_484 3,4-Dihydroxybenzoic acid meta_484 |
ko00591 | 亚油酸代谢 Linoleic acid metabolism | 2.3376 | 4.1084 | 亚油酸meta_594 Linoleic acid meta_594 |
ko00790 | 叶酸生物合成 Folate biosynthesis | 6.7045 | 3.6467 | 4-羟基苯甲酸meta_423 4-Hydroxybenzoic acid meta_423 |
ko00950 | 异喹啉生物碱的生物合成 Isoquinoline alkaloid biosynthesis | 3.0892 | 4.2224 | 4-羟基肉桂酸meta_517 4-Hydroxycinnamic acid meta_517 |
ko01040 | 不饱和脂肪酸的生物合成 Biosynthesis of unsaturated fatty acids | 2.7263 | 2.9874 | 亚油酸meta_594;山嵛酸meta_681;木质酸meta_714 Linoleic acid meta_594. Behenic acid meta_681.Lignoceric acid meta_714 |
Fig. 4 KEGG pathway enrichment map of differential metabolites A and B are the comparison group of T0-T5 and T0-T15,respectively. The light yellow nodes in the figures refer to pathways,and the small nodes connected to the pathways are the specific metabolites remarked on them. The shade of the color refers to the different multiple based on the log2fold change,the size of circle refers to the value of the effects of differential metabolites on the pathways. Usually,the darker of the color and bigger of the circle refer to the differences of metabolites are more significant and effects on pathways bigger. The figure shows at most 5 pathways
Fig. 7 Relative abundance of different soil bacterial community at phyla,genus and order level in the different continuous-cropping years of the pineapple orchard Picture A,B and C are refer to the levels of bacterial phyla,genus and order,respectively
连作年限 Continuous-cropping year | ACE指数 ACE index | Chao1指数 Chao1 index | Simpson指数 Simpson index | Shannon指数 Shannon index |
---|---|---|---|---|
T5 | 802.89±22.72a | 805.23±20.92a | 0.01±0.00b | 5.39±0.11a |
T15 | 577.27±17.43b | 579.76±16.38b | 0.04±0.00b | 4.66±0.03b |
T0 | 643.25±32.15b | 610.45±39.27b | 0.12±0.03a | 3.90±0.18c |
Table 4 Diversity of relative abundance of different bacterial of soil under different continuous-cropping years of the pineapple orachard
连作年限 Continuous-cropping year | ACE指数 ACE index | Chao1指数 Chao1 index | Simpson指数 Simpson index | Shannon指数 Shannon index |
---|---|---|---|---|
T5 | 802.89±22.72a | 805.23±20.92a | 0.01±0.00b | 5.39±0.11a |
T15 | 577.27±17.43b | 579.76±16.38b | 0.04±0.00b | 4.66±0.03b |
T0 | 643.25±32.15b | 610.45±39.27b | 0.12±0.03a | 3.90±0.18c |
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