不同连作年限菠萝园土壤差异代谢物和细菌群落结构分析

doi:10.13560/j.cnki.biotech.bull.1985.2021-0273

摘要/Abstract

摘要：

为探讨菠萝连作对土壤代谢物和细菌群落结构的影响,本研究采用非靶向代谢组学和细菌16S高通量测序技术,以菠萝园边角地未耕作过的土壤为对照（T0）,对连续种植了5 a（T5）和15 a（T15）菠萝的土壤进行了代谢物与细菌群落差异比较分析。研究结果表明,与T0比较,T5和T15土壤中分别检测到120种和80种差异代谢物;T0-T5和T0-T15组间共筛选出11种具有显著性差异（VIP>1,P<0.05）的代谢物,包括有机酸4种,糖类3种,氨基酸及其衍生物2种,醇类2种;KEGG代谢通路分析表明差异代谢物主要富集在不饱和脂肪酸的生物合成途径等11条关键代谢通路中。细菌16S高通量测序结果表明,与T0相比,T5中土壤细菌群落多样性、丰富度指数均升高;T15中土壤细菌群落多样性指数升高,但丰富度指数下降。T5和T15的土壤细菌群落结构存在明显差异,在门、属水平上T5和T15土壤细菌群落的相对丰度明显高于T0。与T0相比,菠萝园土壤的两个优势菌群变形细菌门（Proteobacteria）和酸杆菌门（Acidobacteriota）在T5中所占比例升高,但在T15中所占比例下降。CCA分析结果表明,土壤pH与连作年限呈负相关。本结果可为菠萝连作对土壤环境的影响相关研究提供参考,对提高菠萝园土壤管理水平有指导意义。

关键词: 菠萝连作, 土壤, 差异代谢物, 细菌群落结构

Abstract:

In order to investigate the effects of continuous-cropping pineapple on the soil metabolites and bacteria community structure,taking the soil of uncultivated land（T0）on the edge of the orchard as the control,untargeted metabolomics and bacterial 16S high-throughput sequencing were combined to compare and analyze the metabolites of the soils continuous-cropping pineapple for 5（T5）and 15（T15）years respectively as well as differences in bacterial community. The results of metabolites analysis indicated that there were totally 120 and 80 differential metabolites in the soil of T5 and T15 when compared with T0. And 11 significantly different metabolites（VIP>1,P<0.05）were detected between T0-T5 group and T0-T15 group in total,including 4 organic acids,3 carbohydrates,2 amino acid derivatives,2 alcohols. KEGG metabolic pathways analysis suggested that the differential metabolites were mostly enriched in 11 key metabolic pathways,e.g.,biosynthesis of unsaturated fatty acids. The bacterial 16S high-throughput sequencing profiling indicated that the bacteria diversity and abundance of the soil in T5 increased when compared with T0. The bacteria diversity increased while the abundance decreased in T15 soil when compared with T0. And there were significant differences in bacterial community structure between T5 and T15. In the levels of phyla and genus,the relative abundances of soil bacteria in T5 and T15 were significantly higher than that in T0. The proportion of two dominant bacteria communities in the soil of the pineapple orchard,Proteobacteria and Acidobacteriota,increased in T5 but decreased in T15 when compared with T0. CCA analysis showed that ccontinuous-cropping years were in significantly negative correlation with pH. This work would contribute to provide a reference for related research on the impacts of ccontinuous-cropping pineapple on the soil environment and contribute to provide the guidance for improving soil management level of pineapple orchard.

Key words: continuous-cropping pineapple, soil, differential metabolites, bacterial community structure

刘传和, 贺涵, 何秀古, 刘开, 邵雪花, 赖多, 匡石滋, 肖维强. 不同连作年限菠萝园土壤差异代谢物和细菌群落结构分析[J]. 生物技术通报, 2021, 37(8): 162-175.

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.

图/表 12

参考文献 43

[1]	尹承苗, 王玫, 王嘉艳, 等. 苹果连作障碍研究进展[J]. 园艺学报, 2017, 44(11):2215-2230.
	Yin CM, Wang M, Wang JY, et al. The research advance on apple replant disease[J]. Acta Hortic Sin, 2017, 44(11):2215-2230.
[2]	张兆波, 毛志泉, 朱树华. 6种酚酸类物质对平邑甜茶幼苗根系线粒体及抗氧化酶活性的影响[J]. 中国农业科学, 2011, 44(15):3177-3184.
	Zhang ZB, Mao ZQ, Zhu SH. Effects of phenolic acids on mitochondria and the activity of antioxidant enzymes in roots of seedlings of Malus hupehensis rehd[J]. Sci Agric Sin, 2011, 44(15):3177-3184.
[3]	侯慧, 董坤, 杨智仙, 等. 连作障碍发生机理研究进展[J]. 土壤, 2016, 48(6):1068-1076.
	Hou H, Dong K, Yang ZX, et al. Advance in mechanism of continuous cropping obstacle[J]. Soils, 2016, 48(6):1068-1076.
[4]	刘晓丽, 赵娜, 贾钰莹, 等. 酚酸类物质在植物-土壤-环境中的作用[J]. 园艺与种苗, 2020, 40(8):56-58, 63.
	Liu XL, Zhao N, Jia YY, et al. The role of phenolic acids in plant-soil-environment system[J]. Hortic Seed, 2020, 40(8):56-58, 63.
[5]	Rao GD, Liu XX, Zha WW, et al. Metabolomics reveals variation and correlation among different tissues of olive(Olea europaea L.)[J]. Biol Open, 2017, 6(9):1317-1323.
[6]	Arapitsas P, Corte AD, Gika H, et al. Studying the effect of storage conditions on the metabolite content of red wine using HILIC LC-MS based metabolomics[J]. Food Chem, 2016, 197 Pt B:1331-1340.
[7]	VanderWeide J, Medina-Meza IG, Frioni T, et al. Enhancement of fruit technological maturity and alteration of the flavonoid metabolomic profile in merlot(Vitis vinifera L.)by early mechanical leaf removal[J]. J Agric Food Chem, 2018, 66(37):9839-9849. doi: 10.1021/acs.jafc.8b02709 URL
[8]	殷继忠, 李亮, 接伟光, 等. 连作对大豆根际土壤细菌菌群结构的影响[J]. 生物技术通报, 2018, 34(1):230-238.
	Yin JZ, Li L, Jie WG, et al. Effects of continuous cropping on bacterial flora structure in soybean rhizosphere soil[J]. Biotechnol Bull, 2018, 34(1):230-238.
[9]	杨旭初, 龙莉, 柳开楼, 等. 连作对烟草根际土壤细菌碳代谢指纹的影响[J]. 河北农业大学学报, 2017, 40(5):38-43.
	Yang XC, Long L, Liu KL, et al. Effect of continuous cropping on carbon metabolic fingerprint of tobacco rhizosphere soil bacteria[J]. J Agric Univ Hebei, 2017, 40(5):38-43.
[10]	Liu HJ, Yang XY, Miao ZQ, et al. Characteristics of soil microflora of Panax notoginseng in different continuous cropping years[J]. Allelopathy J, 2018, 44(2):145-158. doi: 10.26651/AJ URL
[11]	郭睿, 王士华, 赵应勇. 不同种植年限对大叶女贞土壤生态环境的影响[J]. 西部林业科学, 2019, 48(6):86-92.
	Guo R, Wang SH, Zhao YY. Effects of planting years on the soil environment of Ligustrum compactum[J]. J West China For Sci, 2019, 48(6):86-92.
[12]	许广, 王梦姣, 邓百万, 等. 不同植茶年限茶树根际土壤细菌多样性及群落结构研究[J]. 生物技术通报, 2020, 36(3):124-132.
	Xu G, Wang MJ, Deng BW, et al. Bacterial diversity and community structure of rhizosphere soil of tea plants in different years of planting[J]. Biotechnol Bull, 2020, 36(3):124-132.
[13]	董艳辉, 于宇凤, 温鑫, 等. 基于高通量测序的藜麦连作根际土壤微生物多样性研究[J]. 华北农学报, 2019, 34(2):205-211.
	Dong YH, Yu YF, Wen X, et al. Studies on diversity of rhizosphere microorganism in quinoa continuous cropping soil by high throughput sequencing[J]. Acta Agric Boreali Sin, 2019, 34(2):205-211.
[14]	叶雯, 李永春, 喻卫武, 等. 不同种植年限香榧根际土壤微生物多样性[J]. 应用生态学报, 2018, 29(11):3783-3792.
	Ye W, Li YC, Yu WW, et al. Microbial biodiversity in rhizospheric soil of Torreya grandis ‘Merrillii’ relative to cultivation history[J]. Chin J Appl Ecol, 2018, 29(11):3783-3792.
[15]	洪洁, 康建依, 刘一倩, 等. 生菜连作及生菜-菠菜轮作对土壤细菌群落结构的影响[J]. 生物技术通报, 2019, 35(8):17-26.
	Hong J, Kang JY, Liu YQ, et al. Effects of continuous cropping of lettuce and rotation of LettuceSpinach on soil bacterial community structure[J]. Biotechnol Bull, 2019, 35(8):17-26.
[16]	国家统计局农村社会经济调查司. 中国农村统计年鉴-2020[M]. 北京: 中国统计出版社, 2020.
	Department of rural social and economy survey of national bureau of statistics. China rural statistical yearbook-2020[M]. Beijing: China Statistics Press, 2020.
[17]	刘传和, 刘岩, 凡超, 等. 菠萝茎叶还田对土壤理化特性及下茬菠萝生长的调控效应[J]. 热带作物学报, 2012, 33(12):2230-2235.
	Liu CH, Liu Y, Fan C, et al. Effects of stem-leaf returning on physic-chemical properties of soil and growth performances of next-cropping pineapples[J]. Chin J Trop Crops, 2012, 33(12):2230-2235.
[18]	Liu CH, Liu Y, Fan C, et al. The effects of composted pineapple residue return on soil properties and the growth and yield of pineapple[J]. J Soil Sci Plant Nutr, 2013(ahead):.
[19]	Kind T, Wohlgemuth G, Lee DY, et al. FiehnLib:mass spectral and retention index libraries for metabolomics based on quadrupole and time-of-flight gas chromatography/mass spectrometry[J]. Anal Chem, 2009, 81(24):10038-10048. doi: 10.1021/ac9019522 URL
[20]	Dunn WB, Broadhurst D, Begley P, et al. Procedures for large-scale metabolic profiling of serum and plasma using gas chromatography and liquid chromatography coupled to mass spectrometry[J]. Nat Protoc, 2011, 6(7):1060-1083. doi: 10.1038/nprot.2011.335 URL
[21]	Edgar RC. UPARSE:highly accurate OTU sequences from microbial amplicon reads[J]. Nat Methods, 2013, 10(10):996-998. doi: 10.1038/nmeth.2604 URL
[22]	Wang Y, Sheng HF, He Y, et al. Comparison of the levels of bacterial diversity in freshwater, intertidal wetland, and marine sediments by using millions of illumina tags[J]. Appl Environ Microbiol, 2012, 78(23):8264-8271. doi: 10.1128/AEM.01821-12 URL
[23]	Looft T, Johnson TA, Allen HK, et al. In-feed antibiotic effects on the swine intestinal microbiome[J]. PNAS, 2012, 109(5):1691-1696. doi: 10.1073/pnas.1120238109 URL
[24]	Lindström ES, Kamst-Van Agterveld MP, Zwart G. Distribution of typical freshwater bacterial groups is associated with pH, temperature, and lake water retention time[J]. Appl Environ Microbiol, 2005, 71(12):8201-8206. doi: 10.1128/AEM.71.12.8201-8206.2005 URL
[25]	贾新民, 宋义军, 娄战胜, 等. 大豆连作条件下土壤多糖组成的研究[J]. 黑龙江八一农垦大学学报, 1999, 11(4):12-17.
	Jia XM, Song YJ, Lou ZS, et al. Research of polysaccharides composition in soils in continues soybean cropping[J]. J Heilongjinag August First Land Reclam Univ, 1999, 11(4):12-17.
[26]	于方玲, 孙冰玉, 元野, 等. 连作对烤烟叶片淀粉和还原糖含量的影响[J]. 安徽农业科学, 2010, 38(4):1824-1825.
	Yu FL, Sun BY, Yuan Y, et al. Effects of continuous cropping on contents of starch and reducing sugar in the leaves of flue-cured tobacco[J]. J Anhui Agric Sci, 2010, 38(4):1824-1825.
[27]	王海斌, 陈晓婷, 丁力, 等. 连作茶树根际土壤自毒潜力、酶活性及微生物群落功能多样性分析[J]. 热带作物学报, 2018, 39(5):852-857.
	Wang HB, Chen XT, Ding L, et al. Analysis on autotoxic potential, enzyme activity and microbial community function diversity of the rhizosphere soils from tea plants with continuous cropping years[J]. Chin J Trop Crops, 2018, 39(5):852-857.
[28]	王艳芳, 潘凤兵, 展星, 等. 连作苹果土壤酚酸对平邑甜茶幼苗的影响[J]. 生态学报, 2015, 35(19):6566-6573.
	Wang YF, Pan FB, Zhan X, et al. Effects of five kinds of phenolic acid on the function of mitochondria and antioxidant systems in roots of Malus hupehensis Rehd. seedlings[J]. Acta Ecol Sin, 2015, 35(19):6566-6573.
[29]	尹承苗, 胡艳丽, 王功帅, 等. 苹果连作土壤中主要酚酸类物质对平邑甜茶幼苗根系的影响[J]. 中国农业科学, 2016, 49(5):961-969.
	Yin CM, Hu YL, Wang GS, et al. Effect of main phenolic acids of the apple replanted soil on the roots of Malus hupehensis Rehd. Seedlings[J]. Sci Agric Sin, 2016, 49(5):961-969.
[30]	谢星光, 陈晏, 卜元卿, 等. 酚酸类物质的化感作用研究进展[J]. 生态学报, 2014, 34(22):6417-6428.
	Xie XG, Chen Y, Bu YQ, et al. A review of allelopathic researches on phenolic acids[J]. Acta Ecol Sin, 2014, 34(22):6417-6428.
[31]	李华玮, 赵绪永, 李鹏坤. 作物连做障碍中酚酸类物质的生物降解研究[J]. 中国农学通报, 2011, 27(18):168-173.
	Li HW, Zhao XY, Li PK. Research on biodegradation effect to phenolic acid in even cook obstacles[J]. Chin Agric Sci Bull, 2011, 27(18):168-173.
[32]	Chen Y, Peng Y, Dai CC, et al. Biodegradation of 4-hydroxybenzoic acid by Phomopsis liquidambari[J]. Appl Soil Ecol, 2011, 51:102-110. doi: 10.1016/j.apsoil.2011.09.004 URL
[33]	Chen SL, Zhou BL, Lin SS, et al. Accumulation of cinnamic acid and vanillin in eggplant root exudates and the relationship with continuous cropping obstacle[J]. Afr J Biotechnol, 2011, 10(14):2659-2665. doi: 10.5897/AJB URL
[34]	李培栋, 王兴祥, 李奕林, 等. 连作花生土壤中酚酸类物质的检测及其对花生的化感作用[J]. 生态学报, 2010, 30(8):2128-2134.
	Li PD, Wang XX, Li YL, et al. The contents of phenolic acids in continuous cropping peanut and their allelopathy[J]. Acta Ecol Sin, 2010, 30(8):2128-2134.
[35]	Kelderer M, Manici LM, Caputo F, et al. Planting in the ‘inter-row’ to overcome replant disease in apple orchards:a study on the effectiveness of the practice based on microbial indicators[J]. Plant Soil, 2012, 357(1/2):381-393. doi: 10.1007/s11104-012-1172-0 URL
[36]	李丽, 蒋景龙. 基于代谢组学分析根腐病西洋参根际土壤特异代谢物[J]. 分子植物育种, 2020.
	Li L, Jiang JL. Analysis of specific metabolites in rhizosphere soil of panax quinquefolius l. with root rot diseases based on metabolomics[J]. Molecular Plant Breeding, 2020.
[37]	卢维宏, 张乃明, 包立, 等. 我国设施栽培连作障碍特征与成因及防治措施的研究进展[J]. 土壤, 2020, 52(4):651-658.
	Lu WH, Zhang NM, Bao L, et al. Study advances on characteristics, causes and control measures of continuous cropping obstacles of facility cultivation in China[J]. Soils, 2020, 52(4):651-658.
[38]	刘株秀, 刘俊杰, 徐艳霞, 等. 不同大豆连作年限对黑土细菌群落结构的影响[J]. 生态学报, 2019, 39(12):4337-4346.
	Liu ZX, Liu JJ, Xu YX, et al. Effects of continuous cropping years of soybean on the bacterial community structure in black soil[J]. Acta Ecol Sin, 2019, 39(12):4337-4346.
[39]	纳小凡, 郑国琦, 彭励, 等. 不同种植年限宁夏枸杞根际微生物多样性变化[J]. 土壤学报, 2016, 53(1):241-252.
	Na XF, Zheng GQ, Peng L, et al. Microbial biodiversity in rhizosphere of Lycium bararum L. relative to cultivation history[J]. Acta Pedol Sin, 2016, 53(1):241-252.
[40]	Shen C, Ni Y, Liang W, et al. Distinct soil bacterial communities along a small-scale elevational gradient in alpine tundra[J]. Front Microbiol, 2015, 6:582.
[41]	Zhang B, Liang C, He H, et al. Variations in soil microbial communities and residues along an altitude gradient on the northern slope of Changbai mountain, China[J]. PLoS One, 2013, 8(6):e66184. doi: 10.1371/journal.pone.0066184 URL
[42]	De Feudis M, Cardelli V, Massaccesi L, et al. Altitude affects the quality of the water-extractable organic matter(WEOM)from rhizosphere and bulk soil in European beech forests[J]. Geoderma, 2017, 302:6-13. doi: 10.1016/j.geoderma.2017.04.015 URL
[43]	胡菏, 吴宪, 赵建宁, 等. 有机-无机肥配施对麦玉轮作土壤中细菌氮循环功能基因的影响[J]. 农业环境科学学报, 2021, 40(1):144-154.
	Hu H, Wu X, Zhao JN, et al. The effects of combined organic and inorganic fertilizer on the bacterial nitrogen cycling functional genes in wheat and maize soils by PICRUSt functional prediction[J]. J Agro Environ Sci, 2021, 40(1):144-154.

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

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

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
代谢物序号 Metabolite ID	代谢物名称 Metabolite name	分类Classification	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
代谢物序号 Metabolite ID	代谢物名称 Metabolite name	分类Classification	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
代谢通路ID Metabolic pathway ID	代谢通路名称 Pathway name	T0-T5	T0-T15	代谢物序号 Metabolite ID
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