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多环芳烃(PAHs)是一类具有致癌、致畸、致突变效应的有机污染物,可在水体、土壤或沉积物中积累,被植物吸收而进入食物链中,通过生物富集作用对人类健康产生极大威胁[1-2]。在PAHs污染的诸多修复方法中,生物修复具有绿色环保、成本低、大面积应用等优点[3-4],是目前最具潜力的方法之一。一般来说,PAHs的环数越高,其化学结构越稳定、疏水性越高,单一利用植物或白腐真菌对PAHs去除能力有限[5-7]。近来的研究表明,植物-白腐真菌联合修复是一种高效的PAHs污染土壤的生物修复方法,能有效去除高分子量PAHs[8]。但目前植物-白腐真菌联合修复策略的作用机理尚不完全清楚。
虽然植物从土壤中吸收和积累PAHs的能力有限,但植物能促进根际土壤环境中PAHs的降解,学者推测可能是植物和根际微生物相互协作的结果[9]。据报道,植物约20%的光合产物用于根部合成有机分子,并以根系分泌物的形式释放到土壤中[10]。根系分泌物主要成分有低分子量有机酸(LWMOAs)、氨基酸、糖、酰胺、脂族酸、芳香酸、醇、酮和烯烃等,这些代谢物为植物根际土壤微生物提供碳源和氮源,包括PAHs降解菌,增加微生物的生物量和活性[11-13],从而提高PAHs的降解率。此外,LWMOAs、氨基酸、糖类均在PAHs脱附方面有显著效果,从而提高了PAHs的溶解度和生物可利用度,加速土壤中PAHs的降解[14-16]。研究表明[17],低分子量PAHs处理下玉米的光合作用增强,玉米根系分泌物组分及其含量也趋于增加。目前尚不清楚接种平滑白蛋巢菌对蒿柳根系分泌物的影响,后者是影响根际PAHs降解菌生长的重要因素。
近来,在环境学科中使用的非靶向代谢组学引起了生物学者们的广泛关注,代谢组技术的进步使研究人员能够在短时间内分析一个样品中的数百种化合物。植物产生的代谢物范围比其他任何一类生物体都要广,代谢组学在植物科学中的重要性也相应更高[18]。利用非靶向代谢组学进行植物组织内代谢产物的研究日渐增多,但针对植物根系分泌物的代谢组学研究相对较少[13, 19-22]。根际代谢组学的关键在于如何收集原生状态的根系分泌物。大多研究利用霍格兰营养液、石英砂等基质对植物进行培养并收集根系分泌物,这些方法的好处在于有效消除土壤环境的噪音干扰,但生长介质能影响根系形态、分泌的方式和代谢物组分[23]。同时,由于考虑到污染土壤修复实际情况,以及需要满足白腐真菌的定殖、生长和产酶条件时,需选择土壤作为培养基质。
基于此,本研究在温室中设置了为期60天的盆栽试验,选择蒿柳(Salix viminalis L.)和平滑白蛋巢菌(Crucibulum laeve)作为试验材料,试验材料的选择以及该组合体系的研究价值在先前研究[24]中有详细表述,利用自然衰减(NA),真菌修复(M),植物修复(P)和植物-微生物联合修复(PMR)等4种策略修复PAHs污染土壤,利用非靶向代谢组学定性和定量分析蒿柳的根际土壤代谢物,从根系分泌物的角度研究了PAHs污染土壤中蒿柳对平滑白蛋巢菌刺激的应答机理,以期为植物-白腐真菌联合修复的作用机理研究提供理论依据。
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试验第60天时,通过POS模式检测并鉴定了各处理样品中881种化合物,通过NEG模式检测并鉴定了各处理样品中828种化合物。将所有代谢物进行数据min-max标准化后进行聚类分析并绘制热图(图1),可直观展示各处理样品的代谢物组成和含量有明显差别。已鉴定的化合物包括丙酸、琥珀酸、苯甲酸、戊酸、庚酸、癸酸、山楂酸等有机酸,半胱氨酸等氨基酸,松二糖、蔗糖、葡萄糖、海藻糖等糖类,15-棕榈酸甲酯、5-羟基水杨酸酯等酯类;此外还有醇、酮、腺苷、酰胺、生物碱、醛等小分子代谢物。
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采用主成分分析法(PCA)和正交偏最小二乘判别分析法(OPLS-DA),将已鉴定的代谢物进行聚类,以区分各处理间代谢物的变化,并鉴定潜在的根系分泌物组分。本研究利用PCA对P处理、NA处理样品的代谢表型进行了聚类,在PCA评分图(图2)中,每个数据点代表一个样品,聚集在一起的点具有比分开的点更相似的生化组成。图2A显示POS模式下P处理和NA处理间的样品能显著区分,表明蒿柳显著影响了根际土壤代谢物的组分和含量。随后应用OPLS-DA最大化各处理样品间差异,如图2所示,在POS模式和NEG模式下P处理和NA处理的样品均有明显的分离。应用OPLS-DA生成的载荷图和VIP评分(表1)来解释代谢模式,在POS和NEG模式下总共筛选到18种化合物并将其假定为根系分泌物组分,包括丙二酸、乙醛酸、邻苯二甲酸、4-甲基-2-氧戊酸、癸酸、十五烷酸、十六烷二酸、蓖麻油酸、棕榈油酸、反式亚油酸等有机酸,半胱氨酸等氨基酸,葡萄糖、海藻糖、松二糖等糖类,5-羟基水杨酸酯等酯类,此外还有脯氨酸甜菜碱、L-抗坏血酸2-磷酸钠、5′-甲硫腺苷等小分子代谢物。
图 2 蒿柳根际土壤代谢物的多元分析。POS模式下的(A)PCA评分图,(C)OPLS-DA评分图;NEG模式下的(B)PCA评分图,(D)OPLS-DA评分图
Figure 2. Multivariate analysis of rhizosphere soil metabolites from S. viminalis. (A) PCA scores plots and (C) OPLS-DA scores plots of OPLS-DA under POS mode; (B) PCA scores plots and (D) OPLS-DA scores plots of OPLS-DA under NEG mode
表 1 NA和P处理条件下土壤样品间差异显著的潜在根系分泌物组分
Table 1. Potential root exudate components of soil samples between NA and P treatment (n = 6)
化合物
Compounds模式
ModeNA处理
NA treatmentP处理
P treatmentVIP p值
p-value蓖麻油酸 Ricinoleic acid NEG 1.00 ± 0.10 2.49 ± 0.30 1.89 8.45e-04 海藻糖 Trehalose NEG 1.00 ± 0.04 2.70 ± 0.45 4.62 3.46e-03 十五烷酸 Pentadecanoic acid NEG 1.00 ± 0.03 3.24 ± 0.26 4.81 5.91e-06 棕榈油酸(Z)-hexadec-9-enoic acid NEG 1.00 ± 0.13 4.62 ± 0.47 6.52 3.34e-03 癸酸 Decanoic acid NEG 1.00 ± 0.08 2.54 ± 0.29 2.20 4.94e-04 丙二酸 Propanedioic acid NEG 1.00 ± 0.17 1.98 ± 0.31 1.02 2.00e-02 4-甲基-2-氧戊酸 4-methyl-2-oxopentanoic acid NEG 1.00 ± 0.13 3.91 ± 0.54 3.18 4.05e-04 十六烷二酸 Hexadecanedioic acid NEG 1.00 ± 0.05 4.85 ± 0.95 1.11 2.38e-03 半胱氨酸 Cysteine NEG 1.00 ± 0.85 13.77 ± 2.91 1.34 1.81e-03 反式亚油酸 Linoelaidic acid NEG 1.00 ± 0.06 9.98 ± 1.60 10.87 2.22e-04 L-抗坏血酸2-磷酸钠 Sodium L-ascorbic acid 2-phosphate NEG 1.00 ± 0.06 2.00 ± 0.21 3.38 1.10e-03 5-羟水杨酸酯 5-Hydroxyisourate NEG 1.00 ± 0.52 48.48 ± 10.95 4.29 1.48e-03 葡萄糖 Glucose NEG 1.00 ± 0.10 3.08 ± 0.45 2.67 1.08e-03 乙醛酸 Glyoxylate NEG 1.00 ± 0.04 1.12 ± 0.04 1.30 4.94e-02 邻苯二甲酸 Phthalic acid NEG 1.00 ± 0.01 1.11 ± 0.03 1.04 7.20e-03 松二糖 Turanose POS 1.00 ± 0.07 2.25 ± 0.35 2.71 5.33e-03 脯氨酸甜菜碱 Proline betaine POS 1.00 ± 0.12 3.96 ± 0.82 4.53 5.06e-03 5′-甲硫腺苷 5′-Methylthioadenosine POS 1.00 ± 0.08 2.19 ± 0.27 1.31 1.89e-03 -
采用PCA对NA、M、P、PMR处理样品的代谢表型进行分类,图3显示POS模式和NEG模式下各处理间的样品均能明显区分,表明各处理对土壤代谢物组分和含量均有不同影响。随后,采用OPLS-DA对各处理间样品两两比较生成的载荷图,结合VIP评分筛选差异代谢物。由图4可知,在POS模式下M处理、P处理相较于NA处理相对含量上调和下调的差异代谢物数量相当(相差 < 15),但在NEG模式下相对含量上调的代谢物数量远大于(相差 > 150)下调的代谢物数量;两种模式下,M处理相较于P处理样品相对含量上调的代谢物数量大于(相差 ≥ 50)下调的代谢物数量;两种模式下,PMR处理相比其它处理样品相对含量下调的代谢物数量都大于(相差32-186)上调的代谢物数量。
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各处理对18种潜在根系分泌物组分的影响如图5所示。相比NA处理,M处理提高了十六烷二酸、半胱氨酸、5-羟基水杨酸酯、葡萄糖、4-甲基-2-氧戊酸、5′-甲硫腺苷、反式亚油酸、邻苯二甲酸、十五烷酸、棕榈油酸、癸酸、乙醛酸、蓖麻油酸、海藻糖、松二糖、L-抗坏血酸2-磷酸钠等16种代谢物的相对含量;PMR处理提高了十六烷二酸、半胱氨酸、5-羟基水杨酸酯、葡萄糖、4-甲基-2-氧戊酸、5′-甲硫腺苷、反式亚油酸、邻苯二甲酸、十五烷酸、棕榈油酸、癸酸等11种代谢物的相对含量,而其它代谢物的相对含量下降或变化不明显。PMR处理相较于P处理仅提高了葡萄糖和十五烷酸等2种代谢物的相对含量,相较于M处理提高了十六烷二酸、半胱氨酸、5-羟基水杨酸酯、葡萄糖、十五烷酸、棕榈油酸、癸酸和丙二酸等8种代谢物的相对含量。
接种平滑白蛋巢菌对蒿柳根系分泌物代谢组的影响
Effects of Crucibulum laeve Inoculation on Metabolome in Root Exudate from Salix viminalis L.
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摘要:
目的 虽然已有研究表明植物-白腐真菌联合修复是一种更高效的多环芳烃(PAHs)污染土壤修复策略,但目前尚不清楚接种白腐真菌对植物根系分泌物的影响,后者与根际PAHs降解菌的生长和PAHs的生物降解密切相关。基于此,我们开展PAHs污染土壤中接种白腐真菌对植物根际土壤代谢组的影响研究。 方法 在温室中设置了PAHs污染土壤的盆栽修复实验,以蒿柳(Salix viminalis L.)作为植物修复树种,基于LC-MS检测,重点比较PAHs污染土壤中接种平滑白蛋巢菌(Crucibulum laeve)对蒿柳根际土壤代谢谱的影响。 结果 分别在正离子模式(POS)和(NEG)模式下鉴定出881和828种代谢物,不同处理之间代谢物的组分及其含量存在显著差异。比较分析了差异代谢物并鉴定出18个潜在根系分泌物组分,接种平滑白蛋巢菌显著减少了蒿柳根际土壤代谢物的组分和含量,其中16种根系分泌物的含量明显降低。 结论 接种白腐真菌促进了植物根系对广谱土壤化合物的吸收能力,推测植物根系对白腐真菌的刺激作用做出应答,引发了植物PAHs提取能力提高,对揭示植物-白腐真菌联合修复的作用机理有重要意义。 Abstract:Objective To study the effects of plant/white rot fungi (WRF) inoculation on rhizosphere soil metabolome in root exudate of plants cultivated in PAH-contaminated soil. Method A pot experiment was conducted in greenhouse for bioremediation of PAH-contaminated soils and Salix viminalis L. was used as phytoremediation materials. Based on liquid chromatography-mass spectrometry (LC-MS) and metabolomics method, the test was focused on comparing the effect of inoculating Crucibulum laeve on metabolic profiling of rhizosphere PAH-contaminated soil of S. viminalis. Result Under POS and NEG mode, 881 and 823 compounds were detected and identified in metabolic profiling, respectively. Among them, 18 compounds were identified and assumed to be potential root exudates components. The component variety and content of rhizosphere metabolites were remarkably reduced by C. laeve inoculating, which including 16 root exudate components. Conclusion In this study, it is showed that inoculation with WRF can promote the uptake capability of roots to a broad spectrum of soil compounds. It is speculated that the response of plant roots to WRF stimulating leads to the improvement of PAHs phytoextraction capacity. This is of great significance for revealing the mechanism of plant/WRF combination remediation of PAH-contaminated soils. -
Key words:
- Salix viminalis
- / PAHs contamination
- / Crucibulum laeve
- / rhizosphere soil
- / metabolome
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图 2 蒿柳根际土壤代谢物的多元分析。POS模式下的(A)PCA评分图,(C)OPLS-DA评分图;NEG模式下的(B)PCA评分图,(D)OPLS-DA评分图
Figure 2. Multivariate analysis of rhizosphere soil metabolites from S. viminalis. (A) PCA scores plots and (C) OPLS-DA scores plots of OPLS-DA under POS mode; (B) PCA scores plots and (D) OPLS-DA scores plots of OPLS-DA under NEG mode
表 1 NA和P处理条件下土壤样品间差异显著的潜在根系分泌物组分
Table 1. Potential root exudate components of soil samples between NA and P treatment (n = 6)
化合物
Compounds模式
ModeNA处理
NA treatmentP处理
P treatmentVIP p值
p-value蓖麻油酸 Ricinoleic acid NEG 1.00 ± 0.10 2.49 ± 0.30 1.89 8.45e-04 海藻糖 Trehalose NEG 1.00 ± 0.04 2.70 ± 0.45 4.62 3.46e-03 十五烷酸 Pentadecanoic acid NEG 1.00 ± 0.03 3.24 ± 0.26 4.81 5.91e-06 棕榈油酸(Z)-hexadec-9-enoic acid NEG 1.00 ± 0.13 4.62 ± 0.47 6.52 3.34e-03 癸酸 Decanoic acid NEG 1.00 ± 0.08 2.54 ± 0.29 2.20 4.94e-04 丙二酸 Propanedioic acid NEG 1.00 ± 0.17 1.98 ± 0.31 1.02 2.00e-02 4-甲基-2-氧戊酸 4-methyl-2-oxopentanoic acid NEG 1.00 ± 0.13 3.91 ± 0.54 3.18 4.05e-04 十六烷二酸 Hexadecanedioic acid NEG 1.00 ± 0.05 4.85 ± 0.95 1.11 2.38e-03 半胱氨酸 Cysteine NEG 1.00 ± 0.85 13.77 ± 2.91 1.34 1.81e-03 反式亚油酸 Linoelaidic acid NEG 1.00 ± 0.06 9.98 ± 1.60 10.87 2.22e-04 L-抗坏血酸2-磷酸钠 Sodium L-ascorbic acid 2-phosphate NEG 1.00 ± 0.06 2.00 ± 0.21 3.38 1.10e-03 5-羟水杨酸酯 5-Hydroxyisourate NEG 1.00 ± 0.52 48.48 ± 10.95 4.29 1.48e-03 葡萄糖 Glucose NEG 1.00 ± 0.10 3.08 ± 0.45 2.67 1.08e-03 乙醛酸 Glyoxylate NEG 1.00 ± 0.04 1.12 ± 0.04 1.30 4.94e-02 邻苯二甲酸 Phthalic acid NEG 1.00 ± 0.01 1.11 ± 0.03 1.04 7.20e-03 松二糖 Turanose POS 1.00 ± 0.07 2.25 ± 0.35 2.71 5.33e-03 脯氨酸甜菜碱 Proline betaine POS 1.00 ± 0.12 3.96 ± 0.82 4.53 5.06e-03 5′-甲硫腺苷 5′-Methylthioadenosine POS 1.00 ± 0.08 2.19 ± 0.27 1.31 1.89e-03 -
[1] Buonanno G, Giovinco G, Morawska L, et al. Lung cancer risk of airborne particles for Italian population[J]. Environmental Research, 2015, 142: 443-451. doi: 10.1016/j.envres.2015.07.019 [2] Joner E J, Leyval C, Colpaert J V, et al. Ectomycorrhizas impede phytoremediation of polycyclic aromatic hydrocarbons (PAHs) both within and beyond the rhizosphere[J]. Environmental Pollution, 2006, 142(1): 34-38. doi: 10.1016/j.envpol.2005.09.007 [3] Agnello A C, Bagard M, van Hullebusch E D, et al. Comparative bioremediation of heavy metals and petroleum hydrocarbons co-contaminated soil by natural attenuation, phytoremediation, bioaugmentation and bioaugmentation-assisted phytoremediation[J]. Science of the Total Environment, 2016, 563-564: 693-703. doi: 10.1016/j.scitotenv.2015.10.061 [4] Kuppusamy S, Thavamani P, Venkateswarlu K, et al. Remediation approaches for polycyclic aromatic hydrocarbons (PAHs) contaminated soils: Technological constraints, emerging trends and future directions[J]. Chemosphere, 2017, 168: 944-968. doi: 10.1016/j.chemosphere.2016.10.115 [5] IARC Working Group on the Evaluation of Carcinogenic Risks to Humans. Some non-heterocyclic polycyclic aromatic hydrocarbons and some related exposures[C]. Lyons: International Agency for Research on Cancer, 2010. [6] Bamforth S M, Singleton I. Bioremediation of polycyclic aromatic hydrocarbons: current knowledge and future directions[J]. Journal of Chemical Technology & Biotechnology, 2005, 80(7): 723-736. [7] Ghosal D, Ghosh S, Dutta T K, et al. Current state of knowledge in microbial degradation of polycyclic aromatic hydrocarbons (PAHs): a review[J]. Frontiersin Microbiology, 2016, 7: 1369. [8] García-Sánchez M, Kosnar Z, Mercl F, et al. A comparative study to evaluate natural attenuation, mycoaugmentation, phytoremediation, and microbial-assisted phytoremediation strategies for the bioremediation of an aged PAH-polluted soil[J]. Ecotoxicology and Environmental Safety, 2018, 147: 165-174. doi: 10.1016/j.ecoenv.2017.08.012 [9] Alagić S Č, Maluckov B S, Radojičić V B, et al. How can plants manage polycyclic aromatic hydrocarbons? May these effects represent a useful tool for an effective soil remediation? A review[J]. Clean Technologies and Environmental Policy, 2015, 17(3): 597-614. doi: 10.1007/s10098-014-0840-6 [10] Huang A C, Jiang T, Liu Y X, et al. A specialized metabolic network selectively modulates Arabidopsis root microbiota[J]. Science, 2019, 364: 6440. [11] Marmiroli M, Pietrini F, Maestri E, et al. Growth, physiological and molecular traits in Salicaceae trees investigated for phytoremediation of heavy metals and organics[J]. Tree Physiology, 2011, 31(12): 1319-1334. doi: 10.1093/treephys/tpr090 [12] Carvalhais L C, Dennis P G, Fedoseyenko D, et al. Root exudation of sugars, amino acids, and organic acids by maize as affected by nitrogen, phosphorus, potassium, and iron deficiency[J]. Journal of Plant Nutrition and Soil Science, 2011, 174(1): 3-11. doi: 10.1002/jpln.201000085 [13] Luo Q, Wang S, Sun L N, et al. Metabolic profiling of root exudates from two ecotypes of Sedum alfredii treated with Pb based on GC-MS[J]. Scientific Reports, 2017, 7: 39878. doi: 10.1038/srep39878 [14] Guo M, Gong Z, Miao R, et al. The influence of root exudates of maize and soybean on polycyclic aromatic hydrocarbons degradation and soil bacterial community structure[J]. Ecological Engineering, 2017, 99: 22-30. doi: 10.1016/j.ecoleng.2016.11.018 [15] Tian W, Zhao J, Zhou Y, et al. Effects of root exudates on gel-beads/reeds combination remediation of high molecular weight polycyclic aromatic hydrocarbons[J]. Ecotoxicology and Environmental Safety, 2017, 135: 158-164. doi: 10.1016/j.ecoenv.2016.09.021 [16] Jiang S, Xie F, Lu H, et al. Response of low-molecular-weight organic acids in mangrove root exudates to exposure of polycyclic aromatic hydrocarbons[J]. Environmental Science and Pollution Research, 2017, 24(13): 12484-12493. doi: 10.1007/s11356-017-8845-4 [17] Lapie C, Sterckeman T, Paris C, et al. Impact of phenanthrene on primary metabolite profiling in root exudates and maize mucilage[J]. Environmental Science and Pollution Research, 2020, 27(3): 3124-3142. doi: 10.1007/s11356-019-07298-x [18] Kawamoto K, Oashi T, Oami K, et al. Perfluorooctanoic acid (PFOA) but not perfluorooctane sulfonate (PFOS) showed DNA damage in comet assay on Paramecium caudatum.[J]. The Journal of Toxicological Sciences, 2010, 35(6): 835-841. doi: 10.2131/jts.35.835 [19] Sivaram A K, Subashchandrabose S R, Logeshwaran P, et al. Metabolomics reveals defensive mechanisms adapted by maize on exposure to high molecular weight polycyclic aromatic hydrocarbons[J]. Chemosphere, 2019, 214: 771-780. doi: 10.1016/j.chemosphere.2018.09.170 [20] Yuan J, Zhao J, Wen T, et al. Root exudates drive the soil-borne legacy of aboveground pathogen infection[J]. Microbiome, 2018, 6(1): 156. doi: 10.1186/s40168-018-0537-x [21] Swenson T L, Karaoz U, Swenson J M, et al. Linking soil biology and chemistry in biological soil crust using isolate exometabolomics[J]. Nature Communications, 2018, 9(1): 19. doi: 10.1038/s41467-017-02356-9 [22] Luo F, Wang Q, Yin C, et al. Differential metabolic responses of Beauveria bassiana cultured in pupae extracts, root exudates and its interactions with insect and plant[J]. Journal of Invertebrate Pathology, 2015, 130: 154-164. doi: 10.1016/j.jip.2015.01.003 [23] 彭钰洁, 程 楠, 李佳佳, 等. 氮肥减施对玉米幼苗根系分泌物影响的根际代谢组学分析[J]. 中国生态农业学报, 2018, 26(164):21-28. [24] 马晓东, 李 霞, 刘俊祥, 等. 多环芳烃(PAHs)污染土壤中接种平滑白蛋巢菌对蒿柳光合作用的影响[J]. 北京林业大学学报, 2020, 42(5):80-87. doi: 10.12171/j.1000-1522.20190340 [25] Reina R, Liers C, Ocampo J A, et al. Solid state fermentation of olive mill residues by wood- and dung-dwelling Agaricomycetes: effects on peroxidase production, biomass development and phenol phytotoxicity[J]. Chemosphere, 2013, 93(7): 1406-1412. doi: 10.1016/j.chemosphere.2013.07.006 [26] Wang J, Zhang T, Shen X, et al. Serum metabolomics for early diagnosis of esophageal squamous cell carcinoma by UHPLC-QTOF/MS[J]. Metabolomics, 2016, 12(7): 116. doi: 10.1007/s11306-016-1050-5 [27] Want E J, O′Maille G, Abagyan R, et al. XCMS: Processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching, and identification[J]. Analytical Chemistry, 2006, 78(3): 779-787. doi: 10.1021/ac051437y [28] Saccenti E, Hoefsloot H C J, Smilde A K, et al. Reflections on univariate and multivariate analysis of metabolomics data[J]. Metabolomics, 2014, 10(3): 361-374. doi: 10.1007/s11306-013-0598-6 [29] Bais B P, Weir T L, Perry L G, et al. The role of root exudates in rhizosphere interactions with plants and other organisms[J]. Annual Review of Plant Biology, 2006, 57: 233-266. doi: 10.1146/annurev.arplant.57.032905.105159 [30] Mhlongo M I, Piater L A, Madala N E, et al. The chemistry of plant-microbe interactions in the rhizosphere and the potential for metabolomics to reveal signaling related to defense priming and induced systemic resistance[J]. Frontiers in Plant Science, 2018, 9: 112. doi: 10.3389/fpls.2018.00112 [31] Smith K E, Schwab A P, Banks M K, et al. Dissipation of PAHs in saturated, dredged sediments: A field trial[J]. Chemosphere, 2008, 72(10): 1614-1619. doi: 10.1016/j.chemosphere.2008.03.020 [32] 刘世亮, 骆永明, 吴龙华, 等. 污染土壤中苯并[a]芘的微生物共代谢修复研究[J]. 土壤学报, 2010, 47(2):364-369. doi: 10.11766/trxb2010470223 [33] 吴 颖, 梁月荣. S-腺苷甲硫氨酸在茶树生理代谢中的研究现状[J]. 茶叶, 2005, 2:18-20. [34] Shen Y, Li J, Gu R, et al. Carotenoid and superoxide dismutase are the most effective antioxidants participating in ROS scavenging in phenanthrene accumulated wheat leaf[J]. Chemosphere, 2018, 197: 513-525. doi: 10.1016/j.chemosphere.2018.01.036 [35] 张建锋, 李吉跃, 宋玉民, 等. 植物耐盐机理与耐盐植物选育研究进展[J]. 世界林业研究, 2003, 16(2):16-22. doi: 10.3969/j.issn.1001-4241.2003.02.004 [36] Ma X, Li X, Liu J, et al. Enhancing Salix viminalis L. –mediated phytoremediation of polycyclic aromatic hydrocarbon–contaminated soil by inoculation with Crucibulum laeve (white-rot fungus). Environmental Science and Pollution Research , 2020 , 27: 41326 – 41341. DOI: 10.1007/s11356-020-10125-3 [37] Ma X, Li X, Liu J, et al. Soil microbial community succession and interactions during combined plant/white-rot fungus remediation of polycyclic aromatic hydrocarbons. Science of The Total Environment , 2021 , 752: 142224. DOI: 10.1016/j.scitotenv.2020.142224