Phytomanagement of a metal(loid)-contaminated agricultural site using aromatic and medicinal plants to produce essential oils: analysis of the metal(loid) fate in the value chain.
Alexandre Perlein, Isabelle Zdanevitch, Rodolphe Gaucher, Brett Robinson, Arnaud Papin, Anissa Lounes-Hadj Sahraoui, Valérie Bert
Alexandre Perlein, Isabelle Zdanevitch, Rodolphe Gaucher, Brett Robinson, Arnaud Papin, et al.. Phytomanagement of a metal(loid)-contaminated agricultural site using aromatic and medicinal plants to produce essential oils: analysis of the metal(loid) fate in the value chain.. Environmental Science and Pollution Research, Springer Verlag, 2021, 10.1007/s11356-021-15045-4. ineris-03310658
Alexandre Perlein, Isabelle Zdanevitch, Rodolphe Gaucher, Brett Robinson, Arnaud Papin, et al.. 利用芳香植物和药用植物生产精油对金属（类金属）污染农业场地进行植物管理：价值链中金属（类金属）归趋分析. Environmental Science and Pollution Research, Springer Verlag, 2021, 10.1007/s11356-021-15045-4. ineris-03310658
HAL Id: ineris-03310658
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Submitted on 19 Oct 2021
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Title: Phytomanagement of a metal(loid)-contaminated agricultural site using aromatic and medicinal plants to produce essential oils: analysis of the metal(loid) fate in the value chain.
题目：利用芳香植物和药用植物生产精油对金属（类金属）污染农业场地进行植物管理：价值链中金属（类金属）归趋分析
Alexandre PERLEINa,b, Isabelle ZDANEVITCHa, Rodolphe GAUCHERa, Brett ROBINSONc, Arnaud PAPINd,
Alexandre PERLEINa,b, Isabelle ZDANEVITCHa, Rodolphe GAUCHERa, Brett ROBINSONc, Arnaud PAPINd,
Anissa LOUNES-HADJ SAHRAOUIe, Valérie BERTa*Anissa LOUNES-HADJ SAHRAOUIe, Valérie BERTa*
a Unité Technologies Propres et Economie Circulaire, INERIS, Parc Technologique Alata, BP2, 60550 Verneuilen-Halatte, France b Department of Green Chemistry and Technology, Ghent University, Ghent, Belgium c School of Physical and Chemical Sciences, University of Canterbury, Christchurch 8041, New Zealand d Analytical methods and developments for the environment, INERIS, Parc Technologique Alata BP 2, 60550
a Unité Technologies Propres et Economie Circulaire, INERIS, Parc Technologique Alata, BP2, 60550 Verneuilen-Halatte, 法国 b 根特大学绿色化学与技术系，Ghent，比利时 c 坎特伯雷大学物理与化学科学学院，基督城 8041，新西兰 d 环境分析方法与开发部，INERIS， Parc Technologique Alata BP 2, 60550
Verneuil en Halatte, FranceVerneuil en Halatte，法国
e Université du Littoral Côte d’Opale, Unité de Chimie Environnementale et Interactions sur le Vivant (UCEIV, UR 4492), SFR Condorcet FR CNRS 3417, 50 rue Ferdinand Buisson, 62228 Calais cedex, France
e Université du Littoral Côte d’Opale, Unité de Chimie Environnementale et Interactions sur le Vivant (UCEIV, UR 4492), SFR Condorcet FR CNRS 3417, 50 rue Ferdinand Buisson, 62228 Calais cedex，法国
*Corresponding author: valerie.bert@ineris.fr, +33 3 44 55 63 82
1Abstract摘要
Phytomanagement uses plants and soil conditioners to create value on contaminated land while minimising environmental risk. This work was carried out on a metal(loid)-contaminated site and aimed at assessing the suitability of Salvia sclarea L. (sage) and Coriandrum sativum L. (coriander) combined with an Arbuscular Mycorrhizal Fungus (AMF) inoculant to immobilise metal(loid)s and produce essential oils (EO). The effect of the inoculant on the transfer of metal(loid)s (ML, i.e. Cd, Cu, Pb, Zn, As, Ni, and Sb) to plants and the ML soil mobility were investigated. The ML concentrations in EO from both plant species and the valorisation options for the distillation residues (soil conditioner, animal fodder, and anaerobic digestion) were studied. Sage was a suitable candidate for this value chain because it presents an excluder phenotype and the residues of oil extraction could be used as a soil conditioner. The metal concentrations in the sage EO were similar to those obtained from plants cultivated on an uncontaminated soil. These results indicate the suitability of sage harvested on the contaminated soil according to the ML fate in the whole value chain. Like the EO of sage, ML concentrations in the coriander
植物管理利用植物和土壤调理剂在污染土地上创造价值，同时将环境风险降至最低。本研究在一处金属（类金属）污染场地开展，旨在评估Salvia sclarea L.（鼠尾草）和Coriandrum sativum L.（芫荽）结合丛枝菌根真菌（AMF）接种剂固定金属（类金属）并生产精油（EO）的适宜性。研究了接种剂对金属（类金属）（ML，即Cd、Cu、Pb、Zn、As、Ni和Sb）向植物转移的影响以及ML在土壤中的迁移性。同时研究了两种植物精油中的ML浓度以及蒸馏残渣的资源化利用途径（土壤调理剂、动物饲料和厌氧消化）。鼠尾草表现出排斥型表型，且精油提取残渣可用作土壤调理剂，因此是该价值链的适宜候选植物。鼠尾草精油中的金属浓度与未污染土壤上栽培植物所得精油相当。这些结果表明，从整个价值链中ML归趋的角度看，在污染土壤上收获的鼠尾草是适宜的。与鼠尾草精油类似，芫荽
EO did not differ from those in the commercial EO that were obtained from plants grown on uncontaminated soil. However, the use of distillation residues of coriander was limited by their relatively elevated Cd concentrations. The use of a mycorrhizal inoculum did not decrease the Cd mobility in soil for the coriander.
精油中的ML浓度与由未污染土壤上生长的植物所得商品精油无差异。然而，芫荽蒸馏残渣中Cd浓度相对较高，限制了其利用。对于芫荽，施用菌根接种剂并未降低土壤中Cd的迁移性。
Keywords Salvia sclarea, Coriandrum sativum, Cadmium, Arbuscular mycorrhizal fungi, Aided phytostabilisation, Biomass valorisation, Distillation residues
关键词 Salvia sclarea，Coriandrum sativum，镉，丛枝菌根真菌，辅助植物稳定，生物质资源化，蒸馏残渣
Ethics approval and consent to participate Not applicable.
伦理批准与参与同意 不适用。
Consent for publication Not applicable.
发表同意 不适用。
Availability of data and materials Not applicable.
数据与材料可获得性 不适用。
Competing interests The authors declare that they have no competing interests.
利益冲突 作者声明不存在利益冲突。
Funding information This work was supported by national public funds through PhyTEO (ADEME – Agence De la Transition Ecologique, convention n° 1772C0020, 2017-2020).
资助信息 本研究由PhyTEO项目国家公共资金资助（ADEME – Agence De la Transition Ecologique，协议号1772C0020，2017-2020）。
Authors’ contributions This collaboration work was carried out among all the authors. AP designed outlines and wrote the draft, performed the statistical analyses and prepared the figures and the tables of the manuscript. VB proposed original idea, wrote some part of the manuscript and reviewed the scientific contents of the manuscript. IZ wrote some part of the manuscript and reviewed the manuscript. RG reviewed the manuscript. AP was responsible for the validation of the data quality obtained by ICP and wrote some part of the manuscript. BR reviewed the manuscript and improve the text quality. AL was responsible for the funding acquisition (PhyTEO), coordinated the project and reviewed the manuscript.
作者贡献 本研究由全体作者合作完成。AP设计提纲并撰写初稿，进行统计分析并制作稿件中的图表。VB提出最初构想，撰写部分稿件并审阅稿件的科学内容。IZ撰写部分稿件并审阅稿件。RG审阅稿件。AP负责ICP所得数据的质量验证并撰写部分稿件。BR审阅稿件并提高了文字质量。AL负责获取资助（PhyTEO），协调项目并审阅稿件。
Acknowledgments The authors thank Fabrice Richez, Yohann Baillon, Samuel Teillaud, Robin Raveau and Joël Fontaine for their technical contribution during the sampling campaigns and/or chemical analysis and fruitful discussions. Commercial EO of coriander and sage were kindly provided by Ferrand PHE. Ferrand PHE is also thanked for the production and the providing of non-commercial EOs used in this work. We thank the anonymous reviewers for their comments, which greatly helped us to improve the manuscript.
致谢 作者感谢Fabrice Richez、Yohann Baillon、Samuel Teillaud、Robin Raveau和Joël Fontaine在采样和/或化学分析中提供的技术支持以及富有成效的讨论。芫荽和鼠尾草商品精油由Ferrand PHE惠赠。同时感谢Ferrand PHE生产并提供本研究所用的非商品精油。感谢匿名审稿人的意见，这些意见对改进本文帮助很大。
21) Introduction1）引言
Soil contamination is a worldwide concern. In Europe, industrial and commercial activities have contaminated 342,000 sites to the extent that they pose a potential risk for human health and the environment (van Liedekerke et al. 2014). Among these sites, one third were contaminated by metal(loid)s (ML), which cause persistent degradation of soil quality. To manage the risk due to ML contamination, civil engineering remediation techniques such as excavation are used to remove the contaminated soil. Although these techniques are efficient in removing the contamination from the site, they are prohibitively expensive over large areas. They also produce waste that needs to be managed and they often destroy local ecosystems (Ali et al. 2013; Cundy et al. 2016; Hooper et al. 2015). In contrast, plant-based technologies (phytotechnologies) may represent a low-cost and long-term approach to improving contaminated soil. Ideally suited to large areas of contamination, the aim of these phytotechnologies is to manage environmental risk and exposure by decreasing pollutant transfers in the environment (phytostabilisation) or by the partial removal of pollutants in the soil (phytoextraction or bioavailable contaminant stripping) (Kidd et al. 2015; Robinson et al. 2015). Phytomanagement aims to generate financial returns from contaminated land via the introduction of biomass into a sustainable value chain (Robinson et al. 2009; Evangelou and Deram, 2014; Burges et al. 2018). Since 2010, much research has focused on the conversion of biomass into bioenergy and raw materials. Biomass could be valorised through combustion, pyrolysis, gasification, anaerobic digestion (Delplanque et al. 2013; Chalot et al. 2012; Bert et al. 2017; Meers et al. 2010) or prepared to obtain high value products such as platform-chemicals, fibers and ecomaterials (e.g. Deyris et al. 2018; Ziegler-Devin et al. 2019; Jeannin et al. 2020; Dias and Waldmann 2020).
土壤污染是一个全球性问题。在欧洲，工业和商业活动已污染了342,000处场地，其污染程度对人体健康和环境构成潜在风险（van Liedekerke et al. 2014）。其中三分之一的场地受到金属（类金属）（ML）污染，导致土壤质量持续退化。为管控ML污染带来的风险，人们采用开挖等土木工程修复技术移除污染土壤。尽管这些技术能有效清除场地污染，但在大面积区域应用成本过高。此外，这些技术会产生需要处置的废弃物，并常常破坏当地生态系统（Ali et al. 2013; Cundy et al. 2016; Hooper et al. 2015）。相比之下，基于植物的技术（植物技术）可能是一种低成本、长期改良污染土壤的途径。植物技术尤其适用于大面积污染区域，其目的是通过减少污染物在环境中的迁移（植物稳定）或部分去除土壤中的污染物（植物提取或生物有效态污染物剥离）来管控环境风险和暴露（Kidd et al. 2015; Robinson et al. 2015）。植物管理旨在通过将生物质引入可持续价值链，从污染土地上获得经济收益（Robinson et al. 2009; Evangelou and Deram, 2014; Burges et al. 2018）。自2010年以来，大量研究聚焦于将生物质转化为生物能源和原材料。生物质可通过燃烧、热解、气化、厌氧消化实现资源化（Delplanque et al. 2013; Chalot et al. 2012; Bert et al. 2017; Meers et al. 2010），或加工制备平台化学品、纤维和生态材料等高价值产品（如Deyris et al. 2018; Ziegler-Devin et al. 2019; Jeannin et al. 2020; Dias and Waldmann 2020）。
Essential Oils (EOs) distilled from aromatic and medicinal plants (AMP) are used in foods, personal care products, cosmetics, aromatherapy sectors, and in products such as detergents, biological pesticides, soaps, concentrates, extracts, additives, etc. (Barbieri and Borsotto, 2018; Eikani et al. 2007; Jisha et al. 2017; El-Alam et al. 2020; Raveau et al. 2020). High-value EOs are traded worldwide and the market is expected to grow (Barbieri and Borsotto, 2018).
由芳香植物和药用植物（AMP）蒸馏得到的精油（EO）被用于食品、个人护理产品、化妆品和芳香疗法等领域，以及洗涤剂、生物农药、肥皂、浓缩物、提取物、添加剂等产品中（Barbieri and Borsotto, 2018; Eikani et al. 2007; Jisha et al. 2017; El-Alam et al. 2020; Raveau et al. 2020）。高价值精油在全球范围内交易，其市场预计将持续增长（Barbieri and Borsotto, 2018）。
Few studies have reported the use of AMP grown on polluted soils to produce EOs (Zheljazkov et al. 2008; Angelova et al. 2016; Seyedalikhani et al. 2020). The soil-plant transfer of Cd, Pb and Cu in polluted plots was low and the authors concluded that AMP such as Ocimum basilicum L. (basil), Mentha x piperita L. (peppermint), coriander and Salvia officinalis L. did not accumulate Cd, Pb and Cu in aerial parts and that the hydrodistillation of basil, peppermint and sage led to a limited transfer of Cu, Cd and Pb in EO, which suggested that these EO could be marketed (Zheljazkov et al,. 2008; Seyedalikhani et al. 2019).
利用生长在污染土壤上的AMP生产精油的研究报道较少（Zheljazkov et al. 2008; Angelova et al. 2016; Seyedalikhani et al. 2020）。在污染样地中，Cd、Pb和Cu的土壤-植物转移较低。作者得出结论：Ocimum basilicum L.（罗勒）、Mentha x piperita L.（薄荷）、芫荽和Salvia officinalis L.等AMP的地上部不积累Cd、Pb和Cu，且罗勒、薄荷和鼠尾草经水蒸馏后Cu、Cd和Pb向精油中的转移有限，表明这些精油可以上市销售（Zheljazkov et al,. 2008; Seyedalikhani et al. 2019）。
Two review papers proposed using AMP in a phytomanagement context based on their composition leading to EO and their tolerance to ML in soil (Jisha et al. 2017; Ait Elallem et al. 2020). They suggested integrating distillation residues for bioenergy production. To our knowledge, amongst AMP, only a few were studied in this context. For instance, Angelova et al. (2016) performed in situ experiments with sage according to different levels of ML pollution and found no signs of toxicity up to 3000 and 60 mg kg-1 foliar Zn and Cd concentrations, respectively. Mycorrhizal inoculation improves the ML stress tolerance of plants and decreases the ML transfer in aerial plant parts and participates in the reduction of the ML mobility in soil by different mechanisms (Leung et al. 2013; Phanthavongsa et al. 2017). Mycorrhizal inoculation of coriander in a pot experiment with a Pb salt showed a reduction of toxicity signs and foliar Pb concentration at 1500 mg Pb kg-1 in the soil (Fatemi et al. 2020). In contrast, the mycorrhizal inoculation of sage increased the shoot Cd and Pb concentrations and the authors reported the limited tolerance of the arbuscular mycorrhizal fungi (AMF) to Cd and Pb (Nowak, 2007).
两篇综述论文基于AMP可产生精油的成分组成及其对土壤中ML的耐受性，提出在植物管理背景下利用AMP（Jisha et al. 2017; Ait Elallem et al. 2020），并建议将蒸馏残渣用于生物能源生产。据我们所知，在此背景下开展研究的AMP为数不多。例如，Angelova et al.（2016）在不同ML污染水平下对鼠尾草开展了原位试验，发现叶片Zn和Cd浓度分别高达3000和60 mg kg-1时仍未出现毒害症状。菌根接种可提高植物对ML胁迫的耐受性，减少ML向植物地上部的转移，并通过不同机制降低ML在土壤中的迁移性（Leung et al. 2013; Phanthavongsa et al. 2017）。在添加Pb盐的盆栽试验中，对芫荽进行菌根接种后，在土壤Pb为1500 mg Pb kg-1时毒害症状和叶片Pb浓度均有所降低（Fatemi et al. 2020）。相反，对鼠尾草进行菌根接种却提高了地上部Cd和Pb浓度，作者指出丛枝菌根真菌（AMF）对Cd和Pb的耐受性有限（Nowak, 2007）。
Amongst the AMP, many produce a high aerial biomass of which only a part is used for EO production. The residues of distillation as well as the remaining plant parts could be integrated to the value chain if they are beneficially reused. On uncontaminated soil, biomass residues are commonly used as animal fodder, composts and for energy production (Jisha et al. 2017). To our knowledge, on polluted soils, the feasibility of such an overall value chain, i.e. including the production of EO and the valorisation of the distillation residues, based on the ML fate has not been studied.
许多AMP地上生物量较高，但只有一部分用于精油生产。蒸馏残渣及其余植物部分若能得到有益再利用，便可纳入价值链。在未污染土壤上，生物质残渣通常用作动物饲料、堆肥和能源生产原料（Jisha et al. 2017）。据我们所知，在污染土壤上，基于ML归趋的这种完整价值链（即包括精油生产和蒸馏残渣资源化）的可行性尚未得到研究。
This work aimed to determine the feasibility of phytomanaging a metal-contaminated site using sage and coriander to produce EOs. Both plant species are considered as relevant candidates in this value chain (Zheljazkov et al. 2008; Angelova et al. 2016; Cui et al. 2015; Mandal and Mandal, 2015; Matasyoh et al. 2009). A second aim was to assess the effect of an AMF inoculant on (i) the ML transfer to aerial plant parts and (ii) the solubility of the ML in soil. Other specific objectives were to investigate the EO quality obtained from both plant species regarding ML and the valorisation options for the distillation residues (soil improver, animal fodder and anaerobic digestion). This study was carried out in the frame of the PhytEO project in which other plant parameters such as plant growth, development and biomass production, as well as the techno-economic feasibility of the phytomanagement strategy were studied (Raveau, 2020).
本研究旨在确定利用鼠尾草和芫荽生产精油对金属污染场地进行植物管理的可行性。这两种植物均被认为是该价值链的合适候选物种（Zheljazkov et al. 2008; Angelova et al. 2016; Cui et al. 2015; Mandal and Mandal, 2015; Matasyoh et al. 2009）。第二个目的是评估AMF接种剂对（i）ML向植物地上部转移和（ii）ML在土壤中溶解性的影响。其他具体目标包括：从ML角度考察两种植物所得精油的品质，以及蒸馏残渣的资源化途径（土壤改良剂、动物饲料和厌氧消化）。本研究在PhytEO项目框架下开展，该项目还研究了植物生长、发育和生物量产量等其他植物参数，以及该植物管理策略的技术经济可行性（Raveau, 2020）。
32) Materials and Methods2）材料与方法
2.1) Site description and experimental design2.1）场地概况与试验设计
Experiments were performed in the north of France on an agricultural area (Evin-Malmaison; 1.3 ha; 50°25’55.5’’ N, 3°02’25.5’’ E, Fig.1) contaminated by past industrial emissions of a Pb/Zn smelter that led to ML contamination of the topsoil with, notably, Cd, Pb, Cu and Zn (Sterckeman et al. 2002). Two AMP species, the biennial clary sage (Salvia sclarea L.) and the annual coriander (Coriandrum sativum L.), were cultivated on the contaminated site. Half of the site was devoted to sage whereas the other part was devoted to coriander. Each half was further divided according to inoculation carried out with a commercial AMF inoculant (Rhizophagus irregularis; 12000 viable spores per gram of product; AGTIV® Specialty Crops; PremierTech). After soil preparation realized with conventional tillage (0.1 m -depth ploughing), the powdery inoculant was introduced during sowing of certified sage and coriander seeds (https://www.iteipmai.fr/) at a rate of 125 g ha-1. This division led to inoculated (I, 2000m²) and non-inoculated (NI, 4000m²) plots. Sowing occurred in April 2017 for both species (300 000 seeds per hectare, 3-4 cm depth). The annual coriander was resown in April 2018 for a second crop.
试验在法国北部的一处农业区（Evin-Malmaison；1.3 ha；50°25’55.5’’ N，3°02’25.5’’ E，图1）进行，该区域受到一座Pb/Zn冶炼厂历史工业排放的污染，表层土壤受到ML污染，尤其是Cd、Pb、Cu和Zn（Sterckeman et al. 2002）。在该污染场地上种植了两种AMP：二年生的香紫苏（Salvia sclarea L.）和一年生的芫荽（Coriandrum sativum L.）。场地的一半种植鼠尾草，另一半种植芫荽。每一半再根据是否施用商品AMF接种剂（Rhizophagus irregularis; 每克产品含12000个活孢子；AGTIV® Specialty Crops；PremierTech）进一步划分。采用常规耕作（0.1 m深翻耕）整地后，在播种经认证的鼠尾草和芫荽种子（https://www.iteipmai.fr/）时施入粉状接种剂，施用量为125 g ha-1。由此划分为接种（I，2000m²）和未接种（NI，4000m²）样地。两种植物均于2017年4月播种（每公顷300 000粒种子，播深3-4 cm）。一年生芫荽于2018年4月重新播种，进行第二茬种植。
A similar experimental design was set up on an uncontaminated soil (1.4 ha - 50°49'55.7" N, 1°55'46.9" E). This site was used as a reference for comparing the metal-concentrations in the plant parts (see section 2.3). Plants were processed the same as those from the contaminated site. We report the concentrations obtained in the EO and the distillation residues from this site for comparison with those obtained from the contaminated site (see sections 2.4 and 2.5).
在一处未污染土壤上（1.4 ha - 50°49'55.7" N，1°55'46.9" E）设置了类似的试验设计。该场地作为比较植物各部位金属浓度的对照（见2.3节）。植物的处理方式与污染场地相同。本文报告了该场地精油和蒸馏残渣中的浓度，用于与污染场地所得结果进行比较（见2.4节和2.5节）。
2.2) Soil sampling and agro-physicochemical analyses2.2）土壤采样与农业理化分析
In April 2017, soil sample composites were made from 19 top-soil samples randomly collected with a hand auger on the whole site (Table 1). The soil physicochemical properties measured on these samples are presented in Table 2 and reported as initial state (IS) data in the further text.
2017年4月，用手动土钻在整个场地随机采集19个表层土壤样品，制成混合土样（表1）。这些样品的土壤理化性质测定结果见表2，下文称为初始状态（IS）数据。
Once the plots were designed, soil samples were taken before sowing and harvesting. In July and September 2017, 12 samples were taken from the inoculated plots (I) and 18 samples were taken from the non-inoculated ones (NI). After geo-localisation, all top-soil samples (0-20 cm depth) were dried at 40 °C in a forced air oven to a constant weight, ground with a grinder (agate mortar, Retsch BB51) and sieved to < 2 mm (Retsch AS 200 digit). Soil pH was measured following the NF ISO 10390 (2005) standard. Five g of 2 mm sieved soils were mixed with 25 mL of distilled water and shaken for 2 hours. Water pH was measured after a 1 h rest. Total concentrations of ML were analysed in the soil samples collected in July and September 2017 and digested (Mars Xpress CEM) following the NF-EN-13656. Briefly, 0.5 g soil were digested at 180 °C for 20 min with 2 mL of nitric acid (HNO3 67%), 6 mL of hydrochloric acid (HCl 36%) and 2 mL of hydrofluoric acid (HF 48%) and filtered using 0.45 µm Whatman® filter paper. To estimate the easily soluble fraction (water-soluble compounds, exchangeable ions, and readily soluble metal complexes) of ML in these soils, a selective extraction was performed with 1 M ammonium nitrate (NH4NO3) solution following the NF ISO 19730:2008(E).
样地划定后，在播种前和收获前采集土壤样品。2017年7月和9月，在接种样地（I）采集12个样品，在未接种样地（NI）采集18个样品。经地理定位后，所有表层土壤样品（0-20 cm深度）在40 °C强制通风烘箱中烘干至恒重，用研磨机（玛瑙研钵，Retsch BB51）研磨，并过< 2 mm筛（Retsch AS 200 digit）。土壤pH按NF ISO 10390（2005）标准测定：取5 g过2 mm筛的土壤与25 mL蒸馏水混合，振荡2小时，静置1 h后测定水浸pH。对2017年7月和9月采集的土壤样品按NF-EN-13656进行消解（Mars Xpress CEM），分析ML全量。简言之，取0.5 g土壤，加入2 mL硝酸（HNO3 67%）、6 mL盐酸（HCl 36%）和2 mL氢氟酸（HF 48%），于180 °C消解20 min，并用0.45 µm Whatman®滤纸过滤。为估算这些土壤中ML的易溶部分（水溶性化合物、可交换离子和易溶金属络合物），按NF ISO 19730:2008(E)采用1 M硝酸铵（NH4NO3）溶液进行选择性提取。
After 2 h shaking on an agitation table, the soil-extractant mixtures were passed through a Millipore filter (Ø 0.45µm) and leachates acidified to a pH of 2 and stored for analysis. The ML concentrations in the eluates and solutions were analysed either by ICP-OES (Agilent 700, Agilent 5100) or ICP-MS (Agilent 7500) depending on sample concentrations. Recoveries are presented in supplemental table 1. As no upper critical threshold ML concentrations exist in France to define a contaminated soil (as it is a site-specific risk assessment accounting for soil series and land use), all total soil ML (Table 2) were compared to background ML concentrations in French soils (Baize, 2000). The total topsoil Cd, Cu, Pb and Zn were all above the highest values (0.45, 20, 50 and 100 µg g-1 dry soil (DS), respectively), thereby indicating soil ML contamination. Due to their inordinately high concentrations and their mobility (Denaix et al. 2001), the results section of this paper focuses on Cd, Cu, Pb and Zn.
在振荡台上振荡2 h后，土壤-提取剂混合物经Millipore滤膜（Ø 0.45µm）过滤，滤液酸化至pH 2后保存待测。根据样品浓度，采用ICP-OES（Agilent 700，Agilent 5100）或ICP-MS（Agilent 7500）测定洗脱液和溶液中的ML浓度。回收率见补充表1。由于法国没有界定污染土壤的ML浓度上限临界阈值（因其属于考虑土壤系列和土地利用的场地特异性风险评估），所有土壤ML全量（表2）均与法国土壤ML背景浓度（Baize, 2000）进行比较。表层土壤Cd、Cu、Pb和Zn全量均高于其最高背景值（分别为0.45、20、50和100 µg g-1干土（DS）），表明土壤受到ML污染。鉴于Cd、Cu、Pb和Zn浓度异常高且迁移性强（Denaix et al. 2001），本文结果部分重点讨论这四种元素。
2.3) Plant sampling and metal(loid) analyses2.3）植物采样与金属（类金属）分析
The plant sampling strategy occurred in accordance with the soil sampling strategy (Table 1), i.e. 12 and 18 plant samples were collected for the I and the NI plots, respectively. For both, the harvest was different according to the species and the plant stage (Table 1). Samples of sage inflorescence were either washed or unwashed to determine the effect of washing on inflorescence ML concentration (Table 1). Plant washing could be important if soil dust containing ML are deposited on aerial plant parts (Hibben et al. 1984). In case of washing, plant materials were carefully washed with tap and deionised water before drying at 40 °C until constant weight. ML were analyzed in all plant samples (0.5 g) after digestion at 180 °C for 20 min in 10 mL of nitric acid (HNO3 69% AnalaR Normapur) and 3 mL of ultra-pure water using a microwave digester (Mars Xpress CEM) by ICP-OES (Agilent 5100) or ICP- MS (Agilent 7500). To get ML-reference values, coriander and sage grown on the uncontaminated site (Table 2) were collected at the same stage and date and analysed as described above. Recovery rates for ML are presented in the supplemental table 1.
植物采样策略与土壤采样策略一致（表1），即在I和NI样地分别采集12和18个植物样品。两类样地的收获部位均因物种和植物生育期而异（表1）。鼠尾草花序样品分为清洗和未清洗两类，以确定清洗对花序ML浓度的影响（表1）。若含ML的土壤粉尘沉积在植物地上部，植物清洗可能很重要（Hibben et al. 1984）。需清洗时，植物材料先用自来水和去离子水仔细清洗，再于40 °C烘干至恒重。所有植物样品（0.5 g）用微波消解仪（Mars Xpress CEM）在10 mL硝酸（HNO3 69% AnalaR Normapur）和3 mL超纯水中于180 °C消解20 min后，采用ICP-OES（Agilent 5100）或ICP- MS（Agilent 7500）测定ML。为获得ML参考值，在相同生育期和日期采集未污染场地（表2）上生长的芫荽和鼠尾草，并按上述方法进行分析。ML回收率见补充表1。
2.4) Essential oil sampling and metal(loid) analyses2.4）精油采样与金属（类金属）分析
After harvest, the plant material from the I and NI plots were separately processed using water vapor (80°C) to obtain the essential oils (EO). The distillation process was performed in the equipment of an EO industrial producer (Ferrand P.H.E., https://www.ferrantphe.com/production-de-concentres/ Rodelinghem, Hauts-de-France, France). EO samples (5 mL) obtained from each plant material were sent to INERIS for ML analysis. Each sample was divided in three sub-samples and digested following the plant procedure digestion described in §2.3. ML concentrations in all EOs (0.5 g) were measured by ICP-OES (Agilent 5100) or ICP-MS (Agilent 7500). MLs in EO produced from coriander and sage grown on the uncontaminated site as well as commercial EO of both plant species were measured for comparison. Recovery rates for MLs are presented in the supplemental Table 1.
收获后，I和NI样地的植物材料分别采用水蒸气（80°C）处理以获得精油（EO）。蒸馏过程在一家精油工业生产商的设备中进行（Ferrand P.H.E.，https://www.ferrantphe.com/production-de-concentres/ Rodelinghem，Hauts-de-France，法国）。每种植物材料所得精油样品（5 mL）送至INERIS进行ML分析。每个样品分为三个子样，按§2.3所述的植物消解程序进行消解。所有精油（0.5 g）中的ML浓度采用ICP-OES（Agilent 5100）或ICP-MS（Agilent 7500）测定。同时测定了未污染场地上生长的芫荽和鼠尾草所产精油以及两种植物商品精油中的ML，以作比较。ML回收率见补充表1。
2.5) Sampling of distillation residues and metal(loid) analyses2.5）蒸馏残渣采样与金属（类金属）分析
The distillation process resulted in significant quantities of plant residues. Residue samples were oven-dried (40°C, 48h) and homogenised after grinding to get a representative sample. Three sub-samples were selected to analyse the ML concentrations after digestion and analysis (as described in §2.3). MLs in plant residues produced from coriander and sage grown on the uncontaminated site were measured for comparison.
蒸馏过程产生了大量植物残渣。残渣样品经烘箱干燥（40°C，48h），研磨后均质化以获得代表性样品。选取三个子样进行消解和分析（如§2.3所述），测定ML浓度。同时测定了未污染场地上生长的芫荽和鼠尾草所产植物残渣中的ML，以作比较。
2.6) Biomethane potential of distillation residues2.6）蒸馏残渣的生物甲烷潜力
The measurement of the biomethane potential (BMP) of coriander and sage residues of the distillation performed in 2017 was subcontracted (https://www6.montpellier.inrae.fr/it-e, France). Briefly, the distillation residues as substrates and the microbial inoculum were put in a batch reactor for 23 days at 35 °C. The test was performed in duplicates. BMP was expressed in normal conditions of temperature and pressure (0 °C and 1013 hPa).
2017年芫荽和鼠尾草蒸馏残渣的生物甲烷潜力（BMP）测定委托外部机构完成（https://www6.montpellier.inrae.fr/it-e，法国）。简言之，将作为底物的蒸馏残渣和微生物接种物置于批式反应器中，在35 °C下培养23天。试验设两个重复。BMP以标准温度和压力条件（0 °C和1013 hPa）表示。
2.7) Statistical analysis2.7）统计分析
The statistical analysis of the values and the boxplots were produced using R 3.5.2 (12/20/2018) and R studio. Before performing the variance homogeneity test (Bartlett test or Fligner-Killeen test), the normality of the data (Shappiro test) was checked. Means were compared by ANOVA when the data were parametric and by Kruskal- Wallis test when they were not. A post hoc test (Tukey HSD test or pairwise t test) was applied when a statistical difference at 5 percent risk was found. A Pearson correlation was used to determine correlation between variables.
数值统计分析和箱线图采用R 3.5.2（12/20/2018）和R studio完成。在进行方差齐性检验（Bartlett检验或Fligner-Killeen检验）之前，先检验数据的正态性（Shapiro检验）。数据符合参数检验条件时采用ANOVA比较均值，否则采用Kruskal- Wallis检验。在5%风险水平下存在统计学差异时，进行事后检验（Tukey HSD检验或成对t检验）。采用Pearson相关分析确定变量间的相关性。
2.8 ) Calculations2.8）计算
Bioconcentration factors (BCF) were calculated as follows:
生物富集系数（BCF）按下式计算：
BCF ext = [ML] plant / [ML] extractable soil BCF tot = [ML] plant / [ML] total soil
BCF ext = [ML] plant / [ML] extractable soil BCF tot = [ML] plant / [ML] total soil
ML mobility was calculated as follows:
ML迁移性按下式计算：
% mobility = [ML] extractable soil / [ML] total soil * 100
% mobility = [ML] extractable soil / [ML] total soil * 100
In the calculations, [ML] means ML concentration; [ML] plant means ML concentration in either the shoots or the seeds, depending on the type of plant; [ML] extractable soil means 1M NH4NO3-extractable soil ML; [ML] total soil means total soil ML.
计算中，[ML]表示ML浓度；[ML] plant表示地上部或种子中的ML浓度，视植物类型而定；[ML] extractable soil表示1M NH4NO3可提取态土壤ML；[ML] total soil表示土壤ML全量。
43) Results and Discussion3）结果与讨论
3.1) Mycorrhizal inoculation and plant effects on soil pH3.1）菌根接种和植物对土壤pH的影响
The initial soil pH was alkaline (7.86 ± 0.13; Table 2). As coriander is an annual plant, it was cultivated twice leading to two sowing and two harvests. Fig. 2A presents the soil pH values from each harvest. At the end of the 1srt harvest, the soil pH was similar to the initial sampling, indicating no plant effect. There was no statistical difference (p > 0.05) between the I and NI plots, suggesting no inoculation effect. Before the new sowing, a significant soil pH increase was measured in the case of the I plot indicates an inoculation effect. When symbiosis is effective between the AMF and the root system, a significant reduction in acidity (an increase of one pH unit occurs due to N uptake. Root or hyphae NO3- uptake leads to a release of OH- which increases the pH (Bago et al. 1996; Bago and Azcon-Aguilar 1997; Li and Christie 2001). However, this effect was small (0.5 pH unit) and did not last. Factors other than an inoculation effect, for example climatic conditions and the soil tillage, could explain this result. Bai et al. (2018) showed that the tillage had no long-term impact on pH which is consistent with our hypothesis. In contrast, the significant decrease of the soil pH measured for I and NI plots at the end of the second culture clearly showed a plant effect, as a result of cation uptake and root exudation of organic acids. Microbial respiration may also acidify the soil (Hinsinger et al. 2003).
初始土壤pH呈碱性（7.86 ± 0.13；表2）。芫荽为一年生植物，共种植两次，即两次播种和两次收获。图2A给出了每次收获时的土壤pH值。第1次收获结束时，土壤pH与初始采样相近，表明不存在植物效应。I和NI样地之间无统计学差异（p > 0.05），表明不存在接种效应。在重新播种前，I样地土壤pH显著升高，表明存在接种效应。当AMF与根系之间形成有效共生时，由于氮的吸收，酸度显著降低（pH升高一个单位）。根系或菌丝吸收NO3-会导致OH-释放，从而使pH升高（Bago et al. 1996; Bago and Azcon-Aguilar 1997; Li and Christie 2001）。然而，这一效应较小（0.5个pH单位）且未能持续。气候条件和土壤耕作等接种效应以外的因素可能解释这一结果。Bai et al.（2018）表明耕作对pH无长期影响，这与我们的假设一致。相反，第二茬结束时I和NI样地土壤pH均显著下降，清楚地表明存在植物效应，这是阳离子吸收和根系分泌有机酸的结果。微生物呼吸也可能使土壤酸化（Hinsinger et al. 2003）。
In contrast to coriander, sage is a biennial species. It was sown once and grown for two years. The pH was thus measured in the soil at two different stages of the plant development, i.e. Year 1 (leafy stage) and Year 2 (inflorescence stage) (Fig. 2B). A mycorrhizal inoculation effect leading to a significant increase in pH (0.2 pH units) occurred during the 1st year (p < 0.05) that might result from soil microorganism respiration and root cations/anions uptake modification such as an uptake of NO3- (Hinsinger et al. 2003; Bago et al. 1996). The pH decreased during the second year in both I and NI plots may be attributed to differential root uptake of NH4+ versus NO3-, the release of OH-/H+ and the production of organic acids (Hinsinger et al. 2003).
与芫荽不同，鼠尾草是二年生物种，播种一次后生长两年。因此，在植物发育的两个不同阶段测定了土壤pH，即第1年（叶丛期）和第2年（花序期）（图2B）。第1年出现了菌根接种效应，导致pH显著升高（0.2个pH单位）（p < 0.05），这可能源于土壤微生物呼吸以及根系阳离子/阴离子吸收的改变，如NO3-的吸收（Hinsinger et al. 2003; Bago et al. 1996）。第二年I和NI样地pH均下降，可能归因于根系对NH4+和NO3-的差异吸收、OH-/H+的释放以及有机酸的产生（Hinsinger et al. 2003）。
3.2) Mycorrhizal inoculation and plant effects on NH4NO3 extractable fraction in soil3.2）菌根接种和植物对土壤NH4NO3可提取态的影响
The extractable ML concentrations that are frequently encountered in soil are reported in the ISO 19730:2008 (E). These are: Zn (0.25 µg g-1 dry soil), Cd (0.005 µg g-1 dry soil), Pb (0.02 µg g-1 dry soil) and Cu (0.25 µg g-1 dry soil). The (1M-NH4NO3) extractable metal soil values are used as indicative values in this paper. There are no references for extractable ML in French soils and the extractable ML values of the uncontaminated soil could not be used as a reference for comparison with the contaminated one because it clearly showed different agro-physicochemical properties (Table 2). The mean values of the extractable ML concentrations for both the initial state (IS) of the soil, i.e. before the set-up of the experimental design (Table 2) and the soil once the plots were designed (Fig. 3, Fig. 4) were higher than the indicative values for Zn, Cd, Pb and Cu found in the ISO 19730:2008 (E) indicating that the extractable ML concentrations in the contaminated soil are higher than those frequently measured in soil. The ML mobility, which refers to the extractable ML concentration over the total ML concentration in soil, was ranked in the following order: Cd > Cu=Sb >> Zn=Ni >>> Pb=As=Cr (Supplemental Table 2), i.e. the Cd had the highest mobility whereas the Pb had the lowest.
ISO 19730:2008 (E)中报告了土壤中常见的可提取态ML浓度，分别为：Zn（0.25 µg g-1干土）、Cd（0.005 µg g-1干土）、Pb（0.02 µg g-1干土）和Cu（0.25 µg g-1干土）。本文将这些（1M-NH4NO3）可提取态土壤金属值作为指示值。法国土壤可提取态ML尚无参考值，且未污染土壤的农业理化性质与污染土壤明显不同（表2），因此其可提取态ML值不能作为比较的参考。土壤初始状态（IS，即试验设计建立之前，表2）以及样地划定后土壤（图3，图4）的可提取态ML浓度均值均高于ISO 19730:2008 (E)中Zn、Cd、Pb和Cu的指示值，表明污染土壤中可提取态ML浓度高于土壤中的常见测定值。ML迁移性即土壤中可提取态ML浓度与ML全量之比，其排序如下：Cd > Cu=Sb >> Zn=Ni >>> Pb=As=Cr（补充表2），即Cd迁移性最高，而Pb最低。
An increase in the extractable soil Zn was observed after one harvest in the I and NI plots compared to the IS value (Fig. 3; 2 and 4 times highest, respectively). This increase may be due to a plant effect and not to an inoculation effect (p > 0.05) as shown in Fig. 3A. The cultivation of the coriander for a second year showed neither an inoculation effect nor a plant effect (p > 0.05) on the extractable soil Zn (Fig. 3A).
与IS值相比，收获一次后I和NI样地土壤可提取态Zn均有所增加（图3；分别为2倍和4倍）。如图3A所示，这一增加可能源于植物效应而非接种效应（p > 0.05）。第二年种植芫荽对土壤可提取态Zn既无接种效应也无植物效应（p > 0.05）（图3A）。
The Zn decrease between both cultures (Fig. 3A) could not be explained by the differences in the soil pH nor the extractable soil Zn (p > 0.05), whereas such a correlation would be expected; soil pH being pivotal for Zn solubility (Gartler et al. 2013). Other soil parameters such as the Cation Exchange Capacity (CEC) (Table 2) combined with the climatic conditions could explain this decrease (Kabata-Pendias 2011). Meite et al. (2018) showed that the alternation of drought (April to July 2017 and April to July 2018) and waterlogging (August 2017 to April 2018), which occurred before the first sowing of the coriander and during the coriander monitoring may have led to variations in the leaching of the water-soluble Zn and in extractable Zn concentrations.
两茬之间Zn的下降（图3A）无法用土壤pH或土壤可提取态Zn的差异来解释（p > 0.05），尽管预期存在此类相关性，因为土壤pH对Zn的溶解性起关键作用（Gartler et al. 2013）。阳离子交换量（CEC）（表2）等其他土壤参数与气候条件共同作用可能解释这一下降（Kabata-Pendias 2011）。Meite et al.（2018）表明，芫荽首次播种前及芫荽监测期间发生的干旱（2017年4月至7月和2018年4月至7月）与渍水（2017年8月至2018年4月）交替，可能导致水溶性Zn淋溶和可提取态Zn浓度的变化。
At the end of the 2nd culture before harvest, the extractable soil Zn did not differ from the IS mean (Fig. 3), i.e. before the experimental set up, showing that the addition of the inoculum combined with coriander did not reach the objective of aided phytostabilisation, i.e. a reduction of the initial extractable soil Zn .
在第2茬收获前结束时，土壤可提取态Zn与IS均值（即试验建立之前）无差异（图3），表明接种剂与芫荽相结合未达到辅助植物稳定的目标，即降低初始土壤可提取态Zn。
Over both cultures (Fig. 3B), the mean extractable soil Cd concentrations were similar to the IS value (Fig. 3). There was no effect of the inoculation or the crop during either culture (p > 0.05). As a result, the extractable soil Cd was unchanged by the coriander and the inoculum.
在两茬期间（图3B），土壤可提取态Cd浓度均值与IS值相近（图3）。两茬期间均无接种效应或作物效应（p > 0.05）。因此，芫荽和接种剂未改变土壤可提取态Cd。
No inoculation and no plant effects were shown on the extractable soil Pb and Cu (Fig. 3C and D). As with Cd, the combination of the inoculum and coriander did not decrease these soil fractions. Moreover, the extractable soil fraction did not differ from the IS mean for both metals.
土壤可提取态Pb和Cu均未表现出接种效应和植物效应（图3C和D）。与Cd相同，接种剂与芫荽相结合未能降低这些土壤组分。此外，两种金属的土壤可提取态均与IS均值无差异。
All the extractable soil fractions (Zn, Cd, Pb, and Cu) remained higher than the indicative values (from 22 to 28 times in the case of Cd) (Fig. 3).
所有土壤可提取态组分（Zn、Cd、Pb和Cu）均仍高于指示值（Cd为指示值的22至28倍）（图3）。
The ML mobility calculated as the ratio of the extractable soil ML over the total soil ML agreed with the results on extractable soil ML (Supplemental Table 2). The ML mobility was similar to the initial hypothesis, namely Cd > Cu >> Zn >>> Pb. These results confirmed that the combination of coriander and R. irregularis was not efficient in decreasing ML mobility in our soil conditions. The absence of an inoculum effect could be explained by either an inefficient symbiosis establishment between the root system of coriander and the AMF species or because the hyphae of the AMF did not interact with the extractable ML fraction by binding or sequestering ML or excreting glomalin that creates ML insoluble complexes (Leung et al. 2013). This result is consistent with another study conducted in controlled conditions where the decrease of extractable ML in an aged contaminated soil by the AMF addition could not be shown due to the interaction between glomalin produced by endogenous AMF and soluble ML fraction (Qiao et al. 2015). Concerning, the first hypothesis, it was shown that in Europe, soil moisture was the main factor that influences the successful establishment of R. irregularis, a wet soil presenting a favourable ecological niche for the fungus (Kivlin et al. 2017). Thus, the low amount of precipitation observed during the experiment could have constrained the AMF life cycle and its establishment.
以土壤可提取态ML与土壤ML全量之比计算的ML迁移性与土壤可提取态ML的结果一致（补充表2）。ML迁移性与初始假设相近，即Cd > Cu >> Zn >>> Pb。这些结果证实，在本研究土壤条件下，芫荽与R. irregularis相结合不能有效降低ML迁移性。接种效应缺失的原因可能是芫荽根系与该AMF物种之间未能有效建立共生关系，或是AMF菌丝未通过结合或固存ML、或分泌可与ML形成不溶性络合物的球囊霉素而与可提取态ML组分发生作用（Leung et al. 2013）。这一结果与另一项在控制条件下开展的研究一致：在老化污染土壤中，由于土著AMF产生的球囊霉素与可溶性ML组分之间的相互作用，未能证明添加AMF可降低可提取态ML（Qiao et al. 2015）。关于第一种假设，已有研究表明，在欧洲，土壤湿度是影响R. irregularis能否成功定殖的主要因素，湿润土壤为该真菌提供了有利的生态位（Kivlin et al. 2017）。因此，试验期间降水量偏少可能制约了AMF的生活史及其定殖。
Compared to the IS Zn value (Fig. 4A), the plants increased the extractable soil Zn and this increase was offset by mycorrhizal inoculation (p < 0.05). Nevertheless, the reduction caused by mycorrhizal inoculation only occurred during the 1st year. On the NI plot, there was a significant decrease (p < 0.05) between the extractable soil Zn concentrations measured in the years 1 and 2 indicating a plant effect. This effect was occurred at the inflorescence stage of the sage, which could be related to its biennial status and the fact that Zn is an essential element for plant growth. Overall, this reduction led to a value similar to the IS, showing a negligible impact of the sage and inoculum combination on the solubility of Zn
与IS的Zn值相比（图4A），植物提高了土壤可提取态Zn，而菌根接种抵消了这一增加（p < 0.05）。然而，菌根接种引起的降低仅出现在第1年。在NI样地，第1年和第2年测得的土壤可提取态Zn浓度之间存在显著下降（p < 0.05），表明存在植物效应。这一效应出现在鼠尾草花序期，可能与其二年生特性以及Zn是植物生长必需元素有关。总体而言，这一下降使其值与IS相近，表明鼠尾草与接种剂相结合对Zn溶解性的影响可忽略不计
The extractable soil Cd during the 1st year did not differ from the IS value (Fig. 4B). No plant and no inoculation effects were found. In contrast, during the 2nd year, inoculation reduced extractable Cd (p < 0.05), while there was an increase due to plant growth (p < 0.05). The sage at the inflorescence stage (Year 2 NI) increased the extractable soil Cd. These results are in accordance with the high soil Cd mobility (Supplemental Table 2), which may be explained by the negative correlation between soil pH and the extractable soil Cd (p < 0.001, r = - 0.48) indicating that sage acidifies the soil and consequently increases Cd mobility.
第1年土壤可提取态Cd与IS值无差异（图4B），未发现植物效应和接种效应。相反，在第2年，接种降低了可提取态Cd（p < 0.05），而植物生长则使其增加（p < 0.05）。花序期鼠尾草（第2年NI）提高了土壤可提取态Cd。这些结果与土壤Cd的高迁移性相符（补充表2），这可以用土壤pH与土壤可提取态Cd之间的负相关（p < 0.001，r = - 0.48）来解释，表明鼠尾草使土壤酸化，从而提高了Cd的迁移性。
The inoculation increased (P < 0.05) the extractable Pb during the 2nd year (Fig. 4C). In contrast to Cd, the Pb mobility was extremely low (Supplemental Table 2). Sage did not change soil Pb mobility, in contrast to the inoculum which increased it (Supplemental Table 2). Consequently, the extractable soil Pb on the I plot was higher than the IS value at the end of the second culture.
第2年，接种提高了可提取态Pb（P < 0.05）（图4C）。与Cd相反，Pb的迁移性极低（补充表2）。鼠尾草未改变土壤Pb迁移性，而接种剂则提高了其迁移性（补充表2）。因此，第二茬结束时I样地土壤可提取态Pb高于IS值。
There was a significant decrease in extractable Cu (p < 0.05) during the 2nd year, which may be due to the biennial status of the plant and the fact that, as with Zn, Cu is an essential element (Fig. 4D). Nevertheless, the extractable soil Cu remained similar to the IS value for this element. The aided phytostabilisation strategy applied to the site did not change soil Cu mobility, which is around 1% of the total Cu (Supplemental Table 2).
第2年可提取态Cu显著下降（p < 0.05），这可能与该植物的二年生特性以及Cu与Zn一样同为必需元素有关（图4D）。然而，土壤可提取态Cu仍与该元素的IS值相近。该场地所采用的辅助植物稳定策略未改变土壤Cu的迁移性，其约为Cu全量的1%（补充表2）。
Most of the inoculation treatments did not affect the extractable soil Zn, Cd, Pb and Cu and when there was an effect, it did either not persist for the whole plant harvest cycle or it was insufficient to cause a decrease in
大多数接种处理未影响土壤可提取态Zn、Cd、Pb和Cu；即使存在影响，也未能在整个植物收获周期内持续，或不足以降低
ML mobility. Similarly, Phanthavongsa et al. (2017) reported inconsistent inoculation effects on extractable soil ML with either increased extractability due to the exudation of organic acids by the roots or a decrease due to the production of glomalin by the AMF.
ML迁移性。同样，Phanthavongsa et al.（2017）报道了接种对土壤可提取态ML的影响不一致：根系分泌有机酸会提高其可提取性，而AMF产生球囊霉素则会使其降低。
With the exception of Cu, the extractable soil fractions of Zn, Pb and Cd remained higher than the indicative values (from 24 to 30 times in the case of Cd) (Fig. 4).
除Cu外，土壤中Zn、Pb和Cd的可提取态组分均仍高于指示值（Cd为指示值的24至30倍）（图4）。
3.3 ) Metal(loid) concentrations in plant and mycorrhizal inoculation effect3.3）植物中金属（类金属）浓度及菌根接种效应
The Zn, Cd, Pb and, Cu concentrations were measured in the shoots and seeds of coriander grown on the contaminated soil to assess the pattern of the plant regarding these ML (Fig. 5). They were all above reported values on uncontaminated soils (Ozkutlu 2008; Singh and Garg 2006; Zheljazkov et al. 2008). In addition, they exceeded those measured in the shoots of coriander collected on the uncontaminated site (Zn: 35; Cd: 0.76; Cu: 8.5; Pb: 0.2 µg g-1 DW) (Fig. 5). The mean concentrations measured in the shoots were, respectively, 25, 9, 2.5 and 1.4 times above the Pb, Cd, Zn, and Cu concentrations measured in uncontaminated conditions.
测定了污染土壤上生长的芫荽地上部和种子中的Zn、Cd、Pb和Cu浓度，以评估该植物对这些ML的响应模式（图5）。这些浓度均高于未污染土壤上的报道值（Ozkutlu 2008; Singh and Garg 2006; Zheljazkov et al. 2008），并且超过了在未污染场地采集的芫荽地上部中的测定值（Zn：35；Cd：0.76；Cu：8.5；Pb：0.2 µg g-1 DW）（图5）。地上部Pb、Cd、Zn和Cu浓度均值分别为未污染条件下测定值的25、9、2.5和1.4倍。
In agreement with Zheljazkov et al. (2008), the BCF tot calculated with the mean shoot Cd concentrations (BCF tot > 1; Table 3), indicated an accumulator response for this element. The results of the BCF tot demonstrated an excluder response for the other ML (Zn, Pb, Cu, As, Cr, Ni and Sb) (Table 3). The Cd accumulator pattern for coriander shoots is in accordance with the fact that the mean Cd shoot concentration measured on uncontaminated soil was already two times above the ordinary Cd foliar lettuce concentration (Kabata-Pendias, 2011).
与Zheljazkov et al.（2008）一致，以地上部Cd浓度均值计算的BCF tot（BCF tot > 1；表3）表明芫荽对该元素呈积累型响应。BCF tot结果表明其对其他ML（Zn、Pb、Cu、As、Cr、Ni和Sb）呈排斥型响应（表3）。芫荽地上部的Cd积累模式与以下事实相符：在未污染土壤上测得的地上部Cd浓度均值已是莴苣叶片常见Cd浓度的两倍（Kabata-Pendias, 2011）。
The mean concentrations of Cd and Pb in the seeds of coriander were higher than those in the shoots of the plant (Fig. 5B, C and D). These results are consistent with previous reports (Smolders 2001; Walker and Waters 2011) which stated that these ML are not translocated into the seeds.
芫荽种子中Cd和Pb浓度均值高于植物地上部（图5B、C和D）。这些结果与此前的报道一致（Smolders 2001; Walker and Waters 2011），即这些ML不会转运至种子中。
Unlike our study, Redon et al. (2008) reported consistent decreases in the shoot Cd concentrations of Medicago truncatula Gaertn. inoculated with Rhizophagus (formerly Glomus) irregularis . One explanation could be that the mycorrhizal inoculum used in this study was not isolated from a metal-contaminated site, which implies that it was less tolerant to the soil contamination than the indigenous mycorrhizal fungi (Malcova et al. 2003, Orlowska et al. 2005; Hildebrandt et al. 2007). Similarly, excessive ML concentrations may have impeded AMF germination and growth leading to a weak root colonisation by the fungi (Nowak 2007; Sanchez-Castro et al. 2017).
与本研究不同，Redon et al.（2008）报道，接种Rhizophagus（原Glomus）irregularis的Medicago truncatula Gaertn.地上部Cd浓度持续降低。一种解释是，本研究所用的菌根接种剂并非分离自金属污染场地，这意味着其对土壤污染的耐受性低于土著菌根真菌（Malcova et al. 2003, Orlowska et al. 2005; Hildebrandt et al. 2007）。同样，过高的ML浓度可能抑制了AMF的萌发和生长，导致真菌对根系的侵染较弱（Nowak 2007; Sanchez-Castro et al. 2017）。
At the inflorescence stage (Year 2), the relevance of using sage in the production of EO without washing the inflorescences was evaluated. In agreement with the literature (Hibben et al. 1984), the results indicated that there was no washing effect (p > 0.05, not shown) on the ML concentrations in the inflorescences of sage, indicating that no additional washing step in the whole distillation process was necessary for this plant. In addition, this result indicated that the dense sage cover observed on the site could reduce the dispersion of ML-contaminated dust, reducing contamination of the aerial portions of the plants. Consequently, the ML concentrations measured for both washed and unwashed inflorescences were pooled and presented all together in the further figures and tables (Fig. 6; Table 4).
在花序期（第2年），评估了花序不经清洗直接用于生产精油的适宜性。与文献一致（Hibben et al. 1984），结果表明清洗对鼠尾草花序中ML浓度无影响（p > 0.05，数据未列出），说明对该植物而言，整个蒸馏流程中无需额外的清洗步骤。此外，这一结果表明场地上观察到的鼠尾草茂密覆盖可减少ML污染粉尘的扩散，从而减少植物地上部的污染。因此，清洗和未清洗花序的ML浓度数据合并后在后续图表中一并列出（图6；表4）。
In contrast to Cu, the mean Zn, Cd and Pb concentrations measured in the leaves (Year 1) of sage collected on the contaminated site were all above those measured in the leaves of sage from the uncontaminated site (Zn: 31; Cd: 0.075; Pb: 0.6 µg g-1 DW) (Fig. 6). An excluder response of the sage for all the ML studied in this work was indicated by the values of the BCFtot (BCF <1, Table 4). The BCF tot for Cu ranged between 0.4 and 0.5 (Table 4). For Zn, there was a positive correlation between the extractable Zn concentration and the Zn concentrations in leaves and inflorescences of sage (p < 0.001, r = 0.59) (Fig. 4), suggesting that the sage has taken up the soluble Zn. On the contrary, the BCF tot for Pb was <<< 1 (Table 4), suggesting a low uptake of the soluble Pb. This assumption was supported by the moderate negative correlation (p < 0.01, r = -0.29) between the Pb concentrations in the leaves and inflorescences of sage and Pb extractable concentration. Irrespective of the inoculation, the inflorescences (Year 2) of the sage concentrated significantly less Zn and Pb than its leaves (Year 1) (p < 0.05).
与Cu不同，在污染场地采集的鼠尾草叶片（第1年）中Zn、Cd和Pb浓度均值均高于未污染场地鼠尾草叶片的测定值（Zn：31；Cd：0.075；Pb：0.6 µg g-1 DW）（图6）。BCFtot值（BCF <1，表4）表明鼠尾草对本研究所涉及的全部ML均呈排斥型响应。Cu的BCF tot在0.4至0.5之间（表4）。对于Zn，可提取态Zn浓度与鼠尾草叶片和花序中Zn浓度之间呈正相关（p < 0.001，r = 0.59）（图4），表明鼠尾草吸收了可溶性Zn。相反，Pb的BCF tot <<< 1（表4），表明其对可溶性Pb的吸收较低。鼠尾草叶片和花序中Pb浓度与可提取态Pb浓度之间呈中度负相关（p < 0.01，r = -0.29），支持了这一推断。无论接种与否，鼠尾草花序（第2年）富集的Zn和Pb均显著少于叶片（第1年）（p < 0.05）。
The inoculation led to a reduction of the mean Zn, Pb and Cd concentrations in the leaves of sage (Year 1) (p < 0.05) (Fig. 6A, B and C). A similar effect on the Pb concentration in maize shoots was shown in laboratory conditions in presence of a R. irregularis inoculum at low Pb soil concentration (Malcova et al. 2003). For Cd, the inoculation effect persisted into Year 2 leading to a reduction of the mean Cd concentrations in the inflorescences of sage (Fig. 6B, p < 0.05), as has already reported in M. truncatula shoots inoculated with R. irregularis (Redon et al. 2008).
接种使鼠尾草叶片（第1年）中Zn、Pb和Cd浓度均值降低（p < 0.05）（图6A、B和C）。在实验室条件下，低土壤Pb浓度时接种R. irregularis对玉米地上部Pb浓度也表现出类似效应（Malcova et al. 2003）。对于Cd，接种效应持续到第2年，使鼠尾草花序中Cd浓度均值降低（图6B，p < 0.05），这与已报道的接种R. irregularis的M. truncatula地上部结果相同（Redon et al. 2008）。
Despite the reduction of shoot Cd following inoculation, the mean Cd concentrations in the shoots and in the inflorescences were manifold higher than the Cd concentrations measured in the shoots of the sage collected from the uncontaminated site. Heggo and Angle (1990) showed that the same AMF species could have the opposite effect on ML uptake in soybeans depending on the ML extractable concentration in soil. Given that the AMF reduced the ML uptake by the sage root system during the 1st year, it is likely that there was successful root colonisation of sage.
尽管接种后地上部Cd有所降低，但地上部和花序中Cd浓度均值仍比未污染场地采集的鼠尾草地上部Cd浓度高出数倍。Heggo and Angle（1990）表明，同一AMF物种对大豆吸收ML的影响可能因土壤中可提取态ML浓度不同而截然相反。鉴于AMF在第1年降低了鼠尾草根系对ML的吸收，鼠尾草根系很可能已被成功侵染。
3.4) Metal(loid) concentrations in the residues of distillation and valorisation options3.4）蒸馏残渣中金属（类金属）浓度及资源化途径
Table 5 presents the ML concentrations measured in the residues of coriander after the distillation process. Except for Cu, seed residues had lower ML concentrations than the other aerial parts of the plant. This is in agreement with the results obtained for shoots and seeds metal-concentrations previously reported (Fig. 5). The ML concentrations in the residues of the shoots were higher than the ML concentrations in non-extracted shoots whereas the ML concentrations in the seed residues were similar to the non-extracted seeds (Fig. 5). This may indicate that there is a distillation process effect related to the distilled organ. All ML concentrations in the residues of the shoots and seeds from the contaminated site were above those from the uncontaminated site (unpublished data) (Fig. 5).表5给出了蒸馏后芫荽残渣中的ML浓度。除Cu外，种子残渣中的ML浓度低于植物其他地上部位。这与前文报道的地上部和种子金属浓度结果一致（图5）。地上部残渣中的ML浓度高于未提取的地上部，而种子残渣中的ML浓度与未提取的种子相近（图5）。这可能表明存在与蒸馏器官相关的蒸馏过程效应。污染场地地上部和种子残渣中的所有ML浓度均高于未污染场地（未发表数据）（图5）。
The ML concentrations in the coriander residues were compared with the French ML limit values for use as organic soil conditioners (NF U 44-051:2019). Both types of residues from the uncontaminated site could be used as soil conditioners, whereas the residues from the contaminated site exceeded the limit value for Cd (Table 5). As composting results both in a loss of water and a loss of organic matter of up to 50% (Oudart, 2013), concentrations in ML are proportionally increased in compost. Therefore, use of both shoots and seeds as inputs for producing compost will not be possible. Another option could be the use of the coriander residues in animal fodder. In contrast to the residues from the uncontaminated site, both types of residues from the contaminated site presented Cd concentrations above the Cd limit value set in the Directive on animal fodder (Directive 2002/32/CE of the 7 May 2002 on animal fodder) (Table 5). Even if the coriander residues would have relevant nutritional parameters, their use in animal fodder would be forbidden due to exceedance of the Cd limit. The concentrations of Pb indicate that this element could also limit reuse.
将芫荽残渣中的ML浓度与法国有机土壤调理剂限值（NF U 44-051:2019）进行了比较。来自未污染场地的两类残渣均可用作土壤调理剂，而来自污染场地的残渣Cd含量超过了限值（表5）。由于堆肥过程会造成水分损失以及高达50%的有机质损失（Oudart, 2013），堆肥中ML浓度会相应升高。因此，将地上部和种子用作堆肥原料均不可行。另一种可选方案是将芫荽残渣用作动物饲料。与未污染场地的残渣不同，污染场地的两类残渣Cd浓度均高于动物饲料指令（2002年5月7日关于动物饲料的第2002/32/CE号指令）所规定的Cd限值（表5）。即使芫荽残渣具有相应的营养指标，由于Cd超标，其也不得用作动物饲料。Pb浓度表明，该元素同样可能限制残渣的再利用。
Biomethane potentials (BMP) of the shoot residue of coriander from the uncontaminated site was of interest (Table 5) as it was similar to that determined for roadside grass (up to 203 NlCH4 kg-1 VS in André et al. 2019). BMP obtained from the shoot residue of coriander from the contaminated soil were the lowest (26% less than the BMP obtained from the shoot residue of coriander from the uncontaminated soil) and could be due to the presence of ML in such plant material which could have decreased the production of biomethane by the methanogen microorganisms. No inoculation effect was observed. No previous studies have been conducted on plant distillation residue with biogas production as endpoint. In contrast, a few studies have discussed the possibility to use ML-enriched plant biomass due to phytotechnologies in anaerobic digestion (Thewys et al. 2010a; b; Witters et al. 2012; Cao et al. 2014). ML may be stimulatory or inhibitory to anaerobic microorganisms according to the concentration and speciation of the ML, as well as process-related factors such as pH and redox potential (Chen et al. 2008). Some results suggested that ML increased gas yield at low concentrations but inhibited gas yield when they were present at high concentrations (Wong and Cheung, 1995). Verma et al. (2007) conducted an experiment on water hyacinth and water chestnut grown in unpolluted water and water with different doses of brass industry effluents that contained increasing concentrations of Cu and Cr. Biogas yield and biogas rate were increased in presence of brass industry effluents, compared to control conditions but were similar to control conditions at the highest brass industry effluent dose, suggested a stimulatory effect of metals on anaerobic digestion at low-medium dose and an inhibitory effect of metals at high dose. Witters et al. (2012) reported no difference in biogas production potential between maize grown on contaminated soil versus maize grown on uncontaminated soils. Cao et al. (2015) reported that, compared to normal plants with low Cu content, the plants used in remediation with increased Cu levels (100 mg kg -1) promoted anaerobic digestion and increased the methane content in biogas.
未污染场地芫荽地上部残渣的生物甲烷潜力（BMP）具有利用价值（表5），与路边草的BMP相近（André et al. 2019中最高达203 NlCH4 kg-1 VS）。污染土壤芫荽地上部残渣的BMP最低（较未污染土壤芫荽地上部残渣的BMP低26%），这可能是由于该植物材料中存在ML，从而降低了产甲烷微生物的生物甲烷产量。未观察到接种效应。此前尚无以沼气产量为终点的植物蒸馏残渣研究。相比之下，少数研究探讨了将植物修复技术产生的富含ML的植物生物质用于厌氧消化的可能性（Thewys et al. 2010a; b; Witters et al. 2012; Cao et al. 2014）。ML对厌氧微生物可能具有促进或抑制作用，这取决于ML的浓度和形态，以及pH和氧化还原电位等工艺相关因素（Chen et al. 2008）。部分结果表明，ML在低浓度下可提高产气量，而在高浓度下则抑制产气（Wong and Cheung, 1995）。Verma et al. (2007) 以生长于未污染水体及添加不同剂量黄铜工业废水（Cu和Cr浓度递增）水体中的水葫芦和菱角为对象开展了试验。与对照相比，黄铜工业废水的存在提高了沼气产量和产气速率，但在最高废水剂量下与对照相近，表明金属在中低剂量下对厌氧消化具有促进作用，而在高剂量下具有抑制作用。Witters et al. (2012) 报道，污染土壤与未污染土壤上种植的玉米在沼气生产潜力上无差异。Cao et al. (2015) 报道，与Cu含量较低的普通植物相比，用于修复且Cu含量升高（100 mg kg -1）的植物促进了厌氧消化，并提高了沼气中的甲烷含量。
Due to the low BMP of the coriander residue from the metal-contaminated soil, its use in anaerobic digestion could only be of interest if the distance to the AD platform is around 30km to make this option environmentally and economically viable (Zdanevitch et al. 2018). In addition, these residues could complement the inputs in anaerobic digestion if this practice complies with the local regulation in particular regarding the nondilution principle (e.g. Arrêté du 6 juin 2018 for France). Small scale tests on roadside grass showed that no more than 10-15 % by weight of this plant material should be added to the digestion reactor (Zdanevitch et al. 2020). Depending on the physical state of the coriander residues after the oil extraction, they could be used with no more preparation. Nevertheless, due to the high Cd content, the use of Cd-rich residue might impede the valorisation of the digestate as a soil improver or fertiliser.
由于金属污染土壤芫荽残渣的BMP较低，仅当其与厌氧消化（AD）设施的距离约为30 km时，将其用于厌氧消化才具有环境和经济上的可行性（Zdanevitch et al. 2018）。此外，若符合当地法规，特别是关于不稀释原则的规定（如法国2018年6月6日法令），这些残渣可作为厌氧消化原料的补充。针对路边草的小规模试验表明，该类植物材料添加至消化反应器的比例不应超过10–15%（按重量计）（Zdanevitch et al. 2020）。根据精油提取后芫荽残渣的物理状态，其可能无需进一步预处理即可使用。然而，由于Cd含量较高，使用富Cd残渣可能会妨碍沼渣作为土壤改良剂或肥料的利用。
Leaves (Year 1) and inflorescences (Year 2) of sage were collected to produce EO, leading to plant residues after distillation. Table 5 shows for the contaminated site that all ML concentrations were lower in the inflorescence residues than in the leaf residues. The distillation process effect suggested for coriander was not visible in the case of sage except for Zn and Pb. Indeed, the Zn and Pb concentrations in the leaf residues of sage were higher than in the leaves (Fig. 6A and C). The Cd, Pb and Zn concentrations in residues of the leaves and inflorescences from the contaminated site were higher than those measured in the uncontaminated conditions.
采集鼠尾草叶片（第1年）和花序（第2年）用于生产精油（EO），蒸馏后产生植物残渣。表5显示，在污染场地，花序残渣中所有ML浓度均低于叶片残渣。除Zn和Pb外，芫荽中所提出的蒸馏过程效应在鼠尾草中并不明显。事实上，鼠尾草叶片残渣中的Zn和Pb浓度高于叶片本身（图6A和C）。污染场地叶片和花序残渣中的Cd、Pb和Zn浓度均高于未污染条件下的测定值。
All the ML concentrations in residues from the contaminated site were well below the ML limit values set both in the French standard NF U 44-051 (2019) and in the EU regulation on fertilizing products (Regulation (EU) 2019/1009 on the making available on the market of EU fertilizing products), suggesting that these residues could be used in the production of soil conditioners. In contrast to coriander, the residues of sage inflorescences could be used as supplement in animal fodder because neither Cd nor Pb concentrations exceeding the limit values set in the Directive (Directive 2002/32/CE of the 7 May 2002 for fodder).
污染场地残渣中所有ML浓度均远低于法国标准NF U 44-051（2019）及欧盟肥料产品法规（关于欧盟肥料产品上市的第(EU) 2019/1009号法规）规定的ML限值，表明这些残渣可用于生产土壤调理剂。与芫荽不同，鼠尾草花序残渣可作为动物饲料补充物，因为其Cd和Pb浓度均未超过指令（2002年5月7日关于饲料的第2002/32/CE号指令）规定的限值。
BMPs were lower for both the contaminated and the uncontaminated samples of sage compared to coriander (Table 5): this may come from a higher content in lignin, which is disadvantageous for anaerobic digestion (Triolo and al. 2011, 2012). The recalcitrance of native lignocellulosic biomass makes it resistant to microbial hydrolysis, which reduces the bioconversion efficiency of organic matter into biogas (Xu et al. 2019). Compared to the BMPs of the roadside grass previously shown, the BMPs of the Year 1 residues of sage from the uncontaminated site appeared relevant to contribute to biomethane production. As with coriander residue, BMPs of the Year 1 residue of sage from the contaminated site were even lower (38% less than the BMP obtained from the leaf residue of sage from the uncontaminated soil), probably due the presence of ML. No inoculation effect was observed. As there is theoretically no restriction due to ML contents, use of these plants for anaerobic digestion could be relevant, especially in areas where the increasing number of anaerobic digestion plants results in resource competition. Kratky and Jirout (2015) suggested that the methane production of the sage residues could be increased by grinding these materials before introduction in the digester, through size reduction, larger surface area and pore size and lower crystallinity. Overall, as the BMP of the sage residue is low, to make this option environmentally and economically viable, the valorisation should be local (Zdanevitch et al. 2018).
与芫荽相比，鼠尾草污染和未污染样品的BMP均较低（表5）：这可能源于其较高的木质素含量，而木质素不利于厌氧消化（Triolo and al. 2011, 2012）。天然木质纤维素生物质的抗降解性使其难以被微生物水解，从而降低了有机质向沼气的生物转化效率（Xu et al. 2019）。与前述路边草的BMP相比，未污染场地鼠尾草第1年残渣的BMP显示其可用于生物甲烷生产。与芫荽残渣类似，污染场地鼠尾草第1年残渣的BMP更低（较未污染土壤鼠尾草叶片残渣的BMP低38%），可能是由于存在ML所致。未观察到接种效应。由于理论上不存在ML含量方面的限制，将这些植物用于厌氧消化可能具有现实意义，特别是在厌氧消化厂数量不断增加、导致原料竞争的地区。Kratky and Jirout (2015) 提出，在投入消化器前对鼠尾草残渣进行粉碎，通过减小粒径、增大比表面积和孔径以及降低结晶度，可提高其甲烷产量。总体而言，由于鼠尾草残渣的BMP较低，为使该方案在环境和经济上可行，应在本地进行资源化利用（Zdanevitch et al. 2018）。
Sage residues could be used for producing compost, where the higher content in lignin is an advantage for both the efficiency of the process (aeration) and the quality of the product (stability, mineralisation of the carbon). Composting is a common practice by producers of EO (personal communication) which makes this valorisation option highly relevant.
鼠尾草残渣可用于堆肥生产，其较高的木质素含量对堆肥过程效率（通气）和产品质量（稳定性、碳矿化）均有利。堆肥是精油生产商的常见做法（个人交流），因此该资源化途径具有很强的现实意义。
3.5) Metal(loid) concentrations in essential oils of coriander and sage and mycorrhizal inoculation effect
3.5）芫荽和鼠尾草精油中的类金属/金属浓度及菌根接种效应
Essential oils (EO) of coriander and sage were produced by the industrial producer by steam distillation of the plants at different growth stages once harvested from both contaminated and uncontaminated sites. Before processing, the plants were not washed to mimic industrial practice. The ML concentrations in EO from the biomass produced on the uncontaminated soil were used as reference values, as well as, those for commercial EO (Table 6).
芫荽和鼠尾草精油（EO）由工业生产商对从污染场地和未污染场地收获的不同生长阶段植物进行水蒸气蒸馏制得。为模拟工业实践，加工前未对植物进行清洗。以未污染土壤所产生物质制得精油中的ML浓度以及市售精油中的ML浓度作为参考值（表6）。
For both plants, irrespective of the plant stage, the ML concentrations in EO were either close or equal to the quantification limits (QL) (Table 6). There was no difference (p < 0.05) between the ML concentrations in coriander EO from both sites and between shoots and seeds. The same results were obtained for sage except for As, Cr and Cu at the leaves stage (Year 1) of the I uncontaminated soil, where the concentrations were above those of the contaminated soil. The inoculation did not influence the ML concentrations in coriander EO but did it in the case of sage at leaves stage for Cu (increase on the contaminated site, Table 6) and Cr (decrease on the contaminated site, Table 6).
对于两种植物，无论处于何种生长阶段，精油中的ML浓度均接近或等于定量限（QL）（表6）。两个场地之间以及地上部与种子之间，芫荽精油中的ML浓度均无差异（p < 0.05）。鼠尾草的结果相同，但接种（I）未污染土壤叶片阶段（第1年）的As、Cr和Cu除外，其浓度高于污染土壤。接种未影响芫荽精油中的ML浓度，但影响了鼠尾草叶片阶段的Cu（污染场地升高，表6）和Cr（污染场地降低，表6）浓度。
For both plant species, the lowest ML concentrations occurred in the plant parts usually used for distillation (seeds for coriander and inflorescences for sage). Compared to commercial EO of the same plant species, coriander and sage EO from both sites showed similar or lower ML concentrations. Overall, the ML transfer from biomass to EO was limited and ML concentrations in EO were not correlated with the ML concentrations in the initial biomass. In our experimental conditions, the distillation process used was suitable to limit the metal transfer in EO of coriander and sage and produced EO with a quality similar to that obtained commercially or from uncontaminated soil (Angelova et al. 2016). As reported by Ait-Elallem et al. (2020), this result is explained by the process itself: EO and not ML being extracted through the distillation method. The washing of plant samples did not significantly decrease the ML concentration in the sage EO indicating that a supplemental washing step of the plants before processing was not necessary in our case. In accordance with our results, Angelova et al. (2016), using steam distillation, showed little ML transfer from the seeds of unwashed clary sage to the EO and low ML concentrations in EO. They reported a positive correlation between the soil contamination level and the ML concentration in EO. The ML in their EO being higher than ours due to higher soil ML concentrations compared to ours.
对于两种植物，ML浓度最低的均为通常用于蒸馏的植物部位（芫荽为种子，鼠尾草为花序）。与同种植物的市售精油相比，两个场地所产芫荽和鼠尾草精油的ML浓度相近或更低。总体而言，ML从生物质向精油的迁移有限，且精油中的ML浓度与初始生物质中的ML浓度无相关性。在本试验条件下，所采用的蒸馏工艺能够有效限制金属向芫荽和鼠尾草精油中的迁移，所产精油质量与市售精油或未污染土壤所产精油相当（Angelova et al. 2016）。正如Ait-Elallem et al. (2020) 所报道，这一结果可由工艺本身解释：蒸馏法提取的是精油而非ML。清洗植物样品并未显著降低鼠尾草精油中的ML浓度，表明在本研究中加工前无需增加植物清洗步骤。与本研究结果一致，Angelova et al. (2016) 采用水蒸气蒸馏表明，ML从未清洗的香紫苏种子向精油的迁移很少，精油中ML浓度较低。他们报道了土壤污染程度与精油中ML浓度之间存在正相关。由于其土壤ML浓度高于本研究，其精油中的ML也高于本研究。
3.6) Summary of the analysis of the metal(loid) fate in the value chain
3.6）价值链中类金属/金属归趋分析总结
Technical decision trees for phytomanagement of sage and coriander are presented in the figure 7 and the supplemental Figure 1, respectively, based on the results of the analysis of the ML fate in the value chain. We showed that sage could be a suitable candidate for phytostabilisation due to its excluder phenotype (Fig. 7). In addition, the mycorrhizal inoculation reduced the Cd concentration in inflorescences and soil Cd extractable fraction. Moreover, the residues of distillation could be used as soil conditioners or animal fodder, increasing circularity of the agricultural economy (Fig. 7). In contrast, for coriander, the use of a mycorrhizal inoculum affected neither the ML concentrations nor the ML mobility in the soil. Excessive Cd concentrations occurred in the distillation residues, limiting their reuse (Supplemental Fig. 1). Based on ML transfer, EO production on contaminated site with sage and coriander seemed feasible.
基于价值链中ML归趋的分析结果，鼠尾草和芫荽植物管理的技术决策树分别见图7和补充图1。结果表明，鼠尾草因其排斥型表型，可作为植物稳定的适宜候选植物（图7）。此外，菌根接种降低了花序中的Cd浓度及土壤Cd可提取态比例。而且，蒸馏残渣可用作土壤调理剂或动物饲料，从而提高农业经济的循环性（图7）。相比之下，对于芫荽，施用菌根接种剂既未影响ML浓度，也未影响土壤中ML的移动性。蒸馏残渣中Cd浓度过高，限制了其再利用（补充图1）。基于ML迁移情况，在污染场地利用鼠尾草和芫荽生产精油似乎可行。
54) Conclusions4）结论
This study demonstrated the feasibility of harvesting sage and coriander on a metal(loid)-contaminated soil. Based on the fate of Cd, Cu, Pb, and Zn in the whole value chain, only sage fulfilled all phytomanagement objectives. Although our results showed similar metal(loid) concentrations in essential oil as compared to those obtained from plants grown on an uncontaminated soil, the commercial production of essential oil on contaminated soils depends on other crucial factors such as regulation, consumer perception, economic viability, and waste reduction. Future work should test a broader range of soil amendments to decrease the extractable metal(loid) concentrations in soil and in particular reduce plant Cd-uptake.
本研究证明了在类金属/金属污染土壤上收获鼠尾草和芫荽的可行性。基于Cd、Cu、Pb和Zn在整个价值链中的归趋，仅鼠尾草满足了全部植物管理目标。尽管结果显示其精油中类金属/金属浓度与未污染土壤所种植物制得的精油相近，但在污染土壤上进行精油商业化生产还取决于法规、消费者认知、经济可行性和减少废弃物等其他关键因素。未来工作应测试更多种类的土壤改良剂，以降低土壤中可提取态类金属/金属浓度，尤其是减少植物对Cd的吸收。
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REGULATION (EU) 2019/1009 OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 5 June 2019 laying down rules on the making available on the market of EU fertilising products and amending Regulations (EC) No 1069/2009 and (EC) No 1107/2009 and repealing Regulation (EC) No 2003/2003.
Figure captions
Fig. 1. Location, aerial view and experimental design of the field site.图1. 田间试验场地的位置、航拍图及试验设计。
Fig. 2. A and B: Water pH in soil with inoculated (I) and non-inoculated (NI) coriander (A) and sage (B) before harvest of aerial shoots (orange), the second sowing (BS) (green) and harvest of seeds (BH) (chocolate) or before harvest of leaves (Year 1, orange) and inflorescences (Year 2, green) in the case of coriander and sage, respectively. The dotted line separates the first and second harvest in the case of coriander (A). The red line corresponds to the mean of the water pH initially measured in soil – see Table 2. The box plots indicate the median and the 25 and 75 percentiles; mean ± SD; Significant differences between conditions are indicated by different letters at the level of α = 0.05.图2. A和B：接种（I）与未接种（NI）芫荽（A）和鼠尾草（B）土壤的水浸pH，芫荽为收获地上部前（橙色）、第二次播种前（BS）（绿色）及收获种子前（BH）（巧克力色），鼠尾草为收获叶片前（第1年，橙色）及收获花序前（第2年，绿色）。虚线分隔芫荽（A）的第一次和第二次收获。红线表示土壤初始测定的水浸pH均值——见表2。箱线图表示中位数及第25和第75百分位数；均值 ± SD；不同字母表示各处理间在α = 0.05水平上差异显著。
Fig. 3. 1M NH4NO3-extractable concentration of Zn (A), Cd (B), Pb (C) and Cu (D) in soil with inoculated (I) and non-inoculated (NI) coriander before harvest of shoots (orange), before sowing (BS) (green) and before harvest of seeds (BH) (chocolate) (the box plots indicate the median and the 25 and 75 percentiles; mean ± SD; the red line corresponds to the reference value in the ISO 19730:2008(E), the blue dashed line corresponds to the value measured before the experimental set up (Table 2)). Significant differences between conditions are indicated by different letters at the level of α = 0.05.图3. 接种（I）与未接种（NI）芫荽土壤中1M NH4NO3可提取态Zn（A）、Cd（B）、Pb（C）和Cu（D）浓度，分别于收获地上部前（橙色）、播种前（BS）（绿色）及收获种子前（BH）（巧克力色）测定（箱线图表示中位数及第25和第75百分位数；均值 ± SD；红线表示ISO 19730:2008(E)中的参考值，蓝色虚线表示试验布设前的测定值（表2））。不同字母表示各处理间在α = 0.05水平上差异显著。
Fig. 4. 1M NH4NO3-extractable concentration of Zn (A). Cd (B). Pb (C) and Cu (D) in soil with inoculated (I) and non-inoculated (NI) sage before harvest of leaves (Year 1, orange) and before harvest of inflorescences (Year 2, green) (the box plots indicate the median and the 25 and 75 percentiles; mean ± SD; the red line corresponds to the reference value in the ISO 19730:2008(E), the blue dashed line corresponds to the value measured before the experimental set up (Table 2)). Significant differences between conditions are indicated by different letters at the level of α = 0.05.图4. 接种（I）与未接种（NI）鼠尾草土壤中1M NH4NO3可提取态Zn（A）、Cd（B）、Pb（C）和Cu（D）浓度，分别于收获叶片前（第1年，橙色）及收获花序前（第2年，绿色）测定（箱线图表示中位数及第25和第75百分位数；均值 ± SD；红线表示ISO 19730:2008(E)中的参考值，蓝色虚线表示试验布设前的测定值（表2））。不同字母表示各处理间在α = 0.05水平上差异显著。
Fig. 5. Concentration of Zn (A), Cd (B), Pb (C) and Cu (D) in shoots (green) and seeds (orange) of coriander inoculated (I) and non-inoculated (NI) (the box plots indicate the median and the 25 and 75 percentiles; mean ± SD; the red line corresponds to the reference value measured in the uncontaminated site). Significant differences between conditions are indicated by different letters at the level of α = 0.05.图5. 接种（I）与未接种（NI）芫荽地上部（绿色）和种子（橙色）中Zn（A）、Cd（B）、Pb（C）和Cu（D）浓度（箱线图表示中位数及第25和第75百分位数；均值 ± SD；红线表示未污染场地的参考测定值）。不同字母表示各处理间在α = 0.05水平上差异显著。
Fig. 6. Concentration of Zn (A), Cd (B), Pb (C) and Cu (D) in leaves (Year 1, orange) and inflorescences (Year 2, green) of sage inoculated (I) and non-inoculated (NI) (the box plots indicate the median and the 25 and 75 percentiles; mean ± SD; the red line corresponds to the reference value measured in the uncontaminated site). Significant differences between conditions are indicated by different letters at the level of α = 0.05图6. 接种（I）与未接种（NI）鼠尾草叶片（第1年，橙色）和花序（第2年，绿色）中Zn（A）、Cd（B）、Pb（C）和Cu（D）浓度（箱线图表示中位数及第25和第75百分位数；均值 ± SD；红线表示未污染场地的参考测定值）。不同字母表示各处理间在α = 0.05水平上差异显著
Fig. 7. Technical decision tree for phytomanagement of sage (aided phytostabilization and EO production).图7. 鼠尾草植物管理（辅助植物稳定与精油生产）技术决策树。
Supplemental Fig. 1. Technical decision tree for phytomanagement of coriander (aided phytostabilization and EO production).
washed.
| Date of samplings采样日期 | Samples样品 |
|---|---|
| April 2017 | Soil (composites) |
| July and September 2017 | Soil (all plots) |
| July 2017 | Coriandrum sativum L.(shootsW) |
| September 2017 | Salvia sclarea L.(leavesNW) |
| April 2018 | Soil (coriander plots) |
| July 2018 | Soil (sage plots) |
| July 2018 | Salvia sclarea L.(inflorescencesW+ NW) |
| August 2018 | Soil (coriander plots) |
| August 2018 | Coriandrum sativum L.(seedsW) |
| Parameters参数 | Contaminated污染 | Uncontaminated未污染 |
|---|---|---|
| pH-H2O | 7.86 ± 0.13 | 7.1 ± 0.52 |
| Cd | 7 ± 0.4 (0.12 ± 0.04) | 0.4 ± 0.03 (0.01 ±0.006) |
| Zn | 443 ± 12 (0.95 ± 0.33) | 48 ± 2 (0.61 ± 0.35) |
| Cu | 27 ± 3 (0.29 ± 0.11) | 16 ± 0.3 (0.2 ± 0.11) |
| Total Pb | 394 ± 16 (0.07 ± 0.03) | 22 ± 2 (0.03 ± 0.03) |
| (mg kg-1) As | 16 ± 2 (0.01 ± 0.004) | 9 ± 0.6 (0.007±0.002) |
Cr | 59 ± 3 (0.1± 0.1) | 50 ± 3 (0.008±0.003) |
| Ni | 25 ± 1.8 (0.05 ± 0.01) | 16 ± 1 (0.16 ±0.36) |
| Sb | 5 ± 0.3 (0.05 ± 0.01 | 0.7 ± 0.05 (0.006 ± 0) |
| Clay (%) | 26. ± 2 | 14.± 1 |
| Silt (%) | 53 ± 10 | 70 ± 4 |
| Sand (%) | 18 ± 3 | 17 ± 3 |
| Soil texture (USDA) | Silty clay loam | Silt loam |
| Soil type (French classification)* | Fluvisol | Brown soil |
| Total organic carbon (g kg-1) | 17 ± 2 | 10.2 ± 1.4 |
| Organic matter (g kg-1) | 29 ± 4 | 18 ± 2 |
| C/N | 10.5 ± 0.7 | 8.6 ± 0.8 |
| CEC Metson (cmol+kg-1) | 183 ± 50 | 99 ± 3 |
| CaCO3 (%) | 0.48 ± 0.13 | 0.73 ± 0.34 |
| N (g kg-1) | 1.62 ± 0.24 | 1.18 ± 0.05 |
| P available Joret-Hebert (g kg-1) P2O5 | 0.17 ± 0.02 | 0.65 ± 0.26 |
| K available (g kg-1) K2O | 0.26 ± 0 | 0.21 ± 0.04 |
| Mg available (g kg-1) MgO | 0.21 ± 0.02 | 0.18 ± 0.07 |
| Ca available (g kg-1) CaO | 6 ± 0.5 | 3.5 ± 1.3 |
| Na available (g kg-1) Na2O | 0.03 | 0.02 ± 0 |
* Fourrier et al. (2011).
| As | Cd | Cr | Cu | Ni | Pb | Sb | Zn | ||
|---|---|---|---|---|---|---|---|---|---|
| Shoots I | BCFtot | 0.007 (0.003) | 0.99 (0.44)a | 0.022 (0.01) | 0.46 (0.14)a.c | 0.05 (0.015) | 0.012 (0.004)a | 0.006 (0.003)a | 0.19 (0.045) |
| BCFextr | 17 | 61 | 211 | 43 | 28 | 75 | 0.46 | 47 | |
| (6) | (17)A | (94) | (22) | (11)A | (27)A | (0.21)A | (29)A | ||
| BCFtot | 0.009 | 0.94 | 0.017 | 0.42 | 0.04 | 0.011 | 0.005 | 0.18 | |
| Shoots | (0.004) | (0.38)a | (0.012) | (0.102)a | (0.014) | (0.004)a | (0.002)a | (0.05) | |
| NI | BCFextr | 15 | 51 | 169 | 52 | 25 | 59 | 0.55 | 34 |
| (8) | (19) | (104) | (27) | (8)A | (24)A | (0.24) | (21)B | ||
| Seed I | BCFtot | 0.007 (0.005) | 0.42 (0.12)b | 0.024 (0.026) | 0.57 (0.12)b.c | 0.05 (0.023) | 0.004 (0.005)b | 0.011 (0.003)b | 0.2 (0.03) |
| BCFextr | 16 (13) | 24 (8)B | 214 (232) | 50 (12) | 25 (20)B | 21 (20)B | 1.21 (0.51)B | 130 (35)C | |
| Seed NI | BCFtot | 0.008± (0.002) | 0.45 (0.13)b | 0.013 (0.008) | 0.53 (0.107)c | 0.04 (0.013) | 0.004 (0.002)b | 0.010 (0.002)b | 0.2 (0.03) |
| BCFextr | 15 | 26 | 96 | 37 | 15 | 16 | 1.27 | 90 | |
| (9) | (23)B | (63) | (18) | (5)A | (12)B | (0.73)B | (30)C |
| As | Cd | Cr | Cu | Ni | Pb | Sb | Zn | ||
|---|---|---|---|---|---|---|---|---|---|
| BCFtot | 0.017 | 0.11 | 0.04 | 0.47 | 0.05 | 0.02 | 0.02 | 0.24 | |
| (0.004)a | (0.06)a | (0.01)a | (0.05)a.b | (0.01)a | (0.008)a | (0.005)a | (0.05)a | ||
| Leaves I | |||||||||
| BCFextr | 24 (13)A | 6 (2)A | 351 (120)A | 35 (6)A | 37 (11)A | 149 (55)A | 1.4 (0.2) | 70 (36) | |
| BCFtot | 0.017 | 0.19 | 0.034 | 0.51 | 0.05 | 0.03 | 0.02 | 0.32 | |
| Leaves | (0.006)a | (0.06)b | (0.02)a | (0.08)a | (0.02)a | (0.01)b | (0.006)a | (0.07)b | |
| NI | BCFextr | 37 (8)B | 9 (4)B | 316 (79)B | 33 (14)A | 33 (10)A | 171 (68)A | 2 (0.6) | 63 (42) |
| BCFtot | 0.01 | 0.101 | 0.04 | 0.37 | 0.03 | 0.005 | 0.01 | 0.15 | |
| Infloresc | (0.005)b | (0.04)a | (0.02)a | (0.21)b | (0.01)b | (0.002)c | (0.005)b | (0.03)c | |
| ences I | BCFextr | 13 (12)C | 5 (2)A | 216 (135)A | 96 (112)B | 12 (5)B | 13 (10)B | 1.4 (0.7) | 70 (23) |
| BCFtot | 0.006 | 0.13 | 0.02 | 0.41 | 0.03 | 0.004 | 0.01 | 0.12 | |
| Infloresc | (0)c | (0.05)a | (0.01)b | (0.16)a,b | (0.01)b | (0.001)c | (0.004)b | (0.03)c | |
| ences NI | BCFextr | 16 (5)C | 5 (3)A | 194 (114)A | 223 (219)A | 11 (5)B | 17 ( 9)B | 1.7 (0.7) | 56 (27) |
| Coriander芫荽 | As | Cd | Cr | Cu | Ni | Pb | Zn | BMP |
|---|---|---|---|---|---|---|---|---|
| shoots NI | 1.16 | 9.1 | 8.9 | 15.4 | 4.7 | 29.8 | 180 | 132.08 |
| (0.08)a | (1.45)a | (1.36)a | (1.1) a | (0.6)a | (2.3)a | (12)a | ||
| Shoots I | 1.69 | 8.8 | 10.5 | 19.8 | 5.3 | 43.1 | 209 | 145.18 |
| (0.17)b | (1.14)a | (2.8)a | (1.2)b | (1)a | (2.8)b | (26)a | ||
| Seed NI | QL*c | 3.7 | 1.2 | 18.1 | 1.5 | 0.9 | 112 | - |
| (0.44)b | (0.11)b | (2.1)a.b | (0.1)b | (0.07)c | (11)b | |||
| Seed I | QL*c | 3.51 | 1.5 | 18.8 | 1.9 | 0.8 | 103 | - |
| (0.37)b | (0.68)b | (2.3)a.b | (0.2)b | (0.1)c | (19)b | |||
| Shoots (un- contaminated | 0.26 | 0.75 | 6.9 | 10.1 | 4 | 0.6 | 49 | 1958 |
soil) | (0.05) | (0.1) | (2.92) | (1.45) | (1.7) | (0.2) | (7) | . |
| Seed (un- contaminated | 0.79 | 0.39 | 21.2 | 17.4 | 10 | 0.6 | 76.45 | - |
soil) | (0.79) | (0.04) | (22) | (4.2) | (8.7) | (0.5) | ( 30) | |
| Sage | ||||||||
| Leaves I | 0.29 (0.03)a | 1.24 (0.07)a | 2.5 (0.5)a | 11.1 (0.7)a | 1.3 (0.14)a | 13.5 (1.1)a | 164 (10)a | 89.61 |
| Leaves NI | 0.34 | 1.67 | 2.9 | 12.6 | 1.7 | 18.3 | 183 | 7285 |
| (0.09)a | (0.09)b | (1)a | (0.6)b | (0.55)a | (2.7)b | (9)b | . | |
| Inflorescences | 0.11 | 0.69 | 1.6 | 7.6 | 0.7 | 3.3 | 57 | |
| NI | (0.01)b.c | (0.03)c | (0.4)a | (0.6)c | (0.17)b.c | (0.7)c | (3)c | - |
| 0.18 | 0.82 | 1.7 | 7.9 | 0.55 | 3.7 | 67 | ||
| Inflorescences I | - | |||||||
| (0.04)c | (0.18)c | (0.1)a | (0.9)c | (0.03)b | (0.7)c | (5)c |
| Leaves un- contaminated叶片 未污染 | 0.18 (0.01) | 0.19 (0) | 2.9 (0.1) | 8.7 (0.05) | 1.4 (0.04) | 0.37 (0.09) | 33 (0) | 144.5 |
|---|---|---|---|---|---|---|---|---|
| Inflorescences | 0.20 | 0.09 | 3.3 | 7.9 | 1.8 | 0.4 | 29 | |
| un- contaminated | (0.05) | (0.01) | (0.7) | (0.25) | (0.3) | (0.16) | (3) | - |
| NF U 44-051+ | 18 | 3 | 120 | 300 | 60 | 180 | 800 | - |
| EU regulation | ||||||||
| on fertilizing | 40 | 2 | 2(CrVI) | 300 | 50 | 120 | 800 | - |
| products++ | ||||||||
| Animal fodder threshold+++ | 4 | 1 | - | - | - | 40 | - | - |
QL*=0.1; +: NF U 44-051/A3(August 2019) Organic soil improver French standard- Designations, specifications and marking – Additional elements; ++: REGULATION (EU) 2019/1009 OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 5 June 2019 laying down rules on the making available on the market of EU fertilizing products;+++: Directive 2002/32/CE of the 7 May 2002.
(I) and non-inoculated (NI) contaminated and uncontaminated soils. Significant differences between conditions are indicated by the different letters in bold at the level of α = 0.05. Commercial refers to commercial oils. QL = Quantification Limit.
| As | Cd | Cr | Cu | Ni | Pb | Sb | Zn | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| QL (µg g-1) | 0.01 | 0.015 | 0.01 | 0.005 | 0.01 | 0.015 | 0.05 | 0.05 | 0.03 | 0.013 | 0.01 | 0.0125 | 0.01 | 0.005 | 0.01 | 0.05 |
| Coriander | Shoots | Seed | Shoots | Seed | Shoots | Seed | Shoots | Seed | Shoots | Seed | Shoots | Seed | Shoots | Seed | Shoots | Seed |
| 0.01 | 0.036 | 0.022 | 0.056 | 0.06 | ||||||||||||
| Icontaminated | (0.00)a | QL | QL | QL | (0.009)a | (0.009) | (0.1)a | QL | (0.03) | QL | QL | QL | QL | QL | QL | QL |
| NIcontaminated | 0.013 (0.004)a. b | QL | QL | QL | 0.034 (0.03)a | 0.018 (0.003) | QLa | QL | 0.064 (0.04) | QL | 0.073 (0.109) | QL | QL | QL | 0.094 (0.145) | QL |
| 0.014 | 0.017 | 0.017 | 0.081 | 0.072 | ||||||||||||
| Iuncontaminated | (0.002)a. b | QL | QL | QL | QLb | (0.002) | QL a | QL | QL 0 | (0.007) | QL | QL | QL | QL | (0.122) | (0.038) |
| NI | 0.01 | 0.021 | 0.019 | 0.031 | 0.072 | 0.075 | 0.054 | |||||||||
| uncontaminated | (0.002)a | QL | QL | QL | (0.02)a | (0.003) | QL a | QL | (0.001) | QL | (0.106) | QL | QL | QL | (0.111) | (0.007) |
Commercial 0.015QL 0.015QL 0.026 0.862 0.03QL 0.06QL 0.03QL 0.067
7Figure caption图注
The R 3.5.2 software was used to create the boxplot figure (Fig. 2,3,4,5,6). The microsoft powerpoint software was used to create the fig. 1. The Fig. 7 and Supplemental Fig. 1 were created with the microsoft publisher software.
箱线图（图2、3、4、5、6）采用R 3.5.2软件绘制。图1采用Microsoft PowerPoint软件绘制。图7和补充图1采用Microsoft Publisher软件绘制。
Supplemental Fig. 1
Supplemental Table 1. TE recovery values obtained by ICP-OES and MS in the soil, water and plant certified reference materials. Information in brackets refer to paragraph in the text.
补充表1. 采用ICP-OES和MS测定土壤、水和植物有证标准物质中TE的回收率。括号内信息指正文中的相应段落。
| Reference materials标准 物质 | As | Cd | Cr | Cu | Ni | Pb | Sb | Zn |
|---|---|---|---|---|---|---|---|---|
| Soil (§2.2)* | [0.91-0.97] | [0.91-1.13] | [0.94-1] | [0.86-0.99] | [1.21-1.31] | [0.89-1.04] | [0.77-0.9] | [0.85-1.08] |
| Plant (§2.3;§2.5)+ | 0.86 | [0.71-0.99] | - | [0.88-0.92] | [0.77-1.03] | - | [0.84-1.04] | [0.96-1.14] |
| Water (§2.4)++ | 0.9 | 1.07 | 0.96 | 0.94 | 0.95 | 0.98 | 0.88 | 0.86 |
* NIST SRM 2710, Montana soil.
* NIST SRM 2710，Montana土壤。
+ White cabbage « BCR 679 », JRC, community bureau of reference, http://www.irmm.jrc.be/mrm.html. No
+ 白菜 « BCR 679 »，JRC，欧共体标准局，http://www.irmm.jrc.be/mrm.html。无
recovery values were available for Cr and Pb.
Cr和Pb的回收率数据。
++ Environmental matrix water « TMDA – 62.2 », Environnent Canada.
++ 环境基质水样 « TMDA – 62.2 »，加拿大环境部。
Supplemental Table 2. Percent of mobile TE (As, Cd, Cr, Cu, Ni, Pb, Sb, and Zn) calculated as extractable concentrations over total concentrations) in soil with inoculated (I) and non-inoculated (NI) coriander, before harvest of shootsand seeds, and sage, before harvest of leaves (year 1) and inflorescences (year 2) (mean ± SD). Significant differences between conditions (I, NI, year 1 and year 2) are indicated by the different letters in bold at the level of α = 0.05. Percent of mobile TE in the contaminated soil before the set up of the experiment, i.e. initial state, is also shown (mean ± SD).
补充表2. 接种（I）与未接种（NI）芫荽（收获地上部和种子前）及鼠尾草（收获叶片前（第1年）和花序前（第2年））土壤中可移动态TE（As、Cd、Cr、Cu、Ni、Pb、Sb和Zn）百分比（以可提取态浓度与全量浓度之比计算）（均值 ± SD）。粗体不同字母表示各处理间（I、NI、第1年和第2年）在α = 0.05水平上差异显著。同时列出试验布设前污染土壤（即初始状态）中可移动态TE百分比（均值 ± SD）。
| Initial State初始状态 | As | Cd | Cr | Cu | Ni | Pb | Sb | Zn |
|---|---|---|---|---|---|---|---|---|
| Contaminated | 0.06 (0.02) | 1.63 (0.51) | 0.02 (0.02) | 1.08 (0.41) | 0.2 (0.04) | 0.02 (0.006) | 0.97 (0.28) | 0.21 (0.07) |
| Coriander | As | Cd | Cr | Cu | Ni | Pb | Sb | Zn |
| Shoots NI | 0.06 (0.014) | 1.9 (0.74)a | 0.01 (0.001)a | 1.04 (0.48) | 0.17 (0.04) | 0.02 (0.009) | 1.03 (0.28)a | 0.66 (0.32)a |
| Shoots I | 0.05 | 1.63 | 0.01 | 1.98 | 0.18 | 0.016 | 1.21 | 0.56 |
| (0.014) | (0.55)a | (0)a | (2.22) | (0.06) | (0.003) | (0.22) a.b | (0.24)a | |
| Seed NI | 0.069 | 2.27 | 0.014 | 2.02 | 0.28 | 0.03 | 0.876 | 0.27 |
| (0.03) | (0.97)b | (0.003)b | (1.8) | (0.07) | (0.019) | (0.26)b | (0.19)b | |
| Seed I | 0.053 | 1.84 | 0.011 | 1.18 | 0.22 | 0.02 | 0.921 | 0.16 |
| (0.015) | (0.42)a | (0.001)a | (0.26) | (0.03) | (0.006) | (0.15) a.b | (0.04)b | |
| Sage | As | Cd | Cr | Cu | Ni | Pb | Sb | Zn |
| 0.049 | 2.07 | 0.011 | 2.31 | 0.17 | 0.02 | 1.25 | 0.65 | |
| Leaves NI | (0.014)A | (0.7)A.B | (0.001)A | (2.54) | (0.06)A | (0.009)A | ( 0.19)A | (0.39)B |
| 0.09 | 1.78 | 0.011 | 1.36 | 0.15 | 0.013 | 1.59 | 0.44 | |
| Leaves I | (0.05)B | (0.28)A | (0)A.B | (0.2)3 | (0.03)A | (0.002)A | (0.27)B | (0.27)A.B |
| Inflorescences | 0.04 | 2.37 | 0.014 | 0.93 | 0.29 | 0.03 | 0.72 | 0.26 |
| NI | (0.01)A | (0.46)B | (0.004)B | (1.11) | (0.16)B | (0.012)A | (0.18)C | (0.113)A |
| Inflorescences | 0.1 | 2.08 | 0.02 | 1.31 | 0.27 | 0.07 | 0.93 | 0.25 |
| I | (0.04)B | (0.32)A.B | (0.006)C | (2.25) | (0.05)B | (0.04)B | ( 0.19)D | (0.17)A |