Assessing plant uptake of organic contaminants by food crops tomato, wheat, and corn through sap concentration factor.

This study investigated uptake of two organic compounds including hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and exogenous caffeine by tomato (Solanum lycopersicum L.), corn (Zea mays L.), and wheat (Triticum aestivum L.). The plants were grown in a growth chamber under recommended conditions and then were exposed to these compounds for 19 days. The uptake of the compounds was measured by sap concentration factor. The plant samples (stem transpiration stream) and solution in the exposure media were taken and analyzed by high performance liquid chromatography-tandem mass spectrometry. The plant stem samples were analyzed after a freeze-thaw centrifugation process. The average sap concentration factor for the RDX by tomato, wheat, and corn was 0.71, 0.67, and 0.65. The average sap concentration factor for the exogenous caffeine by tomato, wheat, and corn was 0.72, 0.50, and 0.34. These relatively high sap concentration factor values were expected as available predictive models offer high sap concentration factor values for moderately hydrophobic and hydrophilic compounds. The generated sap concentration factor values for the RDX and exogenous caffeine are important for improving the accuracy of previously developed machine learning models predicting the uptake and translocation of emerging contaminants.

The uptake of two organic compounds (RDX and exogenous caffeine) was examined in three crop plants (corn, wheat, and tomato). There have not been any uptake studies on exogenous caffeine and also we do not have good data for the uptake of RDX by these three crop plants. The estimated sap concentration factor from these experiments fills the gap in the data for developing predictive models for uptake of emerging contaminants. A novel rapid freeze­thaw/centrifugation extraction method followed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was used to analyze the samples.

Machine Learning: New Ideas and Tools in Environmental Science and Engineering.

The rapid increase in both the quantity and complexity of data that are being generated daily in the field of environmental science and engineering (ESE) demands accompanied advancement in data analytics. Advanced data analysis approaches, such as machine learning (ML), have become indispensable tools for revealing hidden patterns or deducing correlations for which conventional analytical methods face limitations or challenges. However, ML concepts and practices have not been widely utilized by researchers in ESE. This feature explores the potential of ML to revolutionize data analysis and modeling in the ESE field, and covers the essential knowledge needed for such applications. First, we use five examples to illustrate how ML addresses complex ESE problems. We then summarize four major types of applications of ML in ESE: making predictions; extracting feature importance; detecting anomalies; and discovering new materials or chemicals. Next, we introduce the essential knowledge required and current shortcomings in ML applications in ESE, with a focus on three important but often overlooked components when applying ML: correct model development, proper model interpretation, and sound applicability analysis. Finally, we discuss challenges and future opportunities in the application of ML tools in ESE to highlight the potential of ML in this field.

Amendment-assisted revegetation of mine tailings: improvement of tailings quality and biomass production.

Mining activities have left a legacy of metals containing tailings impoundments. After mine closure, reclamation of mine wastes can be achieved by restoration of a vegetation cover. This study investigated the impact of biochar (BC), biosolids (BS), humic substances (HS), and mycorrhizal fungi (MF) for improving mine tailings fertility and hydraulic properties, supporting plant establishment, tailings revegetation, and enabling growth of energy crops. We conducted a pot trial by growing willow, poplar, and miscanthus in Pb/Zn/Cu mine tailings untreated or amended with two rates of amendments (low or high input). Biosolids resulted in the most significant changes in tailings properties, neutralizing pH and increasing organic carbon, nutrient concentrations, cation exchange capacity, water retention, and saturated hydraulic conductivity. The greatest increase in energy crops production was also observed in BS treatments enabling the financial viability of mine reclamation. Although BC resulted in significant improvements in tailings fertility and hydraulic properties, its impact on biomass was less pronounced, most likely due to lower N and P available concentrations. Increases in willow and miscanthus biomass were observed in HS and MF treatments in spite of their lower nutrient content. A pot experiment is underway to assess synergistic effects of combining BS with BC, HS, or MF.

Phytoremediation removal rates of benzene, toluene, and chlorobenzene.

Phytoremediation is a sustainable remedial approach, although performance efficacy is rarely reported. In this study, we assessed a phytoremediation plot treating benzene, toluene, and chlorobenzene. A comparison of the calculated phytoremediation removal rate with estimates of onsite contaminant mass was used to forecast cleanup periods. The investigation demonstrated that substantial microbial degradation was occurring in the subsurface. Estimates of transpiration indicated that the trees planted were removing approximately 240,000 L of water per year. This large quantity of water removal implies substantial removal of contaminant due to large amounts of contaminants in the groundwater; however, these contaminants extensively sorb to the soil, resulting in large quantities of contaminant mass in the subsurface. The total estimate of subsurface contaminant mass was also complicated by the presence of non-aqueous phase liquids (NAPL), additional contaminant masses that were difficult to quantify. These uncertainties of initial contaminant mass at the site result in large uncertainty in the cleanup period, although mean estimates are on the order of decades. Collectively, the model indicates contaminant removal rates on the order of 10-2-100 kg/tree/year. The benefit of the phytoremediation system is relatively sustainable cleanup over the long periods necessary due to the presence of NAPL.

Tree Sampling as a Method to Assess Vapor Intrusion Potential at a Site Characterized by VOC-Contaminated Groundwater and Soil.

Vapor intrusion (VI) by volatile organic compounds (VOCs) in the built environment presents a threat to human health. Traditional VI assessments are often time-, cost-, and labor-intensive; whereas traditional subsurface methods sample a relatively small volume in the subsurface and are difficult to collect within and near structures. Trees could provide a similar subsurface sample where roots act as the "sampler' and are already onsite. Regression models were developed to assess the relation between PCE concentrations in over 500 tree-core samples with PCE concentrations in over 50 groundwater and 1000 soil samples collected from a tetrachloroethylene- (PCE-) contaminated Superfund site and analyzed using gas chromatography. Results indicate that in planta concentrations are significantly and positively related to PCE concentrations in groundwater samples collected at depths less than 20 m (adjusted R2 values greater than 0.80) and in soil samples (adjusted R2 values greater than 0.90). Results indicate that a 30 cm diameter tree characterizes soil concentrations at depths less than 6 m over an area of 700-1600 m2, the volume of a typical basement. These findings indicate that tree sampling may be an appropriate method to detect contamination at shallow depths at sites with VI.

Contaminant Gradients in Trees: Directional Tree Coring Reveals Boundaries of Soil and Soil-Gas Contamination with Potential Applications in Vapor Intrusion Assessment.

Contaminated sites pose ecological and human-health risks through exposure to contaminated soil and groundwater. Whereas we can readily locate, monitor, and track contaminants in groundwater, it is harder to perform these tasks in the vadose zone. In this study, tree-core samples were collected at a Superfund site to determine if the sample-collection location around a particular tree could reveal the subsurface location, or direction, of soil and soil-gas contaminant plumes. Contaminant-centroid vectors were calculated from tree-core data to reveal contaminant distributions in directional tree samples at a higher resolution, and vectors were correlated with soil-gas characterization collected using conventional methods. Results clearly demonstrated that directional tree coring around tree trunks can indicate gradients in soil and soil-gas contaminant plumes, and the strength of the correlations were directly proportionate to the magnitude of tree-core concentration gradients (spearman's coefficient of -0.61 and -0.55 in soil and tree-core gradients, respectively). Linear regression indicates agreement between the concentration-centroid vectors is significantly affected by in planta and soil concentration gradients and when concentration centroids in soil are closer to trees. Given the existing link between soil-gas and vapor intrusion, this study also indicates that directional tree coring might be applicable in vapor intrusion assessment.

Enhanced Degradation of TCE on a Superfund Site Using Endophyte-Assisted Poplar Tree Phytoremediation.

Trichloroethylene (TCE) is a widespread environmental pollutant common in groundwater plumes associated with industrial manufacturing areas. We had previously isolated and characterized a natural bacterial endophyte, Enterobacter sp. strain PDN3, of poplar trees, that rapidly metabolizes TCE, releasing chloride ion. We now report findings from a successful three-year field trial of endophyte-assisted phytoremediation on the Middlefield-Ellis-Whisman Superfund Study Area TCE plume in the Silicon Valley of California. The inoculated poplar trees exhibited increased growth and reduced TCE phytotoxic effects with a 32% increase in trunk diameter compared to mock-inoculated control poplar trees. The inoculated trees excreted 50% more chloride ion into the rhizosphere, indicative of increased TCE metabolism in planta. Data from tree core analysis of the tree tissues provided further supporting evidence of the enhanced rate of degradation of the chlorinated solvents in the inoculated trees. Test well groundwater analyses demonstrated a marked decrease in concentration of TCE and its derivatives from the tree-associated groundwater plume. The concentration of TCE decreased from 300 µg/L upstream of the planted area to less than 5 µg/L downstream of the planted area. TCE derivatives were similarly removed with cis-1,2-dichloroethene decreasing from 160 µg/L to less than 5 µg/L and trans-1,2-dichloroethene decreasing from 3.1 µg/L to less than 0.5 µg/L downstream of the planted trees. 1,1-dichloroethene and vinyl chloride both decreased from 6.8 and 0.77 µg/L, respectively, to below the reporting limit of 0.5 µg/L providing strong evidence of the ability of the endophytic inoculated trees to effectively remove TCE from affected groundwater. The combination of native pollutant-degrading endophytic bacteria and fast-growing poplar tree systems offers a readily deployable, cost-effective approach for the degradation of TCE, and may help mitigate potential transfer up the food chain, volatilization to the atmosphere, as well as direct phytotoxic impacts to plants used in this type of phytoremediation.

Reducing arsenic accumulation in rice grain through iron oxide amendment.

Effects of soil-arsenic (As), phosphorus and iron oxide on As accumulation in rice grain were investigated. Cultivars that have significantly different sensitivity to As, straighthead-resistant Zhe 733 and straighthead-susceptible Cocodrie, were used to represent different cultivar varieties. The grain accumulation of other elements of concern, selenium (Se), molybdenum (Mo), and cadmium (Cd) was also monitored. Results demonstrated that high soil-As not only resulted in high grain-As, but could also result in high grain-Se, and Zhe 733 had significantly less grain-As than Cocodrie did. However, soil-As did not impact grain-Mo and Cd. Among all elements monitored, iron oxide amendment significantly reduced grain-As for both cultivars, while the phosphate application only reduced grain-Se for Zhe 733. Results also indicated that cultivar type significantly impacted grain accumulation of all monitored trace elements. Therefore, applying iron oxide to As-contaminated land, in addition to choosing appropriate rice cultivar, can effectively reduce the grain accumulation of As.

Phytoscreening with SPME: Variability Analysis.

Phytoscreening has been demonstrated at a variety of sites over the past 15 years as a low-impact, sustainable tool in delineation of shallow groundwater contaminated with chlorinated solvents. Collection of tree cores is rapid and straightforward, but low concentrations in tree tissues requires sensitive analytics. Solid-phase microextraction (SPME) is amenable to the complex matrix while allowing for solvent-less extraction. Accurate quantification requires the absence of competitive sorption, examined here both in laboratory experiments and through comprehensive examination of field data. Analysis of approximately 2,000 trees at numerous field sites also allowed testing of the tree genus and diameter effects on measured tree contaminant concentrations. Collectively, while these variables were found to significantly affect site-adjusted perchloroethylene (PCE) concentrations, the explanatory power of these effects was small (adjusted R(2) = 0.031). 90th quantile chemical concentrations in trees were significantly reduced by increasing Henry's constant and increasing hydrophobicity. Analysis of replicate tree core data showed no correlation between replicate relative standard deviation (RSD) and wood type or tree diameter, with an overall median RSD of 30%. Collectively, these findings indicate SPME is an appropriate technique for sampling and analyzing chlorinated solvents in wood and that phytoscreening is robust against changes in tree type and diameter.

Phytomonitoring of chlorinated ethenes in trees: a four-year study of seasonal chemodynamics in planta.

Long-term monitoring (LTM) of groundwater remedial projects is costly and time-consuming, particularly when using phytoremediation, a long-term remedial approach. The use of trees as sensors of groundwater contamination (i.e., phytoscreening) has been widely described, although the use of trees to provide long-term monitoring of such plumes (phytomonitoring) has been more limited due to unexplained variability of contaminant concentrations in trees. To assess this variability, we developed an in planta sampling method to obtain high-frequency measurements of chlorinated ethenes in oak (Quercus rubra) and baldcypress (Taxodium distichum) trees growing above a contaminated plume during a 4-year trial. The data set revealed that contaminant concentrations increased rapidly with transpiration in the spring and decreased in the fall, resulting in perchloroethene (PCE) and trichloroethene (TCE) sapwood concentrations an order of magnitude higher in late summer as compared to winter. Heartwood PCE and TCE concentrations were more buffered against seasonal effects. Rainfall events caused negligible dilution of contaminant concentrations in trees after precipitation events. Modeling evapotranspiration potential from meteorological data and comparing the modeled uptake and transport with the 4 years of high frequency data provides a foundation to advance the implementation of phytomonitoring and improved understanding of plant contaminant interactions.

Advancing Simultaneous Extraction and Sequential Single-Particle ICP-MS Analysis for Metallic Nanoparticle Mixtures in Plant Tissues.

Engineered nanoparticles (ENPs) have been increasingly used in agricultural operations, leading to an urgent need for robust methods to analyze co-occurring ENPs in plant tissues. In response, this study advanced the simultaneous extraction of coexisting silver, cerium oxide, and copper oxide ENPs in lettuce shoots and roots using macerozyme R-10 and analyzed them by single-particle inductively coupled plasma-mass spectrometry (ICP-MS). Additionally, the standard stock suspensions of the ENPs were stabilized with citrate, and the long-term stability (up to 5 months) was examined for the first time. The method performance results displayed satisfactory accuracies and precisions and achieved low particle concentration and particle size detection limits. Significantly, the oven drying process was proved not to impact the properties of the ENPs; therefore, oven-dried lettuce tissues were used in this study, which markedly expanded the applicability of this method. This robust methodology provides a timely approach to characterize and quantify multiple coexisting ENPs in plants.

Directional phytoscreening: contaminant gradients in trees for plume delineation.

Tree sampling methods have been used in phytoscreening applications to delineate contaminated soil and groundwater, augmenting traditional investigative methods that are time-consuming, resource-intensive, invasive, and costly. In the past decade, contaminant concentrations in tree tissues have been shown to reflect the extent and intensity of subsurface contamination. This paper investigates a new phytoscreening tool: directional tree coring, a concept originating from field data that indicated azimuthal concentrations in tree trunks reflected the concentration gradients in the groundwater around the tree. To experimentally test this hypothesis, large diameter trees were subjected to subsurface contaminant concentration gradients in a greenhouse study. These trees were then analyzed for azimuthal concentration gradients in aboveground tree tissues, revealing contaminant centroids located on the side of the tree nearest the most contaminated groundwater. Tree coring at three field sites revealed sufficiently steep contaminant gradients in trees reflected nearby groundwater contaminant gradients. In practice, trees possessing steep contaminant gradients are indicators of steep subsurface contaminant gradients, providing compass-like information about the contaminant gradient, pointing investigators toward higher concentration regions of the plume.

Fast separation and quantification method for nitroguanidine and 2,4-dinitroanisole by ultrafast liquid chromatography-tandem mass spectrometry.

Explosives are now persistent environmental pollutants that are targets of remediation and monitoring in a wide array of environmental media. Nitroguanidine (NG) and 2,4-dinitroanisole (DNAN) are two insensitive energetic compounds recently used as munitions explosives. To protect our environment and human health, the levels of these compounds in soils and waters need to be monitored. However, no sensitive analytical methods, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS), have been developed for detecting these new compounds at trace levels and to be concurrently applied to monitor the common explosives. In general, the concentrations of explosives in either soil or water samples are very low and widely distributed. Therefore, a fast and sensitive method is required to monitor those compounds and increase our ability to find and address the threats they pose to human health and ecological receptors. In this study, a fast and sensitive analytical method has been developed to quantitatively determine NG and DNAN in soil, tap water, and river water by using ultrafast LC-MS/MS. To make this method a comprehensive analytical technique for other explosives as well, it has included other commonly used explosives in the method development, such as octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), 1,3,5-trinitroper-hydro-1,3,5-triazine (RDX), 2,4,6-trinitrotoluene (TNT), 2-amino-4,6-dinitrotoluene (ADNT), and pentaerythritol tetranitrate (PETN). The method detection limits (MDLs) of these compounds in soil ranged from 0.2 to 5 ppb, and a good linearity was obtained over a concentration range of 0.5-200 ppb. The recoveries of some compounds are equal to or better than the current EPA methods but with much higher sensitivities.

Time-weighted average SPME analysis for in planta determination of cVOCs.

The potential of phytoscreening for plume delineation at contaminated sites has promoted interest in innovative, sensitive contaminant sampling techniques. Solid-phase microextraction (SPME) methods have been developed, offering quick, undemanding, noninvasive sampling without the use of solvents. In this study, time-weighted average SPME (TWA-SPME) sampling was evaluated for in planta quantification of chlorinated solvents. TWA-SPME was found to have increased sensitivity over headspace and equilibrium SPME sampling. Using a variety of chlorinated solvents and a polydimethylsiloxane/carboxen (PDMS/CAR) SPME fiber, most compounds exhibited near linear or linear uptake over the sampling period. Smaller, less hydrophobic compounds exhibited more nonlinearity than larger, more hydrophobic molecules. Using a specifically designed in planta sampler, field sampling was conducted at a site contaminated with chlorinated solvents. Sampling with TWA-SPME produced instrument responses ranging from 5 to over 200 times higher than headspace tree core sampling. This work demonstrates that TWA-SPME can be used for in planta detection of a broad range of chlorinated solvents and methods can likely be applied to other volatile and semivolatile organic compounds.

Dendrochemistry of multiple releases of chlorinated solvents at a former industrial site.

Trees can take up and assimilate contaminants from the soil, subsurface, and groundwater. Contaminants in the transpiration stream can become bound or incorporated into the annual rings formed in trees of the temperate zones. The chemical analysis of precisely dated tree rings, called dendrochemistry, can be used to interpret past plant interactions with contaminants. This investigation demonstrates that dendrochemistry can be used to generate historical scenarios of past contamination of groundwater by chlorinated solvents at a site in Verl, Germany. Increment cores from trees at the Verl site were collected and analyzed by energy-dispersive X-ray fluorescence (EDXRF) line scanning. The EDXRF profiles showed four to six time periods where tree rings had anomalously high concentrations of chlorine (Cl) as an indicator of potential contamination by chlorinated solvents.

Plant tissue analysis for explosive compounds in phytoremediation and phytoforensics.

Plant tissue analysis methods were evaluated for six explosive compounds to assess uptake and phytoforensic methods development to quantify explosives in plant to obtain the plant data for the evaluation of explosive contamination in soil and groundwater. Four different solvent mixtures containing acetonitrile or methanol were tested at variable extraction ratios to compare the extraction efficiency for six explosive compounds: 2,4,6-trinitrotoluene (TNT), pentaerythritoltetranitrate (PETN), hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), 2-amino-4,6-dinitrotoluene (2ADNT), and 2,4-Dinitroanisole (DNAN), in Laurel Willow (Salix pentandra) stem and range grass Big Bluestem (Andropogon gerardii) using LC-MS/MS. Plant tissues were spiked with 500 ng/g of explosives and extracted using ultrasonically-assisted solvent extraction. With the ratio of fresh plant mass to solvent volume of 1:20 for willow and 1:40 for big bluestem grass, results indicated that all explosives in willow except HMX were extracted at higher than 73.3% by using 20 mL of methanol, 50:50 (v/v) methanol:water, or acetonitrile, whereas HMX was extracted with the highest recovery of 61.3% by 20 mL of acetonitrile. In big bluestem grass, the most effective solvents were 20 mL of either methanol or 50:50 (v/v) methanol:water for PETN extraction with a recovery of higher than 101.2% and 20 mL of 50:50 (v/v) methanol:water for HMX, RDX, TNT, 2ADNT, and DNAN extraction with a recovery of 83.8%, 104.4%, 97.5%, 80.7%, and 108.2%, respectively. However, unlike methanol and acetonitrile, 50:50 (v/v) methanol:water provided no problem of leading or split peak in chromatogram; therefore, it was preferred in the test and performed a method validation. Results indicated that 50:50 (v/v) methanol:water provided good repeatability and recovery and method detection limits at 0.5-20 ng/g fresh weight or 8.8-61.3 ng/g dry weight. Overall, results suggested that solvent extraction efficiency of explosives in plant was influenced by plant species and solvent used, and method presented here was believed to provide the preliminary data with respect to the analysis of simultaneous explosives in plants with LC-MS/MS.

Phytomanagement of Pb/Zn/Cu tailings using biosolids-biochar or -humus combinations: Enhancement of bioenergy crop production, substrate functionality, and ecosystem services.

The extreme characteristics of mine tailings generally prohibit microbial processes and natural plant growth. Consequently, vast and numerous tailings sites remain barren for decades and highly susceptible to windblown dust and water erosion. Amendment-assisted phytostabilization is a cost-effective and ecologically productive approach to mitigate the potential transport of residual metals. Due to the contrasting and complementary characteristics of biosolids (BS) and biochar (BC), co-application might be more efficient than individually applied. Studies considering BS and BC co-application for multi-metal tailings revegetation are scarce. As tailings revegetation is a multidimensional issue, clearly notable demand exists for a study that provides a comprehensive understanding on the co-application impact on interrelated properties of physicochemical, biological, mineral nitrogen availability, metal immobilization, water-soil interactions, and impacts on plant cultivation and biomass production. This 8-month greenhouse study aimed at investigating the efficacy of co-application strategies targeting BS and carbon-rich amendments (BC or humic substances (HS)) to phytomanage a slightly alkaline Pb/Zn/Cu tailings with bioenergy crops (poplar, willow, and miscanthus). A complementary assessment linking revegetation effectiveness to ecosystem services (ES) provision was also included. Owing to their rich nutrient and organic matter contents, BS had the most pronounced influence on most of the measured properties including physicochemical, enzyme activities, NH4+-N and NO3--N availability, immobilization of Zn, Cu, and Cd, and biomass production. Co-applying with BC exhibited efficient nutrient release and was more effective than BS alone in reducing metal bioavailability and uptake particularly Pb. Poplar and willow exhibited more superior phytostabilization efficiency compared to miscanthus which caused acidification-induced metal mobilization, yet BC and BS co-application was effective in ameliorating this effect. Enhancement of ES and substrate quality index mirrored the positive effect of amendment co-application and plant cultivation. Co-applying HS with BS resulted in improved nutrient cycling while BC enhanced water purification and contamination control services.

Phytoforensics, dendrochemistry, and phytoscreening: new green tools for delineating contaminants from past and present.

As plants evolved to be extremely proficient in mass transfer with their surroundings and survive as earth's dominant biomass, they also accumulate and store some contaminants from surroundings, acting as passive samplers. Novel applications and analytical methods have been utilized to gain information about a wide range of contaminants in the biosphere soil, water, and air, with information available on both past (dendrochemistry) and present (phytoscreening). Collectively these sampling approaches provide rapid, cheap, ecologically friendly, and overall "green" tools termed "Phytoforensics".

Phytoscreening for chlorinated solvents using rapid in vitro SPME sampling: application to urban plume in Verl, Germany.

Rapid detection and delineation of contaminants in urban settings is critically important in protecting human health. Cores from trees growing above a plume of contaminated groundwater in Verl, Germany, were collected in 1 day, with subsequent analysis and plume mapping completed over several days. Solid-phase microextraction (SPME) analysis was applied to detect tetrachloroethene (PCE) and trichloroethene (TCE) to below nanogram/liter levels in the transpiration stream of the trees. The tree core concentrations showed a clear areal correlation to the distribution of PCE and TCE in the groundwater. Concentrations in tree cores were lower than the underlying groundwater, as anticipated; however, the tree core water retained the PCE:TCE signature of the underlying groundwater in the urban, populated area. The PCE:TCE ratio can indicate areas of differing degradation activity. Therefore, the phytoscreening analysis was capable not only of mapping the spatial distribution of groundwater contamination but also of delineating zones of potentially differing contaminant sources and degradation. The simplicity of tree coring and the ability to collect a large number of samples in a day with minimal disruption or property damage in the urban setting demonstrates that phytoscreening can be a powerful tool for gaining reconnaissance-level information on groundwater contaminated by chlorinated solvents. The use of SPME decreases the detection level considerably and increases the sensitivity of phytoscreening as an assessment, monitoring, and phytoforensic tool. With rapid, inexpensive, and noninvasive methods of detecting and delineating contaminants underlying homes, as in this case, human health can be better protected through screening of broader areas and with far faster response times.

High throughput screening of native species for tailings eco-restoration using novel computer visualization for plant phenotyping.

Historical hard-rock mine activities have resulted in nearly half a million mining-impacted sites scattered around the US. Compared to conventional remediation, (aided) phytostabilization is generally cost-effective and ecologically productive approach, particularly for large-scale sites. Native species act to maintain higher local biodiversity, providing a foundation for natural ecological succession. Due to heterogeneity of mine waste, revegetation strategies are inconsistent in approach, and to avoid failure scenarios, greenhouse screening studies can identify candidate plants and amendment strategies before scaling up. This greenhouse study aimed to concurrently screen a variety of native species for their potential to revegetate Cu/Pb/Zn mine tailings and develop a high throughput and non-destructive approach utilizing computer vision and image-based phenotyping technologies to quantify plant responses. A total number of 34 species were screened in this study, which included: 5 trees, 8 grasses, and 21 forbs and legumes. Most of the species tested were Missouri native and prairie species. Plants were non-destructively imaged, and 15 shape and color phenotypic attributes were extracted utilizing computer vision techniques of PlantCV. Compared to reference soil, all species tested were negatively impacted by the tailings' characteristics, with lowest tolerance generally observed in tree species. However, significant improvement in plant growth and tolerance generally observed with biosolids addition with biomass surpassing reference soil for most legumes. Accumulation of Cu, Pb, and Zn was below Domestic Animal Toxicity Limits in most species. Statistically robust differences in species responses were observed using phenotypic data, such as area, height, width, color, and 9 other morphological attributes. Correlations with destructive data indicated that area displayed the greatest positive correlation with biomass and color the greatest negative correlation with shoot metals. Computer visualization greatly increased the phenotypic data and offers a breakthrough in rapid, high throughput data collection to project site-specific phytostabilization strategies to efficiently restore mine-impacted sites.