Phytotoxicity Assessment of Biofertilizer Produced from Bioreactor Composting Technology Using Lettuce (Lactuca sativa L.) Seeds

Establishing reliable technological information on the safety of biofertilizers produced from a bioreactor composting technique is a must prior to its commercialization. A phytotoxicity study of biofertilizer made from the bioreactor composting technology at Aklan State University, Banga, Aklan, Philippines was conducted for fourteen (14) days using commercially available lettuce seeds (Lactuca sativa L.). Standard phytotoxicity attributes such as hypocotyl length, radicle length, relative germination percentage, and relative radicle growth observed during the germination stage were evaluated. Results revealed no significant difference in the radicle lengths of the germinated lettuce seeds as affected by the varying levels of biofertilizer dilution at H(3) = 10.567, p = 0.061 > 0.05. On the other hand, the hypocotyl length of the lettuce showed significant differences in response to varying levels of biofertilizer dilution with Welch’s F(5, 5.163) = 8.175, p = 0.017 < 0.05. Also, the different levels of biofertilizer affected significantly the germination percentage of lettuce seeds F(5, 12) = 5.822, p = 0.006 < 0.05. All levels of biofertilizer treatments indicated a decrease in relative germination percentage. However, those seeds applied with 10% biofertilizer have the highest reduction of germination percentage, equivalent to 86.9% (RGP = 13.10%). All levels of biofertilizer showed an increase in radicle growth in contrast to the negative control plant except for the one given a 10% level of biofertilizer. Seeds that received 10% biofertilizer showed an extremely high reduction in radicle growth, equivalent to 72.22% (RRG = 27.78%). The study shows that applying low levels of the bioreactor-produced biofertilizer will observably reduce the measure of the germination characteristics of lettuce seeds, but not necessarily low enough to be considered phytotoxic. However, the application of at least 10% bioreactor-produced biofertilizer can presumptively lead to phytotoxicity.

Phytotoxicity assessment of phenanthrene, pyrene and their mixtures by a soil-based seedling emergence test

Seedling emergence tests were conducted in a meadow brown soil using five plant species(i.e., Chinese cabbage, green onion, tomato, turnip and wheat) to determine the phytotoxicity of phenanthrene, pyrene and their mixtures. The soil was amended with up to 1000 mg/kg soil of phenanthrene or 600 mg/kg soil of pyrene. Seedling emergence and root growth were measured. The results indicated that root elongation was more sensitive than seedling emergence. Root length decreased with increasing phenanthrene or pyrene concentrations(p≤0 05). Phenanthrene was more phytotoxic than pyrene. The lowest observable adverse effect concentration(LOAEC) was 10 mg/kg for phenanthrene when tested with green onion, and 50 mg/kg for pyrene when tested with wheat. Among the five species, wheat was found to be the most sensitive. When amended jointly at or below their respective LOAEC, phenanthrene and pyrene produced a synergistic toxic effect.

Bioavailability of bound residue derived from ^(14)C-labeled chlorsulfuron in soil and its mechanism of phytotoxicity

The bioavailability of bound residue(BR) derived from 14 C-labeled chlorsulfuron in soil and effect of the main components of the BR on growth of rape(brassica napus) and rice(Oryza sativa L.) were investigated. The results showed that the BR with the concentration of 0 28 and 0 56 nmol/g air-dried soil, which was calculated by special radioactivity of 14 C-labeled chlorsulfuron parent compound, resulted in significant depression effect on growth of rape seedling. It was assured that the main components(2-amino-4-methoxyl-6-methyl-1,3,5-triazine, 2-amino-4-hydroxyl-6-methyl-1,3,5-triazine, and 2-chloro-benzenesul-f onamide) of the BR did not inhibit the growth of rape and rice. LC-MS analysis demonstrated that the parent compound previously bound to the soil matrix could be again released and transformed into methanol-extractable residue during the course of rape growth. It was concluded that the molecular leading to the phytotoxicity to rape and rice in the BR is still the parent compound.

Alleviation of Soil Acidity and Aluminium Phytotoxicityin Acid Soils by Using Alkaline-Stabilized Biosolids

A pot experiment was catried out to study alleviation of soil acidity and Al toxicity by applying analkaline-stabilised sewage sludge product (biosolids) to an acid clay sandy loam (pH 5.7) and a strongly acidsandy loam (pH 4.5). Barley (Hondeum vulgare L. cv. Forrester) was used as a test crop and was grownin the sewage sludge-amended (33.5 t sludge DM ha-1) and unamended soils. The results showed that thealka1ine biosloids increased soil pH from 5.7 to 6.9 for the clay sandy loam and from 4.5 to 6.0 for the sandyloam. The sludge product decreased KCl-extractable Al from 0.1 to 0.0 cmol kg-1 for the former soil andfrom 4.0 to 0.1 cmol kg-1 for the latter soil. As a result, barley plants grew much better and grain yieldincreased greatly in the amended treatments compared with the unamended controls. These observationsindicate that alkaline-stabilised biosolids can be used as a liming material for remedying Al phytotoxicity instrongly acid soils by increasing soil pH and lowering Al bioavailability.

Phytotoxicity of four herbicides on Ceratophyllum demersum,Vallisneria natans and Elodea nuttallii

The physiological effects of 4 herbicides （butachlor, quinclorac, bensulfuron-methyl and atrazine） on 3 submerged macrophytes （Ceratophyllum demersum, Vallisneria natans and Elodea nuttallii） were tested in laboratory. The variables of the relative growth rate and the photosynthetic pigment content showed that all of the tested herbicides affected the growth of the plants obviously, even at the lowest concentration （0.0001 mg/L）. Except for the C. demersum treated with quinclorac at 0.005 and 0.01 mg/L, the relative growth rates of the plants were inhibited significantly （p 〈 0.01）. Statistical analysis of chlorophyll a （Chl-a） contents was carded out with both the t-test and one-way ANOVA to determine the difference between the treatment and control. The results showed that Chl-a contents of the plants in all treatment groups were affected by herbicides significantly, except for the C. demersum treated with bensulfuron-methyl at 0.0005 mg/L. The decrease in Chl-a content was positively correlated to the dosage of the herbicides in most treatment groups. It was suggested that herbicides in water bodies might potentially affect the growth of aquatic macrophytes. Since the Chl-a content of submerged macrophytes responded to the stress of herbicides sensitively and directly, it could be used as a biomaker in environmental monitoring or in the ecological risk assessment of herbicide contamination.

Effects of two sludge application on fractionation and phytotoxicity of zinc and copper in soil

The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% （by weight） of digested sewage sludge （SS） and composted sludge （SC）. The additions of the both sludges caused a significant raise in all fractions, resulting in that exchangeable （EXCH） and organic matter （OM） became predominance for Zn and organic bound Cu occupied the largest portion. There was more available amount for Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate （CAR） and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to AEXCH and ACAR forms but also in AOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by AEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.

Phytotoxicity of Chromium on Germination, Growth and Biochemical Attributes of <i>Hibiscus esculentus</i>L.

Chromium is found in all phases of the environment, including air, water and soil. The contamination of environment by chromium has become a major area of concern. Chromium effluent is highly toxic to plant and is harmful to their growth and development. In present study, a pot experiment was carried out to assess the phytotoxicity of chromium in Hibiscus esculentus at different concentration (0.5, 2.5, 5, 10, 25, 50 and 100 mg·kg-1) of chromium metal. The phytotoxic effect of chromium was observed on seed germination, seedling growth, seedling vigor index, chlorophyll content and tolerance indices of Hibiscus esculentus. All results when compared with control show that chromium metal adversely affects the growth of Hibiscus esculentus by reducing seed germination and decreasing seedling growth. The toxic effects of chromium metal to seed germination and young seedling are arranged in order of inhibition as: 0.5 > 2.5 > 5 > 10 > 25 > 50 > 100 mg·kg-1 respectively. The toxicity of chromium metal to young seedling and their effects on chlorophyll content were increased with higher concentration of chromium in the soil system. The major inhibitory effect of chromium in Hibiscus esculentus seedling was determined as stress tolerance index (%). The present study represents that the seed and seedling of Hibiscus esculentus has potential to counteract the deleterious effects of chromium metal in soil.

Correlation of Quantitative Structure and Inhibition Phytotoxicity for Aromatic Compounds Using Ab Initio Method

29 aromatic compounds were computed at the HF/6-31G^＊ level. Based on linear solvation energy theory firstly, the parameters of molecular structure were taken as theoretical descriptors, and the corresponding linear solvation energy relationship （LSER） （r^2= 0.8993, q^2=0.8559） between the structural parameters and inhibition phytotoxicity to the seed germination rate of cucumis （-lgGC50） was thus obtained. Then the parameters of molecular structure and thermodynamics were taken as theoretical descriptors, and as a result the other corresponding correlation equation （r^2=0.9268, q^2=0.8960） relating to -lgGC50 was achieved. The two equations obtained in this work by HF/6-31G^＊ are both more advantageous than that from AM 1.

Alleviation Effect of Trace Elements Application on Biuret Phytotoxicity of Maize

A pot experiment was carried out to study the effect of trace elements on the biuret toxicity of maize, with the aim to provide theoretical basis for improving the urea-based compound fertilizer production and reducing the biuret toxicity. The results showed that the compound fertilizer with 2% and 4% biuret caused obvious toxicity on the maize. The plant became short, and the biomass decreased, and the toxicity phenomenon of maize would become more obvious when the biuret leve increased. Application of 5% EDTA-Zn and 0.4% B could reduce the toxicity of bi uret in fertilizers. The toxicity would be eliminated completely when the level of biuret was 2% in compound fertilizers, while the toxicity effects of 4% biuret was just alleviated to a certain extent. In addition, application of 2% amino acids with ureabased compound fertilizer could promote root absorption of biuret, and increase the biuret toxicity. In conclusion, the urea compound fertilizers with high biuret concentrations should be applied with appropriate trace elements to reduce the biuret dam age. It was suggested that the biuret content in the complex fertilizers for maize should be controlled below 2%, and appropriate trace elements should be applied at the same time.

Chemical Basis for the Phytotoxicity of Indoles in Relation to the Allelopathy of Cereals

Phytotoxic activity of an indole series substituted with electron-acceptor and electron-donor groups in the aromatic ring was determined. They are potential decomposition products, of natural indole alkaloids in cereals plants with allelopathic properties. Phytotoxic selectivity was evaluated from antialgal activity against the microalga Chlorella vulgaris, seed germination seeds and biomass weight of seedling of barley, rye, wheat, oat and maize species and the weed Lolium rigidum. Lipophilia character of the compounds was determined by RP-HPLC method. Both, the electronic character of the substituents, evaluated from σp parameter, and the lipophilia character of the molecules measured from logPHPLC parameter, are involved in the phytotoxic activity. The three bio indicators has shown that the compounds with the higher electron-acceptor groups showed the higher level of phytotoxicity and the molecules with electron-donor groups showed the lowest activity, although, in some cases, this behavior is modified by the lipophilic properties of the molecules. These results are rationalized in terms of polarization of N-H bond of heterocyclic ring. Compounds with the higher logPHPLC values showed the higher phytotoxic activity. Further evidence on the role of lipophilicity was obtained from linear regression between the average inhibitions percentages of biomass and logPHPLC values. The activity increased linearly by increasing the lipophilic character of the compounds. Therefore, quantitative effects in the phytotoxic activity of the electronic properties of the substituents in the aromatic ring and lipophilic character of the indoles can be inferred from σp and logPHPLC parameters, respectively. The results strongly suggest that the potential decomposition products of the natural indole alkaloids from cereals or other natural sources may be in connection with the allelopathic phenomenon of plants when are released into the soil.

Effects of zeolite imidazole frameworks on rice seedlings (Oryza sativa L.): Phytotoxicity, transformation, and bioaccumulation

Zeolite imidazole frameworks(ZIFs),a class of the metal organic framework,have been extensively studied in environmental applications.However,their environmental fate and potential ecological impact on plants remain unknown.Here,we investigated the phytotoxicity,transformation,and bioaccumulation processes of two typical ZIFs(ZIF-8 and ZIF-67)in rice(Oryza sativa L.)under hydroponic conditions.ZIF-8 and ZIF-67 in the concentration of 50 mg/L decreased root and shoot dry weight maximally by 55.2%and 27.5%,53.5%and 37.5%,respectively.The scanning electron microscopy(SEM)imaging combined with X-ray diffraction(XRD)patterns revealed that ZIFs on the root surface gradually collapsed and transformed into nanosheets with increasing cultivation time.The fluorescein isothiocyanate(FITC)labeled ZIFs were applied to trace the uptake and translocation of ZIFs in rice.The results demonstrated that the transformed ZIFs were mainly distributed in the intercellular spaces of rice root,while they cannot be transported to culms and leaves.Even so,the Co and Zn contents of rice roots and shoots in the ZIFs treated groups were increased by 1145%and 1259%,145%and 259%,respectively,compared with the control groups.These findings suggested that the phytotoxicity of ZIFs are primarily attributed to the transformed ZIFs and to a less extent,the metal ions and their ligands,and they were internalized by rice root and increased the Co and Zn contents of shoots.This study reported the transformation of ZIFs and their biological effectiveness in rice,highlighting the potential environmental hazards and risks of ZIFs to crop plants.

Mitigating phytotoxicity of tetracycline by metal-free 8-hydroxyquinoline functionalized carbon nitride photocatalyst

Photooxidative removal of pharmaceuticals and organic dyes is an effective way to eliminate growing micropollutants. However, photooxidation often results in byproducts as secondary hazardous substances such as phytotoxins. Herein, we found that photooxidation of common antibiotic tetracycline hydrochloride(TCH) over a metal-free 8-hydroxyquinoline(8-HQ) functionalized carbon nitride(CN) photocatalyst significantly reduces the TCH phytotoxic effect. The phytotoxicity test of photocatalytic treated TCH-solution evaluated towards seed growth of Cicer arietinum plant model endowed natural root and shoot growth.This study highlights the conceptual insights in designing of metal-free photocatalyst for environmental remediation.

Effects of persulfate-assisted hydrothermal treatment of municipal sludge on aqueous phase characteristics and phytotoxicity

Hydrothermal technology (HT) has received much attention in recent years as a process to convert wet organic waste into hydrochar.The aqueous phase (HTAP) produced by this process is still a burden and has become a bottleneck issue for HT process development.In this study,we provide the?rst investigation of the HTAP characteristics,phytotoxicity,and their correlation with persulfate (PS)(PS,2.0 mmol/g TS)-assisted municipal sludge HT.The results showed that PS accelerated the hydrolysis of protein substances and increased the concentration of NH_(4)^(+)by 13.4%to 190.5%and that of PO_(4)^(3-)by 24.2%to 1103.7%in HTAP at hydrothermal temperatures of 120 to 240℃.PS can reduce the phytotoxicity of HTAP by reducing aldehydes,ketones,N heterocyclic compounds,and particle size and by increasing its humi?cation index.The maximum values of the root length and biomass of pakchoi(Brassica chinensis L.) seedlings occurred when electrical conductivity was 0.2 mS/cm of HTAP.This work provided a new strategy for the selection and design of HTAP management strategies.

Phytotoxicity in seven higher plant species exposed to di-n-butyl phthalate or bis （2-ethylhexyl） phthalate

We investigated phytotoxicity in seven plant species exposed to a range of concentrations （0- 500 mg· kg^-1 soil） of di-n-butyl phthalate （DnBP） or his （2- ethylhexyl） phthalate （DEHP）, two representative phthalate esters （PAEs） nominated by USEPA as priority pollutants and known environmental estrogens. We studied seed germination, root elongation, seedling growth, biomass （fresh weight, FW） and malondialdehyde （MDA） content of shoots and roots of wheat （Triticum aestivum L.）, alfalfa （Medicago sativa L.）, perennial ryegrass （Lolium perenne）, radish （Raphanus sativus L.）, cucumber （Cucumis sativus L.）, oat （Avena sativa） and onion （Allium cepa L.）, together with monitoring of plant pigment content （chlorophyll a, b and earotinoids） in alfalfa, radish and onion shoots. Root elongation, seedling growth and biomass of the test species were generally inhibited by DnBP but not by DEHP, indicating a lower level of phytotoxicity of DEHP than of DnBP. MDA contents of four species were promoted by PAE exposure, but not in alfalfa, ryegrass or onion shoots, indicating lower sensitivity of these three species to PAE pollutants. Plant pigment contents were clearly affected under the stress of both pollutants, implying the potential damage to the photosynthetic system of test plants, mainly by decreasing the content of chlorophyll a and b. Results of DnBP and DEHP phytotoxicity to the primary growth of test plants has provided information for the assessment of their environmental risk in the soil and also forms a basis for the further analysis of their toxic effects over the whole growth period of different plant species.

Kinetic study of phytotoxicity induced by foliar lead uptake for vegetables exposed to fine particles and implications for sustainable urban agriculture

At the global scale, foliar metal transfer occurs for consumed vegetables cultivated in numerous urban or industrial areas with a polluted atmosphere. However, the kinetics of metal uptake, translocation and involved phytotoxicity was never jointly studied with vegetables exposed to micronic and sub-micronic particles（PM）. Different leafy vegetables（lettuces and cabbages） cultivated in RHIZOtest？ devices were, therefore, exposed in a greenhouse for 5, 10 and 15 days to various Pb O PM doses. The kinetics of transfer and phytotoxicity was assessed in relation to lead concentration and exposure duration. A significant Pb accumulation in leaves（up to 7392 mg/kg dry weight（DW） in lettuce） with translocation to roots was observed. Lead foliar exposure resulted in significant phytotoxicity, lipid composition change, a decrease of plant shoot growth（up to 68.2% in lettuce） and net photosynthesis（up to 58% in lettuce）. The phytotoxicity results indicated plant adaptation to Pb and a higher sensitivity of lettuce in comparison with cabbage. Air quality needs, therefore, to be considered for the health and quality of vegetables grown in polluted areas, such as certain megacities（in China, Pakistan, Europe, etc.） and furthermore, to assess the health risks associated with their consumption.

The effects of temperature on decomposition and allelopathic phytotoxicity of boneseed litter

Decomposition of plant litter is a fundamental process in ecosystem function, carbon and nutrient cycling and, by extension, climate change. This study aimed to investigate the role of temperature on the decomposition of water soluble phenolics（WSP）, carbon and soil nutrients in conjunction with the phytotoxicity dynamics of Chrysanthemoides monilifera subsp. monilifera（boneseed） litter. Treatments consisted of three factors including decomposition materials（litter alone, litter with soil and soil alone）, decomposition periods and temperatures（5-15, 15-25and 25-35°C（night/day））. Leachates were collected on 0, 5, 10, 20, 40 and 60 th days to analyse physico-chemical parameters and phytotoxicity. Water soluble phenolics and dissolved organic carbon（DOC） increased with increasing temperature while nutrients like SO-24 and NO-13 decreased. Speed of germination, hypocotyl and radical length and weight of Lactuca sativa exposed to leachates were decreased with increasing decomposition temperature. All treatment components had significant effects on these parameters. There had a strong correlation between DOC and WSP, and WSP content of the leachates with radical length of test species. This study identified complex interactivity among temperature, WSP, DOC and soil nutrient dynamics of litter occupied soil and that these factors work together to influence phytotoxicity.

Insights on mechanisms of aluminum phytotoxicity mitigation by canola straw biochars from different regions

To better understand the amendment effects and mechanisms of aluminum(Al(Ⅲ))phytotoxicity mitigation by differ-ent regional crop straw biochars,wheat seedling root elongation trials were conducted.The contributions of liming effect,oxygen-containing surface functional group adsorption,and oxyanions precipitation to Al(Ⅲ)phytotoxicity mitigation by Ca(OH)_(2),pristine and ash-free canola straw biochar were evaluated.The results indicated that biochars derived from canola straw collected from four different regions(Yingtan,Xuancheng,Nanjing,and Huaiyin)caused 22-70%wheat seedling root elongation,which might be linked to liming effect.Incorporation of the corresponding ash-free biochars caused 15-30%elongation,which could be attributed to the surface functional group adsorption.About 0-60%of changes could be explained by Al(Ⅲ)precipitation with inorganic oxyanions.These findings provide new insights into the physicochemical properties,potential applications,efficiencies,and underlying mechanisms of crop straw biochar in alleviating Al(Ⅲ)phytotoxicity,which is dependent on the cultivation soil,and indicate similar application of crop straw biochar for acidic soil amelioration,contaminated soil remediation,and arable soil improvement.

Phytotoxicity and groundwater impacts of leaching from thermal treatment residues in roadways

The use of coal fly ash（CFA）, municipal solid waste incinerator bottom ash（MSWIBA） and flue gas desulfurization residue（FGDR） in road construction has become very common owing to its economical advantages. However, these residues may contain toxic constituents that pose an environmental risk if they leach out and flow through the soil, surface water and groundwater.Therefore, it is necessary to assess the ecotoxicity and groundwater impact of these residues before decisions can be made regarding their utilization for road construction. In this study,the physico-chemical characteristics, leaching and phytotoxicity of these residues were investigated. Specifically, multivariate analyses were used to evaluate the contributions of the leaching constituents of the CFA, MSWIBA and FGDR leachates to the germination index of wheat seeds. B, Ba, Cr, Cu, Fe and Pb were found to be more toxic to the wheat seeds than the other heavy metals. Furthermore, the leached concentrations of the constituents from the CFA, MSWIBA and FGDR were below the regulatory threshold limits of the Chinese identification standard for hazardous wastes. Analyses conducted using a numerical groundwater model（Wisc LEACH） indicated that the predicted field concentrations of metals from the CFA, MSWIBA and FGDR increased with time up to about 30 years at the point of compliance, then decreased with time and distance. Overall, this study demonstrated that the risks resulting from MSWIBA, CFA and FGDR leaching could be assessed before its utilization for road construction, providing crucial information for the adoption of these alternative materials.

A Preliminary Assessment of the Effect of Urban Waste Pollution in the Korle Lagoon Area of Accra, Ghana, on Nutrition and Growth of Maize (Zea mays L.) Plants

Disposal of domestic and industrial waste into the Odaw River and Korle Lagoon in Accra, Ghana, has led to pollution of the lagoon and surrounding soils. This study compared the elemental concentrations of heavy metals (lead and zinc) and essential plant macronutrients (nitrogen, phosphorus and potassium) in soils from the Korle Lagoon Area to those in baseline soils from the University of Ghana Agricultural Farm (UG Farm), also in Accra. A comparative pot experiment, using maize (Zea mays L.) as test plant, was conducted to assess the effect of each soil type on plant growth. Soil samples from the Korle Lagoon Area were significantly higher (P < 0.001) in lead (Pb), zinc (Zn), nitrogen (N) and potassium (K) concentrations than samples from the UG Farm. Mean plant height and mean leaf width of maize plants harvested 7 weeks after planting (WAP) were both significantly higher (P < 0.003) for samples from the Korle Lagoon Area soil compared to those from the UG Farm soil. Higher accumulation of Zn (448.3 ± 45.5) in maize shoot within 3 weeks of planting in the Korle Lagoon Area soil had phytotoxic effects on growth, resulting in shoot growth inhibition and reduced uptake of P and K in 11-week-old plants. The findings indicate that though nutrient enrichment due to sewage disposal into the lagoon may increase crop production, the nutritional quality of the crop produced could be compromised by heavy metal accumulation in the soil and subsequent uptake by the plant.

The effect of ozone on pine forests in South-Eastern France from 2017 to 2019

In South-Eastern forests of France,risks linked to the effects of tropospheric ozone(O_(3))are real;its annual impact has been observed specifically near the coastline and in high altitude mountains during the period 2017-2019.In this study,the risk assessment of O_(3)pollutant was carried out using two approaches based on forest response indicators such as O_(3)specific foliar visible injury and by stomatal O_(3)flux.Phytotoxic O_(3)dose values(POD_(0))were obtained by the DO_(3)SE model.The model requires hourly O_(3)concentration for POD_(0)calculation.A modified approach that uses measurements from passive samplers(monthly average O_(3)concentration)was tested for the calculation of POD_(0)and test results showed good agreement with the POD_(0)calculated using hourly O_(3)data.In the model input file,the average O_(3)concentration is used for POD_(0),and this could be useful for POD_(0)calculation when the active monitor is limited.In this study,a flux-based assessment provided better correlation with O_(3)specific leaf injury,which is also species-specific.Foliar visible injury in response to O_(3)indicates that Pinus cembra and Pinus halepensis are more affected and therefore more sensitive than Pinus sylvestris.The POD_(0)and stomatal conductance(Gsto)seem to be induced by environmental factors,primarily rainfall and the soil water potential(fSWP).The correlation between the O_(3)flux metric and environmental variables with forest response indicators by Spearman rank test confirms P.cembra as one of the most sensitive species to O_(3).