Plants as effective bioindicators for heavy metal pollution monitoring.

This study investigated the bioindicator potential of Amaranthus retroflexus L., Plantago lanceolata L., Rumex acetosa L., and Trifolium pratense L. including the use of Lolium multiflorum L. as a reference species, for heavy metal pollution monitoring, in particular Zinc (Zn), Cadmium (Cd), Nickel (Ni), and Lead (Pb). Controlled heavy metal contamination was applied through irrigation with metal nitrate solutions two levels of contamination (low and high). The study also focused on analyzing heavy metals concentration in plant tissues and related physiological responses. Distinct physiological responses to heavy metal stress were observed among the investigated species, highlighting unique variations in their reactions. Hydrogen peroxide, malondialdehyde content, and enzymatic activities emerged as reliable indicators of plant stress induced by heavy metal solutions. P. lanceolata displayed elevated Zn concentrations in both roots and leaves (3271 ± 337 and 4956 ± 82 mg kg-1). For Pb, L. multiflorum and P. lanceolata showed highest root concentrations (2964 ± 937 and 1605 ± 289 mg kg-1), while R. acetosa had higher leaf concentration (1957 ± 147 mg kg-1). For Ni, L. multiflorum had the highest root concentration (1148 ± 93 mg kg-1), and P. lanceolata exhibited the highest leaf concentration (2492 ± 28 mg kg-1). P. lanceolata consistently demonstrated the highest Cd concentrations in both roots (126 ± 21 mg kg-1) and leaves (163 ± 12 mg kg-1). These results provide valuable insights for selecting effective bioindicator species to establish control strategies for heavy metal pollution.

Determination of Polycyclic Aromatic Hydrocarbon Content in Garden Herbal Plants Using Liquid Chromatographic Analysis (HPLC-FL).

Polycyclic aromatic hydrocarbons (PAHs) are a group of chemical compounds generated as a result of the incomplete combustion of fossil fuels or wood. PAHs are known for their negative effect on living organisms, including teratogenic, carcinogenic and mutagenic activity. The objective of this study is to determine the contamination of three popular herbal species showing pro-health properties, i.e., lavender, parsley and mint, with polycyclic aromatic hydrocarbons, collected from three different backyard gardens in Poland. The concentration of PAHs in plant material was determined by high-performance liquid chromatography with a fluorescence detector (HPLC-FL). The concentration of eleven PAHs in plant material was determined with high-pressure liquid chromatography after extraction using the QuEChERS purification technique. Mint collected within an area of a mining and energy production complex (the city of Konin) was characterized by the highest Σ of 11 PAHs, equaled to 902.35 µg/g FW, with anthracene being the most abundant compound. However, it contained the lowest sum of PAHs, among all tested plants, with high carcinogenicity. Parsley from the city of Poznan showed the highest content of benzo[a]pyrene (BaP), showing the strongest carcinogenicity, while the highest value of BaP equivalent was calculated for mint collected in Konin. The obtained results suggest that the level and profile of plant contamination with PAHs depend on the species and the location of herb cultivation. In particular, mining and energy industry facilities are sources of PAHs, which contaminate plant material for further direct use or as bioactive herbal extracts.

Copper, lead and zinc interactions during phytoextraction using Acer platanoides L.-a pot trial.

Of the many environmental factors that modulate the phytoextraction of elements, little has been learnt about the role of metal interactions. The study aimed to show how different concentrations of Cu, Pb and Zn in the cultivation medium influenced the biomass, plant development and phytoextraction abilities of Acer platanoides L. seedlings. Additionally, the impact on the content and distribution of Ca, K, Mg and Na in plant parts was studied with an analysis of phenols. Plants treated with a mixture of two metals were characterised by lower biomass of leaves and higher major elements content jointly than those grown in the salt of one element. Leaves of A. platanoides cultivated in Pb5 + Zn1, Pb1 + Zn1 and Pb1 + Zn5 experimental systems were characterised by specific browning of their edges. The obtained results suggest higher toxicity to leaves of Pb and Zn present simultaneously in Knop solution than Cu and Pb or Cu and Zn, irrespective of the mutual ratio of the concentrations of these elements. Antagonism of Cu and Zn concerning Pb was clearly shown in whole plant biomass when one of these elements was in higher concentration (5 mmol L-1) in solution. In the lowest concentrations (1 mmol L-1), there was a synergism between Cu and Zn in plant roots. Plants exposed to Zn5, Cu1 + Pb5, Pb5 + Zn1 and Cu1 + Zn1 were characterised by higher total phenolic content than the rest plants. Both the presence and the concentration of other elements in the soil are significant factors that modulate element uptake, total phenolic content, and plant development.

A Comprehensive Study of Lupin Seed Oils and the Roasting Effect on Their Chemical and Biological Activity.

The present investigation aimed to study the impact of roasting on the chemical composition and biological activities of sweet and bitter lupin seed oils. Lupin oils were extracted using petroleum ether (40-60) with ultrasonic assisted method. Lupin Fatty acids, phytosterols, carotenoids, and total phenolic contents were determined. In addition, antioxidant, antimicrobial, and antifungal activities were evaluated. The results showed a ratio between 7.50% to 9.28% of oil content in lupin seed. Unroasted (bitter and sweet) lupin oil contained a high level of oleic acid ω9 (42.65 and 50.87%), followed by linoleic acid ω6 (37.3 and 34.48%) and linolenic acid ω3 (3.35 and 6.58%), respectively. Concerning phytosterols, unroasted (bitter and sweet lupin) seed oil reflected high values (442.59 and 406.18 mg/100 g oil, respectively). Bitter lupin oil contains a high amount of phenolics, although a lower antioxidant potency compared to sweet lupin oil. This phenomenon could be connected with the synergistic effect between phenolics and carotenoids higher in sweet lupin oil. The results reflected a more efficiently bitter lupin oil against anti-toxigenic fungi than sweet lupin oil. The roasting process recorded enhances the antimicrobial activity of bitter and sweet lupin seed oil, which is linked to the increment in bioactive components during the roasting process. These results concluded that lupin oil deems a novel functional ingredient and a valuable dietary fat source. Moreover, lupin oil seemed to have antifungal properties, which recommended its utilization as a carrier for active-antifungal compounds in food products.

The impact of iron coagulant on the behavior and biochemistry of freshwater mussels Anodonta cygnea and Unio tumidus during lake restoration.

Iron (Fe) treatment is one of the most commonly used methods to restore eutrophic lakes and reservoirs. The Fe-based coagulants dosage results in an almost immediate improvement in water quality at a relatively low cost. However, the effects of the application of coagulants are not always predictable, and the scale of the risks is not fully understood. The dosage of coagulants changes the chemical and physical properties of water, thereby affecting aquatic biocenoses. In this study, several laboratory experiments were conducted to evaluate the effects of Fe-based coagulant dosage on two bivalves species: Anodonta anatina and Unio tumidus. Their ability to efficiently filter water and reduce seston makes them a key component of aquatic ecosystems in terms of maintaining proper ecological health and stable functioning. Behavioral response, biochemical parameters, and body chemistry changes in mussels exposed to different doses of coagulant were surveyed. A dose-dependent reduction in filtration activity of both species was observed. As early as 10 g Fe m2 (which is a moderate dose used in lakes restoration), mussels of both species almost completely reduced their filtration activity and remained with closed valves for several subsequent days. Significant Fe accumulation in muscles of bivalves exposed to coagulant was also observed. This was particularly the case when very high doses of coagulant were applied. Then, the iron content in leg muscles of both species increased over fourfold. At the same time, a decrease in muscles calcium and phosphorus content was observed. No symptoms of oxidative stress (TBARS, H2O2) after mussels exposure to coagulants were found. The results suggest that the application of Fe-based coagulant for water ecosystem restoration may be a threat to the mussels population. These findings are significant for decisions on the selection of restoration methods for a specific lake.

The Possibility of Using Paulownia elongata S. Y. Hu × Paulownia fortunei Hybrid for Phytoextraction of Toxic Elements from Post-Industrial Wastes with Biochar.

The potential of the Paulownia hybrid for the uptake and transport of 67 elements along with the physiological response of plants cultivated in highly contaminated post-industrial wastes (flotation tailings-FT, and mining sludge-MS) was investigated. Biochar (BR) was added to substrates to limit metal mobility and facilitate plant survival. Paulownia could effectively uptake and translocate B, Ca, K, P, Rb, Re and Ta. Despite severe growth retardation, chlorophyll biosynthesis was not depleted, while an increased carotenoid content was noted for plants cultivated in waste materials. In Paulownia leaves and roots hydroxybenzoic acids (C6-C1) were dominant phenolics, and hydroxycinnamic acids/phenylpropanoids (C6-C3) and flavonoids (C6-C3-C6) were also detected. Plant cultivation in wastes resulted in quantitative changes in the phenolic fraction, and a significant drop or total inhibition of particular phenolics. Cultivation in waste materials resulted in increased biosynthesis of malic and succinic acids in the roots of FT-cultivated plants, and malic and acetic acids in the case of MS/BR substrate. The obtained results indicate that the addition of biochar can support the adaptation of Paulownia seedlings growing on MS, however, in order to limit unfavorable changes in the plant, an optimal addition of waste is necessary.

Arsenic uptake, speciation and physiological response of tree species (Acer pseudoplatanus, Betula pendula and Quercus robur) treated with dimethylarsinic acid.

The aim of the study was to evaluate the effect of dimethylarsinic acid (DMA) on growth parameters and levels of stress-related metabolites in Acer pseudoplatanus, Betula pendula and Quercus robur. The increase of DMA concentration in the solution led to a notable growth retardation of trees. An intense As accumulation (mainly As(III) and As(V)) expressed as BCF and TF > 1 was recorded only for Q. robur. Generally a decrease in contents of cellulose, hemicellulose and holocellulose with a simultaneous increase in lignin content were recorded. Phenolic composition of leaf extracts was modified by DMA, while root and rhizosphere extracts were poor in phenolics. Toxicity of DMA leads to a significant drop in salicylic acid content in leaves observed at lower doses. Higher DMA levels caused a second, probably ROS-derived depletion of the metabolite accompanied with a severe growth retardation, most pronounced in the case of B. pendula. DMA caused the inhibition of LMWOA biosynthesis in roots of A. pseudoplatanus, B. pendula and their exudation into the rhizosphere, while in Q. robur roots and leaves a stimulation of their accumulation was observed. Disturbances in the activity of enzymatic antioxidants were observed for all the species following the increasing level of DMA.

Synergistic Impact of Bioactive Byproduct Extract Leads to Anti-Fusarium and Anti-Mycotoxin Secretion.

Fruit byproducts are considered a high source of bioactive molecules, which possess antioxidant activities. These antioxidants play principal functions in mycotoxin reduction. This study aimed to evaluate crude mandarin byproduct extract for its chemical interaction with fungal growth and suppression of mycotoxin production, and to illustrate whether the impact was regarding individual molecules or a synergistic antioxidation process. Extract contents were analyzed for their phenolic, flavonoids, and antioxidant activity. The fatty acid composition and volatile components were determined using the GC apparatus. The influence of the extract evaluated versus the standard phenolics of trans-ferulic and hesperidin were evaluated. The liposome technique was applied to prevent the antioxidant properties of the bioactive extract. The anti-mycotoxigenic effects of the liposomal and non-liposomal extract were determined in fungal media against the standard phenolics. The results manifested ferulic (235.54 ± 3.34 mg/100 g) and hesperidin (492.11 ± 1.15 mg/100 g) as high phenolics in the extract. Limonene was the main volatile (67.54 ± 1.74%), as well antioxidant activities determined in considerable values. The crude extract recorded efficiency as an anti-Fusarium agent, but less than the standard hesperidin applied in fungal media. The bioactive extract recorded possessed a reduction influence on mycotoxin production. The impact may be joining with its fungal inhibition or its component activity with the active groups on the mycotoxin molecule. The formation of liposomal extract enhanced its efficacy in mycotoxin reduction. This enhancement may illustrate its protective properties for antioxidant components of the bioactive extract.

Iron-induced behavioural and biochemical responses of charophytes in consequence of phosphates coagulant addition: Threats to lake ecosystems restoration.

The study aimed to evaluate the impact of iron (Fe) on the physiological and behavioural reaction of Chara tomentosa L. Fe was introduced into the environment in the form of iron chloride, the most common coagulants used in the restoration of water bodies. The investigations concerned the oxidative stress comprising phenolic compounds content, antioxidant activity and photosynthetic pigments concentration. Research was conducted as a laboratory microcosm experiment with one-off application of Fe at the level of 26.8 mg dm-3. Coagulant application caused short-term acidification, increased salinity and deterioration of light conditions. The shading resulted initially from the increase of water colour and turbidity and was followed by covering of the charophytes with a precipitated suspension. C. tomentosa did not activate defensive mechanisms to prevent the shading effect such as intensive elongation and elevated concentration of chlorophylls. Neither oxidative stress nor production of stress-specific phenolic metabolites was found. It was a result of iron coagulant toxicity, which led to cell membrane damage and leakage of cell contents to the water environment. Charophyte growth was significantly impaired, and thalli suffered numerous chlorotic and necrotic spots which extended gradually during experiment and finally caused death of specimens.

The Role of Sugars in the Regulation of the Level of Endogenous Signaling Molecules during Defense Response of Yellow Lupine to Fusarium oxysporum.

Soluble sugars such as sucrose, glucose and fructose in plant host cells not only play the role as donors of carbon skeletons, but they may also induce metabolic signals influencing the expression of defense genes. These metabolites function in a complex network with many bioactive molecules, which independently or in dialogue, induce successive defense mechanisms. The aim of this study was to determine the involvement of sucrose and monosaccharides as signaling molecules in the regulation of the levels of phytohormones and hydrogen peroxide participating in the defense responses of Lupinus luteus L. to a hemibiotrophic fungus Fusarium oxysporum Schlecht f. sp. lupini. A positive correlation between the level of sugars and postinfection accumulation of salicylic acid and its glucoside, as well as abscisic acid, was noted. The stimulatory effect of sugars on the production of ethylene was also reported. The protective role of soluble sugars in embryo axes of yellow lupine was seen in the limited development of infection and fusariosis. These results provide evidence for the enhanced generation of signaling molecules both by sugar alone as well as during the crosstalk between sugars and infection caused by F. oxysporum. However, a considerable postinfection increase in the level of these signaling molecules under the influence of sugars was recorded. The duration of the postinfection generation of these molecules in yellow lupine was also variable.

Differences of Acer platanoides L. and Tilia cordata Mill. Response patterns/survival strategies during cultivation in extremely polluted mining sludge - A pot trial.

The study aimed to evaluate the physiological mechanisms underlying differences in metals and metalloid uptake and tolerance of two tree species cultivated in mining waste material. Two-year old Acer platanoides L. and Tilia cordata Mill. were cultivated in mining sludge characterized by high pH, salinity and an extremely high concentration of As. Both species were able to develop leaves from leafless seedlings, however, their total biomass was greatly reduced in comparison to control plants, following the severe disturbances in chlorophyll content. Phytoextraction abilities were observed for T. cordata for Ba, Nb, Rb and Se, and phytostabilisation was stated for Pd, Ru, Sc and Sm for both species, Ba and Nd for A. platonoides and Be for T. cordata only. Metal exclusion was observed for the majority of detected elements indicating an intense limitation of metal transport to photosynthetic tissue. A diversified uptake of elements was accompanied by a species-specific pattern of physiological reaction during the cultivation in sludge. Organic ligands (glutatnione and low-molecular-weight organic acids) were suppressed in A. platanoides, and enhanced biosynthesis of phenolic compounds was observed for both species, being more pronounced in T. cordata. Despite its higher accumulation of key metabolites for plant reaction to oxidative stress, such as phenolic acids, flavonoids and organic ligands, T. cordata exhibited relatively lower tolerance to sludge, probably due to the increased uptake and translocation rate of toxic metal/loids to aerial organs and/or restricted accumulation of salicylic acid which is known to play a decisive role in mechanisms of plant tolerance.

Arsenic forms and their combinations induce differences in phenolic accumulation in Ulmus laevis Pall.

Total phenolics and the profile of phenolic acids and flavonoids were investigated in the roots and leaves of Ulmus laevis cultured on the medium with inorganic and organic arsenic - As(III), As(V) and DMA(V) at 0.06mM and their equimolar combinations. Further, the accumulation of salicylic acid (free and glucoside-bound) and lipid oxidation were assayed following a three-month long experiment. As treatment caused elevated production of phenolics, which was higher in photosynthetic tissue than in roots for all As forms and their combinations, and their overall content was correlated with the accumulation of organic As in roots and As(III) in leaves. The accumulation of organic As strongly induced shikimate-derived protocatechiuc acid in roots. Contrary to this, shikimate-derived phenolics (protocatechuic, gallic acids and 4-HBA) were suppressed in leaves, while the accumulation of C6C3 acids (caffeic, p-coumaric and chlorogenic) was stimulated by As(V) application. Surprisingly, these acids were not detected in the leaves of As(III)-treated plants, and mutually applied As(III) and DMA(V) reduced their content. DMA(V) negatively influenced the level of salicylic acid and its storage mechanism and this effect correlated with elevated MDA content in leaves. Quercetin accumulation was observed in both organs (mainly leaves) of DMA(V)-treated plants thereby proving its function in defensive response of Ulmus laevis to organic forms of As.

The Influence of Lead on Generation of Signalling Molecules and Accumulation of Flavonoids in Pea Seedlings in Response to Pea Aphid Infestation.

The aim of this study was to investigate the effect of an abiotic factor, i.e., lead at various concentrations (low causing a hormesis effect and causing high toxicity effects), on the generation of signalling molecules in pea (Pisum sativum L. cv. Cysterski) seedlings and then during infestation by the pea aphid (Acyrthosiphon pisum Harris). The second objective was to verify whether the presence of lead in pea seedling organs and induction of signalling pathways dependent on the concentration of this metal trigger defense responses to A. pisum. Therefore, the profile of flavonoids and expression levels of genes encoding enzymes of the flavonoid biosynthesis pathway (phenylalanine ammonialyase and chalcone synthase) were determined. A significant accumulation of total salicylic acid (TSA) and abscisic acid (ABA) was recorded in the roots and leaves of pea seedlings growing on lead-supplemented medium and next during infestation by aphids. Increased generation of these phytohormones strongly enhanced the biosynthesis of flavonoids, including a phytoalexin, pisatin. This research provides insights into the cross-talk between the abiotic (lead) and biotic factor (aphid infestation) on the level of the generation of signalling molecules and their role in the induction of flavonoid biosynthesis.

Copper and nickel co-treatment alters metal uptake and stress parameters of Salix purpurea×viminalis.

Simultaneous treatment of Salix purpurea×viminalis with copper (Cu2+) and nickel (Ni2+) altered metal phytoextraction rates in favor of leaves. Still, metal translocation patters remained unaffected (roots≈rods>>leaves≥shoots), reaching ∼20 and 14.5mgkg-1 dry weight in roots for Cu and Ni, respectively. Biometric parameters revealed overall growth inhibition correlated with Cu content in leaves, thus proving its negative effect on photosynthesis. Metal toxicity was strongly affirmed in the case of roots (∼90% loss of root biomass at 3mM), rather than in the above-ground organs. Plant treatment accelerated the accumulation of soluble carbohydrates, phenolics including salicylic acid and glutathione in Salix leaves. However, significant differences in plant reactions to the applied metals were noted. Metal accumulation in leaves was correlated with soluble sugars and elevated glutathione, and also with total phenolics content, in the case of Cu and Ni, respectively. Glutathione synthesis was induced by both metals, and correlated with salicylic acid in leaves of Ni-treated plants.

Differential induction of Pisum sativum defense signaling molecules in response to pea aphid infestation.

This study demonstrates the sequence of enhanced generation of signal molecules such as phytohormones, i.e. jasmonic acid (JA), ethylene (ET), salicylic acid (SA), and a relatively stable free radical, nitric oxide (NO), in response of Pisum sativum L. cv. Cysterski seedling leaves to the infestation of pea aphid Acyrthosiphon pisum (Harris) at a varied population size. In time from 0 to 96h after A. pisum infestation these signal molecules accumulated transiently. Moreover, the convergence of these signaling pathways occurred. JA and its methyl derivative MeJA reached the first maximum of generation at 24th hour of infestation. An increase in ET and NO generation was observed at 48th hour of infestation. The increase in SA, JA/MeJA and ET concentrations in aphid-infested leaves occurred from the 72nd to 96th hour. In parallel, an increase was demonstrated for the activities of enzymes engaged in the biosynthesis of SA, such as phenylalanine ammonia-lyase (PAL) and benzoic acid 2-hydroxylase (BA2H). Additionally, a considerable post-infestation accumulation of transcripts for PAL was observed. An increase in the activity of lipoxygenase (LOX), an important enzyme in the biosynthesis of JA was noted. This complex signaling network may contribute to the coordinated regulation of gene expression leading to specific defence responses.

Aluminum induces cross-resistance of potato to Phytophthora infestans.

The phenomenon of cross-resistance allows plants to acquire resistance to a broad range of stresses after previous exposure to one specific factor. Although this stress-response relationship has been known for decades, the sequence of events that underpin cross-resistance remains unknown. Our experiments revealed that susceptible potato (Solanum tuberosum L. cv. Bintje) undergoing aluminum (Al) stress at the root level showed enhanced defense responses correlated with reduced disease symptoms after leaf inoculation with Phytophthora infestans. The protection capacity of Al to subsequent stress was associated with the local accumulation of H2O2 in roots and systemic activation of salicylic acid (SA) and nitric oxide (NO) dependent pathways. The most crucial Al-mediated changes involved coding of NO message in an enhanced S-nitrosothiol formation in leaves tuned with an abundant SNOs accumulation in the main vein of leaves. Al-induced distal NO generation was correlated with the overexpression of PR-2 and PR-3 at both mRNA and protein activity levels. In turn, after contact with a pathogen we observed early up-regulation of SA-mediated defense genes, e.g. PR1, PR-2, PR-3 and PAL, and subsequent disease limitation. Taken together Al exposure induced distal changes in the biochemical stress imprint, facilitating more effective responses to a subsequent pathogen attack.

Salicylic acid - a potential biomarker of tobacco Bel-W3 cell death developed as a response to ground level ozone under ambient conditions.

Salicylic acid content and benzoic acid 2-hydroxylase (BA2H) activity were investigated in tobacco Bel-W3 and Bel-B leaves after exposure to tropospheric ozone in the conditions of ambient air. Plants were exposed in accordance with a standard methodology for ozone biomonitoring, in a three-year experiment. Free salicylic acid (SA), conjugated with glucose (SAG), and as a product of the BA2H activity was quantified with HPLC. In order to evaluate ozone injuries of leaves, an open source image analysis software was employed. Plants exposure to ambient ozone resulted in enhanced BA2H activity and intensified salicylic acid biosynthesis in leaves of Bel-W3 cultivar showing visible ozone injuries. The BA2H activity significantly correlated with SAG for ozone-exposed Bel-W3 plants. Both injuries and salicylic acid biosynthesis rate depended on the growth phase of leaves and nearly linear correlation between SA content and injuries was found for particular leaves of Bel-W3.

Changes in Salix viminalis L. cv. 'Cannabina' morphology and physiology in response to nickel ions--hydroponic investigations.

The aim of the study was to assess the ability of 'basket willow' to bioaccumulate nickel with simultaneous analysis of biomass parameters and biomarkers of plants physiological reaction to the metal. Cuttings of Salix viminalis L. cv. 'Cannabina' were cultivated in Knop's medium containing nickel at 0-3mM stabilized with quartz sand. Higher nickel contents were observed in Salix rods and roots (~15), and lower for leaves and shoots (~3 mg kg(-1)DW) after 14 days at 3 mM Ni. The strongest inhibition was observed for root biomass, weaker for shoot, root and leaf elongation, and the weakest for the photosynthetic area (4, 24, 36, 55 and 70% of control, respectively). Soluble carbohydrates in leaves reached 340% of control at 3.0mM Ni. Phenolics content increased four-fold at 3mM Ni versus control, while salicylic acid content at 2.5 mM Ni was nearly 68 times higher than for Ni-untreated plants. The exudation of low molecular weight organic acids increased from ~40 to 130 µM kg(-1)DW at 3 mM. The investigated cultivar exhibits sufficient resistance to nickel and can be cultivated at heavily contaminated sites. Nevertheless, strong inhibition of plant growth was observed resulting probably from Ni-induced disturbances in nutrient uptake accompanied by oxidative stress.

Free Radicals, Salicylic Acid and Mycotoxins in Asparagus After Inoculation with Fusarium proliferatum and F. oxysporum.

Electron paramagnetic resonance was used to monitor free radicals and paramagnetic species like Fe, Mn, Cu generation, stability and status in Asparagus officinalis infected by common pathogens Fusarium proliferatum and F. oxysporum. Occurrence of F. proliferatum and F. oxysporum, level of free radicals and other paramagnetic species, as well as salicylic acid and mycotoxins content in roots and stems of seedlings were estimated on the second and fourth week after inoculation. In the first term free and total salicylic acid contents were related to free radicals level in stem (P = 0.010 and P = 0.033, respectively). Concentration of Fe(3+) ions in porphyrin complexes (g = 2.3, g = 2.9) was related to the species of pathogen. There was no significant difference between Mn(2+) concentrations in stem samples; however, the level of free radicals in samples inoculated with F. proliferatum was significantly higher when compared to F. oxysporum.

SIPK signaling controls multiple components of harpin-induced cell death in tobacco.

Harpin from Pseudomonas syringae pv. phaseolicola (HrpZ) elicits a rapid cell death response in tobacco plants. Multiple signaling components, including mitogen-activated protein kinase (MAPK), reactive oxygen species (ROS) and salicylic acid (SA), have been reported to be involved in this cell death process, but the interaction between these molecules is poorly understood. Here we show through utilizing plants manipulated in SIPK expression levels that lack of SIPK results in increased sensitivity to harpin with concomitant accumulation of higher levels of ROS. Conversely, SIPK-overexpressing plants show reduced sensitivity to harpin relative to wild-type plants, and display reduced ROS accumulation. Harpin-induced cell death was found to be conditional on the ability of the plant to accumulate SA, whereas harpin induction of MAPK activation and ROS accumulation are not. However, harpin-induced ROS accumulation is required for activation of SIPK and wound-induced protein kinase. Transcriptional profiling revealed that suppression of SIPK signaling also affects early expression of a range of pathogen- and stress-responsive genes during harpin challenge.