Electron paramagnetic resonance studies of manganese toxicity,tolerance, and amelioration with silicon in snapbean

Plant genotypes within species differ widely in tolerance to excess manganese (Mn) that may occur in acid soils, or in neutral or alkaline soils having poor aeration caused by imperfect drainage or compaction. However, Mn tolerance mechanisms in plants are largely unknown. Silicon (Si) is reported to detoxify Mn within plants, presumably by preventing localized accumulations of Mn associated with lesions on leaves. Because Mn is paramagnetic, electron paramagnetic resonance (EPR) spectroscopy, shows promise as a tool for characterizing toxic and non‐toxic forms of Mn in tolerant and sensitive plants. The objective of our study was to use EPR to: i) determine the chemical/ physical state of Mn in Mn‐tolerant and ‐sensitive snapbean cultivars; and ii) characterize the protective effects of Si against Mn toxicity. Manganese‐sensitive Wonder Crop 2 (WC) and Mn‐tolerant Green Lord (GL) cultivars of snapbean were grown at pH 5.0, in a greenhouse, in a modified Steinberg solution containing: Mn=0.05mg.L^‐1 (optimal); Mn=1.0mgL^‐1 (toxic); Mn=1.0 mg L^‐1 plus Si=4 mg L^‐1; and Mn=0.05 mg L^‐1 plus 4 mg Si L^‐1. All trifoliate leaf samples exhibited a 6‐line EPR signal that is characteristic of hexaaquo Mn²⁺. In both cultivars, a higher EPR Mn²⁺ signal‐intensity generally correlated with lower total leaf mass, higher total Mn concentrations and more pronounced symptoms of toxicity. Tolerance to excess Mn coincided with lower Mn²⁺ signal intensity. Silicon treatments ameliorated Mn toxicity symptoms in both genotypes, decreased total leaf Mn concentrations, and decreased EPR Mn²⁺ signal intensity. Results suggest that Mn toxicity is associated with reduced electron transport and accumulation of oxidation products in leaves. Amelioration of Mn toxicity by Si is regarded as connected with a reduction in this Mn‐induced process. Results indicated that EPR spectroscopy can be useful in investigating the biochemical basis for differential Mn tolerance in plants. The EPR observations might also help plant breeders in developing Mn‐tolerant cultivars.     

Leaf position and genotype differences for mineral element concentrations in sorghum grown on tropical acid soil 1

Mineral element deficiencies and toxicities are common problems associated with sorghum [Sorghum bicolor (L.) Moench] production on acid soils. To better understand some of the mineral element problems and the analysis of plant tissue of sorghum plants grown on acid soils, four sorghum genotypes were grown on an acid Oxisol at Carimagua, Colombia limed with dolomite at 2 and 6 Mg ha^‐1.

Samples for mineral element analyses were obtained from leaves at different positions on the four genotypes. Concentrations of P and Mg were highest in the flag leaf (Leaf No. 1) and decreased as the position on the plant declined from the top of the plant for plants grown at 2 Mg lime ha^‐1. Similar decreases in P, Mg, K, and Zn concentrations occurred in plants grown with 6 Mg lime ha^‐1. Concentrations of Ca, S, Si, Mn, Fe, Cu, and Al increased as leaf position declined from the flag leaf for plants grown at 2 and 6 Mg lime ha^‐1. The higher lime supply enhanced Ca and reduced Mn and Fe concentrations in leaves. Differences in mineral element concentrations for the four genotypes used were fairly extensive. The elements to show the greatest range among genotypes were Al and Si and the elements to show the least range among genotypes were P, K, and S. Care should be used in collecting leaf samples for plant analysis and genotypic differences for accumulation of mineral elements should be considered in interpretation of results.     

Manganese toxicity alleviated by mycorrhizae in soybean

Abstract

Soybean (Glycine max (L.) Merr.) plants nodulated with Bradyrhizobium japonicum. Nitragin strain 61A118, were grown with or without the vesicular‐arbuscular mycorrhizal (VAM) fungus Glomus mosseae (Nicol. Gerd.) Gerd. and Trappe in pot cultures in soil high (40.4 μg/g) in available Mn. Leaves of the nonVAM plants showed severe symptoms of Mn toxicity and had toxic (314 μg/g) concentrations of Mn in the foliage. NonVAM plants had significantly lower dry weights and nodule mass than VAM plants. Concentrations of Mn in the VAM plants were significantly (P<0.05) lower than in the nonVAM plants, and there were no symptoms of Mn toxicity. Both VAM and nonVAM plants had a significant negative correlation between shoot dry mass and leaf Mn concentration. Since levels of Mn increased with increasing VAM‐fungal colonization, we conclude that it was not the VAM condition per se which alleviated Mn toxicity. We suggest that the significantly higher levels of Mn in the leaves (P<0.05) and the roots (P<0.001) of nonVAM plants was due to increased uptake of Mn by the nonVAM plants. This exudation, which are generally observed in nonVAM plants, and to the role of such exudates in solubilizing MnO₂ and chelating the resulting Mn²? for facilitated absorption.     

The relationship of soil and leaf nutrients to rice leaf oranging

A symptom called leaf‐oranging, indicating a deficiency of many nutrients, occurs in paddy rice (Oryzasativa L.) when production expands into some upland soils. Rice (Gui Chou cv.) was grown in culture pots in a flooded, weathered, upland soil (Nacogdoches) and compared to rice growth in a flooded soil currently used for paddy rice production (Dacosta) in Texas to understand the soil and plant factors involved in leaf‐oranging. Fertilizer rates of 0, 10, and 100 mg N/kg as (NH₄)₂SO₄ were applied to each soil along with phosphorus (P) and potassium (K) fertilizer. The orange Leaf Index (OLI), a measure of leaf‐oranging, was determined weekly and increased to 60–70% for plants grown in the upland soil but its progression was delayed by higher N treatments. No leaf‐oranging was observed in the paddy soil. The soil evoking leaf‐oranging was low in silicon (Si) and high in iron (Fe). In addition, analysis of leaves from these plants showed 19–25% higher leaf ammonium‐nitrogen (NH₄‐N), 9–137% higher manganese (Mn) levels and lower total N:NH₄ concentration compared to normal rice leaves four weeks after transplanting. This inferred that leaf‐oranging probably was associated with some degree of NH₄‐N toxicity and antagonism with K. Leaf‐oranging was also associated with low calcium (Ca) assimilation or Ca uptake inhibition because of the heavy Fe‐oxide coating of the roots of the affected rice plants. In this experiment, leaf‐oranging was not associated with toxic levels of Fe or Mn.     

Differential manganese tolerances of cotton genotypes in nutrient solution

Cotton genotypes [Gossypium hirsutum (L.)] C‐310–73,‐307 (307) and C‐Sgl, 70–517 (517), shown previously to differ in tolerance to an acid (pH 5.1), high manganese (Mn) Grenada soil from Arkansas, were grown in nutrient solutions containing variable concentrations of excess Mn to confirm and characterize their postulated differences in Mn tolerance. Based on crinkle leaf symptoms and leaf dry weights, the 307 genotype was significantly more tolerant than 517 to 4, 8, or 16 mg Mn/L at a maintained pH of 4.6 (Experiment 1) and also to 4 or 8 mg Mn/L at an initial pH of 5.0, not subsequently adjusted (Experiment 2). In Experiment 1, the relative leaf dry weight (wt. with no Mn/wt. with 8 mg Mn/L × 100) was 94% for genotype 307 and only 27% for 517. In Experiment 2, the corresponding relative leaf weights were 75% and 26% for 307 and 517, respectively. Plant analytical results indicated that the 307 genotype tolerates a higher concentration of Mn in its leaves than does 517. This failure to correlate Mn tolerance with Mn concentrations in plant shoots agrees with previous findings when these two genotypes were grown in acid Grenada soil. Iron (Fe) concentrations, Fe/Mn ratios, and magnesium (Mg) concentrations were higher in the Mn‐tolerant 307 than in the Mn‐sensitive 517, but concentrations of phosphorus (P), potassium (K), calcium (Ca), copper (Cu), and zinc (Zn) were not related to Mn tolerance. Because differential Mn tolerance in these two genotypes is associated with differential internal tolerance to excess Mn, rather than differential Mn uptake, studies are needed to determine the chemical forms of Mn in tolerant and sensitive plants whose leaves contain comparable concentrations of total Mn. Because both Mn and Fe (closely related elements in the Mn toxicity syndrome) have spin resonances, electron paramagnetic resonance (EPR) offers promise in attacking the problem of differential Mn tolerance in plants.     

Effect of manganese on the resistance of tomato to Pseudocercospora fuligena

Black leaf mold caused by Pseudocercospora fuligena is an important fungal disease of tomato in Southeast Asia. The objective of this study was to evaluate the control of this disease using high manganese (Mn) applied to the root substrate and to evaluate the role of the leaf apoplast in plant response to fungal infection. In a nethouse experiment in Thailand, Mn above the optimum for plant growth but below toxicity increased resistance of tomato plants to black leaf mold. Enhanced resistance caused by Mn was also obtained when tomato plants were grown under controlled conditions in a mist chamber and artificially inoculated with the fungus. Manganese significantly increased plant peroxidases in the leaf apoplast. The highest peroxidase activity was measured when plants were inoculated with Pseudocercospora fuligena. Defense‐related proteins in the leaf apoplast increased when plants were inoculated with Pseudocercospora fuligena but not when treated with high Mn. It is concluded that Mn above the optimum level for plant growth can contribute to the control of Pseudocercospora fuligena in tomato. The Mn effect on disease resistance is associated with the activation of plant peroxidases in the leaf apoplast. A systemic response, possibly mediated by NADH peroxidase activity, also seems to trigger disease resistance in leaves with low Mn concentrations.     

Phytotoxicity and some interactions of the essential trace metals iron,manganese, molybdenum,zinc, copper,and boron

Abstract

The essential trace elements Fe, Mn, Zn, Cu, and B in high concentrations can produce phytotoxicities. Iron toxicity resulted from 5 × 10^‐4 M and 10^‐3 M FeSO₄, but not from equivalent amounts of FeEDDHA (ferric ethylenediamine di (o‐hydroxyphenylacetic acid) ). Leaf concentrations in bush beans of 465 μg Mn/g, 291 μg B/g, and 321 μg Zn/g all on the dry weight basis resulted in 27%, 45%, and 34% reduction in yields of leaves, respectively. Zinc was concentrated in roots while Mn and B concentrated in leaves. Solution concentrations of MnS0₄ of 10^‐3 and 10^‐2 M depressed leaf yields of bush beans by 63% and 83%, respectively, with 5140 and 10780 μg Mn/g dry weight of leaves. Copper concentrations were simultaneously increased and those of Ca were decreased. Bush bean plants grown in Yolo loam soil with 200 μg Cu/g soil had a depression in leaf yield of 26% (with 28. 8 μg Cu/g leaf); plants failed to grow with 500 μg Cu/g soil. A level of 10^‐3 M H₂MoO₄ was toxic to bush beans grown in solution culture. Leaves, stems, and roots, respectively, contained 710, and 1054, and 5920 μg Mo/g dry weight.     

Calcium deficiency and CaCO3 on micronutrient status of plants grown in solution culture

Abstract

Plants were grown in solution culture with different levels of Ca to further evaluate Ca relationships to trace metal uptake and to toxicity of trace metals. When tomato plants (Lycopersicon esculentum L., Tropic) were grown at a low level of Ca, the Zn, Cu, Fe, Mn, Al, and Ti concentrations of leaves, stems, and roots were considerably increased. The use of an excess of CaCO₃ which increased pH did not influence the trace metal concentrations of plants any more than did Ca⁺⁺. In a factorial experiment with bush beans (Phaseolus vulgaris L. C.V. Improved Tendergreen) with Ca (10^‐4,10^‐2, 10^‐2 N) and Ni (0, 2 × 10^‐6 M, 2 X10^‐5 M), Ni phytotoxicity and Ni uptake were decreased somewhat at the highest Ca level. High Ni tended to decrease the Ca concentration in leaves. High Ca and Ni both tended to decrease Fe, Cu, Zn, and Mn concentrations in leaves. The Ni had some interactions on the P concentrations of shoots.     

Manganese toxicity in tomato plants: Effects on cation uptake and distribution

Two experiments are described in which tomato plants (Lycopersicon esculentum L. var Ailsa Craig) were grown in water culture supplied with 10–300 μM Mn. Toxicity symptoms associated with a yield reduction were observed only in treatments in excess of 50 μM Mn indicating that this species is relatively tolerant of high Mn supply. Dark brown/black spots appeared first in the cotyledons. Similar symptoms were observed in the leaves, progressively from the oldest leaf. Manganese concentration in the shoot tissues ranged from 286 to 4240 μg. g^‐1 dry weight. The high Mn concentration values found in the shoot tissues of the toxic plants indicate that Mn was highly mobile in the xylem as confirmed by xylem sap analysis.

The concentrations of both Ca and Mg were lower in the smaller Mn toxic plants. Not only was uptake of Ca and Mg retarded but so also was the distribution of Ca and Mg to the younger tissues as illustrated by measurements of Ca and Mg concentrations along a leaf age sequence. This is in accord with the cation‐anion balance of the xylem exudates collected from decapitated plants.

Higher cation exchange capacity (CEC) was found in the leaf tissues of toxic plants particularly in the older leaves but similar values of C.E.C were recorded for the younger leaf tissues of both control and toxic plants.     

Mechanism of manganese toxicity and tolerance of plants VI. Effect of silicon on alleviation of manganese toxicity of barley

Effect of Si on alleviation of Mn toxicity of barley (Hordeum vulgare L.) seedlings was investigated with special reference to the effect on Mn microdistribution and peroxidase activity. Manganese treatment was conducted by growing the seedlings in nutrient solutions containing different concentrations of Mn. Silicon treatment was conducted by growing the seedlings in the solutions with or without Si supply. Silicon supply alleviated the necrotic browning in the leaves but did not affect the chlorosis caused by Mn toxicity. Silicon treatment did not appreciably alter the uptake of Mn by the plants. Electron probe X‐ray microanalysis revealed that Mn accumulated in high concentration around the necrotic brown spots and that Si supply prevented the uneven distribution of Mn in the tissues. Increase in the level of Mn supply caused an increase in peroxidase activity in the tissues, and Si supply prvented the increase in peroxidase activity.     

Physiology of manganese toxicity and tolerance in Vigna unguiculata (L.) Walp.

In cowpea (Vigna unguiculata (L.) Walp.) tolerance of manganese (Mn) excess depends on genotype, silicon (Si) nutrition, form of nitrogen (N) supply, and leaf age. The physiological mechanisms for improved Mn leaf-tissue tolerance are still poorly understood. On the basis of the density of brown spots per unit of leaf area and the callose content which are sensitive indicators of Mn toxicity, it was confirmed that cultivar (cv.) TVu 1987 was more Mn-tolerant than cv. TVu 91, young leaves were more Mn-tolerant, Si improved Mn tolerance, and NO₃^—-grown plants were more Mn-tolerant than NH₄⁺-grown plants. A close positive relationship existed between the bulk-leaf Mn content and the vacuolar Mn concentration from the same leaves. Since no clear and consistent differences existed between leaf tissues differing in Mn tolerance, the results suggest that accumulation of Mn in the vacuoles and its complexation by organic anions do not play a role in Mn leaf-tissue tolerance in cowpea. A near linear relationship was found between leaf Mn contents and concentrations of free (H₂O-soluble) and exchangeable-bound (BaCl₂-extractable) Mn in the apoplastic washing fluid (AWF) extracted from whole leaves by an infiltration and centrifugation technique. There were no differences in apoplastic Mn concentrations owing to genotype and form of nitrogen nutrition. However, Si decreased the Mn concentration in the AWF. With increasing bulk-leaf Mn contents, concentrations of organic anions in the AWF also increased. The results suggest that complexation of Mn by organic anions in the leaf apoplast contribute to Mn tolerance due to genotype and more clearly due to NO₃-N nutrition. Cell wall-bound peroxidase activity increased with leaf age and was higher in the Mn-sensitive cv. TVu 91 than in cv. TVu 1987. This was in agreement with a higher H₂O₂ production rate in cv. TVu 91. Also, a lower ratio of reduced to oxidized ascorbic acid in the AWF revealed that in Mn-sensitive leaf tissue, the apoplastic reduction capacity was lower than in Mn-tolerant leaf tissue when genotypes and leaves of different age were compared. We interpret our results as strong circumstantial evidence that Mn tolerance depends on the control of the free Mn²⁺concentration and of Mn²⁺-mediated oxidation/reduction reactions in the leaf apoplast.     

Effect of silicon on alleviation of manganese toxicity in pumpkin (Cucurbita moschata Duch cv. Shintosa)

A series of experiments was conducted to investigate the mechanism of Siinduced alleviation of Mn toxicity in pumpkin (Cucurbita moschata Duch cv. Shintosa) which has been used as a bloom-type stock for grafting cultivation of cucumbers. In the first experiment, the effect of Si on Mn toxicity in the Shintosa cultivar was compared with that in the Super unryu cultivar which is used as a bloomless-type stock. Without Si supply, growth depression occurred in both cultivars at the levels of 100 and 500 µM Mn in the nutrient solution. The alleviative effect of Si on the growth depression was observed more distinctly for the Shintosa cultivar than for the Super unryu cultivar, and the effect became more pronounced with increasing Si levels in the nutrient solution. Regardless of the Mn levels, addition of Si did not decrease the Mn content of the plants. The relative shoot growth was reduced to less than 80% when the molar ratio of Si/Mn in the shoots was lower than 4.5. In the second experiment, Mn and Si in the lower leaves of the Shintosa cultivar treated with different levels of Mn with and without Si supply were extracted with 10 mM Tris-HCI buffer (pH 7.4). At the levels of 100 and 250 µM Mn, the amounts of Mn in the buffer-insoluble fraction increased in the presence of Si supply and the buffer-soluble Mn accounted for less than 9% of the total Mn in the leaf. In addition, more than 84% of the total Si was found in the buffer-insoluble fraction at each Mn level. Distribution of Mn and Si on the surface of the lowerú leaves of the Shintosa cultivar treated with 250 µM Mn with and without Si supply was examined by electron probe X-ray microanalysis in the third experiment; Both Mn and Si accumulated at high concentrations around the base of the trichomes in the presence of Si supply. Without Si supply, Mn was detected around the necrotic brown lesions in addition to the base of the trichomes. These results suggested that in the Shintosa cultivar, Si alleviated the Mn toxicity through a localized accumulation of Mn with Si in a metabolically inactive form around the base of the trichomes on the leaf surface.     

Effects of aluminum on the growth and element composition of 20 winter cultivars of Triticum aestivum L. (wheat) grown in solution culture

Seedlings of twenty cultivars of Triticum aestivum L. differing in tolerance to aluminum (Al) were grown with Al (74 uM, 2.0 mg L^‐1 ) and without Al at pH 4.5 to examine the effect of Al on the element composition of leaf and root tissues of juvenile plants. Treatment with Al reduced concentrations of Mg and Mn and increased concentrations of Al, P, Ca, and Fe in roots. Treatment with Al reduced concentrations of P, Ca, Mg, Fe, and Mn in leaves. Concentrations of Mg, Fe, and Mn in leaves were in the range considered to be deficient.

Cultivars differed in the effect of Al on element composition. Concentrations of Ca, Mg, Fe, and Mn in leaves of the 20 cultlvars grown with Al were positively correlated with cultivar tolerance to Al as measured by relative root yield. The variance, however, was relatively high. Leaf P concentrations of seedlings grown with Al were not significantly correlated with tolerance to Al. Differences among cultivars in the effect of Al on element, composition were not likely a primary cause of differential tolerance to Al, but Al‐induced element deficiencies may have a secondary effect on the yield of cultivars grown on sub‐lethal, Al‐toxic substrates.     

Stimulation of phenolic metabolism by silicon contributes to rice resistance to sheath blight

Sheath blight caused by Rhizoctonia solani is a major disease of rice worldwide. Silicon (Si) can enhance rice resistance to sheath blight, but the relation with phenolic metabolism is poorly known. Two rice cultivars with different levels of resistance to R. solani (resistant Teqing and susceptible Ningjing 1) were studied to determine the effects of Si on disease intensity (rated from 0 to 9) and the involvement of phenolic compounds in disease resistance. Variation in the concentrations of phenolics (including total soluble phenolics, flavonoids, and lignin) and in the activities of defense‐related enzymes polyphenoloxidase (PPO) and phenylalanine ammonia‐lyase (PAL) in rice leaf sheaths was investigated. The results show that Si application reduced sheath‐blight disease ratings of Ningjing 1 and Teqing by 2.96 and 0.65, respectively. In uninoculated plants, Si application alone had no significant effects on the concentrations of phenolic compounds or on the activities of PPO and PAL. In inoculated plants, Si application increased phenolics concentrations and PPO and PAL activities only in the susceptible cultivar Ningjing 1. We conclude that Si‐induced enhancement of phenolic metabolism contributed to the improved resistance of rice to sheath blight in the sensitive cultivar.     

Vanadium induced manganese toxicity in bush bean plants grown in solution culture

Seedlings of two bush bean cultivars (Phaseolus vulqaris L. cvs. Mn‐sensitive ‘Wonder Crop 2’ and Mn‐tolerant ‘Green Lord') were grown for 14 days in full strength Hoagland No. 2 nutrient solution containing 0.05 ‐ 2 mg L^‐1 of vanadium (V) as ammonium vanadate.

Increasing V concentration in the solution decreased total dry weight of both cultivars. Plant tops were stunted and leaf color became dark green at 1 ‐ 2 mg L^‐1 V, especially in ‘Green Lord’. Veinal necrosis similar to that of Mn toxicity was observed in the primary leaves of ‘Wonder Crop 2’ at 0.2 mg L^‐1 V or above, but not in those of ‘Green Lord’.

The V concentrations in the roots increased exponentially with increasing V concentration in the solution; however, V concentrations in the leaves and stems were not affected. The Mn concentrations in the primary leaves increased under the higher V treatment in ‘Wonder Crop 2'; but not in ‘Green Lord’. In contrast, Fe concentration in the leaves of ‘Wonder Crop 2’ decreased markedly with increasing V concentration in the solution. Enhanced Mn uptake and greater reduction of Fe uptake by ‘Wonder Crop 2’ may explain the incidence of V‐induced Mn toxicity.     

Differential tolerance of bush bean cultivars to excess manganese in solution and sand culture

Nineteen bush bean cultivars were screened for tolerance to excess Mn in nutrient solution and sand culture experiments. Seven‐day‐old seedlings were treated with full strength Hoagland No. 2 nutrient solution containing different Mn concentrations for 12 days in the greenhouse.

Cultivars showing the greatest sensitivity to Mn toxicity were ‘Wonder Crop 1’ and ‘Wonder Crop 2'; those showing the greatest tolerance were ‘Green Lord’, ‘Red Kidney’ and ‘Edogawa Black Seeded’.

Leaf Mn concentration of plants grown in sand culture was higher than that for plants grown in solution culture. The lowest leaf Mn concentration at which Mn toxicity symptoms developed, was higher in tolerant than in sensitive cultivars. The Fe/Mn ratio in the leaves at which Mn toxicity symptoms developed, was higher in the sensitive cultivars than in the tolerant ones.

We concluded that Mn tolerance in certain bush bean cultivars is due to a greater ability to tolerate a high level of Mn accumulation in the leaves.     

Short‐term effects of manganese toxicity on ribulose 1,5 bisphosphate carboxylase in tobacco chloroplasts

The short‐term effects of manganese (Mn) toxicity on ribulose 1,5 bisphosphate carboxylase EC 4.1.1.39 (Rubisco) activity and concentration in tobacco (Nicotiana tabacum L. ‘KY 17') chloroplasts were examined. The activity of the enzyme from both Mn‐treated and control plants was determined 6,12,18, 24, and 48 h after introduction of Mn (80 mg/L). Enzyme activity was determined by monitoring rates of carbon dioxide (¹⁴CO₂) fixation into acid stable products. A decrease in the enzyme's activity in experimental plants was noted after 48 h of exposure. Visible symptoms, such as chlorosis and decreased leaf size, were also observed after 48 h of Mn exposure in experimental plants. Using Rocket Immunoelectrophoresis, no appreciable differences between Rubisco concentration levels of the experimental plants and the control plants were noted indicating that the effect on Rubisco activity is a post‐translational phenomenon and that Rubisco is not being degraded at an accelerated rate. Manganese accumulated in the experimental plants to concentrations as high as 3282 μg/g dry wt as determined by atomic absorption spectrophotometry. A shuttling mechanism for Mn between young and old leaves was indicated by an observed decrease in the concentration of Mn in the young leaf tissue between 12 and 18 h after treatment.     

Ionic interactions and salinity affect monoterpene and phenolic diterpene composition in rosemary (Rosmarinus officinalis)

In this study, we evaluated how increased cation supply can alleviate the toxic effects of NaCl on plants and how it affects essential oils (EOs) and phenolic diterpene composition in leaves of rosemary (Rosmarinus officinalis L.) plants grown in pots. Two concentrations of the chloride salts KCl, CaCl₂, MgCl₂, and FeCl₃ were used together with 100 mM NaCl to study the effects of these nutrients on plant mineral nutrition and leaf monoterpene, phenolic diterpene, and EO composition. The addition of 100 mM NaCl, which decreased K⁺, Ca²⁺, and Mg²⁺ concentrations with increasing Na⁺ in leaves, significantly altered secondary metabolite accumulation. Addition of MgCl₂ and FeCl₃ altered leaf EO composition in 100 mM NaCl–treated rosemary plants while KCl and CaCl₂ did not. Furthermore, addition of CaCl₂ promoted the accumulation of the major phenolic diterpene, carnosic acid, in the leaves. The carnosol concentration was reduced by the addition of KCl to salt‐stressed plants. It is concluded that different salt applications in combination with NaCl treatment may have a pronounced effect on phenolic diterpene and EO composition in rosemary leaves thus indicating that ionic interactions may be carefully considered in the cultivation of these species to achieve the desired concentrations of these secondary metabolites.     

Phytosiderophore release from manganese‐induced iron deficiency in barley

An experiment was conducted in the phytotron with barley (Hordeum vulgare L. cv. Minorimugi) grown in nutrient solution to compare iron (Fe) deficiency caused by the lack of Fe with manganese (Mn)‐induced Fe deficiency. Dark brown spots on older leaves and stems, and interveinal chlorosis on younger leaves were common symptoms of plants grown in either Mn‐toxic or Fe‐deficient treatments. Dry matter yield was affected similarly by Fe deficiency and Mn toxicity. The Mn toxicity significantly decreased the translocation of Fe from roots to shoots, caused root browning, and inhibited Fe absorption. The rate of Fe translocated from roots to shoots in the 25.0 μM Mn (toxic) treatment was similar to the Fe‐deficient treatment. Manganese toxicity, based on the release of phytosiderophore (PS) from roots, decreased from 25.0>250>2.50 uM Mn. The highest release of PS from roots occurred 7 and 14 days after transplanting (DAT) to Mn‐toxic and Fe‐deficient treatments, respectively; but was always higher in the Fe‐deficient treatment than the Mn‐toxic treatments. The release of PS from roots decreased gradually with plant age and with severity of the Mn toxicity symptoms. The PS content in roots followed the PS release pattern.     

Manganese in substrate clays—harmful for plants?

Mixtures of peat and substrate clays are commonly used as growth media for horticultural plant production. A quality protocol for substrate clays defines a threshold value of active manganese (Mn_act = sum of exchangeable and easily reducible Mn) in substrate clays of < 500 mg kg^–1 to prevent toxic reactions of plants. This threshold value was tested in experiments with peat‐clay blends under various growth conditions, and nutrient solution experiments were additionally conducted to investigate the effects of silicic acid and dissolved organic matter on the occurrence of Mn toxicity. Common bean (Phaseolus vulgaris L.) and hydrangea (Hydrangea macrophylla) plants were cultivated in different peat‐clay substrates and in peat under different moisture and pH levels. The clays varied in their Mn_act content from 4–2354 mg kg^–1. The results of the substrate experiments reveal that a threshold value for Mn in substrate clays is not justified, as plants grown in all peat‐clay substrates did not develop any Mn toxicity even at high substrate moisture or low pH conditions which are known to increase the Mn availability. The extraction of active Mn did not well reflect the Mn concentrations in plant dry matter and substrate solution. As plants tolerated high Mn concentrations in the substrate solution compared to the nutrient solution without toxicity symptoms, the influence of silicic acid and dissolved organic matter (DOM) on Mn toxicity was characterized in a nutrient‐solution experiment. Manganese toxicity was clearly diminished by silicic acid application, but not by DOM. The former effect probably explains the tolerance of bean plants in peat substrates where high silicon concentrations in the substrate solution were observed. Peat‐clay blends even provided up to five times more silicon to plants than pure peat.