Copper and herbivory lead to priming and synergism in phytohormones and plant volatiles in the absence of salicylate-jasmonate antagonism

Abiotic stress factors can interfere with the emission of herbivore-induced plant volatile organic compounds (VOCs) and thus disrupt chemical communication channels between plants and other organisms. We investigated whether copper (Cu) stress alone or in conjunction with insect damage modifies the kinetics of (1) VOCs, (2) the VOC-inducing phytohormone jasmonic acid (JA) and (3) its putative antagonist salicylic acid (SA). Hydroponically grown Zea mays exposed to 10 and 80 µM of Cu showed no increases in JA or VOC levels in the absence of herbivory. However when challenged by herbivores, Cu (80 µM) caused ROS generation in root tissues and primed for increased JA accumulation and VOC emission in leaves. SA synthesis was equally primed but higher concentrations were also apparent before insects started feeding. In contrast, plants grown at 10 µM Cu did not differ from controls. These results show that abiotic and biotic stresses result in concentration-dependent, non-additive defense responses. Further support is given to the notion that JA-SA antagonism is absent in Z. mays.     

Priming by airborne signals boosts direct and indirect resistance in maize

Plants counteract attack by herbivorous insects using a variety of inducible defence mechanisms. The production of toxic proteins and metabolites that instantly affect the herbivore's development are examples of direct induced defence. In addition, plants may release mixtures of volatile organic compounds (VOCs) that indirectly protect the plant by attracting natural enemies of the herbivore. Recent studies suggest that these VOCs can also prime nearby plants for enhanced induction of defence upon future insect attack. However, evidence that this defence priming causes reduced vulnerability to insects is sparse. Here we present molecular, chemical and behavioural evidence that VOC-induced priming leads to improved direct and indirect resistance in maize. A differential hybridization screen for inducible genes upon attack by Spodoptera littoralis caterpillars identified 10 defence-related genes that are responsive to wounding, jasmonic acid (JA), or caterpillar regurgitant. Exposure to VOCs from caterpillar-infested plants did not activate these genes directly, but primed a subset of them for earlier and/or stronger induction upon subsequent defence elicitation. This priming for defence-related gene expression correlated with reduced caterpillar feeding and development. Furthermore, exposure to caterpillar-induced VOCs primed for enhanced emissions of aromatic and terpenoid compounds. At the peak of this VOC emission, primed plants were significantly more attractive to parasitic Cotesia marginiventris waSPS. This study shows that VOC-induced priming targets a specific subset of JA-inducible genes, and links these responses at the molecular level to enhanced levels of direct and indirect resistance against insect attack.     

Dispensing synthetic green leaf volatiles in maize fields increases the release of sesquiterpenes by the plants, but has little effect on the attraction of pest and beneficial insects

Maize plants respond to feeding by arthropod herbivores by producing a number of secondary plant compounds, including volatile organic compounds (VOCs). These herbivore-induced VOCs are not only known to attract natural enemies of the herbivores, but they may also prime inducible defences in neighbouring plants, resulting in stronger and faster defence responses in these VOC-exposed plants. Among the compounds that cause this priming effect, green leaf volatiles (GLVs) have received particular attention, as they are ubiquitous and rapidly emitted upon damage. In this study, we investigated their effects under realistic conditions by applying specially devised dispensers to release four synthetic GLVs at physiologically relevant concentrations in a series of experiments in maize fields. We compared the VOC emission of GLV-exposed maize plants to non-exposed plants and monitored the attraction of herbivores and predators, as well as parasitism of the caterpillar Spodoptera frugiperda, the most common herbivore in the experimental maize fields. We found that maize plants that were exposed to GLVs emitted increased quantities of sesquiterpenes compared to non-exposed plants. In several replicates, herbivorous insects, such as adult Diabrotica beetles and S. frugiperda larvae, were observed more frequently in GLV-treated plots and caused more damage to GLV-exposed plants than to non-exposed plants. Parasitism of S. frugiperda was only weakly affected by GLVs and overall parasitism rates of S. frugiperda were similar in GLV-exposed and non-exposed plots. The effects on insect presence depended on the distance from the GLV-dispensers at which the plants were located. The results are discussed in the context of strategies to improve biological control by enhancing plant-mediated attraction of natural enemies.     

Phytochelatins as biomarkers for heavy metal stress in maize (Zea mays L.) and wheat (Triticum aestivum L.): combined effects of copper and cadmium

Heavy metal contaminated soils often show increased levels of more than one metal, e.g. copper (Cu), cadmium (Cd), zinc (Zn), lead (Pb) or nickel (Ni). In case such soils are used for crop production, prediction of yield reduction or quality decline due to heavy metals in the soil is inadequate when based only on chemical soil analysis. The use of biomarkers such as phytochelatins (PC), non-protein thiols specifically induced in plants upon exposure to heavy metals, may be an additional tool or diagnostic criterion in heavy metal research and in practice. In the present work, Cu and Cd uptake and induction of PC synthesis are studied with hydroponically grown maize and wheat plants exposed to mixtures of the two metals. We observed a close positive relationship between the concentrations of Cd and PC in the plant shoot material. A decreased shoot concentration of Cd after addition of Cu, due to metal competition at common root absorption sites, coincided with lower shoot PC levels. Also differences in metal uptake and xylary metal transport among the two plant species were reflected in corresponding differences in PC concentration. The observed direct relationship between shoot PC concentration and the degree of metal-induced growth inhibition makes the use of PC promising for the purpose tested for.     

铜、镉胁迫下外源NO介导的番茄解毒途径

一氧化氮(NO)作为信号分子,在抵御重金属胁迫中起重要作用,但对不同离子胁迫下的解毒机制尚缺乏研究.本研究采用营养液培养法,研究了铜(Cu)、镉(Cd)单一或复合胁迫下,番茄幼苗对Cu、Cd的吸收转运特性及对外源NO的响应机制.结果表明: 50 μmol·L^-1的Cu²⁺、Cd²⁺均显著抑制番茄植株的生长,其中Cd胁迫对生长的抑制效应远高于Cu胁迫.Cu、Cd单一或复合胁迫均使番茄根系Cu、Cd含量显著升高,但根系对Cu、Cd吸收存在严格选择性.根细胞对必需元素Cu表现出“奢侈吸收”的现象,而对毒性较强的Cd则吸收相对较少,胞内Cd浓度仅为Cu的1/10左右.外源NO处理可不同程度地缓解Cu、Cd胁迫,其中缓解Cd胁迫的效能更强.番茄对被动进入细胞的Cu、Cd具有相似的解毒机制:一方面,Cu、Cd胁迫诱导细胞质中产生谷胱甘肽(GSH)、植物螯合肽(PCs)和金属硫蛋白(MTs),络合过多的Cu、Cd离子,降低其生物毒性;另一方面,过多的Cu、Cd离子或螯合物被转运至液泡区隔化.外源NO通过调控GSH-GSSG(氧化型谷胱甘肽)氧化还原状态及GSH-PCs代谢方向的改变,促进Cu、Cd离子转运至液泡区隔化来缓解胁迫抑制;NO还可诱导植株叶片或根系表达更多的金属硫蛋白、GSH和PCs,而且上述响应普遍存在叠加效应.这可能是NO介导番茄对Cu、Cd胁迫的另一主要解毒途径.     

Feeding by Hessian fly [Mayetiola destructor (Say)] larvae does not induce plant indirect defences

Abstract.  1. Recent research has addressed the function of herbivore-induced plant volatiles in attracting natural enemies of feeding herbivores. While many types of insect herbivory appear to elicit volatile responses, those triggered by gall insects have received little attention. Previous work indicates that at least one gall insect species induces changes in host-plant volatiles, but no other studies appear to have addressed whether gall insects trigger plant indirect defences.
2. The volatile responses of wheat to feeding by larvae of the Hessian fly Mayetiola destructor (Say) (Diptera: Cecidomyiidae) were studied to further explore indirect responses of plants to feeding by gall insects. This specialist gall midge species did not elicit a detectable volatile response from wheat plants, whereas a generalist caterpillar triggered volatile release. Moreover, Hessian fly feeding altered volatile responses to subsequent caterpillar herbivory.
3. These results suggest that Hessian fly larvae exert a degree of control over the defensive responses of their host plants and offer insight into plant-gall insect interactions. Also, the failure of Hessian fly larvae to elicit an indirect defensive response from their host plants may help explain why natural enemies, which often rely on induced volatile cues, fail to inflict significant mortality on M. destructor populations in the field.     

Integrative approach gives new insights into combined Cd/Cu exposure in tobacco

Heavy metals are generally known to induce oxidative stress, but are rarely strategically studied in an embracive manner, taking into account interplay between their various effects. Furthermore, although metals in the environment are present in mixtures and interact with each other, their combined effects to organisms have been much less studied in comparison to individual effects. Here, we present a complete comprehensive study of cadmium (Cd)/copper (Cu) oxidative stress interactions in Nicotiana tabacum seedlings and adult plants. Plants were treated with Cd (10 and 15 μM), Cu (2.5 and 5 μM) and their combinations; seedlings during 1 month period and adult plants during the period of 7 days. Metal accumulation measurements showed that Cd and Cu influence each other uptake, with Cu reducing Cd translocation to shoots. PCA analysis showed that MDA and carbonyls, biomarkers of oxidative stress, as well as ascorbate peroxidase activity, highly correlated across tissues and with Cd content. Majority of toxic effects were caused by Cd-alone, while addition of Cu often resulted in damage alleviation. However, mixture of high concentrations of both Cd and Cu induced most adverse effects. In conclusion, our results indicate that Cu in lower concentration has antagonistic effect to Cd toxicity, while in higher concentration these metals interact additively in tobacco.     

Distribution features and internal relations of heavy metals in soil–maize system of mining area,Anhui Province,Eastern China

A total of 59 topsoil and corresponding maize plants were collected from this study area. The spatial distribution, correlation analysis, and multiple linear regression of heavy metals were researched detailedly in this article. The results showed that distribution characteristics of heavy metals (Pb, Cd, and Ni) in different parts of maize plants (immature stage) accumulated mostly in stems, with Pb mainly accumulated in roots (mature period), and Cd and Ni mostly in leaves. Except for the southeastern local region of this mining area, Mn and Cu possessed roughly similar spatial distribution characteristics. The results of partial correlation analysis indicated that Cu, Cd in the roots of the tested maize plants and Ni in soil may have antagonistic effects, Cu (soil)–Cu (stem) and Ni (soil)–Pb (stem) had a certain promoting effect. Besides, Cu, Pb, and Ni in soil promoted the absorption of Cu, Pb, and Ni in the leaves, whereas Cr and Pb in soil can promote the enrichment of Mn in maize grains. Our findings suggested that the concentrations of heavy metals in maize organs could be predicted accurately using the established models.     

Heavy metal accumulation in iron plaque and growth of rice plants upon exposure to single and combined contamination by copper, cadmium and lead

A pot experiment was conducted to evaluate the bioaccumulation of heavy metals and growth response of rice plants after exposure to single and combined contamination by Cu, Cd and Pb. The results showed that the biomass production was not significantly affected by either single or combined treatment of Cu, Cd and Pb. Adding Cu (Cd, or Pb) separately all increased concentrations of the respective element in root and shoot (p < 0.001). In the combined contamination, Pb promoted both root and shoot absorption of Cu and Cd (p < 0.001), and Cu affected Cd and Pb absorption in the root, but Pb concentrations in both root and shoot were not affected by Cd application. The formation of iron plaques varied obviously with soil types. Heavy metal accumulation in iron plaques was induced by the three elements (p < 0.001). Furthermore, the three heavy metals exhibited an interactive relationship as measured by the Cu, Cd, Pb and Fe concentrations in root surface iron plaques. The iron plaques partially inhibited transfer of Pb to root and shoot, but no such effect was observed for Cu and Cd. This research indicates that the interaction among different heavy metal elements is very complex. It is very important to have a clear understanding on the associated mechanism and the consequential impact on plant growth.     

Heavy metal accumulation in iron plaque and growth of rice plants upon exposure to single and combined contamination by copper, cadmium and lead

A pot experiment was conducted to evaluate the bioaccumulation of heavy metals and growth response of rice plants after exposure to single and combined contamination by Cu, Cd and Pb. The results showed that the biomass production was not significantly affected by either single or combined treatment of Cu, Cd and Pb. Adding Cu (Cd, or Pb) separately all increased concentrations of the respective element in root and shoot (p < 0.001). In the combined contamination, Pb promoted both root and shoot absorption of Cu and Cd (p < 0.001), and Cu affected Cd and Pb absorption in the root, but Pb concentrations in both root and shoot were not affected by Cd application. The formation of iron plaques varied obviously with soil types. Heavy metal accumulation in iron plaques was induced by the three elements (p < 0.001). Furthermore, the three heavy metals exhibited an interactive relationship as measured by the Cu, Cd, Pb and Fe concentrations in root surface iron plaques. The iron plaques partially inhibited transfer of Pb to root and shoot, but no such effect was observed for Cu and Cd. This research indicates that the interaction among different heavy metal elements is very complex. It is very important to have a clear understanding on the associated mechanism and the consequential impact on plant growth.     

Eco-service enhancement in peri-urban area of coal mining city of Huaibei in East China

After 50 years of coal mining, Huaibei Mine, located at 50 km southeast of Xuzhou City in East China, has grown to a middle-size city of 600,000 people from a small village of 2000 farmers. The Zhahe Valley, with 400 km² of a built-up area and more than 100 km² of subsided peri-urban wetland at the city center, is surrounded by eight exhausted old mines and communities. In cooperation with the local city government, an ecological landuse change assessment and eco-city planning project has been carried out with a focus on the assessment, restoration and enhancement of the wetland as an eco-service to the community. The assessment includes relationships to Green House Gas emissions and heat island effects, as well as measures for a livable, workable, affordable and sustainable human settlement development through industrial transition, landscape design and capacity building. This paper will briefly introduce the main ecological approaches and results of the assessment, including measures such as changing the car-dominated transportation network to a rail-dominated network, transforming the coal-oriented high-carbon industry to a service-oriented low-carbon industry, the C-shape urban form to an O-shape with a green–blue core at the center, and the fragmentized collapsed land to integrative eco-service land.     

Accumulation of cadmium, zinc, and copper by Helianthus annuus L.: impact on plant growth and uptake of nutritional elements

We investigated the effects on physiological response, trace elements and nutrients accumulation of sunflower plants grown in soil contaminated with: 5 mg kg(-1) of Cd; 5 and 300 mg kg(-1) of Cd and Zn, respectively; 5, 300, and 400 mg kg(-1) of Cd, Zn, and Cu, respectively. Contaminants applied did not produce large effects on growth, except in Cd-Zn-Cu treatment in which leaf area and total dry matter were reduced, by 15%. The contamination with Cd alone did not affect neither growth nor physiological parameters, despite considerable amounts of Cd accumulated in roots and older leaves, with a high bioconcentration factor from soil to plant. By adding Zn and then Cu to Cd in soil, significant were the toxic effects on chlorophyll content and water relations due to greater accumulation of trace elements in tissues, with imbalances in nutrients uptake. Highly significant was the interaction between shoot elements concentration (Cd, Zn, Cu, Fe, Mg, K, Ca) and treatments. Heavy metals concentrations in roots always exceeded those in stem and leaves, with a lower translocation from roots to shoots, suggesting a strategy of sunflower to compartmentalise the potentially toxic elements in physiologically less active parts in order to preserve younger tissues.     

Parasitoids select plants more heavily infested with their caterpillar hosts: a new approach to aid interpretation of plant headspace volatiles

Plants produce volatile organic compounds (VOCs) in response to herbivore attack, and these VOCs can be used by parasitoids of the herbivore as host location cues. We investigated the behavioural responses of the parasitoid Cotesia vestalis to VOCs from a plant-herbivore complex consisting of cabbage plants (Brassica oleracea) and the parasitoids host caterpillar, Plutella xylostella. A Y-tube olfactometer was used to compare the parasitoids' responses to VOCs produced as a result of different levels of attack by the caterpillar and equivalent levels of mechanical damage. Headspace VOC production by these plant treatments was examined using gas chromatography-mass spectrometry. Cotesia vestalis were able to exploit quantitative and qualitative differences in volatile emissions, from the plant-herbivore complex, produced as a result of different numbers of herbivores feeding. Cotesia vestalis showed a preference for plants with more herbivores and herbivore damage, but did not distinguish between different levels of mechanical damage. Volatile profiles of plants with different levels of herbivores/herbivore damage could also be separated by canonical discriminant analyses. Analyses revealed a number of compounds whose emission increased significantly with herbivore load, and these VOCs may be particularly good indicators of herbivore number, as the parasitoid processes cues from its external environment.     

Iron acquisition by phytosiderophores contributes to cadmium tolerance

Based on the ability of phytosiderophores to chelate other heavy metals besides iron (Fe), phytosiderophores were suggested to prevent graminaceous plants from cadmium (Cd) toxicity. To assess interactions between Cd and phytosiderophore-mediated Fe acquisition, maize (Zea mays) plants were grown hydroponically under limiting Fe supply. Exposure to Cd decreased uptake rates of 59Fe(III)-phytosiderophores and enhanced the expression of the Fe-phytosiderophore transporter gene ZmYS1 in roots as well as the release of the phytosiderophore 2'-deoxymugineic acid (DMA) from roots under Fe deficiency. However, DMA hardly mobilized Cd from soil or from a Cd-loaded resin in comparison to the synthetic chelators diaminetriaminepentaacetic acid and HEDTA. While nano-electrospray-high resolution mass spectrometry revealed the formation of an intact Cd(II)-DMA complex in aqueous solutions, competition studies with Fe(III) and zinc(II) showed that the formed Cd(II)-DMA complex was weak. Unlike HEDTA, DMA did not protect yeast (Saccharomyces cerevisiae) cells from Cd toxicity but improved yeast growth in the presence of Cd when yeast cells expressed ZmYS1. When supplied with Fe-DMA as a Fe source, transgenic Arabidopsis (Arabidopsis thaliana) plants expressing a cauliflower mosaic virus 35S-ZmYS1 gene construct showed less growth depression than wild-type plants in response to Cd. These results indicate that inhibition of ZmYS1-mediated Fe-DMA transport by Cd is not related to Cd-DMA complex formation and that Cd-induced phytosiderophore release cannot protect maize plants from Cd toxicity. Instead, phytosiderophore-mediated Fe acquisition can improve Fe uptake in the presence of Cd and thereby provides an advantage under Cd stress relative to Fe acquisition via ferrous Fe.     

Soil Metal Sorption Characteristics and its Influence on the Comparative Effectiveness of EDTA and Legume Intercrop on the Phytoremediative Abilities of Maize (Zea mays), Mucuna (Mucuna pruriens), Okra (Abelmoschus esculentus), and Kenaf (Hibiscus cannabinus)

Heavy metal pollution in soils and the high costs of remediation necessitate the evaluation of cheaper alternatives. The aim of this experiment was to evaluate Cd, Pb, Zn, and Cu sorption characteristics of three soils and their influence on the comparative effectiveness of EDTA and legume intercrop on the remediative abilities of maize, mucuna, okra, and kenaf. The sorption studies were done using standard procedures. The EDTA-assisted phytoextraction (6 mmol kg^?1) and the cowpea intercrop trials were conducted in triplicate. The metal-spiked soils were planted with maize, kenaf, and mucuna in the EDTA trial and maize, kenaf, and okra were planted in the cowpea intercrop experiment. Cadmium was prefentially sorbed in acid and alkaline soils and Cu in slightly acid soil. Cadmium uptake was significantly lower (P < 0.05) in all the plants. Bioconcentration factors of Pb, Cu, and Zn were higher (P < 0.05) in maize compared with other plants. Phytoremediative ability of the plants in trials were maize > kenaf > mucuna and okra > maize > kenaf, respectively. It was concluded that a legume intercrop can substitute EDTA- assisted phytoextraction to prevent groundwater contamination resulting from high solubility of metals by EDTA.     

Priming of Production in Maize of Volatile Organic Defence Compounds by the Natural Plant Activator cis-Jasmone

cis-Jasmone (CJ) is a natural plant product that activates defence against herbivores in model and crop plants. In this study, we investigated whether CJ could prime defence in maize, Zea mays, against the leafhopper, Cicadulina storeyi, responsible for the transmission of maize streak virus (MSV). Priming occurs when a pre-treatment, in this case CJ, increases the potency and speed of a defence response upon subsequent attack on the plant. Here, we tested insect responses to plant volatile organic compounds (VOCs) using a Y-tube olfactometer bioassay. Our initial experiments showed that, in this system, there was no significant response of the herbivore to CJ itself and no difference in response to VOCs collected from unexposed plants compared to CJ exposed plants, both without insects. VOCs were then collected from C. storeyi-infested maize seedlings with and without CJ pre-treatment. The bioassay revealed a significant preference by this pest for VOCs from infested seedlings without the CJ pre-treatment. A timed series of VOC collections and bioassays showed that the effect was strongest in the first 22 h of insect infestation, i.e. before the insects had themselves induced a change in VOC emission. Chemical analysis showed that treatment of maize seedlings with CJ, followed by exposure to C. storeyi, led to a significant increase in emission of the defensive sesquiterpenes (E)-(1R,9S)-caryophyllene, (E)-α-bergamotene, (E)-β-farnesene and (E)-4,8-dimethyl-1,3,7-nonatriene, known to act as herbivore repellents. The chemical analysis explains the behavioural effects observed in the olfactometer, as the CJ treatment caused plants to emit a blend of VOCs comprising more of the repellent components in the first 22 h of insect infestation than control plants. The speed and potency of VOC emission was increased by the CJ pre-treatment. This is the first indication that CJ can prime plants for enhanced production of defensive VOCs antagonist towards herbivores.     

Metal accumulation and response of antioxidant enzymes in seedlings and adult sunflower mutants with improved metal removal traits on a metal-contaminated soil

Sunflower mutant lines with an enhanced tolerance and metal accumulation capacity obtained by mutation breeding have been proposed for Zn, Cd and Cu removal from metal-contaminated soils in previous studies. However, soils contaminated with trace elements induce various biochemical alterations in plants leading to oxidative stress. There is a lack of knowledge concerning the metal accumulation and antioxidant responses during the growth and development of sunflowers. This study, therefore, aimed to characterise metal accumulation and possible metal detoxification mechanisms in young seedlings and adult sunflowers. Beside the inbred line, two mutant lines with an improved growth and enhanced metal uptake capacity on a metal contaminated soil were investigated in more detail.Sunflowers cultivated on a metal-contaminated soil in the greenhouse showed a decrease in shoot biomass and chlorophyll concentration in two different developmental stages. Adult sunflowers showed a lower sensitivity to metal toxicity than young seedlings, whereas mutant lines were more tolerant to metal stress than the control. Mutant lines also produced a higher amount of carotenoids on a metal-contaminated soil than on the control soil, indicating a possible protective mechanism of sunflower mutants against oxidative stress caused by Cd and excess Zn.Heavy metals primarily increased the activity of antioxidant enzymes involved in the ascorbate–glutathione cycle in sunflower leaves. Activity of dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR) was strongly increased in young seedlings exposed to heavy metals. The enzyme activities were even more pronounced in mutant lines. A significantly increased ascorbate peroxidase (APOX) activity in adult sunflowers exposed to heavy metals indicated an elevated use of ascorbate after a longer exposure to metal stress.An increased antioxidant level corresponded to a high Cd and Zn accumulation in young and adult sunflowers. Metal distribution, zinc translocation in particular, from the root into the shoot tissue obviously increased during sunflower growth and ripening. Altogether, these results suggest that sunflower plants, primarily the mutant lines, possess an efficient defence mechanism against oxidative stress caused by metal toxicity. A good tolerance of sunflowers toward heavy metals coupled with an increased metal accumulation capacity might contribute to an efficient removal of heavy metals from a polluted area.     

菌根植物根际环境对污染土壤中Cu、Zn、Pb、Cd形态的影响

采用根垫法和连续形态分析技术,分析了生长在污灌土壤中菌根小麦和无菌根小麦根际Ｃｕ、Ｚｎ、Ｐｂ、Ｃｄ的形态分布和变化趋势。结果表明,下对照土壤相比,菌根际土壤中交换态Ｃｕ含量显著增加,交换态Ｃｄ呈减少的趋势;与非菌根际相比,Ｃｕ、Ｚｎ、Ｐｂ的有机结合态在菌根根际中显著增加,而４种测定金属２的碳酸盐态和铁锰氧化态都没有显著改变,该结果表明,植物根系能影响根际中金属形态的变化,且菌根比无菌根的影响程度大     

Heavy metal stress and sulfate uptake in maize roots

ZmST1;1, a putative high-affinity sulfate transporter gene expressed in maize (Zea mays) roots, was functionally characterized and its expression patterns were analyzed in roots of plants exposed to different heavy metals (Cd, Zn, and Cu) interfering with thiol metabolism. The ZmST1;1 cDNA was expressed in the yeast (Saccharomyces cerevisiae) sulfate transporter mutant CP154-7A. Kinetic analysis of sulfate uptake isotherm, determined on complemented yeast cells, revealed that ZmST1;1 has a high affinity for sulfate (Km value of 14.6 +/- 0.4 microm). Cd, Zn, and Cu exposure increased both ZmST1;1 expression and root sulfate uptake capacity. The metal-induced sulfate uptakes were accompanied by deep alterations in both thiol metabolism and levels of compounds such as reduced glutathione (GSH), probably involved as signals in sulfate uptake modulation. Cd and Zn exposure strongly increased the level of nonprotein thiols of the roots, indicating the induction of additional sinks for reduced sulfur, but differently affected root GSH contents that decreased or increased following Cd or Zn stress, respectively. Moreover, during Cd stress a clear relation between the ZmST1;1 mRNA abundance increment and the entity of the GSH decrement was impossible to evince. Conversely, Cu stress did not affect nonprotein thiol levels, but resulted in a deep contraction of GSH pools. Our data suggest that during heavy metal stress sulfate uptake by roots may be controlled by both GSH-dependent or -independent signaling pathways. Finally, some evidence suggesting that root sulfate availability in Cd-stressed plants may limit GSH biosynthesis and thus Cd tolerance are discussed.