Non-glandular trichomes of sunflower are important in the absorption and translocation of foliar-applied Zn.

Trichomes are potentially important for absorption of foliar fertilizers. A study has shown that the non-glandular trichromes (NGTs) of sunflower (Helianthus annuus) accumulated high concentrations of foliar-applied zinc (Zn); however, the mechanisms of Zn accumulation in the NGTs and the fate of this Zn are unclear. Here we investigated how foliar-applied Zn accumulates in the NGTs and the subsequent translocation of this Zn. Time-resolved synchrotron-based X-ray fluorescence microscopy and transcriptional analyses were used to probe the movement of Zn in the NGTs, with the cuticle composition of the NGTs examined using confocal Raman microscopy. The accumulation of Zn in the NGTs is both an initial preferential absorption process and a subsequent translocation process. This preferred absorption is likely because the NGT base has a higher hydrophilicity, whilst the subsequent translocation is due to the presence of plasmodesmata, Zn-chelating ligands, and Zn transporters in the NGTs. Furthermore, the Zn sequestered in the NGTs was eventually translocated out of the trichome once the leaf Zn concentration had decreased, suggesting that the NGTs are also important in maintaining leaf Zn homeostasis. This study demonstrates for the first time that trichomes have a key structural and functional role in the absorption and translocation of foliar-applied Zn.

Time-resolved laboratory micro-X-ray fluorescence reveals silicon distribution in relation to manganese toxicity in soybean and sunflower.

BACKGROUND AND AIMS: Synchrotron- and laboratory-based micro-X-ray fluorescence (µ-XRF) is a powerful technique to quantify the distribution of elements in physically large intact samples, including live plants, at room temperature and atmospheric pressure. However, analysis of light elements with atomic number (Z) less than that of phosphorus is challenging due to the need for a vacuum, which of course is not compatible with live plant material, or the availability of a helium environment. METHOD: A new laboratory µ-XRF instrument was used to examine the effects of silicon (Si) on the manganese (Mn) status of soybean (Glycine max) and sunflower (Helianthus annuus) grown at elevated Mn in solution. The use of a helium environment allowed for highly sensitive detection of both Si and Mn to determine their distribution. KEY RESULTS: The µ-XRF analysis revealed that when Si was added to the nutrient solution, the Si also accumulated in the base of the trichomes, being co-located with the Mn and reducing the darkening of the trichomes. The addition of Si did not reduce the concentrations of Mn in accumulations despite seeming to reduce its adverse effects. CONCLUSIONS: The ability to gain information on the dynamics of the metallome or ionome within living plants or excised hydrated tissues can offer valuable insights into their ecophysiology, and laboratory µ-XRF is likely to become available to more plant scientists for use in their research.

Assessing radiation dose limits for X-ray fluorescence microscopy analysis of plant specimens.

BACKGROUND AND AIMS: X-ray fluorescence microscopy (XFM) is a powerful technique to elucidate the distribution of elements within plants. However, accumulated radiation exposure during analysis can lead to structural damage and experimental artefacts including elemental redistribution. To date, acceptable dose limits have not been systematically established for hydrated plant specimens. METHODS: Here we systematically explore acceptable dose rate limits for investigating fresh sunflower (Helianthus annuus) leaf and root samples and investigate the time-dose damage in leaves attached to live plants. KEY RESULTS: We find that dose limits in fresh roots and leaves are comparatively low (4.1 kGy), based on localized disintegration of structures and element-specific redistribution. In contrast, frozen-hydrated samples did not incur any apparent damage even at doses as high as 587 kGy. Furthermore, we find that for living plants subjected to XFM measurement in vivo and grown for a further 9 d before being reimaged with XFM, the leaves display elemental redistribution at doses as low as 0.9 kGy and they continue to develop bleaching and necrosis in the days after exposure. CONCLUSIONS: The suggested radiation dose limits for studies using XFM to examine plants are important for the increasing number of plant scientists undertaking multidimensional measurements such as tomography and repeated imaging using XFM.

Synchrotron-Based X-Ray Fluorescence Microscopy as a Technique for Imaging of Elements in Plants.

Understanding the distribution of elements within plant tissues is important across a range of fields in plant science. In this review, we examine synchrotron-based x-ray fluorescence microscopy (XFM) as an elemental imaging technique in plant sciences, considering both its historical and current uses as well as discussing emerging approaches. XFM offers several unique capabilities of interest to plant scientists, including in vivo analyses at room temperature and pressure, good detection limits (approximately 1-100 mg kg-1), and excellent resolution (down to 50 nm). This has permitted its use in a range of studies, including for functional characterization in molecular biology, examining the distribution of nutrients in food products, understanding the movement of foliar fertilizers, investigating the behavior of engineered nanoparticles, elucidating the toxic effects of metal(loid)s in agronomic plant species, and studying the unique properties of hyperaccumulating plants. We anticipate that continuing technological advances at XFM beamlines also will provide new opportunities moving into the future, such as for high-throughput screening in molecular biology, the use of exotic metal tags for protein localization, and enabling time-resolved, in vivo analyses of living plants. By examining current and potential future applications, we hope to encourage further XFM studies in plant sciences by highlighting the versatility of this approach.

Manganese distribution and speciation help to explain the effects of silicate and phosphate on manganese toxicity in four crop species.

Soil acidity and waterlogging increase manganese (Mn) in leaf tissues to potentially toxic concentrations, an effect reportedly alleviated by increased silicon (Si) and phosphorus (P) supply. Effects of Si and P on Mn toxicity were studied in four plant species using synchrotron-based micro X-ray fluorescence (µ-XRF) and nanoscale secondary ion mass spectrometry (NanoSIMS) to determine Mn distribution in leaf tissues and using synchrotron-based X-ray absorption spectroscopy (XAS) to measure Mn speciation in leaves, stems and roots. A concentration of 30 µM Mn in solution was toxic to cowpea and soybean, with 400 µM Mn toxic to sunflower but not white lupin. Unexpectedly, µ-XRF analysis revealed that 1.4 mM Si in solution decreased Mn toxicity symptoms through increased Mn localization in leaf tissues. NanoSIMS showed Mn and Si co-localized in the apoplast of soybean epidermal cells and basal cells of sunflower trichomes. Concomitantly, added Si decreased oxidation of Mn(II) to Mn(III) and Mn(IV). An increase from 5 to 50 µM P in solution changed some Mn toxicity symptoms but had little effect on Mn distribution or speciation. We conclude that Si increases localized apoplastic sorption of Mn in cowpea, soybean and sunflower leaves thereby decreasing free Mn2+ accumulation in the apoplast or cytoplasm.

Synchrotron-Based Techniques Shed Light on Mechanisms of Plant Sensitivity and Tolerance to High Manganese in the Root Environment.

Plant species differ in response to high available manganese (Mn), but the mechanisms of sensitivity and tolerance are poorly understood. In solution culture, greater than or equal to 30 µm Mn decreased the growth of soybean (Glycine max), but white lupin (Lupinus albus), narrow-leafed lupin (Lupin angustifolius), and sunflower (Helianthus annuus) grew well at 100 µm Mn. Differences in species' tolerance to high Mn could not be explained simply by differences in root, stem, or leaf Mn status, being 8.6, 17.1, 6.8, and 9.5 mmol kg(-1) leaf fresh mass at 100 µm Mn. Furthermore, x-ray absorption near edge structure analyses identified the predominance of Mn(II), bound mostly to malate or citrate, in roots and stems of all four species. Rather, differences in tolerance were due to variations in Mn distribution and speciation within leaves. In Mn-sensitive soybean, in situ analysis of fresh leaves using x-ray fluorescence microscopy combined with x-ray absorption near edge structure showed high Mn in the veins, and manganite [Mn(III)] accumulated in necrotic lesions apparently through low Mn sequestration in vacuoles or other vesicles. In the two lupin species, most Mn accumulated in vacuoles as either soluble Mn(II) malate or citrate. In sunflower, Mn was sequestered as manganite at the base of nonglandular trichomes. Hence, tolerance to high Mn was ascribed to effective sinks for Mn in leaves, as Mn(II) within vacuoles or through oxidation of Mn(II) to Mn(III) in trichomes. These two mechanisms prevented Mn accumulation in the cytoplasm and apoplast, thereby ensuring tolerance to high Mn in the root environment.

Identification of the primary lesion of toxic aluminum in plant roots.

Despite the rhizotoxicity of aluminum (Al) being identified over 100 years ago, there is still no consensus regarding the mechanisms whereby root elongation rate is initially reduced in the approximately 40% of arable soils worldwide that are acidic. We used high-resolution kinematic analyses, molecular biology, rheology, and advanced imaging techniques to examine soybean (Glycine max) roots exposed to Al. Using this multidisciplinary approach, we have conclusively shown that the primary lesion of Al is apoplastic. In particular, it was found that 75 µm Al reduced root growth after only 5 min (or 30 min at 30 µm Al), with Al being toxic by binding to the walls of outer cells, which directly inhibited their loosening in the elongation zone. An alteration in the biosynthesis and distribution of ethylene and auxin was a second, slower effect, causing both a transient decrease in the rate of cell elongation after 1.5 h but also a longer term gradual reduction in the length of the elongation zone. These findings show the importance of focusing on traits related to cell wall composition as well as mechanisms involved in wall loosening to overcome the deleterious effects of soluble Al.

Kinetics and nature of aluminium rhizotoxic effects: a review.

Acid soils with elevated levels of soluble aluminium (Al) comprise ~40% of the world's arable land, but there remains much uncertainty regarding the mechanisms by which Al is rhizotoxic. This review examines the kinetics of the toxic effects of Al on the root elongation rate (RER), its effects on root tissues, and its location at a subcellular level. Depending upon the concentration and plant species, soluble Al decreases the RER in a median time of 73min, but in as little as 5min in soybean. This is initially due to a decreased rate at which cells expand anisotropically in the elongation zone. Thereafter, rhizodermal and outer cortical cells rupture through decreased cell wall relaxation. It is in this region where most Al accumulates in the apoplast. Subsequently, Al impacts root growth at a subcellular level through adverse effects on the plasma membrane (PM), cytoplasm, and nucleus. At the PM, Al alters permeability, fluidity, and integrity in as little as 0.5h, whilst it also depolarizes the PM and reduces H(+)-ATPase activity. The Al potentially crosses the PM within 0.5h where it is able to bind to the nucleus and inhibit cell division; sequestration within the vacuole is required to reduce the toxic effects of Al within the cytoplasm. This review demonstrates the increasing evidence of the importance of the initial Al-induced inhibition of wall loosening, but there is evidence also of the deleterious effects of Al on other cellular processes which are important for long-term root growth and function.

Aluminium effects on mechanical properties of cell wall analogues.

Aluminium (Al) toxicity adversely impacts plant productivity in acid soils by restricting root growth and although several mechanisms are involved the physiological basis of decreased root elongation remains unclear. Understanding the primary mechanisms of Al rhizotoxicity is hindered due to the rapid effects of soluble Al on root growth and the close proximity of many cellular components within the cell wall, plasma membrane, cytosol and nucleus with which Al may react. To overcome some of these difficulties, we report on a novel method for investigating Al interactions with Komagataeibacter xylinus bacterial cellulose (BC)-pectin composites as cell wall analogues. The growth of K. xylinus in the presence of various plant cell wall polysaccharides, such as pectin, has provided a unique in vitro model system with which to investigate the interactions of Al with plant cell wall polysaccharides. The BC-pectin composites reacted in a similar way with Al as do plant cell walls, providing insights into the effects of Al on the mechanical properties of the BC-pectin composites as cell wall analogues. Our findings indicated that there were no significant effects of Al (4-160 µM) on the tensile stress, tensile strain or Young's modulus of the composites. This finding was consistent with cellulose, not pectin, being the major load bearing component in BC-pectin composites, as is also the case in plant cell walls.

Distribution and speciation of Mn in hydrated roots of cowpea at levels inhibiting root growth.

The phytotoxicity of Mn is important globally due to its increased solubility in acid or waterlogged soils. Short-term (≤24 h) solution culture studies with 150 µM Mn were conducted to investigate the in situ distribution and speciation of Mn in apical tissues of hydrated roots of cowpea [Vigna unguiculata (L.) Walp. cv. Red Caloona] using synchrotron-based techniques. Accumulation of Mn was rapid; exposure to 150 µM Mn for only 5 min resulting in substantial Mn accumulation in the root cap and associated mucigel. The highest tissue concentrations of Mn were in the root cap, with linear combination fitting of the data suggesting that ≥80% of this Mn(II) was associated with citrate. Interestingly, although the primary site of Mn toxicity is typically the shoots, concentrations of Mn in the stele of the root were not noticeably higher than in the surrounding cortical tissues in the short-term (≤24 h). The data provided here from the in situ analyses of hydrated roots exposed to excess Mn are, to our knowledge, the first of this type to be reported for Mn and provide important information regarding plant responses to high Mn in the rooting environment.

Translocation of Foliar Absorbed Zn in Sunflower (Helianthus annuus) Leaves.

Foliar zinc (Zn) fertilization is an important approach for overcoming crop Zn deficiency, yet little is known regarding the subsequent translocation of this foliar-applied Zn. Using synchrotron-based X-ray fluorescence microscopy (XFM) and transcriptome analysis, the present study examined the translocation of foliar absorbed Zn in sunflower (Helianthus annuus) leaves. Although bulk analyses showed that there had been minimal translocation of the absorbed Zn out of the leaf within 7 days, in situ analyses showed that the distribution of Zn in the leaf had changed with time. Specifically, when Zn was applied to the leaf for 0.5 h and then removed, Zn primarily accumulated within the upper and lower epidermal layers (when examined after 3 h), but when examined after 24 h, the Zn had moved to the vascular tissues. Transcriptome analyses identified a range of genes involved in stress response, cell wall reinforcement, and binding that were initially upregulated following foliar Zn application, whereas they were downregulated after 24 h. These observations suggest that foliar Zn application caused rapid stress to the leaf, with the initial Zn accumulation in the epidermis as a detoxification strategy, but once this stress decreased, Zn was then moved to the vascular tissues. Overall, this study has shown that despite foliar Zn application causing rapid stress to the leaf and that most of the Zn stayed within the leaf over 7 days, the distribution of Zn in the leaf had changed, with Zn mostly located in the vascular tissues 24 h after the Zn had been applied. Not only do the data presented herein provide new insight for improving the efficiency of foliar Zn fertilizers, but our approach of combining XFM with a transcriptome methodological system provides a novel approach for the study of element translocation in plants.

Separating multiple, short-term, deleterious effects of saline solutions on the growth of cowpea seedlings.

• Reductions in plant growth as a result of salinity are of global importance in natural and agricultural landscapes. • Short-term (48-h) solution culture experiments studied 404 treatments with seedlings of cowpea (Vigna unguiculata cv Caloona) to examine the multiple deleterious effects of calcium (Ca), magnesium (Mg), sodium (Na) or potassium (K). • Growth was poorly related to the ion activities in the bulk solution, but was closely related to the calculated activities at the outer surface of the plasma membrane, {I(z)}0°. The addition of Mg, Na or K may induce Ca deficiency in roots by driving {Ca²+}0° to < 1.6 mM. Shoots were more sensitive than roots to osmolarity. Specific ion toxicities reduced root elongation in the order Ca²+ > Mg²+ > Na+ > K+. The addition of K and, to a lesser extent, Ca alleviated the toxic effects of Na. Thus, Ca is essential but may also be intoxicating or ameliorative. • The data demonstrate that the short-term growth of cowpea seedlings in saline solutions may be limited by Ca deficiency, osmotic effects and specific ion toxicities, and K and Ca alleviate Na toxicity. A multiple regression model related root growth to osmolarity and {I(z)}0° (R²=0.924), allowing the quantification of their effects.

Calculated activity of Mn2+ at the outer surface of the root cell plasma membrane governs Mn nutrition of cowpea seedlings.

Manganese (Mn) is an essential micronutrient for plant growth but is often toxic in acid or waterlogged soils. Using cowpea (Vigna unguiculata L. Walp.) grown with 0.05-1500 µM Mn in solution, two short-term (48 h) solution culture experiments examined if the effects of cations (Ca, Mg, Na, Al, or H) on Mn nutrition are related to the root cells' plasma membrane (PM) surface potential, ψ(0)(0). When grown in solutions containing levels of Mn that were toxic, both relative root elongation rate (RRER) and root tissue Mn concentration were more closely related to the activity of Mn(2+) at the outer surface of the PM, {Mn(2+)}(0)(0) (R(2)=0.812 and 0.871) than to its activity in the bulk solution, {Mn(2+)}(b) (R(2)=0.673 and 0.769). This was also evident at lower levels of Mn (0.05-10 µM) relevant to studies investigating Mn as an essential micronutrient (R(2)=0.791 versus 0.590). In addition, changes in the electrical driving force for ion transport across the PM influenced both RRER and the Mn concentration in roots. The {Mn(2+)}(b) causing a 50% reduction in root growth was found to be c. 500 to >1000 µM (depending upon solution composition), whilst the corresponding value was 3300 µM when related to {Mn(2+)}(0)(0). Although specific effects such as competition are not precluded, the data emphasize the importance of non-specific electrostatic effects in the Mn nutrition of cowpea seedlings over a 1×10(5)-fold range of Mn concentration in solution.

Alleviation of Cu and Pb rhizotoxicities in cowpea (Vigna unguiculata) as related to ion activities at root-cell plasma membrane surface.

Cations, such as Ca and Mg, are generally thought to alleviate toxicities of trace metals through site-specific competition (as incorporated in the biotic ligand model, BLM). Short-term experiments were conducted with cowpea (Vigna unguiculata L. Walp.) seedlings in simple nutrient solutions to examine the alleviation of Cu and Pb toxicities by Al, Ca, H, Mg, and Na. For Cu, the cations depolarized the plasma membrane (PM) and reduced the negativity of ψ(0)(o) (electrical potential at the outer surface of the PM) and thereby decreased {Cu(2+)}(0)(o) (activity of Cu(2+) at the outer surface of the PM). For Pb, root elongation was generally better correlated to the activity of Pb(2+) in the bulk solution than to {Pb(2+)}(0)(o). However, we propose that the addition of cations resulted in a decrease in {Pb(2+)}(0)(o) but a simultaneous increase in the rate of Pb uptake (due to an increase in the negativity of E(m,surf), the difference in potential between the inner and outer surfaces of the PM) thus offsetting the decrease in {Pb(2+)}(0)(o). In addition, Ca was found to alleviate Pb toxicity through a specific effect. Although our data do not preclude site-specific competition (as incorporated in the BLM), we suggest that electrostatic effects have an important role.

Trace metal phytotoxicity in solution culture: a review.

Solution culture has been used extensively to determine the phytotoxic effects of trace metals. A review of the literature from 1975 to 2009 was carried out to evaluate the effects of As(V), Cd(II), Co(II), Cu(II), Hg(II), Mn(II), Ni(II), Pb(II), and Zn(II) on plants grown in solution. A total of 119 studies was selected using criteria that allowed a valid comparison of the results; reported toxic concentrations varied by five orders of magnitude. Across a range of plant species and experimental conditions, the phytotoxicity of the trace metals followed the trend (from most to least toxic): Pb approximately Hg >Cu >Cd approximately As >Co approximately Ni approximately Zn >Mn, with median toxic concentrations of (muM): 0.30 Pb, 0.47 Hg, 2.0 Cu, 5.0 Cd, 9.0 As, 17 Co, 19 Ni, 25 Zn, and 46 Mn. For phytotoxicity studies in solution culture, we suggest (i) plants should be grown in a dilute solution which mimics the soil solution, or that, at a minimum, contains Ca and B, (ii) solution pH should be monitored and reported (as should the concentrations of the trace metal of interest), (iii) assessment should be made of the influence of pH on solution composition and ion speciation, and (iv) both the period of exposure to the trace metal and the plant variable measured should be appropriate. Observing these criteria will potentially lead to reliable data on the relationship between growth depression and the concentration of the toxic metal in solution.

Metal ion effects on hydraulic conductivity of bacterial cellulose-pectin composites used as plant cell wall analogs.

Low concentrations of some trace metals markedly reduce root elongation rate and cause ruptures to root rhizodermal and outer cortical cells in the elongation zone. The interactions between the trace metals and plant components responsible for these effects are not well understood but may be linked to changes in water uptake, cell turgor and cell wall extensibility. An experiment was conducted to investigate the effects of Al, La, Cu, Gd, Sc and Ru on the saturated hydraulic conductivity of bacterial cellulose (BC)-pectin composites, used as plant cell wall analogs. Hydraulic conductivity was reduced to approximately 30% of the initial flow rate by 39 microM Al and 0.6 microM Cu, approximately 40% by 4.6 microM La, 3 microM Sc and 4.4 microM Ru and approximately 55% by 3.4 microM Gd. Scanning electron microscopy (SEM) revealed changes in the ultrastructure of the composites. The results suggest that trace metal binding decreases the hydraulic conductivity through changes in pectin porosity. The experiment illustrates the importance of metal interactions with pectin, and the implications of such an interaction in plant metal toxicity and in normal cell wall processes.

Evaluating effects of iron on manganese toxicity in soybean and sunflower using synchrotron-based X-ray fluorescence microscopy and X-ray absorption spectroscopy.

With similar chemistry, Mn and Fe interact in their many essential roles in plants but the magnitude and mechanisms involved of these interactions are poorly understood. Leaves of soybean (a Mn-sensitive species) developed a mild chlorosis and small dark spots and distorted trifoliate leaves with 30 µM Mn and 0.6 µM Fe in nutrient solution (pH 5.6; 3 mM ionic strength). At 0.6 µM Fe, lower alternate leaves of sunflower (a Mn-tolerant species) were chlorotic at 30 µM Mn and had a pale chlorosis and necrosis at 400 µM Mn. A concentration of 30 and 300 µM Fe in solution alleviated these typical symptoms of Mn toxicity and decreased the concentration of Mn from >3000 to ca. 800 mg kg-1 dry mass (DM) in all leaf tissues. As expected, increased Fe supply increased Fe in leaves from <100 up to 1350 mg Fe kg-1 DM. In situ synchrotron-based X-ray fluorescence microscopy showed that increased Fe supply caused an overall decrease in Mn in the leaf tissue but had little effect on the pattern of its distribution. Similarly, X-ray absorption spectroscopy identified only slight effects of Fe supply on Mn speciation in leaf tissues. Thus, the results of this study indicate that increased Fe supply ameliorated Mn toxicity in soybean and sunflower largely through decreased Mn uptake and translocation to leaf tissues rather than through changes in Mn distribution or speciation within the leaves.

Silver sulfide nanoparticles (Ag2S-NPs) are taken up by plants and are phytotoxic.

Silver nanoparticles (NPs) are used in more consumer products than any other nanomaterial and their release into the environment is unavoidable. Of primary concern is the wastewater stream in which most silver NPs are transformed to silver sulfide NPs (Ag2S-NPs) before being applied to agricultural soils within biosolids. While Ag2S-NPs are assumed to be biologically inert, nothing is known of their effects on terrestrial plants. The phytotoxicity of Ag and its accumulation was examined in short-term (24 h) and longer-term (2-week) solution culture experiments with cowpea (Vigna unguiculata L. Walp.) and wheat (Triticum aestivum L.) exposed to Ag2S-NPs (0-20 mg Ag L(-1)), metallic Ag-NPs (0-1.6 mg Ag L(-1)), or ionic Ag (AgNO3; 0-0.086 mg Ag L(-1)). Although not inducing any effects during 24-h exposure, Ag2S-NPs reduced growth by up to 52% over a 2-week period. This toxicity did not result from their dissolution and release of toxic Ag(+) in the rooting medium, with soluble Ag concentrations remaining below 0.001 mg Ag L(-1). Rather, Ag accumulated as Ag2S in the root and shoot tissues when plants were exposed to Ag2S-NPs, consistent with their direct uptake. Importantly, this differed from the form of Ag present in tissues of plants exposed to AgNO3. For the first time, our findings have shown that Ag2S-NPs exert toxic effects through their direct accumulation in terrestrial plant tissues. These findings need to be considered to ensure high yield of food crops, and to avoid increasing Ag in the food chain.

Inhibition of cell-wall autolysis and pectin degradation by cations.

Modification of cell wall components such as cellulose, hemicellulose and pectin plays an important role in cell expansion. Cell expansion is known to be diminished by cations but it is unknown if this results from cations reacting with pectin or other cell wall components. Autolysis of cell wall material purified from bean root (Phaseolus vulgaris L.) occurred optimally at pH 5.0 and released mainly neutral sugars but very little uronic acid. Autolytic release of neutral sugars and uronic acid was decreased when cell wall material was loaded with Ca, Cu, Sr, Zn, Al or La cations. Results were also extended to a metal-pectate model system, which behaved similarly to cell walls and these cations also inhibited the enzymatic degradation by added polygalacturonase (EC 3.2.1.15). The extent of sugar release from cation-loaded cell wall material and pectate gels was related to the degree of cation saturation of the substrate, but not to the type of cation. The binding strength of the cations was assessed by their influence on the buffer capacity of the cell wall and pectate. The strongly bound cations (Cu, Al or La) resulted in higher cation saturation of the substrate and decreased enzymatic degradability than the weakly held cations (Ca, Sr and Zn). The results indicate that the junction zones between pectin molecules can peel open with weakly held cations, allowing polygalacturonase to cleave the hairy region of pectin, while strongly bound cations or high concentrations of cations force the junction zone closed, minimising enzymatic attack on the pectin backbone.

The rhizotoxicity of metal cations is related to their strength of binding to hard ligands.

Mechanisms whereby metal cations are toxic to plant roots remain largely unknown. Aluminum, for example, has been recognized as rhizotoxic for approximately 100 yr, but there is no consensus on its mode of action. The authors contend that the primary mechanism of rhizotoxicity of many metal cations is nonspecific and that the magnitude of toxic effects is positively related to the strength with which they bind to hard ligands, especially carboxylate ligands of the cell-wall pectic matrix. Specifically, the authors propose that metal cations have a common toxic mechanism through inhibiting the controlled relaxation of the cell wall as required for elongation. Metal cations such as Al(3+) and Hg(2+), which bind strongly to hard ligands, are toxic at relatively low concentrations because they bind strongly to the walls of cells in the rhizodermis and outer cortex of the root elongation zone with little movement into the inner tissues. In contrast, metal cations such as Ca(2+), Na(+), Mn(2+), and Zn(2+) , which bind weakly to hard ligands, bind only weakly to the cell wall and move farther into the root cylinder. Only at high concentrations is their weak binding sufficient to inhibit the relaxation of the cell wall. Finally, different mechanisms would explain why certain metal cations (for example, Tl(+), Ag(+), Cs(+), and Cu(2+)) are sometimes more toxic than expected through binding to hard ligands. The data presented in the present study demonstrate the importance of strength of binding to hard ligands in influencing a range of important physiological processes within roots through nonspecific mechanisms.