Effect of physiological activities on aluminum uptake in carrot (Daucus carota L.) cells in suspension culture

Responses to aluminum (AI) of carrot (Daucus carota L.) cells in suspension culture (hereafter referred as “carrot suspension cells”) were investigated under different physiological activities. The respiration rate of the cells at the logarithmic growth phase decreased when the external Al concentration increased. A similar tendency was observed for the ATP content of the cells when the external Al concentration of the medium increased. Decrease of about 50% of the respiration rate and of about 35% of the ATP content of the cells was. observed at 300 µm AlCl₃ at 24 h, respectively. The Al content (total and citrate-insoluble AI) in cells increased when the external Al concentration increased. Both the respiration rate and the ATP content in cells were reduced when the culture temperature decreased or ATP synthesis inhibitors, namely CCCP (carbonylcyanide-m-chlorophenylhydrazone) and DNP (2,4-dinitrophenol) were added. The addition of SHAM (salicylhydroxamic acid) did not affect the ATP content in cells. The Al content of the cells treated with 50 µm Al together with SHAM was similar to that of the cells treated with only AI. However, the Al content of the cells increased when the cells were treated with 50 µm Al together with CCCP, DNP, or a low temperature (15°C). The content of citrate Al content in cells increased rapidly when ATP synthesis inhibitors were present or the culture temperature was low. There was a negative linear correlation between the Al content and the ATP content in cells. These results suggest that the cells containing a large amount of ATP and showing normal functions of the plasma membranes may display an apparent tolerance to Al by suppressing Al absorption into their cytoplasm at a cell suspension level.     

Response of cotton cultivars to aluminum in solutions with varying silicon concentrations

Abstract

Cotton (Gossypium hirsutum L.) is extremely sensitive to Al toxicity. Increasing Si concentration in solution has been reported to alleviate AI toxicity. In this investigation the effects of varying Si concentrations (700, 1400, and 2800 μM Si) on reactive Al (defined as Al reactive with aluminon during 10‐s reaction time, without acidification and heating) was studied in solutions containing either 50, 100 or 200 μM Al during 50 d of aging. An increase in Si concentration had negligible effects on the reactive Al in solutions with 50 or 100 μM Al. However, in solutions with 200 μM Al the reactive Al decreased by 6 to 15% with an increase in Si concentration from 0 to 2800 μM.

The effects of either 700, 1400 or 2800 μM Si on root growth of Coker 208, Coker 315, DPL 90, McNair 235, Stoneville 506 and Tifcot 56 cotton cultivars were investigated in solutions containing either 0, 10, 20 or 40 μM Al with 500 μM Ca at pH 4.5. In solutions containing no Al, addition of 700 μM Si improved root growth by 69–87% in Coker 315, DPL 90 and McNair 235 cultivars but not in the other cultivars. In solutions containing 10 μM Al, an increase in Si concentration from 0 to 2800 μM improved the root growth by 15–17% in DPL 90 and McNair 235 cultivars only. An increase in Si additions failed to improve root growth of any of the cultivars in solutions with 20 or 40 μM Al.     

Tea plant (Camellia sinensis L.) roots secrete oxalic acid and caffeine into medium containing aluminum

We examined the response of the tea plant (Camellia sinensis L.) to aluminum (Al) exposure under sterile conditions, focusing specifically on the secretion of low molecular mass organic compounds from roots. After germination in agar medium, tea seedlings together with medium were placed on agar containing 0.4?mM Al with 0.2% hematoxyline (hematoxylin-Al medium). The purple color of the hematoxylin-Al medium was observed to fade gradually, until none of the color remained 6 days later. The tea seedlings were then treated with simple calcium solution (0.2?mM, at pH 4.2) containing AlCl₃, which ranged in concentration from 0 to 0.8?mM, for 24?hrs. The amount of oxalate secreted into the medium increased as the external Al concentration increased, while the concentrations of malate and citrate in the medium remained unchanged. Oxalate secretion started within 30?min after Al exposure and increased linearly thereafter. The findings demonstrated that oxalate was a key compound in the Al-tolerance mechanism employed by the tea plant, which detoxifies Al³⁺ externally in the rhizosphere. In addition to oxalate, caffeine was also secreted by tea roots in response to Al exposure. It is possible that caffeine excretion from the roots of tea plants may stimulate root growth through the inhibition of callose deposition in root tips.     

Mineral composition of leaves and bark in aluminum accumulators in a tropical rain forest in Indonesia

Mineral composition including AI, Ca, Mg, P, S, and Si and relationships between Al and other elements such as Ca, Mg, P, S, and Si in leaves and bark of trees in a tropical rain forest in West Sumatra were studied. Sixty five tree species and 12 unidentified trees were referred to as AI accumulators based on Chenery\s's definition (more than 1 g kg^-1 Al in leaves). For most of the Al accumulators, Al concentration in leaves was higher than in bark. However, some members of Euphorbiaceae, Melastomataceae, and Ulmaceae families showed a reverse trend. Most of the non-accumulators also showed a higher Al concentration in bark than in leaves. These results indicated that there was a difference in the mechanism of Al accumulation in tree bodies. Some of the Al accumulators showed an extremely high Al concentration (more than 10 g kg^-1) not only in the mature leaves, but also in the new leaves. Analysis of the relationships between the concentration of Al and the other 5 elements in leaves, revealed that Al accumulators could be separated into two groups at the Al concentration of 3 g kg^-1. This finding suggested that new criteria based on Al concentration (23 g kg^-1) or Al/Ca ratio in leaves could be proposed in order to define Al accumulators, apart from Chenery's criterion. Aluminium accumulators with an Al concentration in leaves lower than 3 g kg^-1 (AI accumulators <3 g kg^-1) showed the same trend as the non-accumulators in terms of these elemental relationships, while Al accumulators with an Al concentration in leaves higher than 3 g kg^-1 (AI accumulators 23 g kg^-1) showed a different trend from the non-accumulators. The Al accumulators 23 g kg^-1 and the other trees (AI accumulators < 3 g kg^-1 and non-accumulators) showed separately positive correlations between the concentrations of AI and Ca (or Mg) in the leaves. This observation seems to be opposite to general findings in plant nutrition, i.e. inhibition of Ca or Mg uptake by AI. A positive correlation between Al and S was also observed for all the trees. The Al accumulators ≥3 g kg^-1 showed positive correlations between the concentrations of Al and P (or Si) in the leaves, unlike the other trees. These findings suggested that Al stimulated P, S, or Si accumulation in leaves or Al was transported with P, S, or Si for the Al accumulators ≥3 g kg^-1. No negative relationships between Al and the other 5 elements in the leaves were observed for the Al accumulators ≥3 g kg^-1.     

Aluminum toxicity in tomato. Part 2. leaf gas exchange,chlorophyll content,and invertase activity

The effect of aluminum (Al) toxicity on leaf gas exchange, leaf chlorophyll content, and sucrose metabolizing enzyme activity of two tomato cultivars (Lycopersicon esculentum Mill. ‘Mountain Pride’ and ‘Floramerica') was studied to determine the mechanism of growth reduction observed in a related study (Simon et al., 1994, Part 1). Plants were grown in diluted nutrient solution (pH 4.0) with 0, 10, 25, or 50 μM. Al for 16 days. Leaf gas exchange was reduced 2–3 fold in both cultivars as Al concentration increased. Gas exchange of ‘Mountain Pride’ was more sensitive to Al toxicity than ‘Floramerica’, agreeing with growth responses observed. Reductions in carbon dioxide (CO₂) assimilation rate appeared to be due to nonstomatal factors in ‘Floramerica’, but stomatal and non‐stomatal limitations in ‘Mountain Pride’. Chlorophyll content of leaves was not affected by Al. Acid invertase (AI) and neutral invertase (NI) activity of roots responded consistently to Al concentration in both cultivars. Root AI and NI activity decreased to a greater extent for ‘Mountain Pride’ than for ‘Floramerica’.     

Aluminium-Toleranz von Ackerbohne (Vicia faba), Lupine (Lupinus luteus), Gerste (Hordeum vulgare) und Roggen (Secale cereale). I. Sproß- und Wurzelwachstum in Abhängigkeit vom Aluminium-Angebot

AI tolerance of horse bean, yellow lupin, barley and rye. I. Shoot and root growth as affected by Al supply In solution culture considerable differences existed in Al tolerance between the plant species horse bean (Vicia faba ?Herz Freya”?), yellow lupin (Lupinus luteus ?Schwako”?), barley (Hordeum vulgare ?Roland”?) and rye (Secale cereale ?Kustro”?): compared to barley (0.05 μg Al l^?1 = 1.85 μM Al) an 80 fold higher concentration of Al was necessary for lupin and rye for comparable growth depression and for horse bean a 5 times higher Al concentration. Injury by Al after 7 days of Al treatment was most effectively and sensitively characterized by an inhibition of elongation of seminal and especially of lateral roots. Numbers of laterals were also reduced. Dry matter production of roots and shoots was less affected by Al. In lupin, low Al supply even slightly increased the dry weight. The high Al tolerance of rye and yellow lupin in solution culture during the seedling stage is in good agreement with their adaptation to acid mineral soils.     

Effect of Aluminum and Silicon in Solution on the Growth of Rice (Oryza sativa L.) uspension Cells

Cells of two indica rice (Oryza sativa L.) varieties, one aluminum-tolerant (IAC3), and the other aluminum-sensitive (IR45), were grown in suspension culture in a medium with or without Al (0-800 µM), Si (0-2,000 µM), or Al and Si in combination as well as in R2 medium as control. The influence of Al or Si on cell growth was evaluated based on the growth of the cells in the treatment medium as well as in the R2 medium and compared with the effect on rice seedlings. Similar responses were observed for the cells and the seedlings; Al treatment resulted in a significant reduction of cell growth in both varieties, and the growth was reduced with the increase of the Al uptake by the cells. IAC3 was more tolerant to Al and it absorbed more Al than IR45. On the contrary, Si alone had almost no effect or exerted a negligible effect on the cell growth in both varieties and Si was hardly absorbed by the cells. There was a slight alleviative effect of Si on Al toxicity in the suspension cells of IR45, whereas, a considerable ameliorative effect in the seedlings. The content of total Al and of AI insoluble in citric acid in the cells were higher in the sole Al (800 µM) treatment than in the Al-Si combination treatment. This tendency was also observed in aluminon staining. The differences in the alleviative effects of Si and the uptake of Al and Si between the suspension cells and seedlings may have resulted from the presence of roots and morphological differences in the apoplastic tissues.     

Difference in response to aluminum among Japanese coniferous species

Seedlings of Chamaecyparis obtusa, Cryptomeria japonica D. Don, and Abies firma Sieb. et Zucco were grown hydroponically for 4 weeks in the presence or absence of aluminum (Al) and with or without reduced pH. Under exposure to AI, root and shoot growth of C. obtusa was enhanced. A. firma showed the same tendency as C. obtusa, though not significantly. Only in C. japonica, growth was reduced with Al, especially shoot growth. In all the species, callose production in the root tips was observed in the presence of Al. A positive correlation was observed between the relative root callose content and relative root growth (r = 0.83), and significant root elongation with AI treatment was observed in all the species. Therefore, it is considered that callose deposition in the root tips of these species may not indicate the Al-induced root cell injury causing root growth inhibition. The highest callose content in the root tip and strong callose fluorescence in the epidermis and zones of cell contact were observed in C. obtusa. Since the Al translocation rate from roots to leaves was the lowest in C. obtusa and since significant growth enhancement was observed in the presence of Al, it is possible that the accumulation of callose in the root epidermis and in the zones of cell contact is related to Al-resistance in C. obtusa.     

Impact of Boron on Biomass Production and Nutrition of Aluminum‐Stressed Apple Rootstocks

Abstract

The aim of the experiment was to examine the effect of boron (B) on biomass production and nutrition of aluminum (Al)‐stressed apple (Malus sp.) rootstocks. The study was carried out under greenhouse conditions on Polish rootstock (P22) and Malling 26 (M.26) planted singly into 1‐L plastic pots filled with perlite and supplied with Hoagland's medium at pH 4.5 without or with Al (100 µM as AlCl₃). Boron was added into the Al‐containing medium at 20, 40, or 60 µM whereas into the medium without Al only at 20 µM as boric acid. The results showed that the presence of Al in the medium reduced biomass production of P22 and M.26 rootstocks by 22% and 41%, respectively. Rates of uptake and translocation of phosphorus (P), magnesium (Mg), and calcium (Ca) to aerial plant parts were decreased for Al‐treated rootstocks. Aluminum‐stressed P22 rootstocks grown in nutrient solution at 40 and 60 µM B had higher dry weight of leaves and roots, and also higher ability to take up P, Mg, and Ca and lower Al than those grown in the presence of Al at 20 µM B in the medium. Rates of absorption and transport of B to aerial plant parts corresponded with B level in the medium. These results suggest that on acid soils with high Al availability, supra‐optimal B concentrations in soil solution (40–60 µM) can prevent/alleviate Al toxicity in apple trees grafted on P22 rootstocks.     

铝改性秸秆生物质炭去除水体中的大肠杆菌

用一次平衡实验和柱淋溶实验研究了秸秆生物质炭和铝改性秸秆生物质炭对水中大肠杆菌的吸附量和去除率。结果表明,未改性秸秆生物质炭对大肠杆菌的吸附量很低,铝改性秸秆生物质炭对大肠杆菌有很高的吸附容量和去除率,0.6 mol/L Al(Ⅲ)改性生物质炭对大肠杆菌的去除效果优于0.3 mol/L Al(Ⅲ)改性生物质炭。当大肠杆菌浓度低于0.63 mg/ml时,铝改性大豆秸秆炭对大肠杆菌的去除率达100%,铝改性花生秸秆炭和铝改性稻草炭对大肠杆菌的去除率在96%和90%以上。0.5 mg/ml的大肠杆菌悬液通过装有2 g铝改性稻草炭的淋溶柱,每次收集10 ml淋出液,共收集152次,大部分淋出液中大肠杆菌浓度小于0.02 mg/ml,大肠杆菌去除率在95%以上。因此,铝改性生物质炭可用于水体中大肠杆菌的去除。     

Calcium alleviation of hydrogen and aluminum inhibition of soybean root extension from limed soil into acid subsurface solutions

Alleviation by calcium (Ca) of inhibition of soybean [Glycine max (L.) Merr. cv. ‘Ransom'] root elongation by hydrogen (H) and aluminum (Al) was evaluated in a vertical split‐root system. Roots extending from a limed and fertilized soil compartment grew for 12 days into a subsurface compartment containing nutrient solution with treatments consisting of factorial combinations of either pH (4.0, 4.6, and 5.5) and Ca (0.2, 2.0, 10, and 20 mM), Al (7.5, 15, and 30 μM) and Ca (2.0,10, and 20 mM) at pH 4.6, or Ca (2, 7, and 12 mM) levels and counter ions (SO₄ and Cl) at pH 4.6 and 15 μM Al. Length of tap roots and their laterals increased with solution Ca concentration and pH value, but decreased with increasing Al level. Length of both tap and lateral roots were greater when Ca was supplied as CaSO₄ than as CaCl₂, but increasing Ca concentration from 2 to 12 mM had a greater effect on alleviating Al toxicity than Ca source. In the absence of Al, relative root length (RRL) of tap and lateral roots among pH and Ca treatments was related to the Ca:H molar activity ratio of solutions (R²≥0.82). Tap and lateral RRL among solutions with variable concentrations of Al and Ca at pH 4.6 were related to both the sum of the predicted activities of monomeric Al (R²≥0.92) and a log‐transformed and valence‐weighted balance between activities of Ca and selected monomeric Al species (R²≥0.95). In solutions with 15 μM Al at pH 4.6, response of tap and lateral RRL to variable concentrations of CaSO₄ and CaCl₂ were related to predicted molar activity ratios of both Ca:Al³⁺ (R²≥0.89) and Ca:3 monomeric Al (R²≥0.90), provided that AISO₄ and AI(SO₄)₂ species were excluded from the latter index. In all experiments H and Al inhibited length of lateral roots more than tap roots, and a greater Ca:H or Ca:Al concentration ratio was required in solutions to achieve similar RRL values as tap roots.     

Responses to aluminium of suspension-cultured tobacco cells in a simple calcium solution

In a simple solution containing 3 mM CaCl₂ and 3% sucrose (pH 4.5), tobacco cells (Nicotiana tabacum L.) at the logarithmic phase of growth remained viable at least for 24 h. In this medium, the toxic effect of aluminium (Al) on the plasma membrane was investigated for up to 24 h. After the addition of Al to the cell suspension, Al started to accumulate immediately in the cells, and a maximum value was observed at 9 h. Al induced callose deposition, but did not enhance significantly the uptake of Evans blue (a nonpermeating dye), the per oxidation of lipids and the leakage of potassium (K) ions. Furthermore, the AI-treated cells were stained with fluorescein diacetate (FDA) as much as untreated control cells. These results suggest that the accumulation of Al does not damage the membrane. The addition of Fe(II) to the cells which had been exposed to Al for 12 h resulted in immediate lipid peroxidation and Evans blue uptake several hours later. A combination of Al and Fe(II) caused the K leakage, and enhanced the deposition of callose more than Al alone. These results suggest that the accumulation of Al sensitizes the membrane to the Fe(II)-mediated peroxidation of lipids, and that the Al-enhanced per oxidation of lipids is a direct cause of the loss of integrity of the plasma membrane (or cell death) in the Ca medium.     

油茶根系吸收铝导致生长介质酸化

选用油茶为试验材料,采用固体琼脂培养和溶液培养两种试验体系,研究了油茶根系吸收铝离子与生长介质pH变化的关系。琼脂培养试验结果表明,在钙离子和铝离子介质中,油茶根际区域均发生了明显的酸化;溶液培养试验结果进一步表明,阳离子的吸收尤其是铝离子的吸收是介质酸化的原因,铝与钙同时存在时,溶液酸化主要是由铝离子的吸收引起的。另外,随着溶液铝浓度的增加,溶液pH下降幅度增加,二者之间呈现极显著正相关关系,且油茶培养时间越长,溶液pH下降越多。所有这些结果表明油茶吸收铝会降低生长介质pH。     

Effect of Glomus etunicatum inoculation on aluminum,phosphorus, calcium,and magnesium uptake of two barley genotypes with different aluminum tolerance

Abstract

This study was conducted to evaluate the effect of vesicular‐arbuscular mycorrhizal (VAM) fungus Glomus etunicatum on growth, absorption, and distribution of calcium (Ca), magnesium (Mg), phosphorus (P), and aluminum (Al) in one Al‐tolerant and one Al‐sensitive barley cultivar. The plants were grown in sand daily irrigated with nutrient solution containing 0 or 600 μM Al at pH 4.8. Significant interaction (P=0.05) among variety, mycorrhiza, and aluminum (VxMxAl) were noted for both shoot and root dry matter (DM); shoot concentration and content of Al, P, Ca, and Mg; root concentration of Al, P, and Mg; and root content of Al, P, Ca, and Mg. With VAM inoculation: i) root colonization degree was about 50% in all treatment, ii) shoot DM yield increased between 30 and 70%, iii) Al concentration and content decrease down to a half both in shoots and roots of sensitive barley, iv) Ca concentration in shoots of sensitive barley showed a high increase at 600 μM Al, and v) P concentration and content in shoots of both varieties increased significantly.     

Effect of nodulation with Bradyrhizobium japonicum and Shinorhizobium fredii on xylem sap composition of Peking (Glycine max L. Merr )

Five barley cultivars were grown together in complete, low-P·low-pH and high-Al medium containing only NO₃, only NH₄ or both NO₃ and NH₄ as N sources, respectively using an automatic control system of pH for water culture, and the relationship between the differential Al tolerance and the plant-induced pH change of medium among the barley cultivars was investigated.

The pH of the medium containing only NO₃ as N source tended to increase, whereas the pH of the other media containing only NH₄ or both NO₃ and NH₄ as N sources tended to decrease, but the fluctuations of the medium pH could be maintained within the value of 0.2 pH in the complete medium and within the value of 0.1 pH in the high-Al medium.

Barley cultivars still differed in their Al tolerance in the medium which was continuously stirred and circulated at a constant pH. The pattern of Al tolerance was not affected by the N sources in the medium. The plant-induced pH change of medium for each cultivar was influenced by the N sources in the medium, and was not correlated positively with Al tolerance. The contents of Al and Ca or other nutrient cations in roots were positively correlated with Al tolerance and positive correlations were recognized also between the contents of Al and Ca or some other nutrient cations in the roots.

In conclusion, the following mechanisms are proposed. Al tolerant barley cultivars exclude Al actively outside the plasmalemma of the root cells, and the excluded Al may polymerize and or react with P to form Al precipitates. Consequently, in the Al tolerant barley cultivars the Al content may be low in the root protoplasts, high in the whole root tissues and the contents of Ca or other nutrients may be high in the roots. The plant-induced pH change of medium is not considered to be the cause of the differential Al tolerance among barley cultivars.     

Cell death process initiated by a combination of aluminium and iron in suspension-cultured tobacco cells (Nicotiana tabacum): Apoptosis-like cell death mediated by calcium and proteinase

Aluminium (AI) enhances ferrous ion (Fe²⁺) -mediated lipid per oxidation which is the primary factor leading to cell death in nutrient medium in tobacco cells. Under these conditions, cell death processes were examined. Cells which had been treated with AI and Fe²⁺ together exhibited cell shrinkage and DNA ladders which were detected by agarose-gel electrophoresis in the form of bands corresponding to increasing multiples of ～150 bp fragments. In the calcium (Ca)-free medium, a combination of Al and Fe2+ enhanced only partly the per oxidation of lipids and hardly promoted cell death, indicating that Ca²⁺ further stimulates the per oxidation of lipids enhanced by a combination of Al and Fe²⁺ and is required for cell death. N-Tosyl-L-phenylalanylchloromethyl ketone, an inhibitor of some cysteine and serine proteinases, had no effect on the per oxidation of lipids but almost completely blocked cell death enhanced by a combination of AI and Fe²⁺, suggesting that proteinase acts downstream from the per oxidation of lipids. On the basis of these findings, we conclude that cell death initiated by a combination of Al and Fe²⁺ is involved in the apoptosislike cell death program mediated by extracellular Ca²⁺ and endogenous proteinase.     

Aluminium toxicity and multiplication of Rhizobium trifolii in a defined growth medium

The responses of six strains of R. trifolii to pH, Ca, Al and phosphate were studied in the defined medium of Munns and Keyser (1981) supplemented with vitamins. Wild-type strains responded in the same way as rifampicin-resistant mutant strains. Multiplication was inhibited by acidity alone at pH4.3, and by 50 μM Al at pH 5.5 (despite the precipitation of Al). This inhibitory effect of Al was overcome by increasing the pH above 6.0, or by increasing the phosphate concentration from 10 to 100 μm. Ca concentration had no effect, nor did holding the medium at 22°C for 36 weeks before inoculation. There was variation between strains in their responses to concentrations of A1 < 50 μM. These responses indicate that the defined medium is a satisfactory model for the white clover rhizosphere.Results from these studies and from chemical analyses indicate that the inhibitory effect of Al under partially-neutralized conditions is not due to a direct effect of monomeric A1, nor to an indirect effect of polymeric A1 in reducing the concentration of phosphate, but is due to a direct effect of polymeric A1.     

Aluminum status of synthetic Al–humic substance complexes and their influence on plant root growth

Abstract

Aluminum (Al)–humus complexes are abundant in the A horizons of non-allophanic Andosols and contribute to the unique properties of volcanic ash soils, such as high reactivity with phosphate ions and a low bulk density. Natural non-allophanic Andosols commonly show Al toxicity to plant roots. There have been very few studies examining the contribution of Al–humus complexes to the Al toxicity of plant roots, although the complexes are the probable source of the toxic Al. We extracted humic substances from the A horizon of a non-allophanic Andosol using NaOH solution and reacted the humic substances and partially neutralized the AlCl₃ solution at three pH conditions (pH 4.0, 4.5 and 5.5) to prepare pure Al–humic substance complexes. The Al solubility study (equilibrium study in 10^?2 mol L^?1 CaCl₂) and the Al release study (a stirred-flow method using 10^?3 mol L^?1 acetate buffer solution adjusted to pH 3.5) indicated that all the synthetic complexes easily and rapidly release monomeric Al into the liquid phase with slight changes in pH and ion strength, although the Al contents and their extent of polymerization are considerably different among the complexes. A plant growth test was conducted using a medium containing the Al–humic substance complexes and perlite mixture. Root growth in burdock (Arctium lappa) and barley (Hordeum vulgare L.) was reduced equally by all three complex media, and the roots showed the typical injury symptoms of Al toxicity. These results indicate that in soils dominated by Al–humus complexes the Al released from the Al–humus complexes, as well as the exchangeable Al adsorbed by soil minerals, is definitely toxic to plant roots.     

Sucrose transporter NtSUT1 confers aluminum tolerance on cultured cells of tobacco (Nicotiana tabacum L.)

The role of plasma membrane-localized sucrose transporter (NtSUT1) was investigated using cultured tobacco cell (Nicotiana tabacum L.) line BY-2. The wild type (WT) cells were first transformed with the NtSUT1 gene or its fragments cloned from tobacco cell line SL to form the over-expression (OX) and suppression (RNAi) cell lines, respectively. Using OX and RNAi transgenics, the role of NtSUT1 in growth capacity of actively growing cells and in aluminum (Al)-treated cells was examined. During the logarithmic phase of growth in nutrient medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), both the rate of sucrose uptake measured with radio-tracer and the content of soluble sugars were higher in OX and lower in RNAi cell lines compared to WT. Overall, the content of soluble sugars negatively correlated with the time necessary for doubling mass (fresh weight). When cells were treated without (control) or with Al in a simple medium containing calcium, sucrose and 2-(N-morpholino)ethanesulfonic acid (MES; pH 5.0) for up to 18 h, the expression of NtSUT1 under its native promoter, or under the control of strong constitutive cauliflower mosaic virus (CaMV) 35S promoter, was strongly dependent on the presence of 2,4-D. Thereafter, the cells were preferentially treated in the presence of 2,4-D. During 6 h after a start of the control treatment, sucrose uptake rates were, compared to WT, slightly higher and lower in OX and RNAi lines respectively. The addition of Al reduced the sucrose uptake rates of OX and WT to the level of RNAi line, indicating that Al inhibits sucrose uptake via NtSUT1. During the post-Al culture of control and Al-treated cells in a nutrient medium, sucrose uptake rates were much higher in OX compared to WT and RNAi lines, which closely and positively correlated with the growth capacity of the cells. Judging from the growth capacity of Al-treated cells relative to that of control cells, OX cells were more tolerant to Al than WT and RNAi. In summary, we conclude that over-expression of NtSUT1 confers higher growth capacity in actively growing cells as well as in Al-treated cells.     

In vitro evidence of aluminum effects on solution movement through root cell walls

Little is known about the primary effects of aluminum (Al) in reducing root growth. However, the sorption of Al by the root cell wall, particularly by calcium (Ca) pectate, has been suggested as being important in the expression of Al toxicity in plants. To overcome problems arising from the close proximity of root cell components that may react with Al, a synthetic Ca pectate membrane was prepared as a model system for Al studies. Solution containing 1 mM Ca (as CaCl2) was passed through the membrane, and the flow rate measured. Solution containing 29 μM Al (as AlCl3) and 1 mM Ca reduced solution flow rate by > 80% from c. 3.5 to c. 0.6 mL/min within 2 min, with a further slight decline over the next 4 min. The Al concentration in solution proximate to the inlet side of the membrane decreased to 15 μM within 10 min, and only 3 μM Al was measured in solution that had passed through the membrane. These results suggest that an important primary effect of toxic Al is a reduction in water movement into the root, with consequent effects on water relations in the plant.