Methylene Blue Stainability of Root-Tip Protoplasts as an Indicator of Aluminum Tolerance in a Wide Range of Plant Species,Cultivars and Lines

We previously developed a new simple technique of methylene blue (MB) staining for the discrimination of aluminum (Al)-tolerant protoplasts from 4 plant species (rice, oats, maize and pea). The objectives of the present study were to confirm the applicability of this technique to a wider range of plant species, cultivars and lines, and to identify a common strategy for the early stage of Al tolerance. A total of 10 plant species, two Brachiaria spp., two Oryza spp., buckwheat, maize, pea, triticale, wheat and barley, corresponding to 18 different plant samples (species, cultivars, and lines), were used. Al tolerance (relative net root elongation of the longest root), which was screened at 20 μM AICI₃ in 0.2 mM CaCl₂ (pH 4.9) for 24 h, ranged widely from 10 to 88. Among cultivars and lines within the same species, Al accumulation in the root tip portion was higher in Al-sensitive plants, corresponding to more severe permeabilization of the plasma membranes (PM). Protoplasts isolated mainly from the epidermis, and outer and central parts of the cortex were stained to different degrees by MB, and the blue color was observed both on the surface and inside the protoplasts. Color pictures obtained after staining for 3 min with 0.1% MB were analyzed by Image Hyper II. The ratio of the heavily stained area at threshold 95 to the entire area stained with MB at threshold 125 was defined as MB stainability. MB stainability was negatively correlated with Al tolerance (y= 48.6e^?0.02x , R ²= 0.676**) suggesting the common importance of permeation characteristics of PM, in addition to PM negativity for Al tolerance in a wide range of plant species, cultivars and lines. Analysis of the PM lipid composition was proposed as an important topic for future studies on the negativity and permeation of PM.     

The plasma membrane intactness of root-tip cells is a primary factor for Al-tolerance in cultivars of five species

We investigated the role of the cell wall and plasma membrane (PM) of root-tip cells in Al tolerance in Al-tolerant and Al-sensitive cultivars of five plant species (rice, maize, pea, wheat, and sorghum). No correlation was found between the differences in Al tolerance and the cation exchange capacity of cell walls isolated from root-tips (0–1 em). Preliminary exposure to Al for 1 h was sufficient to inhibit subsequent root re-elongation in an Al-free solution, and the inhibitory effect was more pronounced in the Al-sensitive cultivars than in the Al-tolerant ones. Together with the inhibition of root re-elongation, the PM of the root-tip cells of all the Al-sensitive cultivars was more permeabilized than that of the Al-tolerant cultivars, based on the FDA-PI fluorescence staining technique. Exposure for 30 min to Al treatment at 100 µM significantly increased the PM permeability of protoplasts isolated from the root-tips for the Al-sensitive pea cultivar placed in a moderately hypotonic medium. Protoplasts from root-tip portions of all the Al-sensitive cultivars took up more Al than those of the Al-tolerant ones when treated with 100 pM Al under isotonic conditions for 30 min. The co-existence of DNP or hypotonic conditions led to a larger increase of Al uptake by the protoplasts from Al-sensitive maize cultivars. These results suggest that Al ions rapidly alter the PM of the root-tip portion in the Al-sensitive cultivars, irrespective of plant species, resulting in an increase of the PM permeability.     

A New and Simple Technique for the Isolation of Plasma Membrane Lipids from Root-Tips

It has been suggested that plasma membrane (PM) lipids play a major role in aluminum (Al) tolerance; however, no direct investigations have been carried out using PM lipids from root-tips. Here we report a new technique for PM isolation as an alternative to the laborious two-polymer phase partitioning method that is commonly applied, as follows: 1) separation of protoplasts from 1-cm root-tip portions by enzymatic digestion, 2) attachment of the purified protoplasts to glass plates coated with polylysine, 3) preparation of PM ghosts by successive burst of the attached protoplasts using three separate buffer solutions (25 mM PIPES, 5 mM EDTA, and 2 mM MgCl₂, at pH 7.0) with slow stirring for 60 s. The PMs were confirmed to be devoid of organelle membranes by fluorescence microscopy, thin layer chromatography (TLC) and western blot analysis. The PM lipids obtained were found to be useful for studies on their differential permeability and lipid composition between lines of triticale or cultivars of maize under Al stress.     

Aluminum tolerance associated with enhancement of plasma membrane H+-ATPase in the root apex of soybean

Seventeen soybean cultivars were screened to discern differences in aluminum (Al) sensitivity. The Sowon (Al-tolerant) and Poongsan (Al-sensitive) cultivars were selected for further study by simple growth measurement. Aluminum-induced root growth inhibition was significantly higher in the Poongsan cultivar than in the Sowon cultivar, although the differences depended on the Al concentration (0, 25, 50, 75 or 100 μmol L^–1) and the amount of exposure (0, 3, 6, 12 or 24 h). Damage occurred preferentially in the root apex. High-sensitivity growth measurements using India ink implicated the central elongation zone located 2–3 mm from the root apex. The Al content was lower 0–5 mm from the root apices in the Sowon cultivar than in the apices of the Poongsan cultivar when exposed to 50 μmol L^–1 Al for 12 h. Furthermore, the citric acid exudation rate was more than twofold higher in the Sowon cultivar. Protein production of plasma membrane (PM) H⁺-ATPase from the root apices (0–5 mm) was upregulated in the presence of Al for 24 h in both cultivars. This activity, however, decreased in both cultivars treated with Al and the Poongsan cultivar was more severely affected. We propose that Al-induced growth inhibition is correlated with changes in PM H⁺-ATPase activity, which is linked to the exudation of citric acid in the root apex.     

Plasma Membrane Lipids Are the Powerful Components for Early Stage Aluminum Tolerance in Triticale

The order of aluminum (Al) tolerance in triticale lines (ST2>ST22) after re-elongation in an Al-free 0.2 mM calcium (Ca) solution for 9 h (Ca period) following 1 h pretreatment with 20 μM Al (Al period) agreed with that after 24 h of Al treatment. Permeability of the plasma membrane (PM) of root-tip cells after the Ca period was significantly increased in Al-sensitive ST22. Al was accumulated more heavily in the root-tip portion of ST22 than in that of ST2, although similar amounts of malic and citric acid anions were released from both triticale lines. We established a new system examining lipid permeability using synthesized nylon-2,8 ultrathin and porous capsules trapped previously with 0.1% (w/v) methylene blue solution and coated thereafter with PM lipid isolated from root tips by a newly developed technique. Permeability of the PM lipid measured with time in 0.2 mM Ca with or without 50 μM Al photometrically ( A ₆₈₀) was significantly greater in Al-sensitive ST22 after 5 min of Al treatment. This is the first report to directly show the primary and early role of PM lipid in Al tolerance in triticale.     

豌豆不同耐铝品种根尖细胞壁果胶及其甲基酯化度的差异

【目的】研究豌豆不同品种耐铝性和根尖根段耐铝性与果胶及其甲基酯化间的关系,为进一步揭示植物耐铝机理以及耐铝性状的遗传改良提供依据。【方法】以豌豆品种Hyogo和Alaska为试验材料,采用Hoagland培养方式,测定了不同品种不同根段果胶含量、 果胶甲基酯化度和果胶甲酯酶活性,研究了其差异及原因。【结果】在15和30 μmol/L铝浓度胁迫条件下,豌豆品种Alaska根相对伸长率均显著高于品种Hyogo,同时有根尖0~5 mm和5~10 mm段有更少的胼胝质生成和累积,在30 μmol/L浓度下不同根段间均达到显著差异,同时品种Hyogo根尖0~2.5 mm和2.5~5.0 mm段铝含量均显著高于品种Alaska,说明品种Alaska和品种Hyogo间存在耐铝性差异,其中品种Alaska耐铝性高于品种Hyogo,即品种Hyogo为铝敏感品种,品种Alaska是耐铝品种。比较两者不同根段(0~2.5 mm、 2.5~5.0 mm和5.0~10.0 mm)的铝含量与果胶含量、 果胶甲基酯化度、 PME活性间的关系,发现耐铝品种不同根段中的铝含量均小于敏感品种,并且在0~2.5 mm和2.5~5.0 mm段间达到显著性差异; 根尖不同根段果胶糖醛酸含量大小依次为0~2.5＞2.5~5.0＞5.0~10.0 mm,耐铝品种Alaska根尖细胞壁果胶和未甲酯化果胶含量均显著低于Hyogo,并且0~2.5 mm根段差异最大。根尖不同根段果胶甲基酯化度从根尖向上逐渐降低,并且耐铝品种Alaska高于铝敏感品种Hyogo,其中0~2.5 mm段间的差异达到显著水平;在对两个品种果胶甲基酯化酶(PME)活性进一步分析发现,PME活性大小依次为0~2.5＞2.5~5.0＞5.0~10.0 mm,两品种0~2.5 mm和2.5~5.0 mm根段间均达到显著差异。【结论】铝敏感品种Hyogo在0~2.5 mm和2.5~5.0 mm根段具有较高 PME活性和较低果胶甲基酯化程度。豌豆根尖果胶含量和甲基酯化度尤其是0~2.5 mm根段是豌豆耐铝性差异的重要原因;Alaska根尖细胞壁的果胶含量低和果胶甲基酯化度高(尤其是0~2.5 mm段)是其耐铝的重要机制。     

Aluminum tolerance associated with enhancement of plasma membrane H+-ATPase in the root apex of soybean

Abstract

Seventeen soybean cultivars were screened to discern differences in aluminum (Al) sensitivity. The Sowon (Al-tolerant) and Poongsan (Al-sensitive) cultivars were selected for further study by simple growth measurement. Aluminum-induced root growth inhibition was significantly higher in the Poongsan cultivar than in the Sowon cultivar, although the differences depended on the Al concentration (0, 25, 50, 75 or 100?μmol?L^–1) and the amount of exposure (0, 3, 6, 12 or 24?h). Damage occurred preferentially in the root apex. High-sensitivity growth measurements using India ink implicated the central elongation zone located 2–3?mm from the root apex. The Al content was lower 0–5?mm from the root apices in the Sowon cultivar than in the apices of the Poongsan cultivar when exposed to 50?μmol?L^–1 Al for 12?h. Furthermore, the citric acid exudation rate was more than twofold higher in the Sowon cultivar. Protein production of plasma membrane (PM) H⁺-ATPase from the root apices (0–5?mm) was upregulated in the presence of Al for 24?h in both cultivars. This activity, however, decreased in both cultivars treated with Al and the Poongsan cultivar was more severely affected. We propose that Al-induced growth inhibition is correlated with changes in PM H⁺-ATPase activity, which is linked to the exudation of citric acid in the root apex.     

Analysis of Potassium Uptake by Rice Roots Treated with Aluminum Using a Positron Emitting Nuclide, 38K

We investigate the effect of Al on K⁺ uptake by rice roots. Potassium-38 (³⁸K), a positron emitting nuclide (the half-life: 7.61 min), was used to trace K⁺ behavior. When a rice root was treated with 10μM Al for 24 h, the uptake of ³⁸K in the root was increased in the range of 1 to 2 cm from the root tip compared with that of the control sample. Because the root continued to grow without showing any damage of plasma membrane during the Al treatment, it was suggested that the ³⁸K uptake was not occurred through diffusion into the cells. The uptake of ³⁸K in both treatments, with/without Al, was decreased by VO₄^3- (inhibitor of H⁺-ATPase on plasma membrane) and DNP (H⁺ ionophore) treatment, which suggested that the K⁺ uptake was performed through an active transport, such as H⁺:K⁺ transport or H⁺ gradient promoted by an Al treatment.     

铵态氮和硝态氮调节盐胁迫豌豆幼苗生长和根系呼吸的作用

【目的】 土壤盐渍化在干旱和半干旱灌溉区是制约农业生产的非生物因素之一,合理的调控措施可以减轻盐渍化对植物的危害,本文探讨了氮源调节豆科植物盐胁迫的生理生态机制。 【方法】 采用砂培试验,以3个豌豆品种 (银豌1号、S5001-1和Ha) 为供试材料,设定三个盐分浓度(0、50、100 mmol/L),分别供应铵态氮和硝态氮4 mmol/L,每个品种均设六个处理。培养29天后对豌豆幼苗生物量、根系生长参数、根系呼吸及植株盐分离子含量进行测定。 【结果】 1) 三个盐分浓度相比,50 mmol/L NaCl处理下的3个豌豆品种幼苗的地上生物量和根系生长指标(根干重、根长和根表面积)显著高于0和100 mmol/L NaCl处理,且硝态氮处理显著高于铵态氮;2) 与无NaCl处理相比,3个豌豆品种植株含水量在100 mmol/L NaCl处理下明显降低,且硝态氮处理的显著低于铵态氮处理;3)豌豆根系呼吸速率均随着盐分浓度的增加和培养时间的延长总体呈降低趋势。3个豌豆品种根系呼吸速率对硝态氮和铵态氮的反应不同,相同盐分水平下,银豌1号铵态氮处理的高于硝态氮,Ha品种则相反,而S5001-1品种在两种氮源间差异不大。在50 mmol/L NaCl胁迫下,豌豆品种S5001-1与Ha硝态氮处理的根系呼吸累积量明显高于铵态氮,而银豌1号则相反;100 mmol/L NaCl胁迫下,豌豆品种Ha硝态氮处理的根系呼吸累积量显著高于铵态氮,其他两个品种在不同氮源处理间无差异。相同盐分胁迫水平下,银碗1号铵态氮处理的根系呼吸累积量明显高于品种S5001-1和Ha,而硝态氮处理下,品种Ha的根系呼吸累积量最高。4) 3个豌豆品种幼苗地上部Na+和Cl–含量均随盐浓度的增加而增加,而不同氮源对Na+在豌豆体内累积的影响因豌豆品种而异。 【结论】 在中度盐分胁迫下,施氮肥可缓解盐分胁迫对豌豆幼苗生长的影响,硝态氮缓解能力高于铵态氮,但在重度盐分胁迫下,盐胁迫是影响植物生长和离子吸收的主导因子,氮源调节作用变弱。尽管不同豌豆品种的根系呼吸对NH₄+-N与NO₃–-N的反应不同,但NO₃–-N缓解盐胁迫的效果总体上好于NH₄+-N。      

Responses of Kentucky bluegrass cultivars to excess aluminum in nutrient solutions

Kentucky bluegrass, Poa pratensis L., is generally regarded as an acid‐soil‐sensitive species. However, previous studies in our laboratory showed that cultivars within the species differed widely in tolerance to acid Tatum subsoil (pH 4.6) which is used routinely to screen plants for aluminum (Al) tolerance. In the early studies, specific differential Al tolerance was not demonstrated. The objective of the current study was to test the hypothesis of differential Al tolerance more precisely in nutrient solutions. In one experiment, acid‐soil‐tolerant Victa and Fylking and acid‐soil‐sensitive Windsor and Kenblue cultivars were grown for 35 days in nutrient solutions containing 0, 2, 4, 6, 12, and 24 mg Al L^‐1, at initial pH 4.5, with no subsequent adjustment. In a second experiment, Victa and Windsor were grown for 30 days in solutions containing 0, 4, and 6 mg Al L^‐1, at initial pH 4.5, with no further adjustment. For Victa and Windsor, tolerance to Al in nutrient solution corresponded with tolerance to acid Tatum subsoil, however, the cultivar difference in tolerance, based on relative root dry weight, was only about 2‐fold, compared with 20‐fold in acid Tatum subsoil. Fylking and Kenblue cultivars, which showed a wide difference in tolerance to acid Tatum subsoil, did not show distinct differences in tolerance to Al in nutrient solutions. Possible reasons for this discrepancy are discussed. Superior Al tolerance of Victa (compared with Windsor) was associated with a greater plant‐induced increase in the pH of its nutrient solutions and a corresponding decrease in concentrations of soluble Al in the filtered solutions at the end of the experiments. Greater Al sensitivity in Windsor (compared with Victa) was not related to reduced uptake of phosphorus (P) or excessive uptake of Al; neither cultivar accumulated appreciable Al concentrations in its shoots. The observed differential acid soil and Al tolerance among bluegrass cultivars appears worthy of further study. Improved understanding of Al tolerance mechanisms would contribute to fundamental knowledge of plant mineral nutrition and could aid plant breeders in tailoring plants for greater tolerance to acid subsoils.     

不同参数评价植物耐铝性的研究

利用相对根伸长率、根尖Al含量和根尖胼胝质含量等三项参数结合苏木精染色法,研究Al胁迫下,各参数在评价植物耐Al性上的地位,以及各参数间的相互关系。结果表明,不同植物和品种间三个参数变异幅度很大,相互间大都达到显著差异水平。各参数以及苏木精染色法在衡量同一植物不同品种间的耐Al性差异时是一致的,衡量不同植物耐Al性时则有所差别。通过相关性分析,确定根尖Al含量和根尖胼胝质含量都可以在一定程度上反映植物或品种间的耐Al性差异,在需要时可以作为替代相对根伸长率或作为补充对植物的耐Al性差异进行评价。     

Use of agricultural by-products to study the pH effects in an acid tea garden soil

The amelioration of an acid Alfisol from a tea garden was studied by incorporating various plant materials: canola straw, wheat straw, rice straw, corn straw, soybean straw, peanut straw, faba bean straw, Chinese milk vetch shoot and pea straw prior to incubation for a maximum of 65 days. Soil pH increased after incubation with all the incorporated materials with the legumes causing the largest increases. The final soil pH was correlated with ash alkalinity ( r ² = 0.73), base cations ( r ² = 0.74) and N content ( r ² = 0.93) of the applied materials. It was assumed that the incubation released the base cations in plant materials as they decomposed which ultimately increased the base cation saturation of the soil. Similarly, soil exchangeable Al was also decreased with the incorporation of the legume plant materials and corn straw and rice straw. Our investigation demonstrated that legumes are the preferred choice for controlling the soil acidity and also for reducing the toxicity of Al in acid soils.     

The membrane lipid bilayer as a regulated barrier to cope with detrimental ionic conditions: making new tolerant plant lines with altered membrane lipid bilayer

Several detrimental ionic conditions can occur in crop fields: soil acidity, salinity, heavy metal toxicity, and/or nutrient deficiency. Crop plants tolerant to these detrimental ionic conditions have excellent strategies that are related to external and/or internal mechanisms. Recently, we proposed a new concept of aluminum (Al) tolerance in plants; specifically, a plasma membrane (PM) lipid bilayer mechanism. This mechanism is defined as the retardation of Al permeation through the PM lipid bilayer based on the specific composition of the lipid molecules in the PM. The molecular basis of a less negatively charged PM lipid bilayer is smaller proportions of phospholipids and greater proportions of galactolipids and sterols. This leads to reduced adsorbability of detrimental ions onto the PM lipid bilayer, resulting in less permeabilization. Phenolics and carotenoids have two moieties; a hydrophilic moiety and a hydrophobic moiety. The hydrophobic moieties of these compounds can occlude the permeabilized spaces in the PM lipid bilayer, thereby reinforcing it. Another strategy to retard the permeability of the PM to detrimental ions is to decrease the proportion of stigmasterol, which has been shown to have no ability to reduce water permeability. The beneficial or harmful effects of various organic materials (organic fertilizers, soil organic matter, agrochemicals, or organic pollutants) on the productivity or quality of crop plants in relation to changes in the PM lipid bilayer are discussed. Modulation of the PM lipid bilayer is a promising strategy to produce new crop lines tolerant to detrimental ionic conditions.     

Rhizosphere pH Profile of Rice Plant Influenced by Al Treatment

To study the mechanisms of Al tolerance in rice, we focused on the change of rhizosphere pH. The 4-d seedlings were treated with Al solution (0, 10, 50 mM) for 24 h. Then each sample was put on an agarose gel including bromocresol green, so that the color of the gel indicated pH change. During 2-h contact, the pH of rhizosphere was decreased gradually, especially for Al-treated samples, showing the specific pH profiles along the root axis. Pretreatment of sample plants with a decoupling reagent 2,4-dinitrophenol (DNP) or a plasma membrane H⁺-ATPase inhibitor Na₃VO₄ did not decrease rhizosphere pH. Therefore, it was suggested that the H⁺ secretion activity was involved with Al-tolerant mechanisms of rice.     

Tolerances of wheat germplasm to acid subsoil

Aluminum toxicity is a major growth limiting factor for plants in many acid soils of the world. Correcting the problem by conventional liming is not always economically feasible, particularly in subsoils. Aluminum tolerant plants provide an alternative and long‐term supplemental solution to the problem. The genetic approach requires the identification of Al tolerance sources that can be transferred to cultivars already having desirable traits. Thirty‐five cultivars and experimental lines of wheat (Triticum aestivum L. em. Thell) were screened for Al tolerance on acid Tatum soil (clayey, mixed thermic, typic Hapludult) receiving either 0 or 3500 mg CaCO₃/kg (pH 4.1 vs. pH 7.1). Entries showed a wide range of tolerance to the acid soil. On unlimed soil at pH 4.3, absolute shoot dry weights differed by 5‐fold, absolute root dry weights by 6.5‐fold, relative shoot weights (wt. at pH 4.3/wt. at pH 7.1 %) by 4.7‐fold and relative root dry weights by 7‐fold. Superior acid soil (Al) tolerance of ‘BH‐1146’ from Brazil and extreme sensitivities of cultivars ‘Redcoat’ (Indiana, USA) and ‘Sonora 63’ (Mexico) were confirmed. Seven experimental (CNT) lines from Brazil showed a range of acid soil tolerance but were generally more tolerant than germplasm from Mexico and the USA. One line, ‘CNT‐1’, was equal to BH‐1146 in tolerance and may be useful in transferring Al tolerance to existing or new cultivars. Five durum cultivars (Triticum, durum, Desf.) were extremely sensitive to the acid Tatum subsoil at pH 4.3 compared with pH 7.1.     

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.     

不同耐密性玉米品种地上部与根系性状的协同效应

【目的】阐明不同玉米品种在增密种植条件下地上部性状和根系构型的协同响应,为耐密性玉米的遗传改良提供理论支撑。【方法】以我国18个主栽玉米品种为试材,设置2个种植密度(6万株/hm²和7.5万株/hm²),分别在吐丝期和成熟期测定14个地上部农艺性状和8个根系构型性状,利用方差分析与回归分析等统计方法解析耐密高产品种的地上地下协同关系。【结果】随着种植密度的增加,玉米单株地上部和根系生物量及籽粒产量等指标下降,群体地上部生物量和籽粒产量显著提高。根据两个种植密度下的群体产量,受试品种中6个被划分为高低密度下均高产的双高型(DH);3个品种为仅高密度下高产的高密高产型(HH);7个品种为高低密度下均低产的双低型(DL);2个品种为仅低密度下高产的低密高产型(HL),供试品种主要为双高型(DH)和双低型(DL)。在高密度下,DH品种比DL品种具有更多的吐丝前干物质累积量和更高的收获指数,DH品种在减少根系干重、节根数和根系宽度的同时,保持了较高的根系表面积与总根长。综合两个种植密度下地上部与根系性状对产量的贡献,发现吐丝期茎秆干物质、成熟期籽粒干物...     

Response of leguminosae to cadmium exposure

Bush bean and pea plants grown in a sandy substrate and treated daily with nutrient solutions containing either 50 and 125 pM cadmium (Cd), added as cadmium nitrate [Cd(NO₃)2], were analyzed for dry matter production, total Cd content, and extractable Cd. Cadmium depressed dry matter production of both plant species. Bush bean plants accumulated larger amounts of Cd in tissues and displayed lower Cd tolerance than pea plants. The high accumulation of Cd in roots of bush bean does not seem to prevent Cd translocation. Pea plants show a higher exclusion capacity at the root level, suggesting that membrane selectivity rather than apoplastic compartmentation may act as a defence mechanism against Cd toxicity. Gel‐permeation chromatography and voltammetric analyses showed that part of water‐soluble Cd extracted from tissues of pea and bush bean was as free metal ion (Cd²⁺). In addition, Cd into the nutrient solution induced progressively the synthesis of water‐soluble proteins at low molecular weigth in tissues of both plant specie. In root extracts of pea and bush bean, Cd was also associated with “like‐protein”; fraction with apparent molecular weight >30 KDa.     

Aluminum toxicity of synthetic aluminum–humus complexes derived from non-allophanic and allophanic Andosols and its amelioration with allophanic materials

Natural non-allophanic Andosols often show aluminum (Al) toxicity to Al-sensitive plant roots. The significance of Al–humus complexes to Al toxicity has been emphasized. Allophanic Andosols also possess Al–humus complexes, but they rarely show any toxicity. In the present study, using model substances, we tested the toxicity of Al–humus complexes and its amelioration with allophanic materials. We extracted humic substances from the A horizons of a non-allophanic Andosol and an allophanic Andosol using a NaOH solution, and reacted the humic substances and partially neutralized AlCl₃ solution at pH 4. Allophanic material was purified from commercial Kanuma pumice. Plant growth tests were conducted using a medium containing the Al–humus complexes (50 g kg⁻¹), the allophanic material (0, 90, 180 and 360 g kg⁻¹) and perlite. The root growth of barley ( Hordeum vulgare L.) and burdock ( Arctium lappa ) was reduced in the media containing the Al–humus complexes derived from both the non-allophanic and allophanic Andosols when the allophanic material was not added. With the addition of the allophanic materials, particularly in the 360 g kg⁻¹ treatment, the growth of the barley roots was improved markedly. Although the root growth of the burdock tended to improve with allophanic materials, the effect was weaker than that for barley. Monomeric Al in a solution of the medium was not detected (< 0.05 mg L⁻¹) following the addition of 360 g kg⁻¹ of allophanic materials, whereas 0.8–1.7 mg L⁻¹ Al was recorded without the allophanic material.     

The Function of a Maize-Derived Phosphoenol pyruvate Carboxylase (PEPC) in Phosphorus-Deficient Transgenic Rice

Transgenic rice ( Oryza sativa L., a C₃ plant) lines carrying a complete phospho enol pyruvate carboxylase (PEPC) gene from maize (a C₄ plant) were tested for their performance in terms of organic acid synthesis and organic acid exudation into the rhizosphere under phosphorus (P)-deficient conditions. High PEPC activity increased the fraction of photosynthetically fixed carbon allocated to the organic acid pool, and P deficiency enhanced oxalate exudation from the roots of the transgenic plants. There was no evidence that the transformed PEPC was involved in internal P recycling in the plant. However, the root PEPC activity was positively correlated with the oxalate exudation and negatively correlated with the root P concentration, and a higher root PEPC activity led to a higher oxalate exudation. Thus, it is suggested that C₄-PEPC transgenic rice plants had acquired the ability to exude oxalate, which enhanced their capacity to adapt to low P soil conditions.