The in vivo and in vitro effects of Ca2+ and Al3+ upon ATPases from barley roots

Experiments were performed to test the effect of Ca²⁺ and Mg²⁺ upon ATPases (E.C. 3.6.1.3) from barley roots (Hordeum distichon L. cv. MULTUM) that had or had not taken up Al³⁺. Furthermore, the effect of the uptake period was examined. With longer duration of the Al³⁺ uptake, the activity of the ATPases dropped, independently of whether they were activated by Ca²⁺ or by Mg²⁺. Mg²⁺ stimulated the activity of the ATPases if no Al³⁺ had been taken up with lacking Ca²⁺ in the assay. If Al³⁺ had been taken up no additional activation by Mg²⁺ to the Ca²⁺ stimulation could be observed, and in some cases Mg²⁺ decreased the Ca²⁺ stimulation.     

Stability constant of Fe2+ chelates with soluble ligands from incubated soils

Summary The stability constants (log K) of Fe²⁺ chelates were determined on the basis of the shift in peak potential during the reduction of Fe²⁺ by a consortium of soluble ligands from incubated soils. Log K values ranged from 2.6 to 4.5. On average a change in pH of 1 unit induced a change in log K of 0.92 units. Aeration of the anaerobic decomposition products increased log K. The log K for Fe²⁺ chelates was about 0.8 units larger than that for Mn²⁺ chelates. It is considered that the chelation of ferrous iron plays an important role in the mobility and availability of iron to plants.     

Suppression of Methane Production via the Promotion of Fe2+ Oxidation in Paddy Fields

ABSTRACT

Methane is a greenhouse gas, mainly generated from paddy fields and lakes by methanogens using hydrogen and acetic acid as substrates. In anaerobic environments with adequate Fe³⁺, iron-reducing microorganisms utilize these substrates, thus suppressing methane generation. We promoted Fe²⁺ oxidation to Fe³⁺ by physically stirring paddy soil and using a chelating agent (nitrilotriacetic acid; NTA) to evaluate the feasibility of the suppression of methane generation using Fe³⁺-reducing bacteria. Under anaerobic conditions, Fe³⁺ reduction to Fe²⁺ began immediately in the slurry made by adding water into air-dried paddy soil. Methane generation began on the 6th day when most Fe³⁺ was reduced. Under anaerobic conditions for 10 days followed by aerobic conditions, Fe²⁺ oxidation hardly progressed under static conditions. On stirring the slurry, Fe²⁺ oxidation progressed over 12 h (67% Fe²⁺ oxidized to Fe³⁺). When NTA was added under anaerobic conditions followed by stirring under aerobic conditions, Fe²⁺ oxidation was promoted further. The idea of physical stirring of paddy soil in the actual environment was derived from the effects of paddy soil stirring by ducks in interrelated rice–duck farming. In such farming, paddy soil contains more Fe³⁺ in its surface water compared with normal farming, resulting in suppressed methane generation.     

Characterizing nicotianamine aminotransferase: Improving its assay system and details of the regulation of its activity by Fe nutrition status

Under iron deficient conditions, graminaceous plants secrete mugineic acid family phytosiderophores (MAs) from their roots to dissolve sparingly soluble iron compounds in the rhizosphere, and take up iron in the form of an Fe³⁺-MAs complex (Takagi 1976). A good correlation has been reported between the tolerance of Fe-deficiency and the amount of secreted MAs (Takagi 1993). Therefore, by using the genes involved in MAs biosynthesis, molecular breeding might produce transgenic plants tolerant to Fe-deficiency with a high level of MAs secretion. The biosynthetic pathway of MAs from L-methionine has been clarified (Fig. 1) and the enzymes participating in this process are now being investigated to isolate the genes responsible. Nicotianamine aminotransferase (NAAT) catalyzes the amino group transfer between nicotianamine (NA) and 2-oxoglutaric acid (Fig. 1). In order to purify NAAT, enzyme assay methods for NAAT have been developed and modified (Shojima et al. 1990; Ohata et al. 1993; Kanazawa et al. 1994). Some characteristics of NAAT have been reported using these enzyme assay methods (Kanazawa et al. 1994, 1995). Here, we further investigate some characteristics of this enzyme to improve the enzyme assay method, namely 1) the effect of K⁺ and Mg²⁺ on NAAT activity in vitro, and 2) the direct influence of MAs, Fe³⁺, and Fe²⁺ on NAAT activity. In addition, based on these results, the induction of enzyme activity by Fe-deficiency and suppression of the activity by Fe-resupply was investigated, by applying the new enzyme assay method.     

NO-3胁迫下K+、Ca2+对黄瓜幼苗膜质过氧化及活性氧清除酶系统的影响

通过水培试验探讨了NO^-₃胁迫下K⁺、Ca²⁺对黄瓜幼苗膜质过氧化及活性氧清除酶系统的影响。结果表明，在相同NO^-₃浓度胁迫7d后， Ca²⁺浓度越大，膜脂过氧化产物丙二醛(MDA)含量越高，而K⁺浓度越大，电解质相对渗透率越高，由此说明K⁺、Ca²⁺对细胞膜造成伤害的机理不同。黄瓜幼苗活性氧清除酶系统对K⁺、Ca²⁺的响应亦不同，在一定程度上，K⁺和Ca²⁺ 可提高SOD、POD和CAT活性，保护植物免受自由基伤害，继而可增强植物对逆境的适应能力。     

Effects of Cd on the fluxes of K+ and H+ in excised roots

As a result of Cd treatment, K concentrations decreased in Cd sensItive maize and kidney bean calli (Obata et al. 1994) and in intact roots of kidney bean plants (Obata et al. unpublished). Potassium may be extruded from the roots or the absorption of K may be depressed by the Cd treatment in these Cd sensitive plants. Obata et al. (1996) observed that Cd inhibited both the efflux of H⁺ and influx of K⁺ following K⁺ addition in intact roots of bean. Thus Cd may affect the activity of proteins essential to ion movement., i.e. ioncarriers, channels and ATPase embedded in the membranes and/or may affect the permeability of the lipids of the membrane.     

Factors related to iron uptake by dicotyledonous and monocotyledonous plants. II. The reduction of Fe3+ as influenced by roots and inhibitors

In a companion paper (10), varieties of four plant species [two monocotyledons (oats and corn) and two dicotyledons (soybeans and tomato)] were shown to differ widely in their ability to respond to Fe‐stress. The ability of the more Fe‐efficient varieties was manifested by a lowering of the pH of the ambient medium of the root and/or by loss of reductants from the root. Both effects can enhance uptake of Fe by the roots, since Fe is taken up primarily, if not entirely, as Fe²⁺ ions. Thus, a given stressed plant has a means, under some degree of metabolic control, for modifying the root environment and, thereby, alleviating its chlorotic condition.

The present investigation deals with environmental factors, particularly chemical inhibitors, modifying the effectiveness of the stress response. Without inhibitors, excised root samples of the four species exhibited a wide range of abilities to reduce Fe³⁺ to Fe²⁺. Roots of the dicotyledonous species reduced about twice as much Fe³⁺ as did equal weights of the monocotyledonous species. Iron‐efficient tomato, soybean, and oat roots reduced more Fe³⁺ than did roots of the Fe‐inefficient varieties. The two corn varieties were about equal in their effectiveness.

Comparable samples of roots were also exposed to chemicals that induce or aggravate Fe chlorosis. Those found to be very effective inhibitors of Fe³⁺ reduction by the roots included: hydroxide, orthophosphate, pyrophosphate, Cu²⁺ and Ni²⁺. Other ions (includ ing Mn²⁺, Zn²⁺ and molybdate) and ethyl ammonium phosphate also inhibited Fe³⁺ reduction but to a lesser degree. Citrate, however, enhanced Fe³⁺ reduction. The degree of inhibition or enhancement differed for each of the varieties. In general, the Fe‐efficient plants were best able to reduce Fe³⁺ in spite of the inhibitory influence of the imposed treatments. Thus, our findings indicated that inhibition of the Fe³⁺ ‐reduction process at, or near, the periphery of the root is an apparent cause of Fe chlorosis.     

Factors related to iron uptake by dicotyledonous and monocotyledonous plants III. Competition between root and external factors for Fe

In comparison studies (11, 12), monocotyledonous corn (Zea mays L.) and oats (Avena byzantina C. Koch) did not respond to Fe stress as effectively nor to the same degree as the dicotyledonous soybeans (Glycine max (L.) Merr.) or tomatoes (Lycopersicon esculentum Mill.). Both the Fe‐inefficient and Fe‐efficient corn and oats developed Fe chlorosis; the Fe‐efficient dicotyledonous plants were green. In the present study, the method of inducing Fe stress was changed to make it less severe. Instead of using only NO₃‐N and no Fe to induce Fe stress (11, 12), both NH₄‐N and NO₃‐N were used along with varied concentrations of Fe. Iron stress was induced with BPDS (4,7‐diphenyl‐l, 10‐phenan‐throline disulfonic acid) and phosphate; both competed with the plant for Fe. Phosphate also inhibits reduction of Fe³⁺ to Fe²⁺ (12). This method of inducing Fe stress in the plants was less severe than using only NO₃‐N and no Fe in the nutrient solutions and we were able to measure a difference in Fe‐stress response for all four plant species (Fe‐inefficient and Fe‐efficient). At the lower Fe treatments, the roots of Fe‐efficient plants usually reduced more Fe³⁺ to Fe²⁺ than did the roots of Fe‐inefficient plants. The ‘inefficient’ ys₁ corn and TAM 0–312 oat roots did not compete with BPDS or phosphate for Fe as effectively as did the ‘efficient’ WF9 corn and Coker 227 oat roots. The same type mechanism for solubilization, absorption, and transport of Fe seems to function in both monocotyledenous and dicotyledenous plants but it is more effective and more readily detected in the dicot than in the monocot plants. The reactions involved in reduction of Fe³⁺ to Fe²⁺ seemed to be confined inside or at the root surface for the inefficient genotypes; the efficient genotypes alter the ambient medium to a greater degree.     

Determination of Fe2+ in Rice Leaves (Oryza sativa L.) by Using the Chelator BPDS Alone or Combined with the Chelator EDTA

Abstract

Iron toxicity is a problem in many areas of wetland rice. Since Fe²⁺ is considered to be the toxic form of iron, the objective of this research was to determine the Fe²⁺ concentration in rice leaves using the chelator bathophenanthroline disulfonate (BPDS), disodium salt alone or combined with the chelator ethylenediaminetetraacetate (EDTA), disodium salt, where BPDS should solely chelate the Fe²⁺ and EDTA chelate only Fe³⁺. Thus, the combination of these chelators should stabilize the Fe oxidation states. It was also tested whether the chelators BPDS and EDTA could stabilize the oxidation states of Fe during the extraction of rice leaves. Extractions of rice leaves were carried out using an 1 mM BPDS or BPDS‐EDTA extractant solution. To test the stabilization of the Fe oxidation states by the combination of BPDS with EDTA, the extraction solution for one part of the samples contained 0.07 mM Fe³⁺. An extraction without plant material as control was also taken into consideration. The results indicated that the chelators were able to stabilize the oxidation states of Fe in the control (extraction without plant material). However, in the presence of plant material, Fe³⁺ was partly reduced to Fe²⁺, i.e., the chelators could not stabilize the oxidation states of Fe. Accordingly, we concluded that the BPDS‐EDTA method may function for the Fe²⁺ determination in water and soil, but it is apparently not suited for rice leaves.     

Characteristics of ammonium and nitrate fluxes along the roots of Picea asperata

Nitrogen (N), ammonium (NH₄⁺) and nitrate (NO₃^?), is one of the key determinants for plant growth. The interaction of both ions displays a significant effect on their uptake in some species. In the current study, net fluxes of NH₄⁺ and NO₃^? along the roots of Picea asperata were determined using a Non-invasive Micro-test Technology (NMT). Besides, we examined the interaction of NH₄⁺ and NO₃^? on the fluxes of both ions, and the plasma membrane (PM) H⁺-ATPases and nitrate reductase (NR) were taken into account as well. The results demonstrated that the maximal net NH₄⁺ and NO₃^? influxes were detected at 13–15?mm and 8–10.5?mm from the root apex, respectively. Net NH₄⁺ influx was significantly stimulated with the presence of NO₃^?, whereas NH₄⁺ exhibited a markedly negative effect on NO₃^? uptake in the roots of P. asperata. Also, our results indicated that PM H⁺-ATPases and NR play a key role in the control of N uptake.     

Effect of supplied phytosiderophore on 59Fe absorption and translocation in Fe-deficient barley grown hydroponically in low phosphorus media

Abstract

Hydroponically grown barley plants (Hordeum vulgare L. cv. Minorimugi) under iron-deficient (–Fe) and high phosphorus (P) conditions (500 µmol L^?1) showed Fe chlorosis and lower growth compared with plants grown in –Fe and low P conditions (50, 5 and 0.5 µmol L^?1). To understand the physiological role of P in regulating the growth of plants in –Fe medium, we carried out an Fe feeding experiment using four P levels (500, 50, 5 and 0.5 µmol L^?1) and phytosiderophores (PS), mugineic acid. Our results suggest that plants grown in a high P medium had higher absorption activity of ⁵⁹Fe compared with plants grown in low P media, irrespective of the presence or absence of added PS. Translocation of ⁵⁹Fe from roots to shoots was not affected by the P level. The relative translocation rate of ⁵⁹Fe increased with decreasing levels of P in the medium. In general, the addition of PS enhanced the absorption of ⁵⁹Fe and its translocation. Taken together these results suggest that the lower relative translocation rate of Fe in high P plants may be induced by the physiological inactivation of Fe in the roots, and the higher absorption activity of Fe in high P conditions possibly results from the response of barley plants to Fe deficiency.     

Factors related to iron uptake by dicotyledonous and monocotyledonous plants. I. pH and reductant

Some plants respond to Fe‐deficiency stress by inducing Fe‐solubilizing reactions at or near the root surface. In their ability to solubilize Fe, dicotyledonous plants are more effective than monocotyledonous plants. In this study we determined how representative plants differ in their response when subjected to Fe‐deficiency stress in a calcareous soil and in nutrient solutions. Iron‐inefficient genotypes of tomato, soybean, oats, and corn all developed Fe chlorosis when grown in soil, whereas Fe‐efficient genotypes of these same species remained green. The same genotypes were grown in complete nutrient solutions and then transferred to nutrient solutions containing N (as NO₃ ^‐) and no Fe.

The T3238 FER tomato (Lycopersican esculentum Mill.) Fe‐efficient) was the only genotype that released significant amounts of H from the roots (the pH was lowered to 3.9) and concomitantly released reductants. Under similar conditions, Hawkeye soyhean [Glycine max (L.) Merr.] released reductants but the solution pH was not lowered. Both Fe‐inefficient and Fe‐efficient genotypes of oats (Avena sativa L.) and corn (Zea mays L.) released insufficient H or reductant from their roots to solubilize Fe; as a result, each of these genotypes developed Fe‐deficiency (chlorosis).

The marked differences observed among these genotypes illustrate the genetic variability inherent within many plant species. A given species or genotype may accordingly not be adapted to a particular soil. Conversely, a given species or genotype may be found (or developed) that is precisely suited for a particular soil. In this event, the need for soil amendments may be reduced or eliminated.     

羟基磷灰石对铅锌矿区土壤吸附Zn2+、Cd2+的影响

为探究羟基磷灰石(HAP)对矿区土壤重金属的固化效果,采用吸附试验,研究施加HAP的铅锌矿区土壤对Cd~(2+)、Zn~(2+)的动力学吸附和等温吸附效果。结果表明:土壤对Cd~(2+)、Zn~(2+)的吸附量随Cd~(2+)、Zn~(2+)初始浓度的增加而增加;在酸性条件下,其吸附量随pH上升而上升;准二级动力学方程能很好地描述两者的吸附过程,土壤吸附能力随HAP的添加量增大而增强;在Zn—Cd共存体系中,当初始浓度为20mg/L时,土壤对Zn~(2+)、Cd~(2+)的吸附无明显差异,2种金属离子竞争力度小,随着初始浓度上升,竞争明显,对Zn~(2+)的最大吸附量能达到单一体系中的79%～87%,而Cd~(2+)的最大吸附量只有单一体系中的57%～72%,Zn~(2+)的竞争力优于Cd~(2+),Zn~(2+)对Cd~(2+)吸附产生严重的抑制。综上可知,HAP能提高矿区土壤的吸附性能,在Zn、Cd污染土壤中,更能提升土壤对Zn~(2+)的吸附固持能力。     

水稻根尖边缘细胞对铁毒的形态生理响应

以水稻(Oryza sativa L.)品种Azucena（铁耐性）和IR64（铁敏感）为材料,研究了Fe2+毒胁迫下附着于根尖边缘细胞(即原位边缘细胞)的数目、存活率,根尖细胞形态结构、根尖保护酶活性的变化。结果显示,Fe2+ 毒对根边缘细胞的产生有抑制作用。相对于敏感性品种而言,一定浓度Fe2+(100~200 μmol/L)有利于耐性品种边缘细胞的产生;Fe2+ 毒对边缘细胞有致死效应,随Fe2+浓度的提升,边缘细胞的存活率呈下降趋势,根尖外围细胞壁增厚,并出现细胞程序性死亡特征(敏感性品种)。同时,Fe2+ 毒对根尖保护酶活性有一定的影响,200~400 μmol/L Fe2+处理下,耐性品种POD、CAT、SOD活性都超过对照;敏感品种只有SOD活性超过对照。说明Fe2+毒胁迫下,水稻根尖通过增加边缘细胞数目、提高细胞拒铁作用,维持较高水平的POD、CAT和SOD活性来对抗Fe2+毒,缓解铁毒害。     

水稻土Fe2+氧化耦合硝酸根异化还原成铵(DNRA)及其对氧气存在和碳源添加的响应

以江苏常熟和湖南桃源水稻土为研究对象,通过室内15N示踪实验研究水稻土中Fe²⁺氧化耦合硝酸根异化还原成铵(Dissimilatory nitrate reduction to ammonium,DNRA)过程及其对氧气存在和碳源添加的响应。结果表明,两种水稻土中均存在Fe²⁺氧化耦合DNRA过程,常熟和桃源水稻土中DNRA的速率分别为0.38±0.15和0.36±0.21nmol·g^–1·h^–1(以N计),当体系中Fe²⁺浓度为500μmol·L^–1时,DNRA速率有所提升但并不显著,当Fe²⁺浓度为800μmol·L^–1时,DNRA速率提升显著(P <0.05),分别提升至2.35±0.30和0.81±0.22 nmol·g^–1·h^–1。在800μmol·L^–1 Fe²⁺浓度下,常熟水稻土中Fe²⁺     

外源Ca2+和GA3对骆驼蓬生物碱抑制黄瓜种子萌发的缓解作用

采用室内水培方法,研究了外源Ca2 和GA3对骆驼蓬生物碱抑制下黄瓜种子萌发的影响.结果表明,Ca2 和GA3单一及复合处理可促进骆驼蓬生物碱处理的黄瓜种子的萌发,使黄瓜萌发率、发芽指数和活力指数提高,萌发种子的α-淀粉酶、蛋白酶、脂肪酶和异柠檬酸裂解酶活性及赤霉素(GA)和钙调素(CaM)含量增加,烟酰胺腺嘌呤二核苷酸激酶(NADK)和Ca2 -ATP酶活性增强,脱落酸(ABA)含量降低;且Ca2 和GA3复合处理对GA含量的提高和ABA含量的降低与Ca22 单一处理比较呈显著的加成作用.表明外源Ca22 和GA3对骆驼蓬生物碱抑制黄瓜种子的萌发具有缓解效应.     

钙和NO对NaCl胁迫下黄瓜幼苗生长和活性氧代谢的影响

采用营养液培养的方法,研究了Ca2+对外源一氧化氮(Nitric oxide,NO)所诱导的NaCl胁迫下黄瓜幼苗生长、活性氧代谢的影响。结果表明,添加外源NO或Ca2+显著缓解了NaCl胁迫对黄瓜幼苗生长的抑制,叶片和根系超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性较单独NaCl胁迫处理显著提高,丙二醛(MDA)和过氧化氢(H2O2)的含量、超氧阴离子(O.-2)产生速率明显下降;添加NO的同时添加Ca2+通道抑制剂La3+抑制了NO的这些调节作用。结果表明Ca2+对NO诱导的NaCl胁迫下黄瓜幼苗植株活性氧清除能力的提高起重要作用,NO的作用可能依赖于胞浆Ca2+。     

外源精胺对NO_3~-胁迫下黄瓜幼苗抗氧化酶活性及光合作用的影响

采用营养液培养方法,研究了添加不同浓度的精胺(Spm)对NO3-胁迫下黄瓜幼苗生长、叶片抗氧化酶活性及光合作用的影响。结果表明,140 mmol/L NO3-胁迫下,外加1 mmol/L Spm,10 d后,黄瓜幼苗叶片超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)活性显著增加,电解质渗漏率和丙二醛(MDA)含量显著降低;气孔导度(Gs)、胞间CO2浓度(Ci)和净光合速率(Pn)显著升高,气孔限制值(Ls)显著降低。说明1 mmol/L Spm处理能增强黄瓜幼苗对活性氧的清除能力,降低膜脂过氧化程度;降低气孔关闭,改善叶片的气体交换,幼苗生长势增加,对高浓度NO3-胁迫的抗性增强。当Spm浓度高达1.5~2 mmol/L时,与1mmol/L Spm相比,SOD、POD、APX、CAT活性均开始降低,电解质渗漏率和MDA含量增加,Gs、Ci和Pn显著降低,黄瓜幼苗生长受到抑制。可见,外加一定浓度的Spm可通过提高抗氧化酶活性、降低膜脂过氧化程度及改善光合作用来缓解NO3-胁迫对黄瓜幼苗的影响。     

Adsorption and degradation of 2,4-dichlorophenoxyacetic acid in spiked soil with Fe0 nanoparticles supported by biochar

This work studies the adsorption and degradation of 2,4-dichlorophenoxyacetic (2,4-D) in spiked soil with nanoscale Fe⁰ particles (nFe⁰) and biochar derived from maize straw. When biochar concentration was high, the adsorption capacity of soil was enhanced. Furthermore, 2,4-D degraded completely at loading rates of 0.33 and 0.17 g/L nFe⁰ plus biochar (initial 2,4-D concentration of 10 mg/g) within 40 h, according to equilibrium data. Additionally, the theoretical concentration of chloridion was approximately 84%. Further analysis indicated that the effect of nFe⁰ on 2,4-D degradation was weaker in soil columns than that in soil slurry. By contrast, 2,4-D degradation is positively influenced by biochar application, which prevented the aggregation and corrosion of Fe nanoparticles. Although the enhanced capacity for 2,4-D adsorption on the soil decelerated dechlorination rate, long-term nFe⁰ activity was generated. After 72 h, the efficiency of 2,4-D degradation was approximately 53.2% in the soil columns with biochar support.     

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.