記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：TAYLOR & FRANCIS LTD  

We aimed to investigate the role of the plasma membrane-localized sucrose/H+ symporter gene (NtSUT1) in root elongation growth of tobacco seedlings (Nicotiana tabacum L. cv. Samsun NN) under aluminum (Al) stress, by comparative study between wild-type (WT) tobacco and constructed overexpression (OX) and suppression (RNAi) lines of NtSUT1, in which the overexpression or suppression of NtSUT1 was controlled by the cauliflower mosaic virus 35S promoter. Seedlings of each line were grown in a nutrient medium (pH 4.5) without or with aluminum chloride (AlCl3). The roots were analyzed to determine the degree of root elongation and the transcript levels of NtSUT1, soluble sugar content, and cell survival at the root apex. The transcript level of NtSUT1 at the root apex was negatively affected by Al. Compared with WT, OX lines and RNAi lines had higher and lower contents of soluble sugars at the root apex, respectively. Under a 16-h light/8-h dark photoperiod and in the absence of Al, the root elongation rate was similar in OX and WT, but was significantly decreased in RNAi. In the presence of Al, the degree of Al-induced inhibition of root elongation was lower in OX and higher in RNAi than in WT. At the root apex, the initiation of cell death during a 24-h Al treatment was decelerated in OX and accelerated in RNAi, compared with that in WT. The number of live cells remaining at the root apex after the 24-h Al treatment was higher in OX and lower in RNAi than in WT. In the dark, root growth was supported by sucrose supplied in the medium as a substitute for photoassimilate. Under these conditions in the absence or presence of Al, there were strong positive correlations between the transcript level of NtSUT1, the soluble sugar content at the root apex and the root elongation rate. Together, these results indicate that NtSUT1 at the root apex primarily supports sucrose uptake via the apoplastic pathway, and that sucrose positively affects root elongation. We conclude that overexpression of NtSUT1 at the root apex increases the soluble sugar content, resulting in greater cell survival and root elongation under Al stress, leading to the Al-tolerance phenotype.

DOI： 10.1080/00380768.2017.1283646

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担当区分：筆頭著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

The aluminum-activated malate transporter (ALMT) family of proteins transports malate and/or inorganic anions across plant membranes. To demonstrate the possible role of ALMT genes in tomato fruit development, we focused on SlALMT4 and SlALMT5, the two major genes expressed during fruit development. Predicted proteins were classified into clade 2 of the family, many members of which localize to endomembranes. Tissue-specific gene expression was determined using transgenic tomato expressing the beta-glucuronidase reporter gene controlled by their own promoters. Both the genes were expressed in vascular bundles connecting to developing seeds in fruit and in the embryo of mature seeds. Further, SlALMT5 was expressed in embryo in developing seeds in fruit. Subcellular localization of both proteins to the endoplasmic reticulum (ER) was established by transiently expressing the green fluorescent protein fusions in plant protoplasts. SlALMT5 probably localized to other endomembranes as well. Localization of SlALMT5 to the ER was also confirmed by immunoblot analysis. The transport function of both SlALMT proteins was investigated electrophysiologically in Xenopus oocytes. SlALMT5 transported malate and inorganic anions such as nitrate and chloride, but not citrate. SlALMT4 also transported malate, but the results were less consistent perhaps because it did not localize strongly to the plasma membrane. To elucidate the physiological role of SlALMT5 further, we over-expressed SlALMT5 in tomato. Compared with the wild type, overexpressors exhibited higher malate and citrate contents in mature seeds, but not in fruit. We conclude that the malate transport function of SlALMT5 expressed in developing fruit influences the organic acid contents in mature seeds.

DOI： 10.1093/pcp/pcw157

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担当区分：筆頭著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

TaALMT1 from wheat (Triticum aestivum) and AtALMT1 from Arabidopsis thaliana encode aluminum (Al)-activated malate transporters, which confer acid-soil tolerance by releasing malate from roots. Chimeric proteins from TaALMT1 and AtALMT1 (Ta::At, At::Ta) were previously analyzed in Xenopus laevis oocytes. Those studies showed that Al could activate malate efflux from the Ta::At chimera but not from At::Ta. Here, functions of TaALMT1, AtALMT1 and the chimeric protein Ta::At were compared in cultured tobacco BY-2 cells. We focused on the sensitivity and specificity of their activation by trivalent cations. The activation of malate efflux by Al was at least two-fold greater in the chimera than the native proteins. All proteins were also activated by lanthanides (erbium, ytterbium, gadolinium, and lanthanum), but the chimera again released more malate than TaALMT1 or AtALMT1. In Xenopus oocytes, Al, ytterbium, and erbium activated inward currents from the native TaALMT1 and the chimeric protein, but gadolinium only activated currents from the chimera. Lanthanum inhibited currents from both proteins. These results demonstrated that function of the chimera protein was altered compared to the native proteins and was more responsive to a range of trivalent cations when expressed in plant cells. (C) 2016 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.bbamem.2016.03.026

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：AMER PHYTOPATHOLOGICAL SOC  

Legume plants can establish symbiosis with soil bacteria called rhizobia to obtain nitrogen as a nutrient directly from atmospheric N-2 via symbiotic nitrogen fixation. Legumes and rhizobia form nodules, symbiotic organs in which fixed-nitrogen and photosynthetic products are exchanged between rhizobia and plant cells. The photosynthetic products supplied to rhizobia are thought to be dicarboxylates but little is known about the movement of dicarboxylates in the nodules. In terms of dicarboxylate transporters, an aluminum-activated malate transporter (ALMT) family is a strong candidate responsible for the membrane transport of carboxylates in nodules. Among the seven ALMT genes in the Lotus japonicus genome, only one, LjALMT4, shows a high expression in the nodules. LjALMT4 showed transport activity in a Xenopus oocyte system, with LjALMT4 mediating the efflux of dicarboxylates including malate, succinate, and fumarate, but not tricarboxylates such as citrate. LjALMT4 also mediated the influx of several inorganic anions. Organ-specific gene expression analysis showed LjALMT4 mRNA mainly in the parenchyma cells of nodule vascular bundles. These results suggest that LjALMT4 may not be involved in the direct supply of dicarboxylates to rhizobia in infected cells but is responsible for supplying malate as well as several anions necessary for symbiotic nitrogen fixation, via nodule vasculatures.

DOI： 10.1094/MPMI-04-16-0071-R

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担当区分：筆頭著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Wheat and Arabidopsis plants respond to aluminum (Al) ions by releasing malate from their root apices via Al-activated malate transporter. Malate anions bind with the toxic Al ions and contribute to the Al tolerance of these species. The genes encoding the transporters in wheat and Arabidopsis, TaALMT1 and AtALMT1, respectively, were expressed in Xenopus laevis oocytes and characterized electrophysiologically using the two-electrode voltage clamp system. The Al-activated currents generated by malate efflux were detected for TaALMT1 but not for AtALMT1. Chimeric proteins were generated by swapping the N- and C-terminal halves of TaALMT1 and AtALMT1 (Ta::Atand At:: Ta). When these chimeras were characterized in oocytes, Al-activated malate efflux was detected for the Ta:: At chimera but not for At:: Ta, suggesting that the N-terminal half of TaALMT1 is necessary for function in oocytes. An additional chimera, Ta(48):: At, generated by swapping 17 residues from the N-terminus of AtALMT1 with the equivalent 48 residues from TaALMT1, was sufficient to support transport activity. This 48 residue region includes a helical region with a putative transmembrane domain which is absent in AtALMT1. The deletion of this domain from Ta(48):: At led to the complete loss of transport activity. Furthermore, truncations and a deletion at the C-terminal end of TaALMT1 indicated that a putative helical structure in this region was also required for transport function. This study provides insights into the structure-function relationships of Al-activated ALMT proteins by identifying specific domains on the N- and C-termini of TaALMT1 that are critical for basal transport function and Al responsiveness in oocytes.

DOI： 10.1093/pcp/pcu143

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：TAYLOR & FRANCIS LTD  

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.

DOI： 10.1080/00380768.2013.830230

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The role of vacuole in the cell death mechanism induced by aluminium (Al) was investigated in tobacco (Nicotiana tabacum L.) cell line BY-2. Cells at logarithmic phase of growth were treated without (control) or with Al (up to 150 μM) in a treatment medium containing CaCl2, sucrose and 2-(N-morpholino) ethanesulfonic acid (MES) buffer (pH 5.0). After 18 h treatment, both the integrity of the plasma membrane (estimated by Evans blue uptake) and growth capacity (estimated by post-Al treatment growth in nutrient medium) were decreased, while the activity of vacuolar processing enzyme (VPE) was increased, in the Al dose-dependent manner. The activity of the vacuole (estimated by neutral red uptake) was slightly increased at 50 μM then decreased with an increase in Al concentration. Direct observation of morphological changes of vacuole in a transgenic BY-2 expressing GFP-AtVam3p fusion protein localized on tonoplast indicated Al-induced collapse of vacuole. Time-course experiments indicated that both an increase in VPE activity and a loss of growth capacity were clearly observed at 6 h of the treatment time, prior to the loss of plasma membrane integrity. The presence of Ac-YVAD-CHO (an inhibitor effective to VPE) during Al treatment suppressed a loss of plasma membrane integrity. The expression of VPE genes (VPE-1a, VPE-1b) were significantly enhanced by Al treatment. Taken together, we conclude that an enhancement of VPE activity by Al is controlled at transcriptional level, and is a key factor leading to a loss of integrity of the plasma membrane and a loss of growth capacity. © 2013 Elsevier Inc.

DOI： 10.1016/j.jinorgbio.2013.07.001

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：TAYLOR & FRANCIS LTD  

Methylglyoxal (MG) is a reactive aldehyde derived by glycolysis. In Arabidopsis, MG inhibited light-induced stomatal opening in a dose-dependent manner. It significantly inhibited both inward-rectifying potassium (K-in) channels in guard-cell protoplasts and an Arabidopsis K-in channel, KAT1, heterologously expressed in Xenopus oocytes. Thus it appears that MG inhibition of stomatal opening involves MG inhibition of K+ influx into guard cells.

DOI： 10.1271/bbb.110885

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記述言語：英語   出版者・発行元：OXFORD UNIV PRESS  

Acid soils restrict plant production around the world. One of the major limitations to plant growth on acid soils is the prevalence of soluble aluminium (Al(3+)) ions which can inhibit root growth at micromolar concentrations. Species that show a natural resistance to Al(3+) toxicity perform better on acid soils. Our understanding of the physiology of Al(3+) resistance in important crop plants has increased greatly over the past 20 years, largely due to the application of genetics and molecular biology. Fourteen genes from seven different species are known to contribute to Al(3+) tolerance and resistance and several additional candidates have been identified. Some of these genes account for genotypic variation within species and others do not. One mechanism of resistance which has now been identified in a range of species relies on the efflux of organic anions such as malate and citrate from roots. The genes controlling this trait are members of the ALMT and MATE families which encode membrane proteins that facilitate organic anion efflux across the plasma membrane. Identification of these and other resistance genes provides opportunities for enhancing the Al(3+) resistance of plants by marker-assisted breeding and through biotechnology. Most attempts to enhance Al(3+) resistance in plants with genetic engineering have targeted genes that are induced by Al(3+) stress or that are likely to increase organic anion efflux. In the latter case, studies have either enhanced organic anion synthesis or increased organic anion transport across the plasma membrane. Recent developments in this area are summarized and the structure-function of the TaALMT1 protein from wheat is discussed.

DOI： 10.1093/jxb/erq272

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL PUBLISHING, INC  

Acid soils limit plant production worldwide because their high concentrations of soluble aluminium cations (Al(3+)) inhibit root growth. Major food crops such as wheat (Triticum aestivum L.) have evolved mechanisms to resist Al(3+) toxicity, thus enabling wider distribution. The origins of Al(3+) resistance in wheat are perplexing because all progenitors of this hexaploid species are reportedly sensitive to Al(3+) stress. The large genotypic variation for Al(3+) resistance in wheat is largely controlled by expression of an anion channel, TaALMT1, which releases malate anions from the root apices. A current hypothesis proposes that the malate anions protect this sensitive growth zone by binding to Al(3+) in the apoplasm. We investigated the evolution of this trait in wheat, and demonstrated that it has multiple independent origins that enhance Al(3+) resistance by increasing TaALMT1 expression. One origin is likely to be Aegilops tauschii while other origins occurred more recently from a series of cis mutations that have generated tandemly repeated elements in the TaALMT1 promoter. We generated transgenic plants to directly compare these promoter alleles and demonstrate that the tandemly repeated elements act to enhance gene expression. This study provides an example from higher eukaryotes in which perfect tandem repeats are linked with transcriptional regulation and phenotypic change in the context of evolutionary adaptation to a major abiotic stress.

DOI： 10.1111/j.1365-313X.2010.04338.x

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

P>Al3+-resistant cultivars of wheat (Triticum aestivum L.) release malate through the Al3+-activated anion transport protein Triticum aestivum aluminum-activated malate transporter 1 (TaALMT1). Expression of TaALMT1 in Xenopus oocytes and tobacco suspension cells enhances the basal transport activity (inward and outward currents present in the absence of external Al3+), and generates the same Al3+-activated currents (reflecting the Al3+-dependent transport function) as observed in wheat cells. We investigated the amino acid residues involved in this Al3+-dependent transport activity by generating a series of mutations to the TaALMT1 protein. We targeted the acidic residues on the hydrophilic C-terminal domain of TaALMT1 and changed them to uncharged residues by site-directed mutagenesis. These mutant proteins were expressed in Xenopus oocytes and their transport activity was measured before and after Al3+ addition. Three mutations (E274Q, D275N and E284Q) abolished the Al3+-activated transport activity without affecting the basal transport activity. Truncation of the hydrophilic C-terminal domain abolished both basal and Al3+-activated transport activities. Al3+-dependent transport activity was recovered by fusing the N-terminal region of TaALMT1 with the C-terminal region of AtALMT1, a homolog from Arabidopsis. These findings demonstrate that the extracellular C-terminal domain is required for both basal and Al3+-dependent TaALMT1 activity. Furthermore, we identified three acidic amino acids within this domain that are specifically required for the activation of transport function by external Al3+.

DOI： 10.1111/j.1365-313X.2010.04309.x

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担当区分：筆頭著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Plant stomata limit both carbon dioxide uptake and water loss; hence, stomatal aperture is carefully set as the environment fluctuates. Aperture area is known to be regulated in part by ion transport, but few of the transporters have been characterized. Here we report that AtALMT12 (At4g17970), a homolog of the aluminum-activated malate transporter (ALMT) of wheat, is expressed in guard cells of Arabidopsis thaliana. Loss-of-function mutations in AtALMT12 impair stomatal closure induced by ABA, calcium and darkness, but do not abolish either the rapidly activated or the slowly activated anion currents previously identified as being important for stomatal closure. Expressed in Xenopus oocytes, AtALMT12 facilitates chloride and nitrate currents, but not those of organic solutes. Therefore, we conclude that AtALMT12 is a novel class of anion transporter involved in stomatal closure.

DOI： 10.1093/pcp/pcq016

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL PUBLISHING, INC  

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 mu mol 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 mu mol 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.

DOI： 10.1111/j.1747-0765.2009.00437.x

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記述言語：英語   出版者・発行元：SPRINGER  

Photosystem II is vulnerable to various abiotic stresses such as strong visible light and heat. Under both stresses, the damage seems to be triggered by reactive oxygen species, and the most critical damage occurs in the reaction center-binding D1 protein. Recent progress has been made in identifying the protease involved in the degradation of the photo- or heat-damaged D1 protein, the ATP-dependent metalloprotease FtsH. Another important result has been the discovery that the damaged D1 protein aggregates with nearby polypeptides such as the D2 protein and the antenna chlorophyll-binding protein CP43. The degradation and aggregation of the D1 protein occur simultaneously, but the relationship between the two is not known. We suggest that phosphorylation and dephosphorylation of the D1 protein, as well as the binding of the extrinsic PsbO protein to Photosystem II, play regulatory roles in directing the damaged D1 protein to the two alternative pathways.

DOI： 10.1007/s11120-008-9372-4

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

TaALMT1 encodes a putative transport protein associated with Al(3+)-activated efflux of malate from wheat root apices. We expressed TaALMT1 in Nicotiana tabacum L. suspension cells and conducted a detailed functional analysis. Protoplasts were isolated for patch-clamping from cells expressing TaALMT1 and from control cells (empty vector transformed). With malate(2-) as the permeant anion in the protoplast, an inward current (anion efflux) that reversed at positive potentials was observed in protoplasts expressing TaALMT1 in the absence of Al(3+). This current was sensitive to the anion channel antagonist niflumate, but insensitive to Gd(3+). External AlCl(3) (50 mu M), but not La(3+) and Gd(3+), increased the inward current in TaALMT1-transformed protoplasts. The inward current was highly selective to malate over nitrate and chloride (P(mal) >> P(NO3) >= P(Cl), P(mal)/P(Cl) >= 18, +/-Al(3+)), under conditions with higher anion concentration internally than externally. The anion currents displayed a voltage and time dependent deactivation at negative voltages. Voltage ramps revealed that inward rectification was caused by the imposed anion gradients. Single channels with conductances between 10 and 17 pS were associated with the deactivation of the current at negative voltages, agreeing with estimates from voltage ramps. This study of the electrophysiological function of the TaALMT1 protein in a plant heterologous expression system provides the first direct evidence that TaALMT1 functions as an Al(3+)-activated malate(2-) channel. We show that the Al(3+)-activated currents measured in TaALMT1-transformed tobacco cells are identical to the Al(3+)-activated currents observed in the root cells of wheat, indicating that TaALMT1 alone is likely to be responsible for those endogenous currents.

DOI： 10.1093/pcp/pcn107

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Allele diversities of four markers specific to intron three, exon four and promoter regions of the aluminum (Al) resistance gene of wheat (Triticum aestivum L.) TaALMT1 were compared in 179 common wheat cultivars used in international wheat breeding programs. In wheat cultivars released during the last 93 years, six different promoter types were identified on the basis of allele size. A previous study showed that Al resistance was not associated with a particular coding allele for TaALMT1 but was correlated with blocks of repeated sequence upstream of the coding sequence. We verified the linkage between these promoter alleles and Al resistance in three doubled haploid and one intercross populations segregating for Al resistance. Molecular and pedigree analysis suggest that Al resistance in modern wheat germplasm is derived from several independent sources. Analysis of a population of 278 landraces and subspecies of wheat showed that most of the promoter alleles associated with Al resistance pre-existed in Europe, the Middle East and Asia prior to dispersal of cultivated germplasm around the world. Furthermore, several new promoter alleles were identified among the landraces surveyed. The TaALMT1 promoter alleles found within the spelt wheats were consistent with the hypothesis that these spelts arose on several independent occasions from hybridisations between non-free-threshing tetraploid wheats and Al-resistant hexaploid bread wheats. The strong correlation between Al resistance and Al-stimulated malate efflux from the root apices of 49 diverse wheat genotypes examined was consistent with the previous finding that Al resistance in wheat is conditioned primarily by malate efflux. These results demonstrate that the markers based on intron, exon and promoter regions of TaALMT1 can trace the inheritance of the Al resistance locus within wheat pedigrees and track Al resistance in breeding programmes.

DOI： 10.1007/s00122-007-0672-4

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Landes Bioscience  

The wheat ALMT1 gene encodes an aluminum (Al)-activated malate transport protein which confers Al-resistance. We investigated the membrane topology of this plasma-membrane localized protein with immunocytochemical techniques. Several green fluorescent protein (GFP)-fused and histidtne (His)-tagged chimeras of ALMT1 were prepared based on a computer-predicted secondary structure and transiently expressed in cultured mammalian cells. Antibodies raised to polypeptide epitopes of ALMT1 were used in conjunction with the antibody to the His-tags to determine the topology of ALMT1. This study shows that the ALMT1 protein contains six transmembrane domains with the amino and carboxyl termini located on the extracellular side of the plasma membrane. ©2007 Landes Bioscience.

DOI： 10.4161/psb.2.6.4801

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Aluminum (Al) resistance in wheat relies on the Al-activated malate efflux from root apices, which appears to be controlled by an Al-activated anion transporter encoded by the ALMT1 gene on chromosome 4DL. Genomic regions upstream and downstream of ALMT1 in 69 wheat lines were characterized to identify patterns that might influence ALMT1 expression. The first 1,000 bp downstream of ALMT1 was conserved among the lines examined apart from the presence of a transposon-like sequence which did not correlate with Al resistance. In contrast, the first 1,000 bp upstream of the ALMT1 coding region was more variable and six different patterns could be discerned (types I-VI). Type I had the simplest structure, while the others had blocks of sequence that were duplicated or triplicated in different arrangements. A pattern emerged among the lines of non-Japanese origin such that the number of repeats in this upstream region was positively correlated with the levels of ALMT1 expression and Al resistance. In contrast, many of the Japanese lines exhibited a large variation in ALMT1 expression and Al resistance despite possessing the same type of upstream region. Although ALMT1 expression was also poorly correlated with Al-activated malate efflux in the Japanese lines, a strong correlation between malate efflux and Al resistance suggested that malate efflux was still the primary mechanism for Al resistance, and that additional genes are involved in the post-transcriptional regulation of ALMT1 function.

DOI： 10.1093/pcp/pcl002

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：NATL ACAD SCIENCES  

Aluminum (Al) tolerance in Arabidopsis is a genetically complex trait, yet it is mediated by a single physiological mechanism based on Al-activated root malate efflux. We investigated a possible molecular determinant for Al tolerance involving a homolog of the wheat Al-activated malate transporter, ALMT1. This gene, named AtALMT1(At1 g08430), was the best candidate from the 14-member AtALMT family to be involved with Al tolerance based on expression patterns and genomic location. Physiological analysis of a transferred DNA knockout mutant for AtALMT1 as well as electro-physiological examination of the protein expressed in Xenopus oocytes showed that AtALMT1 is critical for Arabidopsis Al tolerance and encodes the Al-activated root malate efflux transporter associated with tolerance. However, gene expression and sequence analysis of AtALMT1 alleles from tolerant Columbia (Col), sensitive Landsberg erecta (Ler), and other ecotypes that varied in Al tolerance suggested that variation observed at AtALMT1 is not correlated with the differences observed in Al tolerance among these ecotypes. Genetic complementation experiments indicated that the Ler allele of AtALMT1 is equally effective as the Col allele in conferring Al tolerance and Al-activated malate release. Finally, fine-scale mapping of a quantitative trait locus (QTL) for Al tolerance on chromosome 1 indicated that AtALMT1 is located proximal to this QTL. These results indicate that AtALMT1 is an essential factor for AI tolerance in Arabidopsis but does not represent the major Al tolerance QTL also found on chromosome 1.

DOI： 10.1073/pnas.0602868103

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

The plasma membrane H+-ATPase plays an important role in the plant response to nutrient and environmental stresses. However, the involvement of plant root plasma membrane H+-ATPase in adaptation to phosphate (P) starvation is not yet fully elucidated. In this study, experiments were performed with soybean roots in low-P nutrient solution (10 mu M). Treatment with fusicoccin, an activator of the plasma membrane H+-ATPase, increased P uptake by 35%, while vanadate, an inhibitor of plasma membrane H+-ATPase, severely suppressed it. These results suggested that P uptake might be regulated via the modulation of the activity of plasma membrane H+-ATPase under P starvation. The relationship between P uptake and the activity of plasma membrane H+-ATPase was examined further by using plasma membrane H+-ATPase transgenic Arabidopsis thaliana under low-P conditions. Transgenic plants absorbed more P compared with wild-type Arabidopsis. Results from real-time RT-PCR, western-blotting and immunolocalization analysis indicated that the increase in activity of the plasma membrane H+-ATPase by P starvation was caused by its transcriptional and translational regulation. A higher expression was observed at the translational level than at the transcriptional level. P starvation could induce a transient increase of endogenous indole-3-acetic acid (IAA) in soybean roots. The exogenous application of IAA stimulated the activity of plasma membrane H+-ATPase and P uptake, while naphthylphthalamic acid (NPA), an IAA transport inhibitor, blocked IAA effects. Taken together, these results suggested an involvement of root plasma membrane H+-ATPase in the adaptation of soybean to P starvation. IAA might be involved in signal transduction of P starvation by activating the plasma membrane H+-ATPase in soybean roots.

DOI： 10.1093/jxb/erj111

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：NATL RESEARCH COUNCIL CANADA  

The major aluminum (M) tolerance gene in wheat ALMTI confers an Al-activated efflux of malate from root apices. We determined the genomic structure of the ALMTI gene and found it consists of 6 exons interrupted by 5 introns. Sequencing a range of wheat genotypes identified 3 alleles for ALMT1. I of which was identical to the ALMT1 gene from an Aegilops tauschii accession. The ALMT1 gene was mapped to chromosome 4DL using 'Chinese Spring' deletion lines, and loss of ALMT1 coincided with the loss of both Al tolerance and Al-activated malate efflux. Aluminium tolerance in each of 5 different doubled-haploid populations was found to be conditioned by a single major gene. When ALMT1 was polymorphic between the parental lines, QTL and linkage analyses indicated that ALMT1 mapped to chromosome 4DL and cosegregated with Al tolerance. In 2 populations examined. Al tolerance also segregated with a greater capacity for Al-activated rnalate efflux. Aluminium tolerance was not associated with a particular coding allele for ALMT1, but was significantly correlated with the relative level of ALMT1 expression. These findings suggest that the Al tolerance in a diverse range of wheat genotypes is primarily conditioned by ALMT1.

DOI： 10.1139/G05-054

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記述言語：英語   出版者・発行元：AMER SOC PLANT BIOLOGISTS  

DOI： 10.1104/pp.104.058065

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE INC  

Aluminium (Al) ion limits root growth of plants in acidic soils, and rice exhibits the highest level of Al-tolerance among graminous crops. To elucidate M-tolerance mechanisms in rice, response to Al was compared between rice (Oryza sativa L., cv. Nippon-bare) and wheat (Triticum aestivum L., cv. ET8), focusing on seminal root growth at seedling stage and germination stage. At seedling stage, rice and wheat were similarly sensitive to Al in both dose- and time-dependent manner during a 24-h Al exposure. On the contrary, at germination stage, rice was more tolerant to Al than wheat, and wheat roots displayed the loss of plasma membrane integrity more extensively than rice. A rice mutant exhibiting Al hypersensitivity at germination stage was obtained by screening 42,840 R-2 progeny derived from the regenerated R-0 plants of Nipponbare and thereafter confirmation of the mutant phenotype in R-3 progeny. At germination stage, root growth of the mutant was strongly inhibited in the presence of Al but not in the absence of Al. However, at seedling stage, root growth of the mutant and wild type was similarly tolerant to Al. Taken together, we conclude that rice possesses Al-tolerant function that is under genetic control and specifically operates for root growth at germination stage, making rice more tolerant to Al than wheat. (c) 2005 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.jinorgbio.2005.06.031

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Aluminum (Al)-activated malate transporter (ALMT1) was recently identified from wheat ( Triticum aestivum). Heterologous expression of ALMT1 led to higher malate exudation that is associated with enhanced Al tolerance in transgenic plants. Here, we show the first direct evidence that ALMT1 is localized in the plasma membrane of Al-tolerant wheat. Phase partitioning experiments showed that this transporter was associated with the plasma membrane fraction. ALMT1 was detected in an Al-tolerant wheat line even without Al treatments. Analysis of transient expression of ALMT1:: green fluorescent protein (GFP) in onion and tobacco cells further confirmed this ALMT1 localization.

DOI： 10.1093/pcp/pci083

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：NATL ACAD SCIENCES  

Acidity is a serious limitation to plant production on many of the world's agricultural soils. Toxic aluminium (Al) cations solubilized by the acidity rapidly inhibit root growth and limit subsequent uptake of water and nutrients. Recent work has shown that the ALMT1 gene of wheat (Triticum aestivum) encodes a malate transporter that is associated with malate efflux and Al tolerance. We generated transgenic barley (Hordeum vulgare) plants expressing ALMT1 and assessed their ability to exude malate and withstand Al stress. ALMT1 expression in barley conferred an Al-activated efflux of malate with properties similar to those of Al-tolerant wheat. The transgenic barley showed a high level of Al tolerance when grown in both hydroponic culture and on acid soils. These findings provide additional evidence that ALMT1 is a major Al-tolerance gene and demonstrate its ability to confer effective tolerance to acid soils through a transgenic approach in an important crop species.

DOI： 10.1073/pnas.0406258101

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BLACKWELL PUBLISHING LTD  

The major constraint to plant growth in acid soils is the presence of toxic aluminum (Al) cations, which inhibit root elongation. The enhanced Al tolerance exhibited by some cultivars of wheat is associated with the Al-dependent efflux of malate from root apices. Malate forms a stable complex with Al that is harmless to plants and, therefore, this efflux of malate forms the basis of a hypothesis to explain Al tolerance in wheat. Here, we report on the cloning of a wheat gene, ALMT1 (aluminum-activated malate transporter), that co-segregates with Al tolerance in F-2 and F-3 populations derived from crosses between near-isogenic wheat lines that differ in Al tolerance. The ALMT1 gene encodes a membrane protein, which is constitutively expressed in the root apices of the Al-tolerant line at greater levels than in the near-isogenic but Al-sensitive line. Heterologous expression of ALMT1 in Xenopus oocytes, rice and cultured tobacco cells conferred an Al-activated malate efflux. Additionally, ALMT1 increased the tolerance of tobacco cells to Al treatment. These findings demonstrate that ALMT1 encodes an Al-activated malate transporter that is capable of conferring Al tolerance to plant cells.

DOI： 10.1111/j.1365-313X.2003.01991.x

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

A cDNA clone exclusively induced by aluminum (Al) was isolated from root apices of wheat (Triticum aestivum L.) by the differential display method. The predicted amino acid sequence exhibited homology to the multidrug resistance (MDR) proteins that is known as a member of the ATP-binding cassette (ABC) protein superfamily. Thus this gene was named TaMDR1 (Triticum aestivum MDR). TaMDR1 was induced as a function of Al concentration in the range from 5 to 50 muM, which is in the range of Al content in natural acid soil environment. The concentration required for the induction was lower in the Al-sensitive cultivar than in the Al-tolerant cultivar, indicating that the accumulation of TaMDR1 mRNA was associated with the events caused by Al toxicity rather than Al tolerance. TaMDR1 was significantly induced by the exposure to lanthanum, gadolinium and ruthenium red, which are known as inhibitors of calcium channels. Furthermore, decreasing of calcium ion in growth medium caused stimulation of the gene expression. These results suggested that the induction of TaMDR1 is caused by the breaking of calcium homeostasis which occurred at early stage of Al toxicity.

DOI： 10.1093/pcp/pcf025

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記述言語：英語   出版者・発行元：OXFORD UNIV PRESS  

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：GEORG THIEME VERLAG  

The number and cross-sectional area of vacuoles in Chlorococcum littorale cells visualized with a differential interference fluorescence microscope increased after their transfer from air to 40% CO2. An immunological observation indicated that the level of subunit B of vacuolar H+-ATPase also increased under 40 % CO2 conditions. The activity of nitrate-sensitive ATPase associated with the vacuolar membrane was 2-fold higher in 40 % CO2-grown cells than in air-grown cells. The effects of inhibitors on the ATPase activity confirmed that these activities were derived from vacuolar-type H+-ATPase. These results suggest that vacuole development associated with that of vacuolar H+-ATPase occurred during the acclimatization of C. littorale cells to extremely high CO2 conditions.

DOI： 10.1111/j.1438-8677.1999.tb00710.x

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JAPANESE SOC PLANT PHYSIOLOGISTS  

The marine green alga, Chlorococcum littorale, accumulated iron in its cells and showed high activity of plasma membrane ferric reductase under high-CO2 and iron-deficient conditions. These activities disappeared upon exposure to ordinary air and by adding excess FeSO4. The iron uptake had high affinity for the Fe(II) form (K-m of 0.13 mu M). Carbonylcyanide m-chlorophenylhydrazone and N,N-dicyclohexylcarbodiimide significantly suppressed the iron uptake, suggesting that the Fe(II) uptake was driven by ATPase. These results indicate that high CO2 and iron deficiency cooperatively induce the Fe(II) uptake and cell-surface ferric reductase activity.

DOI： 10.1093/oxfordjournals.pcp.a029383

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JAPANESE SOC PLANT PHYSIOLOGISTS  

Two cDNA clones exclusively induced under an extremely high-CO2 concentration (20%) were isolated from Chlorococcum littorale by differential screening and named HCR (high-CO2 response) 1 and 2, respectively, The amino acid sequence of the protein encoded by HCR2 exhibited homology to the gp91-phox protein, a critical component of a human phagocyte oxidoreductase, and to the yeast ferric reductases, Saccharomyces cerevisiae FRE1 and FRE2 and Schizosaccharomyces pombe Frp1. The induction of both HCR mRNAs required extremely high-CO2 conditions and iron deficiency, being suppressed under air conditions and by iron sufficiency, suggesting that the expression of these two HCR genes required extremely high-CO2 conditions and iron deficiency in combination, The HCR2 protein was detected in the membrane fractions of cells grown under conditions which would favor the induction of HCR2-mRNA and the protein level was lowered when the cells were transferred from iron deficient to 10 mu M FeSO4 conditions (with 20% CO2).

DOI： 10.1093/oxfordjournals.pcp.a029349

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Blackwell Publishing  

The activity of periplasmic arylsulfatase (Ars), which catalyzes the cleavage of sulfate from aromatic sulfur compounds, was detected in cells acclimated to the sulfate-deficient conditions in a unicellular green alga Chlamydomonas reinhardtii Dangeard, but not in Chlorella, Scenedesmus, Dunaliella and Porphyridium. Upon the transfer of cells to sulfate-deficient autotrophic media under high-CO2 conditions, the induction of Ars was observed only in the light, but not in the light with dichlorophenyldimethylurea (DCMU) nor in the dark. However, Ars was induced in the light with DCMU or in the dark when acetate was present as an organic carbon source, but not citrate. Under similar high-CO2 conditions, high-CO2 requiring mutants of cia-3 and cia-5, whose photosynthetic activities are greatly limited under low CO2, showed much lower level of Ars activities than wild type cells. Under Iow-CO2 conditions the induction of Ars was greatly suppressed even in wild type and no induction was observed in both mutants. These results suggest that the stimulation of photosynthetic or respiratory carbon metabolism are necessary for the induction of Ars. In contrast, the induction of periplasmic carbonic anhydrase (CA) which was synthesized de novo specifically under CO2-limited conditions was strongly suppressed by the addition of organic carbon sources, such as acetate and citrate. When cells are subjected to CO2-limitation and sulfate-deficiency simultaneously, the induction of CA was initiated immediately, while that of Ars was initiated following the completion of CA induction with an about 4-h lag. When the concentration of CO2 was suddenly lowered during the induction of Ars, the induction of Ars ceased quickly, and the induction of CA was initiated instead. From these results the induction of CA was suggested to have priority over that of Ars under the dual stress of CO2 and sulfate-deprivation.

DOI： 10.1111/j.1440-1835.1997.tb00077.x

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語  

DOI： 10.14841/jspp.2011.0.0963.0

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記述言語：日本語   掲載種別：研究発表ペーパー・要旨（全国大会，その他学術会議）  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語  

DOI： 10.14841/jspp.2010.0.0891.0

J-GLOBAL

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記述言語：日本語  

DOI： 10.14841/jspp.2010.0.0889.0

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語  

DOI： 10.14841/jspp.2007.0.288.0

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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DOI： 10.1007/s11104-006-9068-5

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語  

J-GLOBAL

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：日本語   出版者・発行元：Japan Society for Bioscience, Biotechnology, and Agrochemistry  

DOI： 10.1271/kagakutoseibutsu1962.43.569

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その他リンク： https://jlc.jst.go.jp/DN/JALC/00256249994?from=CiNii

記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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DOI： 10.1071/FP04091

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：日本語   出版者・発行元：一般社団法人日本土壌肥料学会  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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記述言語：英語   掲載種別：研究発表ペーパー・要旨（国際会議）   出版者・発行元：OXFORD UNIV PRESS  

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