A Simple Hydroponic Culture Method for the Development of a Highly Viable Root System in<i>Arabidopsis thaliana</i>

Toda, Takeshi; Koyama, Hiroyuki; Hara, Tetsuo

doi:10.1271/bbb.63.210

Cited by 50 publications

(38 citation statements)

References 9 publications

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“…Seedlings were precultured for 10 d in 2% MGRL nutrient with 200 mM CaCl 2 at pH 5.6 hydroponically as described by Toda et al (1999). Solutions were renewed every 2 d, and seedlings were kept in a controlled environment (30 mmol quanta m 22 s 21 , 12-h day/night cycle, 25°C).…”

Section: Methodsmentioning

confidence: 99%

STOP1 Regulates Multiple Genes That Protect Arabidopsis from Proton and Aluminum Toxicities

et al. 2009

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The Arabidopsis (Arabidopsis thaliana) mutant stop1 (for sensitive to proton rhizotoxicity1) carries a missense mutation at an essential domain of the histidine-2-cysteine-2 zinc finger protein STOP1. Transcriptome analyses revealed that various genes were down-regulated in the mutant, indicating that STOP1 is involved in signal transduction pathways regulating aluminum (Al)- and H+-responsive gene expression. The Al hypersensitivity of the mutant could be caused by down-regulation of AtALMT1 (for Arabidopsis ALUMINUM-ACTIVATED MALATE TRANSPORTER1) and ALS3 (ALUMINUM-SENSITIVE3). This hypothesis was supported by comparison of Al tolerance among T-DNA insertion lines and a transgenic stop mutant carrying cauliflower mosaic virus 35S∷AtALMT1. All T-DNA insertion lines of STOP1, AtALMT1, and ALS3 were sensitive to Al, but introduction of cauliflower mosaic virus 35S∷AtALMT1 did not completely restore the Al tolerance of the stop1 mutant. Down-regulation of various genes involved in ion homeostasis and pH-regulating metabolism in the mutant was also identified by microarray analyses. CBL-INTERACTING PROTEIN KINASE23, regulating a major K+ transporter, and a sulfate transporter, SULT3;5, were down-regulated in the mutant. In addition, integral profiling of the metabolites and transcripts revealed that pH-regulating metabolic pathways, such as the γ-aminobutyric acid shunt and biochemical pH stat pathways, are down-regulated in the mutant. These changes could explain the H+ hypersensitivity of the mutant and would make the mutant more susceptible in acid soil stress than other Al-hypersensitive T-DNA insertion lines. Finally, we showed that STOP1 is localized to the nucleus, suggesting that the protein regulates the expression of multiple genes that protect Arabidopsis from Al and H+ toxicities, possibly as a transcription factor.

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Section: Methodsmentioning

confidence: 99%

STOP1 Regulates Multiple Genes That Protect Arabidopsis from Proton and Aluminum Toxicities

et al. 2009

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“…Low pH treatment was carried out using the standard 2% modified MGRL medium acidified to a lower pH (4.7). In each replicate experimental treatment, 10 seedlings per line were grown for 7 d, replacing the medium every 2 d. Plants were grown on culture apparatus made from plastic mesh and photo slide mount as described previously (Toda et al, 1999), and the root length was measured by capturing with a micro video camera (Pico Scopemen, Kenis) as described previously. Growth experiments were repeated at least three times and means and SEs were obtained to calculate relative root length (toxic condition/control [%]).…”

Section: Growth Condition and Root Measurementmentioning

confidence: 99%

“…This schema is also applicable for characterization of other rhizotoxic metals, some of which also elicit OA release (Murphy et al, 1999;Toda et al, 1999). In addition, we developed a sensitive method for detecting malate release from the roots (Hoekenga et al, 2003).…”

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Characterization of AtALMT1 Expression in Aluminum-Inducible Malate Release and Its Role for Rhizotoxic Stress Tolerance in Arabidopsis

et al. 2007

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Malate transporters play a critical role in aluminum (Al) tolerance responses for some plant species, such as Arabidopsis (Arabidopsis thaliana). Here, we further characterize AtALMT1, an Arabidopsis aluminum-activated malate transporter, to clarify its specific role in malate release and Al stress responses. Malate excretion from the roots of accession Columbia was sharply induced by Al, which is concomitant with the induction of AtALMT1 gene expression. The malate release was specific for Al among rhizotoxic stressors, namely cadmium, copper, erbium, lanthanum, sodium, and low pH, which accounts for the specific sensitivity of a null mutant to Al stress. Al-specific malate excretion can be explained by a combined regulation of AtALMT1 expression and activation of AtALMT1 protein, which is specific for Al. Although low pH treatment slightly induced gene expression, other treatments did not. In addition, malate excretion in Al-activated seedlings was rapidly stopped by removing Al from the solution. Other rhizotoxic stressors were not effective in maintaining malate release. Protein kinase and phosphatase inhibitor studies indicated that reversible phosphorylation was important for the transcriptional and posttranslational regulation of AtALMT1. AtALMT1 promoter-β-glucuronidase fusion lines revealed that AtALMT1 has restricted expression within the root, such that unnecessary carbon loss is likely minimized. Lastly, a natural nonsense mutation allele of AtALMT1 was identified from the Al-hypersensitive natural accession Warschau-1.

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“…Seedlings were grown on nylon mesh (about 40 mm span) supported by a plastic photo slide mount (Fuji Film;Toda et al, 1999 were grown in 8 L of the same solution (in plastic containers) to minimize environmental differences. Ten seedlings were grown in both Cu toxic and control solutions, and root length was measured using a video microscope as described by Toda et al (1999). Average root length values were calculated using the top three root lengths in the each treatment.…”

Section: Hydroponic Culture and Phenotyping Of Cu Tolerancementioning

confidence: 99%

“…The ability of Cu to mediate high rates of electron transfer is also a potential cause of toxicity for plant cells (especially in growing roots). Less than 5 mM Cu inhibits the growth of roots in wheat (Triticum aestivum; Taylor et al, 1991;Parker et al, 1998) and Arabidopsis (Arabidopsis thaliana; Toda et al, 1999). This sensitivity of growing roots is a potential risk in agriculture that arises from the use of Cu-containing fungicides and fertilizers.…”

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Amino Acid Polymorphisms in Strictly Conserved Domains of a P-Type ATPase HMA5 Are Involved in the Mechanism of Copper Tolerance Variation in Arabidopsis

et al. 2008

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Copper (Cu) is an essential element in plant nutrition, but it inhibits the growth of roots at low concentrations. Accessions of Arabidopsis (Arabidopsis thaliana) vary in their tolerance to Cu. To understand the molecular mechanism of Cu tolerance in Arabidopsis, we performed quantitative trait locus (QTL) analysis and accession studies. One major QTL on chromosome 1 (QTL1) explained 52% of the phenotypic variation in Cu tolerance in roots in a Landsberg erecta/Cape Verde Islands (Ler/Cvi) recombinant inbred population. This QTL regulates Cu translocation capacity and involves a Cu-transporting P1B-1-type ATPase, HMA5. The Cvi allele carries two amino acid substitutions in comparison with the Ler allele and is less functional than the Ler allele in Cu tolerance when judged by complementation assays using a T-DNA insertion mutant. Complementation assays of the ccc2 mutant of yeast using chimeric HMA5 proteins revealed that N923T of the Cvi allele, which was identified in the tightly conserved domain N(x)6YN(x)4P (where the former asparagine was substituted by threonine), is a cause of dysfunction of the Cvi HMA5 allele. Another dysfunctional HMA5 allele was identified in Chisdra-2, which showed Cu sensitivity and low capacity of Cu translocation from roots to shoots. A unique amino acid substitution of Chisdra-2 was identified in another strictly conserved domain, CPC(x)6P, where the latter proline was replaced with leucine. These results indicate that a portion of the variation in Cu tolerance of Arabidopsis is regulated by the functional integrity of the Cu-translocating ATPase, HMA5, and in particular the amino acid sequence in several strictly conserved motifs.

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A Simple Hydroponic Culture Method for the Development of a Highly Viable Root System inArabidopsis thaliana

Cited by 50 publications

References 9 publications

STOP1 Regulates Multiple Genes That Protect Arabidopsis from Proton and Aluminum Toxicities

STOP1 Regulates Multiple Genes That Protect Arabidopsis from Proton and Aluminum Toxicities

Characterization of AtALMT1 Expression in Aluminum-Inducible Malate Release and Its Role for Rhizotoxic Stress Tolerance in Arabidopsis

Amino Acid Polymorphisms in Strictly Conserved Domains of a P-Type ATPase HMA5 Are Involved in the Mechanism of Copper Tolerance Variation in Arabidopsis

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