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Response of plasma membrane H+-ATPase to heavy metal stress in Cucumis sativus roots.

ABSTRACT: The effect of heavy metals on the modification of plasma membrane H(+)-ATPase (EC 3.6.3.14) activity in cucumber roots was studied. In plants stressed for 2 h with 10 microM or 100 microM Cd, Cu or Ni the hydrolytic as well as the transporting activity of H(+)-ATPase in the plasma membranes of root cells was decreased. Transcript levels of Cucumis sativus plasma membrane H(+)-ATPase in roots treated with 10 microM Cd, Cu, or Ni as well as with 100 microM Cu or Ni were similar to the control, indicating that the action of metals did not involve the gene expression level. Only in roots exposed to 100 microM Cd was the level of plasma membrane H(+)-ATPase mRNA markedly decreased. The inhibition of the plasma membrane proton pump caused by 100 microM Cd, Cu and Ni was partially diminished in the presence of cantharidin, a specific inhibitor of protein phosphatases. Western blot analysis with the antibody against phosphothreonine confirmed that decreased activity of plasma membrane H(+)-ATPase under heavy metals resulted from dephosphorylation of the enzyme protein. Taken together, these data strongly indicated that alteration of the enzyme under heavy metal stresses was mainly due to the post-translational modification of its proteins in short-term experiments.

SUBMITTER: Janicka-Russak M

PROVIDER: S-EPMC2561156 | biostudies-literature | 2008

REPOSITORIES: biostudies-literature

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Response of plasma membrane H+-ATPase to heavy metal stress in Cucumis sativus roots.

Janicka-Russak Małgorzata M Kabała Katarzyna K Burzyński Marek M Kłobus Grazyna G

Journal of experimental botany 20080926 13

The effect of heavy metals on the modification of plasma membrane H(+)-ATPase (EC 3.6.3.14) activity in cucumber roots was studied. In plants stressed for 2 h with 10 microM or 100 microM Cd, Cu or Ni the hydrolytic as well as the transporting activity of H(+)-ATPase in the plasma membranes of root cells was decreased. Transcript levels of Cucumis sativus plasma membrane H(+)-ATPase in roots treated with 10 microM Cd, Cu, or Ni as well as with 100 microM Cu or Ni were similar to the control, ind ...[more]

PMID: 18820260

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Project description:BackgroundIron deficiency induces in Strategy I plants physiological, biochemical and molecular modifications capable to increase iron uptake from the rhizosphere. This effort needs a reorganization of metabolic pathways to efficiently sustain activities linked to the acquisition of iron; in fact, carbohydrates and the energetic metabolism has been shown to be involved in these responses. The aim of this work was to find both a confirmation of the already expected change in the enzyme concentrations induced in cucumber root tissue in response to iron deficiency as well as to find new insights on the involvement of other pathways.ResultsThe proteome pattern of soluble cytosolic proteins extracted from roots was obtained by 2-DE. Of about two thousand spots found, only those showing at least a two-fold increase or decrease in the concentration were considered for subsequent identification by mass spectrometry. Fifty-seven proteins showed significant changes, and 44 of them were identified. Twenty-one of them were increased in quantity, whereas 23 were decreased in quantity. Most of the increased proteins belong to glycolysis and nitrogen metabolism in agreement with the biochemical evidence. On the other hand, the proteins being decreased belong to the metabolism of sucrose and complex structural carbohydrates and to structural proteins.ConclusionsThe new available techniques allow to cast new light on the mechanisms involved in the changes occurring in plants under iron deficiency. The data obtained from this proteomic study confirm the metabolic changes occurring in cucumber as a response to Fe deficiency. Two main conclusions may be drawn. The first one is the confirmation of the increase in the glycolytic flux and in the anaerobic metabolism to sustain the energetic effort the Fe-deficient plants must undertake. The second conclusion is, on one hand, the decrease in the amount of enzymes linked to the biosynthesis of complex carbohydrates of the cell wall, and, on the other hand, the increase in enzymes linked to the turnover of proteins.

Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of gene expression profiles of cucumber under short-term chilling stress. The goals of this study are to transcriptome analysis of cucumber leaves under chilling stress. Methods: mRNA profiles of seedlings exposed to an air temperature of 6°C in the absence of light at 0, 2, 6, and 12 h were generated by deep sequencing, in triplicate, using Illumina Hiseq platform. The reference genome and gene model annotation files were downloaded from the genome website （http://cucurbitgenomics.org/）. An index of the reference genome was built using Bowtie v.2.2.3 and paired-end clean reads were aligned to the reference genome using TopHat v.2.0.12. qRT–PCR validation was performed using SYBR Green assays. Results: A total of 55.7 million clean reads was generated. Based on the threshold values of absolute value of log2 ratio ≥ 1 and FDR ≤ 0.05, a total of 2113 DEGs was identified at three time points (2, 6, and 12 h). A total of 30 genes was detected at all time points. The number of DEGs increased with time. In total, 100 TFs from 22 families in three subsets were detected. And 19 kinase families were identified in three subsets. The DEGs identified by RNA sequencing were confirmed by qRT-PCR analysis, indicating that the data were reliable. These findings provide information that can be useful for investigating the molecular mechanisms underlying the response to chilling stress in cucumber and other plants. Conclusions: The results presented here reveal changes in the transcriptome profile of cucumber in response to chilling stress. Exposure to a low temperature induced genes involved in hormone regulation, lipid metabolism, and photosynthesis, including NAC, WRKY, AP2/ERF, ERD, MYB as well as zinc finger TFs and protein kinases such as receptor-like protein kinase, MAPK, and CDK. Most TFs were upregulated whereas CDKs were downregulated. These findings provide information that can be useful for investigating the molecular mechanisms underlying the response to chilling stress in cucumber and other plants.

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Response of plasma membrane H+-ATPase to heavy metal stress in Cucumis sativus roots.

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Response of plasma membrane H+-ATPase to heavy metal stress in Cucumis sativus roots.

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