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Salt stress-induced alterations in the root proteome of barley genotypes with contrasting response towards salinity.

Witzel K, Weidner A, Surabhi GK, Börner A, Mock HP - J. Exp. Bot. (2009)

Bottom Line: Multivariate analysis of the resulting protein patterns revealed cultivar-specific and salt stress-responsive protein expression.Hierarchical clustering was applied to detect similar protein expression patterns.This study emphasizes the role of proteins involved in ROS detoxification during salinity stress, and identified potential candidates for increasing salt tolerance in barley.

View Article: PubMed Central - PubMed

Affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, D-06466 Gatersleben, Germany.

ABSTRACT
In addition to drought and extreme temperatures, soil salinity represents a growing threat to crop productivity. Among the cereal crops, barley is considered as notably salt tolerant, and cultivars show considerable variation for tolerance towards salinity stress. In order to unravel the molecular mechanisms underlying salt stress tolerance and to utilize the natural genetic variation of barley accessions, a series of hydroponics-based salinity stress experiments was conducted using two genetic mapping parents, cvs Steptoe and Morex, which display contrasting levels of salinity tolerance. The proteome of roots from both genotypes was investigated as displayed by two-dimensional gel electrophoresis, and comparisons were made between plants grown under non-saline and saline conditions. Multivariate analysis of the resulting protein patterns revealed cultivar-specific and salt stress-responsive protein expression. Mass spectrometry-based identification was successful for 26 out of 39 selected protein spots. Hierarchical clustering was applied to detect similar protein expression patterns. Among those, two proteins involved in the glutathione-based detoxification of reactive oxygen species (ROS) were more abundant in the tolerant genotype, while proteins involved in iron uptake were expressed at a higher level in the sensitive one. This study emphasizes the role of proteins involved in ROS detoxification during salinity stress, and identified potential candidates for increasing salt tolerance in barley.

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Related in: MedlinePlus

Estimated protein abundance of catalase (689) based on 2-D gel electrophoresis and western blot analysis. (A) Protein expression in one biological experiment using 2-D gel electrophoresis and immunoblotting. (B) Normalized spot volume as revealed by 2-D gels, and western blot signal intensity determined by densitometric measurement of western blot bands. Error bars show the standard deviation of measurements based on three independent biological experiments.
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fig7: Estimated protein abundance of catalase (689) based on 2-D gel electrophoresis and western blot analysis. (A) Protein expression in one biological experiment using 2-D gel electrophoresis and immunoblotting. (B) Normalized spot volume as revealed by 2-D gels, and western blot signal intensity determined by densitometric measurement of western blot bands. Error bars show the standard deviation of measurements based on three independent biological experiments.

Mentions: In order to confirm the protein expression revealed by 2-D gel electrophoresis, western blot analysis was performed on a selected number of proteins. Available antibodies against catalase, lipoxygenase, ascorbate peroxidase, and the stress-inducible protein F23N19.10 were used for immunoblot experiments. Overall, a high degree of commonality in protein expression was found when normalized spot volumes were compared with signals obtained from immunoblotting. An example is given in Fig. 7 for catalase (spot 689). Shown are the normalized spot volumes determined by the image analysis software and the signal intensity of the western band assessed by densitometric measurement. Here, a higher expression was detected by both methods in cv. Steptoe under control conditions. Upon salinity treatment, a reduction in protein abundance was detected by both approaches in cv. Steptoe, whereas in cv. Morex no significant change was observed. Similar tendencies between spot expression and western signal were retrieved using the remaining antisera (data not shown).


Salt stress-induced alterations in the root proteome of barley genotypes with contrasting response towards salinity.

Witzel K, Weidner A, Surabhi GK, Börner A, Mock HP - J. Exp. Bot. (2009)

Estimated protein abundance of catalase (689) based on 2-D gel electrophoresis and western blot analysis. (A) Protein expression in one biological experiment using 2-D gel electrophoresis and immunoblotting. (B) Normalized spot volume as revealed by 2-D gels, and western blot signal intensity determined by densitometric measurement of western blot bands. Error bars show the standard deviation of measurements based on three independent biological experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2724703&req=5

fig7: Estimated protein abundance of catalase (689) based on 2-D gel electrophoresis and western blot analysis. (A) Protein expression in one biological experiment using 2-D gel electrophoresis and immunoblotting. (B) Normalized spot volume as revealed by 2-D gels, and western blot signal intensity determined by densitometric measurement of western blot bands. Error bars show the standard deviation of measurements based on three independent biological experiments.
Mentions: In order to confirm the protein expression revealed by 2-D gel electrophoresis, western blot analysis was performed on a selected number of proteins. Available antibodies against catalase, lipoxygenase, ascorbate peroxidase, and the stress-inducible protein F23N19.10 were used for immunoblot experiments. Overall, a high degree of commonality in protein expression was found when normalized spot volumes were compared with signals obtained from immunoblotting. An example is given in Fig. 7 for catalase (spot 689). Shown are the normalized spot volumes determined by the image analysis software and the signal intensity of the western band assessed by densitometric measurement. Here, a higher expression was detected by both methods in cv. Steptoe under control conditions. Upon salinity treatment, a reduction in protein abundance was detected by both approaches in cv. Steptoe, whereas in cv. Morex no significant change was observed. Similar tendencies between spot expression and western signal were retrieved using the remaining antisera (data not shown).

Bottom Line: Multivariate analysis of the resulting protein patterns revealed cultivar-specific and salt stress-responsive protein expression.Hierarchical clustering was applied to detect similar protein expression patterns.This study emphasizes the role of proteins involved in ROS detoxification during salinity stress, and identified potential candidates for increasing salt tolerance in barley.

View Article: PubMed Central - PubMed

Affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, D-06466 Gatersleben, Germany.

ABSTRACT
In addition to drought and extreme temperatures, soil salinity represents a growing threat to crop productivity. Among the cereal crops, barley is considered as notably salt tolerant, and cultivars show considerable variation for tolerance towards salinity stress. In order to unravel the molecular mechanisms underlying salt stress tolerance and to utilize the natural genetic variation of barley accessions, a series of hydroponics-based salinity stress experiments was conducted using two genetic mapping parents, cvs Steptoe and Morex, which display contrasting levels of salinity tolerance. The proteome of roots from both genotypes was investigated as displayed by two-dimensional gel electrophoresis, and comparisons were made between plants grown under non-saline and saline conditions. Multivariate analysis of the resulting protein patterns revealed cultivar-specific and salt stress-responsive protein expression. Mass spectrometry-based identification was successful for 26 out of 39 selected protein spots. Hierarchical clustering was applied to detect similar protein expression patterns. Among those, two proteins involved in the glutathione-based detoxification of reactive oxygen species (ROS) were more abundant in the tolerant genotype, while proteins involved in iron uptake were expressed at a higher level in the sensitive one. This study emphasizes the role of proteins involved in ROS detoxification during salinity stress, and identified potential candidates for increasing salt tolerance in barley.

Show MeSH
Related in: MedlinePlus