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Comparative proteomics analysis of the root apoplasts of rice seedlings in response to hydrogen peroxide.

Zhou L, Bokhari SA, Dong CJ, Liu JY - PLoS ONE (2011)

Bottom Line: Of these, 54 were successfully identified by PMF or MS/MS as matches to 35 different proteins including known and novel H(2)O(2)-responsive proteins.Almost all of these identities (98%) were indeed apoplast proteins confirmed either by previous experiments or through publicly available prediction programs.Based on the abundant changes in these proteins, together with their putative functions, we proposed a possible protein network that provides new insights into oxidative stress response in the rice root apoplast and clues for the further functional research of target proteins associated with H(2)O(2) response.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Biology, School of Life Sciences, Tsinghua University, Beijing, People's Republic of China.

ABSTRACT

Background: Plant apoplast is the prime site for signal perception and defense response, and of great importance in responding to environmental stresses. Hydrogen peroxide (H(2)O(2)) plays a pivotal role in determining the responsiveness of cells to stress. However, how the apoplast proteome changes under oxidative condition is largely unknown. In this study, we initiated a comparative proteomic analysis to explore H(2)O(2)-responsive proteins in the apoplast of rice seedling roots.

Methodology/principal findings: 14-day-old rice seedlings were treated with low concentrations (300 and 600 µM) of H(2)O(2) for 6 h and the levels of relative electrolyte leakage, malondialdehyde and H(2)O(2) were assayed in roots. The modified vacuum infiltration method was used to extract apoplast proteins of rice seedling roots, and then two-dimensional electrophoresis gel analysis revealed 58 differentially expressed protein spots under low H(2)O(2) conditions. Of these, 54 were successfully identified by PMF or MS/MS as matches to 35 different proteins including known and novel H(2)O(2)-responsive proteins. Almost all of these identities (98%) were indeed apoplast proteins confirmed either by previous experiments or through publicly available prediction programs. These proteins identified are involved in a variety of processes, including redox homeostasis, cell wall modification, signal transduction, cell defense and carbohydrate metabolism, indicating a complex regulative network in the apoplast of seedling roots under H(2)O(2) stress.

Conclusions/significance: The present study is the first apoplast proteome investigation of plant seedlings in response to H(2)O(2) and may be of paramount importance for the understanding of the plant network to environmental stresses. Based on the abundant changes in these proteins, together with their putative functions, we proposed a possible protein network that provides new insights into oxidative stress response in the rice root apoplast and clues for the further functional research of target proteins associated with H(2)O(2) response.

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Close-up of possible isoforms detected by 2-DE (A) and their expression profile patterns (B).All 29 differentially expressed protein spots, matching 10 different proteins, are shown. T1 and T2 represent H2O2 treatments of 300 µM and 600 µM, respectively. ARA-I, α-arabinofuranosidse/β-D-xylosidase isoenzyme; Prx, peroxidase; MDH, malate dehydrogenase; RMC, root meader curling.
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pone-0016723-g004: Close-up of possible isoforms detected by 2-DE (A) and their expression profile patterns (B).All 29 differentially expressed protein spots, matching 10 different proteins, are shown. T1 and T2 represent H2O2 treatments of 300 µM and 600 µM, respectively. ARA-I, α-arabinofuranosidse/β-D-xylosidase isoenzyme; Prx, peroxidase; MDH, malate dehydrogenase; RMC, root meader curling.

Mentions: Moreover, among the 35 differentially expressed proteins, 10 proteins were present as multiple spots on the 2-DE gels, with one spot representing an isoform (Figure 4A). Of these, the isoforms for 8 proteins [α-L-arabinofuranosidase/β-D-xylosidase isoenzyme, enolase, putative α-galactosidases, a putative β-1,3-glucanase, two peroxidases (OsPrx112 and OsPrx125), a malate dehydrogenase and the DUF26 motif-containing protein, OsRMC], representing 20 identities showed similar up- or down-regulated changes in abundance in response to H2O2 treatment. The isoforms for the other two proteins (OsPrx111 and β-1,3-glucanase), representing 9 identities, exhibited opposite expression patterns (Figure 4B). Likewise, similar phenomena have been observed in previous proteomics studies [3], [23], [24], which are probably due to the posttranslational modifications. These results suggest that isoforms of a certain protein may play either the same or different roles in modulating cell responses to H2O2 treatments in rice seedlings.


Comparative proteomics analysis of the root apoplasts of rice seedlings in response to hydrogen peroxide.

Zhou L, Bokhari SA, Dong CJ, Liu JY - PLoS ONE (2011)

Close-up of possible isoforms detected by 2-DE (A) and their expression profile patterns (B).All 29 differentially expressed protein spots, matching 10 different proteins, are shown. T1 and T2 represent H2O2 treatments of 300 µM and 600 µM, respectively. ARA-I, α-arabinofuranosidse/β-D-xylosidase isoenzyme; Prx, peroxidase; MDH, malate dehydrogenase; RMC, root meader curling.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3037377&req=5

pone-0016723-g004: Close-up of possible isoforms detected by 2-DE (A) and their expression profile patterns (B).All 29 differentially expressed protein spots, matching 10 different proteins, are shown. T1 and T2 represent H2O2 treatments of 300 µM and 600 µM, respectively. ARA-I, α-arabinofuranosidse/β-D-xylosidase isoenzyme; Prx, peroxidase; MDH, malate dehydrogenase; RMC, root meader curling.
Mentions: Moreover, among the 35 differentially expressed proteins, 10 proteins were present as multiple spots on the 2-DE gels, with one spot representing an isoform (Figure 4A). Of these, the isoforms for 8 proteins [α-L-arabinofuranosidase/β-D-xylosidase isoenzyme, enolase, putative α-galactosidases, a putative β-1,3-glucanase, two peroxidases (OsPrx112 and OsPrx125), a malate dehydrogenase and the DUF26 motif-containing protein, OsRMC], representing 20 identities showed similar up- or down-regulated changes in abundance in response to H2O2 treatment. The isoforms for the other two proteins (OsPrx111 and β-1,3-glucanase), representing 9 identities, exhibited opposite expression patterns (Figure 4B). Likewise, similar phenomena have been observed in previous proteomics studies [3], [23], [24], which are probably due to the posttranslational modifications. These results suggest that isoforms of a certain protein may play either the same or different roles in modulating cell responses to H2O2 treatments in rice seedlings.

Bottom Line: Of these, 54 were successfully identified by PMF or MS/MS as matches to 35 different proteins including known and novel H(2)O(2)-responsive proteins.Almost all of these identities (98%) were indeed apoplast proteins confirmed either by previous experiments or through publicly available prediction programs.Based on the abundant changes in these proteins, together with their putative functions, we proposed a possible protein network that provides new insights into oxidative stress response in the rice root apoplast and clues for the further functional research of target proteins associated with H(2)O(2) response.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Biology, School of Life Sciences, Tsinghua University, Beijing, People's Republic of China.

ABSTRACT

Background: Plant apoplast is the prime site for signal perception and defense response, and of great importance in responding to environmental stresses. Hydrogen peroxide (H(2)O(2)) plays a pivotal role in determining the responsiveness of cells to stress. However, how the apoplast proteome changes under oxidative condition is largely unknown. In this study, we initiated a comparative proteomic analysis to explore H(2)O(2)-responsive proteins in the apoplast of rice seedling roots.

Methodology/principal findings: 14-day-old rice seedlings were treated with low concentrations (300 and 600 µM) of H(2)O(2) for 6 h and the levels of relative electrolyte leakage, malondialdehyde and H(2)O(2) were assayed in roots. The modified vacuum infiltration method was used to extract apoplast proteins of rice seedling roots, and then two-dimensional electrophoresis gel analysis revealed 58 differentially expressed protein spots under low H(2)O(2) conditions. Of these, 54 were successfully identified by PMF or MS/MS as matches to 35 different proteins including known and novel H(2)O(2)-responsive proteins. Almost all of these identities (98%) were indeed apoplast proteins confirmed either by previous experiments or through publicly available prediction programs. These proteins identified are involved in a variety of processes, including redox homeostasis, cell wall modification, signal transduction, cell defense and carbohydrate metabolism, indicating a complex regulative network in the apoplast of seedling roots under H(2)O(2) stress.

Conclusions/significance: The present study is the first apoplast proteome investigation of plant seedlings in response to H(2)O(2) and may be of paramount importance for the understanding of the plant network to environmental stresses. Based on the abundant changes in these proteins, together with their putative functions, we proposed a possible protein network that provides new insights into oxidative stress response in the rice root apoplast and clues for the further functional research of target proteins associated with H(2)O(2) response.

Show MeSH
Related in: MedlinePlus