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The alternative respiratory pathway is involved in brassinosteroid-induced environmental stress tolerance in Nicotiana benthamiana.

Deng XG, Zhu T, Zhang DW, Lin HH - J. Exp. Bot. (2015)

Bottom Line: Chemical scavenging of H2O2 and virus-induced gene silencing (VIGS) of NbRBOHB compromised the BR-induced alternative respiratory pathway, and this result was further confirmed by NbAOX1 promoter analysis.Furthermore, inhibition of AOX activity by chemical treatment or a VIGS-based approach decreased plant resistance to environmental stresses and compromised BR-induced stress tolerance.Taken together, our results indicate that BR-induced AOX capability might contribute to the avoidance of superfluous reactive oxygen species accumulation and the protection of photosystems under stress conditions in N. benthamiana.

View Article: PubMed Central - PubMed

Affiliation: Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu, 610064, PR China Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, PR China.

No MeSH data available.


AOX modulates ROS balance in BR-induced stress tolerance. (A) Superoxide contents were detected by 0.5mg ml–1 of NBT staining. (B) Quantitative measurements of NADPH oxidase activity. (C) H2O2 levels were detected by 2mg ml–1 of DAB staining. (D) Quantitative measurements of H2O2 content. The alternative pathway was inhibited by 1mM SHAM pre-treatment or NbAOX1 silencing in the BL-treated plants, and these plants were then challenged with cold (4 °C), 16% PEG 6000, or HL (600 μmol m–2 s–1) stress for 3 d. Experiments were repeated three times with similar results. Bars represent mean and standard deviation of values obtained from three biological repeats. Significant differences (P<0.05) are denoted by different lowercase letters. FW, fresh weight. (This figure is available in colour at JXB online.)
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Figure 8: AOX modulates ROS balance in BR-induced stress tolerance. (A) Superoxide contents were detected by 0.5mg ml–1 of NBT staining. (B) Quantitative measurements of NADPH oxidase activity. (C) H2O2 levels were detected by 2mg ml–1 of DAB staining. (D) Quantitative measurements of H2O2 content. The alternative pathway was inhibited by 1mM SHAM pre-treatment or NbAOX1 silencing in the BL-treated plants, and these plants were then challenged with cold (4 °C), 16% PEG 6000, or HL (600 μmol m–2 s–1) stress for 3 d. Experiments were repeated three times with similar results. Bars represent mean and standard deviation of values obtained from three biological repeats. Significant differences (P<0.05) are denoted by different lowercase letters. FW, fresh weight. (This figure is available in colour at JXB online.)

Mentions: Plant responses to various types of stress are associated with the generation of ROS (Baxter et al., 2013). We further detected the accumulation of superoxide and H2O2 using NBT and DAB staining procedures, respectively. Both procedures detected decreased staining in BL-treated leaves relative to that in water-treated leaves, although both increased after 3 d of stress. However, BR-decreased staining was largely inhibited in SHAM-pre-treated or TRV:NbAOX1 leaves (Fig. 8A, B). We further analysed NADPH oxidase activity and H2O2 content. Similarly, in water-treated leaves, NADPH oxidase activity and H2O2 content were significantly higher than those in the BL-treated leaves under stress conditions, while these decreases were largely alleviated in SHAM-pre-treated or NbAOX1-silenced leaves (Fig. 8C, D).


The alternative respiratory pathway is involved in brassinosteroid-induced environmental stress tolerance in Nicotiana benthamiana.

Deng XG, Zhu T, Zhang DW, Lin HH - J. Exp. Bot. (2015)

AOX modulates ROS balance in BR-induced stress tolerance. (A) Superoxide contents were detected by 0.5mg ml–1 of NBT staining. (B) Quantitative measurements of NADPH oxidase activity. (C) H2O2 levels were detected by 2mg ml–1 of DAB staining. (D) Quantitative measurements of H2O2 content. The alternative pathway was inhibited by 1mM SHAM pre-treatment or NbAOX1 silencing in the BL-treated plants, and these plants were then challenged with cold (4 °C), 16% PEG 6000, or HL (600 μmol m–2 s–1) stress for 3 d. Experiments were repeated three times with similar results. Bars represent mean and standard deviation of values obtained from three biological repeats. Significant differences (P<0.05) are denoted by different lowercase letters. FW, fresh weight. (This figure is available in colour at JXB online.)
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Related In: Results  -  Collection

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Figure 8: AOX modulates ROS balance in BR-induced stress tolerance. (A) Superoxide contents were detected by 0.5mg ml–1 of NBT staining. (B) Quantitative measurements of NADPH oxidase activity. (C) H2O2 levels were detected by 2mg ml–1 of DAB staining. (D) Quantitative measurements of H2O2 content. The alternative pathway was inhibited by 1mM SHAM pre-treatment or NbAOX1 silencing in the BL-treated plants, and these plants were then challenged with cold (4 °C), 16% PEG 6000, or HL (600 μmol m–2 s–1) stress for 3 d. Experiments were repeated three times with similar results. Bars represent mean and standard deviation of values obtained from three biological repeats. Significant differences (P<0.05) are denoted by different lowercase letters. FW, fresh weight. (This figure is available in colour at JXB online.)
Mentions: Plant responses to various types of stress are associated with the generation of ROS (Baxter et al., 2013). We further detected the accumulation of superoxide and H2O2 using NBT and DAB staining procedures, respectively. Both procedures detected decreased staining in BL-treated leaves relative to that in water-treated leaves, although both increased after 3 d of stress. However, BR-decreased staining was largely inhibited in SHAM-pre-treated or TRV:NbAOX1 leaves (Fig. 8A, B). We further analysed NADPH oxidase activity and H2O2 content. Similarly, in water-treated leaves, NADPH oxidase activity and H2O2 content were significantly higher than those in the BL-treated leaves under stress conditions, while these decreases were largely alleviated in SHAM-pre-treated or NbAOX1-silenced leaves (Fig. 8C, D).

Bottom Line: Chemical scavenging of H2O2 and virus-induced gene silencing (VIGS) of NbRBOHB compromised the BR-induced alternative respiratory pathway, and this result was further confirmed by NbAOX1 promoter analysis.Furthermore, inhibition of AOX activity by chemical treatment or a VIGS-based approach decreased plant resistance to environmental stresses and compromised BR-induced stress tolerance.Taken together, our results indicate that BR-induced AOX capability might contribute to the avoidance of superfluous reactive oxygen species accumulation and the protection of photosystems under stress conditions in N. benthamiana.

View Article: PubMed Central - PubMed

Affiliation: Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment, College of Life Science, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu, 610064, PR China Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, PR China.

No MeSH data available.