Gamma-aminobutyric acid elicits H2O2 signalling and promotes wheat seed germination under combined salt and heat stress

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Plant Biology

Main article text

 

Introduction

Materials and Methods

Reagent preparation

Seed germination and treatment

Assay of H2O2, ABA, and GABA contents

Enzyme activity assays

Quantitative real-time PCR (qRT−PCR) for antioxidant enzyme encoding genes

Data analysis

Results

Effects of heat stress, salinity, and GABA on germination rate

Effects of combined stress and GABA on the contents of H2O2, ABA, GABA and GAD activities of GAD and NOX

Effects of combined stress and GABA on activities of SOD and CAT activities and MnSOD and CAT expression levels

Discussion

Conclusions

Supplemental Information

Effects of salinity and heat stress on seed germination.

Effects of heat stress (A), salinity (B), combined stress of salinity and heat stress (C), and GABA (D) on final germination rate of wheat seeds. Bars represent standard deviation of the mean (n = 3); means associated with the same letter are not significantly different (p < 0.05). GABA, γ-aminobutyric acid.

DOI: 10.7717/peerj.17907/supp-1

Effects of GABA on seed germination under combined stress.

Effects of 0–10 mM GABA (A), 2 mM GABA + antioxidant (10 mM DMTU, 0.1 mM DPI or 1 mM IMZ) (B), 2 mM GABA + oxidant (10 mM H2O2, 2 mM ATZ or 2 mM DDC) (C), and 2 mM GABA + hormone (0.5 mM ABA, 0.1 mM Flu or 0.5 mM GA) (D) on final GR of wheat seeds under combined stress (30 °C + 50 mM NaCl). Bars represent standard deviation of the mean (n = 3); means associated with the same letter are not significantly different (p < 0.05). ABA, abscisic acid; GA, gibberellin; GABA, γ-aminobutyric acid; DPI, diphenyleneiodonium chloride; DMTU, dimethylthiourea; IMZ, imidazole; DDC, diethyldithiocarbamic acid; ATZ, aminotriazole; Flu, fluridone.

DOI: 10.7717/peerj.17907/supp-2

Contents of H2O2, ABA and GABA, and activities of GAD and NOX.

Effects of GABA and combined stress (30 °C + 50 mM NaCl) on H2O2 content (A), ABA content (B), GABA content (C), GAD activities (D), and NOX activities (E) in germinated wheat seeds and sprouts within the first 22 h. Bars represent standard deviation of the mean (n = 3); means associated with the same letter are not significantly different (p < 0.05). GABA, γ-aminobutyric acid; ABA, abscisic acid; GAD, glutamic decarboxylase.

DOI: 10.7717/peerj.17907/supp-3

Antioxidant enzyme activities and gene expression levels.

Effects of temperature and salinity (50 mM NaCl) alone and in combination on activities of SOD (A) and CAT (B), and expression levels of MnSOD (C) and CAT (D) in germinated wheat seeds and sprouts within the first 22 h. Bars represent standard deviation of the mean (n = 3); means associated with the same letter are not significantly different (p < 0.05). GABA, γ-aminobutyric acid; SOD, superoxide dismutase; CAT, catalase.

DOI: 10.7717/peerj.17907/supp-4

Supporting materials for MIQE Checklist.

DOI: 10.7717/peerj.17907/supp-6

Additional Information and Declarations

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Song Yu conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Zhihan Lian performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Lihe Yu conceived and designed the experiments, authored or reviewed drafts of the article, and approved the final draft.

Wei Guo performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Chunyu Zhang performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Yifei Zhang conceived and designed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Data Availability

The following information was supplied regarding data availability:

The raw measurements are available in the Supplemental Files.

Funding

This work was financially supported by the National Key Research and Development Program of China (Grant number: 2018YFD1000704), the Postdoctoral Science Foundation Funded General Project of Heilongjiang Province (Grant number: LBH-Z19195), and the Scientific Research Project for People Returned after Further Learning and Talent Introduction of Heilongjiang Bayi Agricultural University (Grant number: XYB2014-02). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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