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Increased Cysteine Accumulation is Essential for Salt Stress Tolerance in Arabidopsis Halotolerance 2-Like (AHL)-Overexpressing Transgenic Plants

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Abstract

Cysteine (Cys) regulates plant growth, development, and various abiotic stress responses. However, the knowledge gained from genetic and molecular studies on the role of Cys metabolism in salinity response remains limited. Here, we introduce a sulfate metabolism-related component, Arabidopsis Halotolerance 2-like (AHL), which is involved in salt stress adaptation. AHL expression was experimentally induced under salt stress conditions using AHL promoter-β-glucuronidase transgenic plants. Phenotypic analysis revealed that the survival rate of AHL-overexpressing (AHL-OE) lines was greater than that of wild-type (WT) plants under high-salinity conditions, while ahl mutant and AHL-RNAi seedlings were more sensitive than WT seedlings. Accumulation of Cys and proline was increased in AHL-OE transgenic lines under salinity stress conditions, but malondialdehyde and hydrogen peroxide levels were lower in AHL-OE than in WT, ahl, and AHL-RNAi plants. In addition, the transcription levels of genes associated with Cys biosynthesis and sulfate metabolism were higher in AHL-OE than ahl and AHL-RNAi seedlings after salt stress treatment. Moreover, exogenous application of Cys could rescue the salt stress-induced sensitive phenotypes of ahl and AHL-RNAi lines, and return them to the WT phenotype. Taken together, our findings indicate that AHL positively regulates salinity response in Arabidopsis seedlings by modulating a Cys-dependent pathway.

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Abbreviations

Cys:

Cysteine

Pro:

Proline

qPCR:

Quantitative real-time polymerase chain reaction

WT:

Wild-type

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Acknowledgements

This work was supported by a National Research Foundation (NRF) of Korea grant funded by the Korean government (Ministry of Science and ICT) (2021R1A2C1009748).

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Authors and Affiliations

  1. Department of Applied Biology, Chonnam National University, Gwangju, 61186, Republic of Korea

    Tinh Van Nguyen, Jeong-In Kim, Cho-Rong Park & Cheol Soo Kim

  2. Department of Basic Science, Buon Ma Thuot University, Buon Ma Thuot, Dak Lak, 630000, Vietnam

    Tinh Van Nguyen

  3. Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup, Jeollabuk, 56212, Republic of Korea

    Moon-Soo Chung

Authors
  1. Tinh Van Nguyen
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  2. Jeong-In Kim
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  3. Cho-Rong Park
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  4. Moon-Soo Chung
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  5. Cheol Soo Kim
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Contributions

TVN and CSK designed the experiments; TVN, JIK, CRP, MSC, and CSK performed the experiments and interpreted the data; TVN, MSC, and CSK wrote the manuscript.

Corresponding author

Correspondence to Cheol Soo Kim.

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The authors declare that there are no conflicts of interest.

Supplementary Information

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12374_2021_9320_MOESM1_ESM.tif

Expression level of AHL in WT, ahl, and AHL transgenic plants. A and B mRNA level of AHL was analyzed by RT-PCR (A) and qPCR (B) using total RNA extracted from 10-day-old WT, ahl, AHL-RNAi (ri2-3, ri5-2), and AHL-overexpressing (OE2-2, OE5-1) transgenic plants. Actin 1 (ACT1) was used as internal control. The bars represent the mean values (± SD) of three biological replicates. Different letters above the bars indicate statistically significant differences at P < 0.05 by Duncan’s multiple-range test (TIF 1746 kb)

Primers used for RT-PCR or qPCR analysis (DOCX 40 kb)

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Van Nguyen, T., Kim, JI., Park, CR. et al. Increased Cysteine Accumulation is Essential for Salt Stress Tolerance in Arabidopsis Halotolerance 2-Like (AHL)-Overexpressing Transgenic Plants. J. Plant Biol. 64, 475–485 (2021). https://doi.org/10.1007/s12374-021-09320-7

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  • DOI: https://doi.org/10.1007/s12374-021-09320-7

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