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Overexpression of CrCOMT from Carex rigescens increases salt stress and modulates melatonin synthesis in Arabidopsis thaliana

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Abstract

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CrCOMT, a COMT gene in Carex rigescens, was verified to enhance salt stress tolerance in transgenic Arabidopsis.

Abstract

High salinity severely restricts plant growth and development while melatonin can alleviate salt damage. Caffeic acid O-methyltransferase (COMT) plays an important role in regulating plant growth, development, and stress responses. COMT could also participate in melatonin biosynthesis. The objective of this study was to identify CrCOMT from Carex rigescens (Franch.) V. Krecz, a stress-tolerant grass species with a widespread distribution in north China, and to determine its physiological functions and regulatory mechanisms that impart tolerance to salt stress. The results showed that the transcription of CrCOMT exhibited different expression patterns under salt, drought, and ABA treatments. Transgenic Arabidopsis with the overexpression of CrCOMT exhibited improved growth and physiological performance under salt stress, such as higher lateral root numbers, proline level, and chlorophyll content, than in the wild type (WT). Overexpression of CrCOMT also increased dehydration tolerance in Arabidopsis. The transcription of salt response genes was more highly activated in transgenic plants than in the WT under salt stress conditions. In addition, the melatonin content in transgenic plants was higher than that in the WT after stress treatment. Taken together, our results indicated that CrCOMT may positively regulate stress responses and melatonin synthesis under salt stress.

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Notes

  1. https://web.expasy.org/cgi-bin/protscale/protscale.pl/.

  2. https://web.expasy.org/cgi-bin/protscale/protscale.pl/.

  3. http://bioinformatics.psb.ugent.be/webtools/plantcare/html/.

  4. http://www.ncbi.nlm.nih.gov/tools/primer-blast/.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant nos. 31872996 and 31472139). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. College of Grassland Science and Technology, China Agricultural University, Bei**g, 100193, People’s Republic of China

    Kun Zhang, Huiting Cui, Shihao Cao, Li Yan, Mingna Li & Yan Sun

  2. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Bei**g, 100193, People’s Republic of China

    Mingna Li

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  1. Kun Zhang
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  4. Li Yan
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Contributions

KZ and YS carried out the experimental design. KZ, HC, SC, LY, and ML performed the experiments and prepared the manuscript and coordinated its revision. ML and YS read and revised the manuscript. All authors provided helpful discussions and approved its final version.

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Correspondence to Mingna Li or Yan Sun.

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299_2019_2461_MOESM1_ESM.tif

Supplementary material S1. Transcription levels of CrCOMT in twelve different overexpression Arabidopsis lines. qRT-PCR analysis of CrCOMT expression in twelve T3 transgenic Arabidopsis. The Atactin7 gene was used as internal controls. OE9 and OE15 performed the high expression level. All the data represent mean ± SE for three biological replicates (TIFF 137 kb)

299_2019_2461_MOESM2_ESM.tif

Supplementary material S2. PCR product in agarose gel electrophoresis of RACE results. (a) The CrCOMT amplified RACE products were showed with agarose gel electrophoresis. (b) The full length of CrCOMT amplified product was showed with agarose gel electrophoresis. Marker:2000bp (TIFF 1528 kb)

299_2019_2461_MOESM3_ESM.tif

Supplementary material S3. Protein hydropathy, isoelectric point and molecular mass prediction of CrCOMT. (a) CrCOMT protein hydropathy was determined by constructing hydropathy plots with the Kyte and Doolittle algorithm (https://web.expasy.org/cgi-bin/protscale/protscale.pl). (b) CrCOMT protein isoelectric point and molecular mass prediction were estimated using the calculate pI/Mw tool (https://web.expasy.org/cgi-bin/protscale/protscale.pl) (TIFF 478 kb)

Supplementary material 4 (XLSX 10 kb)

299_2019_2461_MOESM5_ESM.docx

Supplementary material 5 Table S2. The cis-acting element analysis of COMT promoter in different species. The cis-element analysis of COMT promoters in four sequenced species (Arabidopsis thaliana, Glycine max, Oryza sativa and Zea mays) and C. rigescens (1-1999 from the transcription initiation site) was showed using the Plant CARE website. (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/). (DOCX 17 kb)

Supplementary material 6 (DOCX 19 kb)

Supplementary material 7 (DOCX 14 kb)

Supplementary material 8 (DOCX 17 kb)

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Zhang, K., Cui, H., Cao, S. et al. Overexpression of CrCOMT from Carex rigescens increases salt stress and modulates melatonin synthesis in Arabidopsis thaliana. Plant Cell Rep 38, 1501–1514 (2019). https://doi.org/10.1007/s00299-019-02461-7

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