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High accumulation of anthocyanins via the ectopic expression of AtDFR confers significant salt stress tolerance in Brassica napus L.

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Abstract

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The ectopic expression of AtDFR results in increased accumulation of anthocyanins leading to enhanced salinity and drought stress tolerance in B. napus plants.

Abstract

Flavonoids with antioxidant effects confer many additional benefits to plants. Evidence indicates that flavonoids, including anthocyanins, protect tissues against oxidative stress from various abiotic stressors. We determined whether increases in anthocyanins increased abiotic stress tolerance in Brassica napus, because the values of B. napus L. and its cultivation area are increasing worldwide. We overexpressed Arabidopsis dihydroflavonol-4-reductase (DFR) in B. napus. Increased DFR transcript levels for AtDFR-OX B. shoots correlated with higher anthocyanin accumulation. AtDFR-OX Brassica shoots exhibited lower reactive oxygen species (ROS) accumulation than wild-type (WT) shoots under high NaCl and mannitol concentrations. This was corroborated by 3,3-diaminobenzidine staining for ROS scavenging activity in 1,1-diphenyl-2-picryl-hydrazyl assays. Shoots of the AtDFR-OX B. napus lines grown in a high salt medium exhibited enhanced salt tolerance and higher chlorophyll content than similarly grown WT plants. Our observations suggested that the AtDFR gene can be effectively manipulated to modulate salinity and drought stress tolerance by directing to high accumulation of anthocyanins in oilseed plants.

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Acknowledgements

This work was supported by a Grant from the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (to H.L., 2016; Grant #2016-116118-3).

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Authors and Affiliations

  1. Department of Biosystems and Biotechnology, College of Life Sciences and Biotechnology, Korea University, Anam-dong 5-ga, Seongbuk-Gu, Seoul, 136-713, Republic of Korea

    Jihye Kim, Won Je Lee, Tien Thanh Vu, Chan Young Jeong & Hojoung Lee

  2. Institute of Life Science and Natural Resources, Korea University, Seoul, 136-713, Republic of Korea

    Hojoung Lee

  3. Department of Molecular Biotechnology, College of Agriculture and Life Sciences, Bioenergy Research Center, Chonnam National University, Gwangju, Republic of Korea

    Suk-Whan Hong

Author notes

  1. Jihye Kim and Won Je Lee are co-first authors.

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    1. Jihye Kim
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    Correspondence to Hojoung Lee.

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    Additional information

    Communicated by Qiao Zhao.

    Electronic supplementary material

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    299_2017_2147_MOESM1_ESM.pdf

    Figure S1 Growth performance of AtDFR-OX Brassica transgenic plants. To examine the growth performance of AtDFR-OX Brassica transgenic plants in normal growth condition, each transgenic shoot was excised and transferred to normal growth medium and allowed to grow for 2 more weeks before picture were taken (PDF 52 kb) (PDF 52 kb)

    299_2017_2147_MOESM2_ESM.pdf

    Figure S2 The analysis of DFR cDNA sequence similarity between Brassica napus L. and Arabidopsis thaliana. DFR cDNA sequence similarity between B. napus L. and A. thaliana (PDF 39 kb) (PDF 39 kb)

    Supplementary material 3 (PDF 65 kb)

    299_2017_2147_MOESM4_ESM.pdf

    Figure S3 Expression of structure genes related to the flavonol pathway in wild-type plants and AtDFR-OX transgenic plants. (A) RNA was extracted from the 2-week-old leaves. The control condition was supplemented with MS liquid media only. BnActin was used as a control, because it is a housekeeping gene. The sequence of the primers is in Supporting Table 1. Transcript levels were measured using RT-PCR. The leaves were supplemented with sucrose 200 mM and sucrose 300 mM for 6 h. (B) Phenylpropanoid metabolic pathway for the synthesis of flavonoids using the amino-acid phenylalanine (PDF 65 kb) (PDF 83 kb)

    Supporting Table 1 Primers used in this experiment (PDF 22 kb) (PDF 22 kb)

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    Kim, J., Lee, W.J., Vu, T.T. et al. High accumulation of anthocyanins via the ectopic expression of AtDFR confers significant salt stress tolerance in Brassica napus L.. Plant Cell Rep 36, 1215–1224 (2017). https://doi.org/10.1007/s00299-017-2147-7

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