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Arabidopsis ANAC069 binds to C[A/G]CG[T/G] sequences to negatively regulate salt and osmotic stress tolerance

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ANAC069 binds to the DNA sequence of C[A/G]CG[T/G] to regulate the expression of genes, resulting in decreased ROS scavenging capability and proline biosynthesis, which contribute to increased sensitivity to salt and osmotic stress.

Abstract

NAM-ATAF1/2 and CUC2 (NAC) proteins are plant-specific transcription factors that play important roles in abiotic stress responses. In the present study, we characterized the physiological and regulatory roles of Arabidopsis thaliana ANAC069 in response to abiotic stresses. Arabidopsis plants overexpressing ANAC069 displayed increased sensitivity to abscisic acid, salt, and osmotic stress. Conversely, ANAC069 knockdown plants showed enhanced tolerance to salt and osmotic stress, but no change in ABA sensitivity. Further studies showed that ANAC069 inhibits the expression of SOD, POD, GST, and P5CS genes. Consequently, the transcript level of ANAC069 correlated negatively with the reactive oxygen species (ROS) scavenging ability and the proline level. The genes regulated by ANAC069 were further studied using a gene chip on a genome-wide scale, and 339 and 226 genes up- and downregulated by ANAC069 were identified. Analysis of the promoters of the genes affected by ANAC069 suggested that ANAC069 regulates the expression of genes mainly through interacting with the DNA sequence C[A/G]CG[T/G] in response to abiotic stresses. Collectively, our data suggest that ANAC069 could recognize C[A/G]CG[T/G] sequences to regulate the expression of genes that negatively regulates salt and osmotic stress tolerance by decreasing ROS scavenging capability and proline biosynthesis.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 31270703).

Author contributions

YW conceived and designed the experiments; LH carried out the experiments; LH, XS, HYX, YW, KY and YG analyzed the experimental data. LH and YW wrote the manuscript. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. Key Laboratory of Biogeography and Bioresource in Arid Land, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, 830011, Urumqi, China

    Lin He, Xinxin Shi, Yanmin Wang, Yong Guo & Yucheng Wang

  2. Agricultural College, Heilongjiang Bayi Agricultural University, 163319, Daqing, China

    Lin He & Kejun Yang

  3. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 26 Hexing Road, 150040, Harbin, China

    Xinxin Shi, Yanmin Wang, Yong Guo & Yucheng Wang

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  1. Lin He
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  2. Xinxin Shi
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  3. Yanmin Wang
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  4. Yong Guo
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  5. Kejun Yang
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  6. Yucheng Wang
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Correspondence to Yucheng Wang.

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He, L., Shi, X., Wang, Y. et al. Arabidopsis ANAC069 binds to C[A/G]CG[T/G] sequences to negatively regulate salt and osmotic stress tolerance. Plant Mol Biol 93, 369–387 (2017). https://doi.org/10.1007/s11103-016-0567-3

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