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Arabidopsis thaliana WRKY25, WRKY26, and WRKY33 coordinate induction of plant thermotolerance

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Abstract

Limited information is available regarding the exact function of specific WRKY transcription factors in plant responses to heat stress. We analyzed the roles of WRKY25, WRKY26, and WRKY33, three types of group I WRKY proteins, in the regulation of resistance to heat stress. Expression of WRKY25 and WRKY26 was induced upon treatment with high temperature, whereas WRKY33 expression was repressed. Heat-treated WRKY single mutants exhibited small responses, while wrky25wrky26 and wrky25wrky33 double mutants and the wrky25wrky26wrky33 triple mutants showed substantially increased susceptibility to heat stress, showing reduced germination, decreased survival, and elevated electrolyte leakage, compared with wild-type plants. In contrast, constitutive expression of WRKY25, WRKY26, or WRKY33 enhanced resistance to heat stress. Expression studies of selected heat-defense genes in single, double, and triple mutants, as well as in over-expressing lines, were correlated with their thermotolerance phenotypes and demonstrated that the three WRKY transcription factors modulate transcriptional changes of heat-inducible genes in response to heat treatment. In addition, our findings provided evidence that WRKY25, WRKY26, and WRKY33 were involved in regulation of the heat-induced ethylene-dependent response and demonstrated positive cross-regulation within these three genes. Together, these results indicate that WRKY25, WRKY26, and WRKY33 positively regulate the cooperation between the ethylene-activated and heat shock proteins-related signaling pathways that mediate responses to heat stress; and that these three proteins interact functionally and play overlapping and synergetic roles in plant thermotolerance.

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Abbreviations

ABA:

Abscisic acid

ACC:

1-Aminocyclopropane-1-carboxylic acid

APX:

Cytosolic ascorbate peroxidase

EL:

Electrolyte leakage

HS:

Heat shock

Hsf:

Heat shock transcription factor

Hsp:

Heat shock protein

MBF1c:

Coactivator multiprotein bridging factor 1c

SA:

Salicylic acid

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Acknowledgments

We thank Dr. Zhixiang Chen (Department of Botany and Plant Physiology, Purdue University, West Lafayette, Indiana, USA) for Arabidopsis wrky33-2, wrky25-1wrky26-2wrky33-1, coi1, ein2, and sid2. This work was supported by the Science Foundation of the Ministry of Agriculture of the People’s Republic of China (2009ZX08009-066B) and the Natural Science Foundation of China (Nos. 90817003 and 30871747).

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Authors and Affiliations

  1. Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Kunming, 650223, People’s Republic of China

    Shujia Li, Qiantang Fu, Ligang Chen & Diqiu Yu

  2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, People’s Republic of China

    Shujia Li & Weidong Huang

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  1. Shujia Li
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  2. Qiantang Fu
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  3. Ligang Chen
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  4. Weidong Huang
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Correspondence to Weidong Huang or Diqiu Yu.

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Chinese Academy of Sciences, People’s Republic of China and China Agricultural University, People’s Republic of China contributed equally to this work.

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Fig. S1 Comparison of WRKY25, WRKY26, and WRKY33 sequences (TIFF 2,753 kb)

Fig. S2 Identification of homozygous mutant lines (TIFF 2,125 kb)

Fig. S3 Northern blot analysis of WRKY26 and WRKY33 expression in wild-type and transgenic plants (TIFF 979 kb)

Fig. S4 Characterization of wrky25wrky26wrky33 and wrky25wrky26-2wrky33 triple mutants (TIFF 2,973 kb)

425_2011_1375_MOESM5_ESM.tif

Fig. S5 Expression of heat defense-related genes in 21-day-old wrky33, wild-type, and 35S:W33 plants during heat treatment (TIFF 1,004 kb)

425_2011_1375_MOESM6_ESM.tif

Fig. S6 Expression of heat defense-related genes in 21-day-old wrky26, wild-type, and 35S:W26 plants during heat treatment (TIFF 3,239 kb)

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Li, S., Fu, Q., Chen, L. et al. Arabidopsis thaliana WRKY25, WRKY26, and WRKY33 coordinate induction of plant thermotolerance. Planta 233, 1237–1252 (2011). https://doi.org/10.1007/s00425-011-1375-2

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