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. 2016 Oct;67(18):5381-5390.
doi: 10.1093/jxb/erw299. Epub 2016 Aug 6.

ABA is required for the accumulation of APX1 and MBF1c during a combination of water deficit and heat stress

Sara I Zandalinas  1 , Damián Balfagón  1 , Vicent Arbona  1 , Aurelio Gómez-Cadenas  1 , Madhuri A Inupakutika  2 , Ron Mittler  3
Affiliations

ABA is required for the accumulation of APX1 and MBF1c during a combination of water deficit and heat stress

Sara I Zandalinas et al. J Exp Bot. 2016 Oct.

Abstract

Abscisic acid (ABA) plays a key role in plant acclimation to abiotic stress. Although recent studies suggested that ABA could also be important for plant acclimation to a combination of abiotic stresses, its role in this response is currently unknown. Here we studied the response of mutants impaired in ABA signalling (abi1-1) and biosynthesis (aba1-1) to a combination of water deficit and heat stress. Both mutants displayed reduced growth, biomass, and survival when subjected to stress combination. Focusing on abi1-1, we found that although its stomata had an impaired response to water deficit, remaining significantly more open than wild type, its stomatal aperture was surprisingly reduced when subjected to the stress combination. Stomatal closure during stress combination in abi1-1 was accompanied by higher levels of H2O2 in leaves, suggesting that H2O2 might play a role in this response. In contrast to the almost wild-type stomatal closure phenotype of abi1-1 during stress combination, the accumulation of ascorbate peroxidase 1 and multiprotein bridging factor 1c proteins, required for acclimation to a combination of water deficit and heat stress, was significantly reduced in abi1-1 Our findings reveal a key function for ABA in regulating the accumulation of essential proteins during a combination of water deficit and heat stress.

Keywords: aba1-1; abi1-1; APX1; MBF1c; abiotic stress; abscisic acid; acclimation; heat stress; stomata; stress combination; water deficit..

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Figures

Fig. 1.
Fig. 1.
Growth, biomass, survival, RWC, and stomatal conductance of wild-type and aba1-1 plants subjected to a combination of water deficit and heat stress. (A) Shoot fresh and dry weight (g; average of five individual rosettes), RWC, rosette diameter, survival, and stomatal conductance (gs) of plants subjected to water deficit (WD), heat stress (HS), and a combination of water deficit and heat stress (WD+HS). * Student’s t-test significant at P < 0.05. Error bars represent SD. (B) Representative images of wild-type and aba1-1 plants subjected to the different stresses. CT, control.
Fig. 2.
Fig. 2.
Growth, biomass, survival, RWC, and stomatal conductance of wild-type and abi1-1 plants subjected to a combination of water deficit and heat stress. (A) Shoot fresh and dry weight (g; average of five individual rosettes), RWC, rosette diameter, survival, and stomatal conductance (gs) of plants subjected to water deficit (WD), heat stress (HS), and a combination of water deficit and heat stress (WD+HS). * Student’s t-test significant at P < 0.05. Error bars represent SD. (B) Representative images of wild-type and abi1-1 plants subjected to the different stresses. CT, control.
Fig. 3.
Fig. 3.
Stomatal aperture of wild-type and abi1-1 plants subjected to a combination of water deficit and heat stress. (A) Representative images of stomata from wild-type and abi1-1 plants subjected to water deficit (WD), heat stress (HS), and a combination of water deficit and heat stress (WD+HS). (B) Measurements of stomatal aperture of wild-type and abi1-1 plants subjected to the different stresses. (C) Stomatal density of wild-type and abi1-1 plants. * Student’s t-test significant at P < 0.05. Error bars represent SD. CT, control.
Fig. 4.
Fig. 4.
Accumulation of ABA, JA, and SA in wild-type and abi1-1 plants subjected to a combination of water deficit and heat stress. ABA (A), SA (B), and JA (C) accumulation in wild-type and abi1-1 plants subjected to water deficit (WD), heat stress (HS), and a combination of water deficit and heat stress (WD+HS). * Student’s t-test significant at P < 0.05. Error bars represent SD. CT, control.
Fig. 5.
Fig. 5.
H2O2 accumulation in wild-type and abi1-1 plants in response to a combination of water deficit and heat stress. (A) H2O2 accumulation in wild-type and abi1-1 plants subjected to water deficit (WD), heat stress (HS), and a combination of water deficit and heat stress (WD+HS). (B) Representative images of stomata of Arabidopsis plants 60min after the application of H2O2 or ABA. (C) Measurements of stomatal aperture of wild-type and abi1-1 plants following application of 1mM H2O2. (D) Measurements of stomatal aperture of wild-type and abi1-1 plants following application of 30 µM ABA. * Student’s t-test significant at P < 0.05. Error bars represent SD. CT, control.
Fig. 6.
Fig. 6.
Accumulation of key acclimation proteins involved in the response of plants to a combination of water deficit and heat stress in wild-type and abi1-1 plants. Protein blot analysis of MBF1c (A), HSP101 (B), and APX1 (C) accumulation in leaves of wild-type and abi1-1 plants subjected to water deficit (WD), heat stress (HS), and a combination of water deficit and heat stress (WD+HS). Top: Quantification bar graphs for fold change in protein accumulation. Bottom: Protein blots and Coomassie-stained gels of total protein. Quantification of protein expression was performed per total protein for three different experiments. * Student’s t-test significant at P < 0.05. Error bars represent SD. CT, control.
Fig. 7.
Fig. 7.
Meta-analysis of transcriptomics data from ABA-treated abi1-1 and wild-type (Ler) plants, and wild-type (Col) plants subjected to a combination of water deficit and heat stress. Top: Venn diagrams showing the overlap between transcripts specifically up-regulated (A) or down-regulated (B) in response to a combination of water deficit and heat stress or to ABA treatment in wild-type plants. Bottom: Venn diagrams showing the overlap between transcripts specifically up-regulated (C) or down-regulated (D) in response to a combination of water deficit and heat stress in wild-type plants and ABA treatment in abi1-1 plants. References used for the meta-analysis are Hoth et al. (2002) and Rizhsky et al. (2004).
Fig. 8.
Fig. 8.
A hypothetical model for the signalling role of JA, SA, ABA, and H2O2 in the regulation of stomatal aperture in abi1-1 during water deficit, heat stress, and a combination of water deficit and heat stress. Dotted lines indicate hypothetical interactions. Solid lines and arrows indicate positive and negative regulation based on published literature, respectively. Abbreviations: ABA, abscisic acid; JA, jasmonic acid; NO, nitric oxide; SA, salicylic acid; SLAC1, SLOW ANION CHANNEL-ASSOCIATED 1.
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