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Phosphatidic acid integrates calcium signaling and microtubule dynamics into regulating ABA-induced stomatal closure in Arabidopsis

Planta volume 239, pages 565–575 (2014)Cite this article

Abstract

Specific cellular components have been identified to function in abscisic acid (ABA) regulation of stomatal apertures, including calcium, the cytoskeleton, and phosphatidic acid. In this study, the regulation and dynamic organization of microtubules during ABA-induced stomatal closure by phospholipase D (PLD) and its product PA were investigated. ABA induced microtubule depolymerization and stomatal closure in wide-type (WT) Arabidopsis, whereas these processes were impaired in PLD mutant (pldα1). The microtubule-disrupting drugs oryzalin or propyzamide induced microtubule depolymerization, but did not affect the stomatal aperture, whereas their co-treatment with ABA resulted in stomatal closure in both WT and pldα1. In contrast, the microtubule-stabilizing drug paclitaxel arrested ABA-induced microtubule depolymerization and inhibited ABA-induced stomatal closure in both WT and pldα1. In pldα1, ABA-induced cytoplasmic Ca2+ ([Ca2+]cyt) elevation was partially blocked, and exogenous Ca2+-induced microtubule depolymerization and stomatal closure were impaired. These results suggested that PLDα1 and PA regulate microtubular organization and Ca2+ increases during ABA-induced stomatal closing and that crosstalk among signaling lipid, Ca2+, and microtubules are essential for ABA signaling.

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Abbreviations

ABA:

Abscisic acid

CDPKs:

Calcium-dependent protein kinases

[Ca2+]cyt :

Cytoplasmic Ca2+

Gα:

Heterotrimeric G protein subunit α

IP3:

Inositol-3-phosphate

NO:

Nitric oxide

PA:

Phosphatidic acid

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PS:

Phosphatidylserine

PLD:

Phospholipase D

PPM:

Propyzamide

ROS:

Reactive oxygen species

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Authors and Affiliations

  1. State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    Yan Jiang, Feng Lin, Yana Qu, Xiaoxiang Liu & Qun Zhang

  2. College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    Kai Wu

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  1. Yan Jiang
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  2. Kai Wu
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Correspondence to Qun Zhang.

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425_2013_1999_MOESM1_ESM.pdf

Supplementary Fig. S1 Fluorescence images of propidium iodide/FDA double-stained guard cells. Propidium iodide stains dead cells (red) and FDA stains live cells (green). The guard cells were still alive and stained by propidium iodide/FDA after the treatments as indicated in Fig. 1. The treatments were as follows: treated with 0.1 % (v/v) DMSO (control), 10 μM ABA, 10 μM oryzalin, and 20 μM propyzamide (PPM), respectively. Bar = 10 μm (PDF 140 kb)

425_2013_1999_MOESM2_ESM.pdf

Supplementary Fig. S2 Effects of detergents on stomatal movement and microtubular organization. a Treatment with Triton X-100, SDS or CHAPS for 2 h did not reduce stomatal closure. Triton X-100 (non-ionic detergent), 500 μM; SDS (ionic detergent), 10 mM; CHAPS (zwitterionic detergent), 10 mM. b Microtubules in guard cells depolymerized into diffuse or dot-like fluorescence after detergent treatment for 2 h. Bar = 10 μm. c Quantification of guard cell microtubules after the treatment is shown in b. The data in a and c are the mean ± SE (n = 50-60) from three independent experiments. Values with different letters are significantly different at P < 0.05 based on Duncan’s multiple range test (PDF 170 kb)

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Jiang, Y., Wu, K., Lin, F. et al. Phosphatidic acid integrates calcium signaling and microtubule dynamics into regulating ABA-induced stomatal closure in Arabidopsis . Planta 239, 565–575 (2014). https://doi.org/10.1007/s00425-013-1999-5

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  • DOI: https://doi.org/10.1007/s00425-013-1999-5

Keywords

  • Abscisic acid
  • Calcium
  • Microtubule
  • Phosphatidic acid
  • Phospholipase D