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Advances in the Molecular Mechanisms of Abscisic Acid and Gibberellins Functions in Plants

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Plant Sciences".

Deadline for manuscript submissions: closed (30 September 2021) | Viewed by 45852

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A printed edition of this Special Issue is available here.

Special Issue Editor

Special Issue Information

Dear Colleagues,

Gibberellins (GA) and abscisic acid (ABA) are two phytohormones that regulate in an antagonistic way, plant growth as well as several developmental processes from seed maturation and germination to flowering time, through hypocotyl elongation and root growth. In general, ABA and GA inhibit and promote respectively, cell elongation and growth. Consequently, this mutual antagonism between GA and ABA governs many developmental decisions in plants.

In addition to its role as growth and development modulator, ABA is primarily known for being a major player in the response and adaptation of plants to diverse abiotic stress conditions, including cold, heat, drought, salinity or flooding. Remarkably, different works have also recently pointed to a function for GA in the control of some biological processes in response to stress.

This Special issue will focus on the most recent advances in ABA and GA functions in the regulation of plant growth, development as well as in the response to abiotic stress. The submission of works reporting ABA and GA crosstalk, as well as the integration of GA and ABA action with other plant hormones and/or environmental cues are especially encouraged. Notwithstanding, contributions on other related topics aimed at understanding the molecular mechanisms of ABA and/or GA action in plants are also welcomed, including reviews and original research articles.

Prof. Dr. Víctor Quesada
Guest Editor

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Keywords

  • gibberellic acid (GA)
  • abscisic acid (ABA)
  • seed dormancy
  • germination
  • plant growth
  • plant development
  • flowering time
  • antagonistic interaction
  • signaling pathway
  • abiotic stress

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Published Papers (9 papers)

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Editorial

Jump to: Research, Review

4 pages, 208 KiB  
Editorial
Advances in the Molecular Mechanisms of Abscisic Acid and Gibberellins Functions in Plants
by Víctor Quesada
Int. J. Mol. Sci. 2021, 22(11), 6080; https://doi.org/10.3390/ijms22116080 - 4 Jun 2021
Cited by 3 | Viewed by 2665
Abstract
In this special issue entitled, “Advances in the Molecular Mechanisms of Abscisic Acid and Gibberellins Functions in Plants”, eight articles are collected, with five reviews and three original research papers, which broadly cover different topics on the abscisic acid (ABA) field [...] Read more.
In this special issue entitled, “Advances in the Molecular Mechanisms of Abscisic Acid and Gibberellins Functions in Plants”, eight articles are collected, with five reviews and three original research papers, which broadly cover different topics on the abscisic acid (ABA) field and, to a lesser extent, on gibberellins (GAs) research [...] Full article

Research

Jump to: Editorial, Review

20 pages, 3713 KiB  
Article
The Arabidopsis RLCK VI_A2 Kinase Controls Seedling and Plant Growth in Parallel with Gibberellin
by Ildikó Valkai, Erzsébet Kénesi, Ildikó Domonkos, Ferhan Ayaydin, Danuše Tarkowská, Miroslav Strnad, Anikó Faragó, László Bodai and Attila Fehér
Int. J. Mol. Sci. 2020, 21(19), 7266; https://doi.org/10.3390/ijms21197266 - 1 Oct 2020
Cited by 3 | Viewed by 3909
Abstract
The plant-specific receptor-like cytoplasmic kinases (RLCKs) form a large, poorly characterized family. Members of the RLCK VI_A class of dicots have a unique characteristic: their activity is regulated by Rho-of-plants (ROP) GTPases. The biological function of one of these kinases was investigated using [...] Read more.
The plant-specific receptor-like cytoplasmic kinases (RLCKs) form a large, poorly characterized family. Members of the RLCK VI_A class of dicots have a unique characteristic: their activity is regulated by Rho-of-plants (ROP) GTPases. The biological function of one of these kinases was investigated using a T-DNA insertion mutant and RNA interference. Loss of RLCK VI_A2 function resulted in restricted cell expansion and seedling growth. Although these phenotypes could be rescued by exogenous gibberellin, the mutant did not exhibit lower levels of active gibberellins nor decreased gibberellin sensitivity. Transcriptome analysis confirmed that gibberellin is not the direct target of the kinase; its absence rather affected the metabolism and signalling of other hormones such as auxin. It is hypothesized that gibberellins and the RLCK VI_A2 kinase act in parallel to regulate cell expansion and plant growth. Gene expression studies also indicated that the kinase might have an overlapping role with the transcription factor circuit (PIF4-BZR1-ARF6) controlling skotomorphogenesis-related hypocotyl/cotyledon elongation. Furthermore, the transcriptomic changes revealed that the loss of RLCK VI_A2 function alters cellular processes that are associated with cell membranes, take place at the cell periphery or in the apoplast, and are related to cellular transport and/or cell wall reorganisation. Full article
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Expression of the receptor-like cytoplasmic kinase (<span class="html-italic">RLCK</span>) <span class="html-italic">VI_A2</span> gene in various Arabidopsis lines used in the study. (<b>a</b>) RT-PCR results using <span class="html-italic">RLCK VI_A2</span> (upper row) and <span class="html-italic">GAPC-2</span> specific (lower row) primers in wild type (<b>1</b>), T-DNA insertion line GABI_435H03 (<b>2</b>), T-DNA insertion line GABI_676D12 (<b>3</b>), and the GABI_435H03 line expressing the <span class="html-italic">RLCK VI_A2</span> cDNA transgene under the control of the 35S promoter (complemented mutant) (<b>4</b>). (<b>b</b>) Site of the T-DNA insertion in the third exon of the At2G18890 gene coding for the AtRLCK VI_A2 kinase in the GABI_435H03 line.</p>
Full article ">Figure 2
<p>The <span class="html-italic">rlck vi_a2</span> mutation affects hypocotyl and cotyledon elongation. Hypocotyl (<b>a</b>,<b>d</b>) and cotyledon (<b>b</b>,<b>e</b>) length were measured for 5-days-old short-day (SD; 8/16h light/dark cycle) and 16-days-old dark-grown (cDark; continuous dark) seedlings. WT—wild type; MUT—T-DNA insertion mutant line; CO—complemented mutant line. Three biological replicates were made with 15–25 plants per line. Averages and standard errors are shown. Corresponding representative images are displayed on (<b>c</b>,<b>f</b>). ** <span class="html-italic">p</span> &lt; 0.005 (Student’s <span class="html-italic">t</span>-test; comparison to WT).</p>
Full article ">Figure 3
<p>Greenhouse-grown mutant (MUT) plants exhibited smaller plant size, as evidenced by measuring the rosette diameter. Normal size of the wild type (WT) plants was restored by expressing the kinase cDNA in the mutant background (complemented, CO line). Rosette diameters in mm are shown in (<b>a</b>), and representative images of the measured 4-weeks-old plants in (<b>b</b>). The plants were grown in short day conditions (SD). Averages and standard errors were calculated and are shown on (<b>a</b>). <span class="html-italic">n</span> = 15–25, ** <span class="html-italic">p</span> &lt; 0.005 (Student’s <span class="html-italic">t</span>-test; comparison to WT).</p>
Full article ">Figure 4
<p>The sizes of hypocotyl (<b>a</b>,<b>b</b>) and cotyledon (<b>c</b>,<b>d</b>) cells are significantly smaller in the mutant (MUT) than in the wild type (WT) plants. Fluorescent (<b>a</b>) and scanning electron microscopic (<b>c</b>) images are shown for the epidermal cells of the hypocotyl (<b>a</b>) and the cotyledon (<b>c</b>), respectively, of 5-day-old seedlings. The white bars indicate 100 μm (<b>a</b>), and 10 μm (<b>b</b>), respectively. For the quantitative comparison of cell size (<b>b</b>,<b>d</b>), 150–200 cells were measured for each of three randomly selected seedlings per line. Averages and standard errors are shown on the histograms. * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.005 (Student’s <span class="html-italic">t</span>-test; comparison to WT).</p>
Full article ">Figure 5
<p>Exogenous gibberellin treatments complemented the mutant phenotypes. Wild type (WT) or <span class="html-italic">rlck vi_a2</span> mutant (MUT) seedlings grown in vitro in short days (SD; 8 h/16 h light/dark) for 6 days (<b>a</b>) or in continuous dark (cDark) for 17 days (<b>b</b>,<b>c</b>) and plants grown at short days in greenhouse (<b>d</b>) were or were not treated with 20 μM gibberellic acid (GA<sub>3</sub>). Hypocotyl length (<b>a</b>,<b>b</b>), cotyledon length (<b>c</b>) or rosette diameter (<b>d</b>) were measured in 15–25 seedlings or plants, respectively, in three repetitions. Averages and standard errors are shown. ** <span class="html-italic">p</span> &lt; 0.05 (Student’s <span class="html-italic">t</span>-test; comparison to WT).</p>
Full article ">Figure 6
<p>Gibberellin content and gibberellin sensitivity of the mutant and the wild type. (<b>a</b>) Endogenous content of active gibberellins was measured in 9-days-old seedlings of wild type (WT), <span class="html-italic">rlck vi_a2</span> mutant (MUT), and complemented mutant (CO) lines. Seedlings were grown in vitro under short day (SD; 8 h/16 h light/dark) conditions in a growth chamber. Samples were collected from three independent experiments. Averaged data are shown with the standard deviations. No statistically significant differences could be observed among the tested Arabidopsis lines (<span class="html-italic">p</span> &lt; 0.05 Student’s <span class="html-italic">t</span>-test; comparison to WT). (<b>b</b>) Relative hypocotyl length was determined in response to a range of GA<sub>3</sub> concentrations (0–100 μM), in the case of wild type and mutant seedlings (9-days-old; grown under low intensity continuous white light).</p>
Full article ">Figure 7
<p>Overlaps of the DEGs of the <span class="html-italic">rlck vi_a2</span> mutant with the DEGs of the <span class="html-italic">ga1-3</span> gibberellin synthesis mutant [<a href="#B28-ijms-21-07266" class="html-bibr">28</a>,<a href="#B29-ijms-21-07266" class="html-bibr">29</a>] (<b>a</b>) and with those genes for which the promoters are direct targets of the cell/hypocotyl elongation regulatory transcription factors PIF4, BZR1, and/or ARF6 [<a href="#B31-ijms-21-07266" class="html-bibr">31</a>] (<b>b</b>).</p>
Full article ">
22 pages, 3231 KiB  
Article
The Role of ABA in Plant Immunity is Mediated through the PYR1 Receptor
by Javier García-Andrade, Beatriz González, Miguel Gonzalez-Guzman, Pedro L. Rodriguez and Pablo Vera
Int. J. Mol. Sci. 2020, 21(16), 5852; https://doi.org/10.3390/ijms21165852 - 14 Aug 2020
Cited by 48 | Viewed by 5361
Abstract
ABA is involved in plant responses to a broad range of pathogens and exhibits complex antagonistic and synergistic relationships with salicylic acid (SA) and ethylene (ET) signaling pathways, respectively. However, the specific receptor of ABA that triggers the positive and negative responses of [...] Read more.
ABA is involved in plant responses to a broad range of pathogens and exhibits complex antagonistic and synergistic relationships with salicylic acid (SA) and ethylene (ET) signaling pathways, respectively. However, the specific receptor of ABA that triggers the positive and negative responses of ABA during immune responses remains unknown. Through a reverse genetic analysis, we identified that PYR1, a member of the family of PYR/PYL/RCAR ABA receptors, is transcriptionally upregulated and specifically perceives ABA during biotic stress, initiating downstream signaling mediated by ABA-activated SnRK2 protein kinases. This exerts a damping effect on SA-mediated signaling, required for resistance to biotrophic pathogens, and simultaneously a positive control over the resistance to necrotrophic pathogens controlled by ET. We demonstrated that PYR1-mediated signaling exerted control on a priori established hormonal cross-talk between SA and ET, thereby redirecting defense outputs. Defects in ABA/PYR1 signaling activated SA biosynthesis and sensitized plants for immune priming by poising SA-responsive genes for enhanced expression. As a trade-off effect, pyr1-mediated activation of the SA pathway blunted ET perception, which is pivotal for the activation of resistance towards fungal necrotrophs. The specific perception of ABA by PYR1 represented a regulatory node, modulating different outcomes in disease resistance. Full article
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Figure 1

Figure 1
<p>Participation of SnRK2s kinases in the response of Arabidopsis plants to infection by the fungal pathogen <span class="html-italic">P. cucumerina</span>. (<b>A</b>) ABA accumulation determined in mock and <span class="html-italic">P</span>. <span class="html-italic">cucumerina</span>-infected Col-0 plants. (<b>B</b>) RT-qPCR of <span class="html-italic">ABI4</span> in mock and in <span class="html-italic">P. cucumerina</span>-infected Col-0. (<b>C</b>,<b>D</b>) <span class="html-italic">P. cucumerina</span>-mediated activation of SnRK2.6 (<b>C</b>) and SnRK2.2 (<b>D</b>). Transgenic Arabidopsis plants expressing HA-tagged versions of the kinases were inoculated with <span class="html-italic">P. cucumerina</span>, or were mocked, and leaf samples were taken at 0, 24, and 48 h.p.i., and the protein extracts were immunoprecipitated with anti-HA antibodies. The immunoprecipitates were incubated with a His-ABF2 fragment (Gly73 to Gln 119; ΔABF2) in the presence of [γ-<sup>32</sup>P]ATP, and the proteins were resolved by SDS-PAGE. Bands corresponding to ΔABF2 fragments and to SnRK2.6 and SnRK2.2 kinases are indicated. Radioactivities of ΔABF2 fragment bands were measured with a phosphoimager, and the values were plotted on the graphs shown at the right of the figures. Error bars indicate S.E.M.; <span class="html-italic">n</span> = 3. (<b>E</b>) Disease resistance towards <span class="html-italic">P. cucumerina</span> of transgenic plants overexpressing SnRK2.6, SnRK2.2, and SnRK2.3 in comparison to Col-0. (<b>F</b>) Disease resistance towards <span class="html-italic">P. cucumerina</span> in the double <span class="html-italic">snrk2.2 snrk2.3</span> mutant and in <span class="html-italic">snrk2.6</span> mutant plants. (<b>G</b>) Representative leaves from each genotype at 12 days following inoculation with <span class="html-italic">P. cucumerina</span>. (<b>H</b>) Disease resistance towards <span class="html-italic">P. cucumerina</span> in the triple PP2C mutants <span class="html-italic">pp2ca1 1hab1 1abi1-2</span> and <span class="html-italic">abi2-2 hab1 abi1-2</span>. For the bioassays with <span class="html-italic">P. cucumerina</span>, lesion diameter of 25 plants per genotype and four leaves per plant were determined 12 d following inoculation with <span class="html-italic">P. cucumerina</span>. Data points represent the average lesion size ± SE of measurements. An ANOVA was conducted to assess significant differences in the activation of SnRKs, ABA accumulation, ABI4 transcript accumulation, and disease symptoms, with a priori <span class="html-italic">p</span> &lt; 0.05 level of significance; the asterisks * above the bars indicate statistically significant differences regarding mock treatments or Col-0 plants. Asterisks above the bars indicate different homogeneous groups with statistically significant differences.</p>
Full article ">Figure 2
<p>PYR1 is required for disease resistance towards <span class="html-italic">P. cucumerina</span>. (<b>A</b>). Disease resistance towards <span class="html-italic">P. cucumerina</span> in Col-0, the resistant <span class="html-italic">ocp3-1 mutant</span>, the susceptible <span class="html-italic">aba2-1</span>, and the triple and quadruple multi-locus mutants <span class="html-italic">pyl4 pyl5 pyl8</span>, <span class="html-italic">pyr1 pyl4 pyl8</span>, <span class="html-italic">pyr1 pyl4 pyl5</span>, and <span class="html-italic">pyr1 pyl1 pyl2 pyl4</span>. (<b>B</b>) Disease resistance in single <span class="html-italic">pyl1</span>, <span class="html-italic">pyr1</span>, and <span class="html-italic">pyl4</span> mutants, in a transgenic line overexpressing PYR1 (PYR1-OE), and in Col-0. Below the graph, the representative leaves from each genotype are shown at 12 days following inoculation with <span class="html-italic">P. cucumerina</span>. (<b>C</b>) Comparative disease resistance towards <span class="html-italic">P. cucumerina</span> among the allelic <span class="html-italic">pyr1-1</span>, <span class="html-italic">pyr1-2</span>, and <span class="html-italic">pyr1-8</span> mutants. Data points represent the average lesion size ± SE of measurements. An ANOVA was conducted to assess significant differences in disease symptoms (<span class="html-italic">p</span> &lt; 0.05); the letters above the bars indicate different homogeneous groups with statistically significant differences.</p>
Full article ">Figure 3
<p>Local activation of <span class="html-italic">PYR1</span> gene expression at pathogen inoculation sites, and the requirement of PYR1 for pathogen-induced callose deposition. (<b>A</b>) Comparative histochemical analysis of GUS activity in rosette leaves from transgenic plants carrying <span class="html-italic">pPYR1::GUS</span>, <span class="html-italic">pPYL1::GUS</span>, and <span class="html-italic">pPYL4::GUS</span> gene constructs and those were either mocked or inoculated <span class="html-italic">P. cucumerina</span>. Leaves were stained for GUS activity at 36 h.p.i. The left panel corresponds to mocked plants. The central and right panels correspond to enlargements of the inoculated leaf sectors. Black arrow points towards leaf tissues proximal to the inoculation point, and white arrows denote tissues that directly received the spore inoculum. Note that <span class="html-italic">pPYR1::GUS</span> is heavily induced in leaf veins within the inoculated sector. (<b>B</b>) Characteristic spore-inoculated leaf sector, similar to those shown in A, stained with aniline blue to detect pathogen-induced callose deposition (top panel), or with trypan blue (lower panel) to identify incipient cell deterioration due to fungal infection at 36 h.p.i. (<b>C</b>) Aniline blue staining and epifluorescence microscopy were applied to visualize callose accumulation. Micrographs indicate <span class="html-italic">P. cucumerina</span> inoculation and infection site in the different Arabidopsis genotypes at 0 h.p.i (right panel), at 24 h.p.i. (central panel), and at 48 h.p.i. (right panel). (<b>D</b>) The number of yellows pixels (corresponding to pathogen-induced callose) per million on digital photographs of infected leaves were used as a means to express arbitrary units (i.e., to quantify the image) at the indicated times. Bars represent mean ± SD, <span class="html-italic">n</span> = 15 independent replicates. An ANOVA was conducted to assess significant differences in callose deposition (<span class="html-italic">p</span> &lt; 0.05); the asterisks * above the bars indicate statistically significant differences regarding Col-0 plants.</p>
Full article ">Figure 4
<p>Response of <span class="html-italic">pyr1</span> plants to infection by <span class="html-italic">P. syringae DC3000</span>. (<b>A</b>) Col-0, <span class="html-italic">aba2-1</span>, <span class="html-italic">pyr1-2</span>, <span class="html-italic">pyl1</span>, and <span class="html-italic">pyl4</span> mutants were inoculated with <span class="html-italic">P. syringae</span> DC3000, and their disease responses were recorded. (<b>B</b>) Col-0 and <span class="html-italic">pyr1</span> plants were pre-treated with 150 μM ABA, applied by drenching, before inoculation with <span class="html-italic">P. syringae</span> DC3000, and the growth of the bacteria was recorded in comparison to mocked plants. Growth of <span class="html-italic">P. syringae</span> DC3000 was measured at 3 d.p.i. Error bars represent standard deviation (<span class="html-italic">n</span> = 12). An ANOVA was conducted to assess significant differences in disease symptoms, with a priori <span class="html-italic">p</span> &lt; 0.05 level of significance; the asterisks *, ** above the bars indicate different homogeneous groups with statistically significant differences.</p>
Full article ">Figure 5
<p>Expression of SA-responsive and ET-responsive genes in <span class="html-italic">pyr1</span> and <span class="html-italic">aba2</span> mutants. (<b>A</b>,<b>B</b>) RT-qPCR analysis showing constitutive expression levels of <span class="html-italic">PR-1</span>, <span class="html-italic">PR-2</span>, <span class="html-italic">PR-4</span>, and <span class="html-italic">PR-5</span> genes in (<b>A</b>) Col-0, <span class="html-italic">aba2-1</span>, <span class="html-italic">pyr1-1</span>, <span class="html-italic">pyl1</span>, and <span class="html-italic">pyl4</span> plants, and (<b>B</b>) their comparative expression levels in the allelic <span class="html-italic">pyr1-1</span>, <span class="html-italic">pyr1-2</span>, and <span class="html-italic">pyr1-8</span> mutants. Data represent mean ± SD; <span class="html-italic">n</span> = 3 replicates. The expression was normalized to the constitutive <span class="html-italic">ACT2</span> and <span class="html-italic">ACT8</span> genes and then to the expression in Col-0 plants. (<b>C</b>–<b>E</b>) Accumulation of free SA, total SA, and total JA in Col-0 and <span class="html-italic">pyr1-2</span> plants. Data represent the average of three biological replicates. An ANOVA was conducted to assess significant differences in RT-qPCR and hormone analysis, with a priori <span class="html-italic">p</span> &lt; 0.05 level of significance; the asterisks * above the bars indicate statistically significant differences regarding Col-0 plants.</p>
Full article ">Figure 6
<p>Loss of PYR1 function confers enhanced mitogen-activated kinase activation and PR-1 protein accumulation following <span class="html-italic">P. syringae</span> DC3000 infection. Western blot with anti-pTEpY and anti-PR-1 antibodies of crude protein extracts derived from Col-0, <span class="html-italic">pyr1-2</span> plants at 0, 24, and 48 h.p.i with <span class="html-italic">P. syringae</span> DC3000. Equal protein loading was check by Ponceau-S staining of the nitrocellulose filter. MPK6 and MPK3 migrating bands are indicated on the right. The experiments were repeated three times with similar results. Scan quantification of protein bands corresponding to MPK3 and PR-1 is shown below the Western blot. Data represent the mean ± SD; <span class="html-italic">n</span> = 3 replicates. An ANOVA was conducted to assess significant differences in RT-qPCR analysis, with a priori <span class="html-italic">p</span> &lt; 0.05 level of significance; the asterisks * above the bars indicate statistically significant differences regarding Col-0 plants.</p>
Full article ">Figure 7
<p>Loss of PYR1 function provokes the setting of hallmarks characteristic of primed immunity. (<b>A</b>) Comparative RT-qPCR of <span class="html-italic">SBT3.3</span>, <span class="html-italic">NRPD2</span>, and <span class="html-italic">NRPE1</span> transcript levels between healthy Col-0 and <span class="html-italic">pyr1-2</span> plants. The expression was normalized to the constitutive <span class="html-italic">ACT2/8</span> gene and then to the expression in Col-0 plants. (<b>B</b>) Chromatin immunoprecipitation (ChIP) and comparison between Col-0 and <span class="html-italic">pyr1-2</span> plants of the level of histone H3 Lys4 trimethylation (H3K4me3) and histone H3 Lys9 acetylation (H3K9ac) on the <span class="html-italic">SBT3.3</span>, <span class="html-italic">PR-1</span>, <span class="html-italic">WRKY6</span>, and <span class="html-italic">WRKY53</span> gene promoters as present in leaf samples. The setting of histone marks in <span class="html-italic">ACTIN2</span> was used as an internal control. Data are standardized for Col-0 histone modification levels. Data represent the mean ± SD; <span class="html-italic">n</span> = 3 biological replicates. An ANOVA was conducted to assess significant differences between MPKs activation and PR1 accumulation (<span class="html-italic">p</span> &lt; 0.05); the asterisks * above the bars indicate statistically significant differences regarding Col-0 plants.</p>
Full article ">Figure 8
<p>Effect of <span class="html-italic">NahG</span> on disease resistance and insensitivity to ACC of <span class="html-italic">pyr1</span> plants and seedlings. (<b>A</b>,<b>B</b>) Comparative disease resistance towards <span class="html-italic">P.s.</span> DC3000 and <span class="html-italic">P. cucumerina</span> among Col-0, <span class="html-italic">pyr1-2</span>, <span class="html-italic">NahG</span>, and <span class="html-italic">pyr1-2NahG</span> plants. Growth of <span class="html-italic">P. syringae</span> DC3000 was measured at 3 d.p.i. Error bars represent standard deviation (<span class="html-italic">n</span> = 12). For <span class="html-italic">P. cucumerina</span>, data points represent the average lesion size ± SE of measurements. An ANOVA was conducted to assess significant differences in disease symptoms (<span class="html-italic">p</span> &lt; 0.05); the letters above the bars indicate different homogeneous groups with statistically significant differences. (<b>C</b>) Apical hook region of the indicated seedlings germinated and grown on MS/2 in the dark for 4 d in the presence of the indicated concentration of ACC. (<b>D</b>) Hypocotyl length of seedlings germinated and grown in the dark for 4 d on MS/2 medium supplemented with the denoted concentrations of ACC. Error bars represent standard deviation (<span class="html-italic">n</span> = 50). An ANOVA was conducted, and no significant differences were observed in hypocotyl length (<span class="html-italic">p</span> &lt; 0.05). (<b>E</b>) Apical hook region of Col-0 seedlings germinated and grown on MS/2 in the dark for 4 d in the presence of the indicated concentration of ACC and SA.</p>
Full article ">
15 pages, 1366 KiB  
Article
Kaolin Reduces ABA Biosynthesis through the Inhibition of Neoxanthin Synthesis in Grapevines under Water Deficit
by Tommaso Frioni, Sergio Tombesi, Paolo Sabbatini, Cecilia Squeri, Nieves Lavado Rodas, Alberto Palliotti and Stefano Poni
Int. J. Mol. Sci. 2020, 21(14), 4950; https://doi.org/10.3390/ijms21144950 - 13 Jul 2020
Cited by 23 | Viewed by 3683
Abstract
In many viticulture regions, multiple summer stresses are occurring with increased frequency and severity because of warming trends. Kaolin-based particle film technology is a technique that can mitigate the negative effects of intense and/or prolonged drought on grapevine physiology. Although a primary mechanism [...] Read more.
In many viticulture regions, multiple summer stresses are occurring with increased frequency and severity because of warming trends. Kaolin-based particle film technology is a technique that can mitigate the negative effects of intense and/or prolonged drought on grapevine physiology. Although a primary mechanism of action of kaolin is the increase of radiation reflection, some indirect effects are the protection of canopy functionality and faster stress recovery by abscisic acid (ABA) regulation. The physiological mechanism underlying the kaolin regulation of canopy functionality under water deficit is still poorly understood. In a dry-down experiment carried out on grapevines, at the peak of stress and when control vines zeroed whole-canopy net CO2 exchange rates/leaf area (NCER/LA), kaolin-treated vines maintained positive NCER/LA (~2 µmol m−2 s−1) and canopy transpiration (E) (0.57 µmol m−2 s−1). Kaolin-coated leaves had a higher violaxanthin (Vx) + antheraxanthin (Ax) + zeaxanthin (Zx) pool and a significantly lower neoxanthin (Nx) content (VAZ) when water deficit became severe. At the peak of water shortage, leaf ABA suddenly increased by 4-fold in control vines, whereas in kaolin-coated leaves the variation of ABA content was limited. Overall, kaolin prevented the biosynthesis of ABA by avoiding the deviation of the VAZ epoxidation/de-epoxidation cycle into the ABA precursor (i.e., Nx) biosynthetic direction. The preservation of the active VAZ cycle and transpiration led to an improved dissipation of exceeding electrons, explaining the higher resilience of canopy functionality expressed by canopies sprayed by kaolin. These results point out the interaction of kaolin with the regulation of the VAZ cycle and the active mechanism of stomatal conductance regulation. Full article
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<p>(<b>A</b>)Trends for air (T<sub>amb</sub>) and leaf (T<sub>leaf</sub>) temperature; (<b>B</b>)midday stem water potential (Ψ<sub>MD</sub>); (<b>C</b>) whole-canopy transpiration (E/LA) and (<b>D</b>) specific whole-canopy net CO<sub>2</sub> exchange rate/leaf area (NCER/LA), according to a progressive water shortage (DOY 209–217) and subsequent re-watering (at DOY 218), in vines subjected to the kaolin treatment and in controls. Bars represents standard error (SE), <span class="html-italic">n</span> = 3. Asterisks indicate dates within which differences among treatment were significant (<span class="html-italic">p</span> &lt; 0.05). DOY: day of the year.</p>
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<p>Correlation between whole-canopy transpiration rate/leaf area (E/LA) and midday stem water potential (Ψ<sub>MD</sub>).</p>
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<p>(<b>A</b>) Trends for leaf violaxanthin (Vx) + antheraxanthin (Ax) + zeaxanthin (Zx) content, (<b>B</b>) de-epoxidation state, (<b>C</b>) neoxanthin (Nx) content and (<b>D</b>) abscisic acid (ABA) concentration, according to a progressive water shortage (DOY 209–217) and subsequent re-watering (at DOY 218), in vines subjected to the kaolin treatment and in controls. Bars represents standard error, <span class="html-italic">n</span> = 3. Asterisks indicate dates within which differences among treatment were significant (<span class="html-italic">p</span> &lt; 0.05). DOY: day of the year.</p>
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<p>(<b>A</b>) Course of leaf violaxanthin (Vx) content at dawn; (<b>B</b>) midday to dawn Vx differences and (<b>C</b>) zeaxanthin (Zx) content at midday over the experiment, according to a progressive water shortage (DOY 209–217) and subsequent re-watering (at DOY 218), in vines subjected to the kaolin treatment and in controls. Bars represents standard error, <span class="html-italic">n</span> = 3. Asterisks indicate dates within which differences among treatment were significant (<span class="html-italic">p</span> &lt; 0.05). DOY: day of the year.</p>
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<p>Correlation between whole-canopy transpiration rate/leaf area (E/LA) and leaf abscisic acid (ABA) content.</p>
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<p>Correlation between leaf abscisic acid (ABA) content and leaf temperature (T<sub>leaf</sub>).</p>
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Review

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10 pages, 849 KiB  
Review
Interplay between Abscisic Acid and Gibberellins, as Related to Ethylene and Sugars, in Regulating Maturation of Non-Climacteric Fruit
by Fernando Alferez, Deived Uilian de Carvalho and Daniel Boakye
Int. J. Mol. Sci. 2021, 22(2), 669; https://doi.org/10.3390/ijms22020669 - 12 Jan 2021
Cited by 38 | Viewed by 5005
Abstract
In this review, we address the interaction between abscisic acid (ABA) and gibberellins (GAs) in regulating non-climacteric fruit development and maturation at the molecular level. We review the interplay of both plant growth regulators in regulating these processes in several fruit of economic [...] Read more.
In this review, we address the interaction between abscisic acid (ABA) and gibberellins (GAs) in regulating non-climacteric fruit development and maturation at the molecular level. We review the interplay of both plant growth regulators in regulating these processes in several fruit of economic importance such as grape berries, strawberry, and citrus, and show how understanding this interaction has resulted in useful agronomic management techniques. We then relate the interplay of both hormones with ethylene and other endogenous factors, such as sugar signaling. We finally review the growing knowledge related to abscisic acid, gibberellins, and the genus Citrus. We illustrate why this woody genus can be considered as an emerging model plant for understanding hormonal circuits in regulating different processes, as most of the finest work on this matter in recent years has been performed by using different Citrus species. Full article
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<p>General scheme of pathways leading to production of different hormones. ABA and GAs share their biosynthetic pathway with other hormones, such as CKs. During fruit maturation, balance among hormone biosynthesis changes, and this involves carotenogenesis. IDP, isopentenyl diphosphate; GDP, geranyl diphosphate; FDP, farnesyl diphosphate; GGDP, geranyilgeranyl diphosphate . Number of arrows illustrate number of biosynthetic steps.</p>
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<p>A time-course model of the interplay between abscisic acid (ABA) and gibberellins (GAs) in modulating non-climacteric fruit development and maturation. This is a reductionistic model, as other players involved are not depicted. These include nutritional and environmental factors. The shape of the elements in the figure illustrates the evolution of each component during fruit maturation. GAs decrease, ABA increase and ethylene production remains steady, whereas ethylene perception increases. The crosstalk with ethylene (dash line pointing an induction in ethylene perception driven by ABA) remains to be demonstrated.</p>
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14 pages, 750 KiB  
Review
Abscisic Acid and Flowering Regulation: Many Targets, Different Places
by Damiano Martignago, Beata Siemiatkowska, Alessandra Lombardi and Lucio Conti
Int. J. Mol. Sci. 2020, 21(24), 9700; https://doi.org/10.3390/ijms21249700 - 18 Dec 2020
Cited by 44 | Viewed by 6074
Abstract
Plants can react to drought stress by anticipating flowering, an adaptive strategy for plant survival in dry climates known as drought escape (DE). In Arabidopsis, the study of DE brought to surface the involvement of abscisic acid (ABA) in controlling the floral transition. [...] Read more.
Plants can react to drought stress by anticipating flowering, an adaptive strategy for plant survival in dry climates known as drought escape (DE). In Arabidopsis, the study of DE brought to surface the involvement of abscisic acid (ABA) in controlling the floral transition. A central question concerns how and in what spatial context can ABA signals affect the floral network. In the leaf, ABA signaling affects flowering genes responsible for the production of the main florigen FLOWERING LOCUS T (FT). At the shoot apex, FD and FD-like transcription factors interact with FT and FT-like proteins to regulate ABA responses. This knowledge will help separate general and specific roles of ABA signaling with potential benefits to both biology and agriculture. Full article
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<p>Abscisic acid (ABA) signaling and flowering regulation. In the leaves (left), ABA controls FLOWERING LOCUS T (FT) transcription acting on GIGANTEA (GI) and CONSTANS (CO); ABA-responsive transcription factors (ABFs) can modulate SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) expression, in turn affecting FT transcription through an indirect mechanism. FT moves to the SAM where it interacts with FD and FD-like basic leucine zippers (bZIPs) to activate floral genes and ABA signaling transcriptome. ABA is transported in the phloem, but its roles at the SAM are not yet known. TERMINAL FLOWER 1 (TFL1) antagonizes FT, repressing transcription. Dashed lines represent indirect or not yet confirmed pathways, while full lines represent known ones.</p>
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18 pages, 4112 KiB  
Review
Transcriptional Regulation of Protein Phosphatase 2C Genes to Modulate Abscisic Acid Signaling
by Choonkyun Jung, Nguyen Hoai Nguyen and Jong-Joo Cheong
Int. J. Mol. Sci. 2020, 21(24), 9517; https://doi.org/10.3390/ijms21249517 - 14 Dec 2020
Cited by 56 | Viewed by 6359
Abstract
The plant hormone abscisic acid (ABA) triggers cellular tolerance responses to osmotic stress caused by drought and salinity. ABA controls the turgor pressure of guard cells in the plant epidermis, leading to stomatal closure to minimize water loss. However, stomatal apertures open to [...] Read more.
The plant hormone abscisic acid (ABA) triggers cellular tolerance responses to osmotic stress caused by drought and salinity. ABA controls the turgor pressure of guard cells in the plant epidermis, leading to stomatal closure to minimize water loss. However, stomatal apertures open to uptake CO2 for photosynthesis even under stress conditions. ABA modulates its signaling pathway via negative feedback regulation to maintain plant homeostasis. In the nuclei of guard cells, the clade A type 2C protein phosphatases (PP2Cs) counteract SnRK2 kinases by physical interaction, and thereby inhibit activation of the transcription factors that mediate ABA-responsive gene expression. Under osmotic stress conditions, PP2Cs bind to soluble ABA receptors to capture ABA and release active SnRK2s. Thus, PP2Cs function as a switch at the center of the ABA signaling network. ABA induces the expression of genes encoding repressors or activators of PP2C gene transcription. These regulators mediate the conversion of PP2C chromatins from a repressive to an active state for gene transcription. The stress-induced chromatin remodeling states of ABA-responsive genes could be memorized and transmitted to plant progeny; i.e., transgenerational epigenetic inheritance. This review focuses on the mechanism by which PP2C gene transcription modulates ABA signaling. Full article
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Graphical abstract

Graphical abstract
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<p>Abscisic acid (ABA) signaling pathway in the nuclei of guard cells. (<b>A</b>) Repression of ABA-responsive gene expression. In the absence, the clade A protein phosphatases (PP2Cs) physically interact with the sucrose non-fermenting 1-related protein kinases (SnRK2s) to reduce kinase activity via dephosphorylation. This inhibits the activity of ABRE-binding (AREB)/ABRE-binding factor (ABF) transcription factors and suppression of ABA-responsive gene transcription. (<b>B</b>) Activation of ABA-responsive gene expression. Under osmotic stress conditions, the interaction with ABA leads to conformational changes in the ABA receptors [PYR (pyrabactin resistance)/PYL (PYR-related)/RCAR (regulatory component of the ABA receptor)], allowing them to interact with PP2Cs. PP2Cs act as a coreceptor to capture ABA, thereby suppressing its phosphatase activity. This sequestrates PP2Cs from SnRK2s, and free SnRK2s phosphorylate the downstream transcription factors AREB/ABFs. The phosphorylated AREB/ABFs trigger the transcription of numerous ABA-responsive genes.</p>
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<p>A working model of chromatin remodeling for regulation of PP2C gene transcription. (<b>A</b>) Repression of PP2C gene transcription. Under normal conditions, enhancer of ABA coreceptor (EAR) motif-containing MYB repressors (AtMYB44 and AtMYB20) interact with a TOPLESS-related corepressor (TPR), which recruits histone deacetylase (HDA) to suppress PP2C gene transcription. The chromatin remodeler, BRM-containing SWI/SNF complex, occupies the promoter and contributes to the repression of PP2C gene transcription. (<b>B</b>) Activation of PP2C gene transcription. Under osmotic stress conditions, the repressor is released from the promoter, and histone acetyltransferases (HATs) that acetylate the histones and relax DNA–histone binding in chromatin. Activator (AREB/ABFs) binds to the open promoter region, and RNA polymerase II (RNAPII) accesses and starts gene transcription.</p>
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16 pages, 2138 KiB  
Review
Abscisic Acid Mediates Drought and Salt Stress Responses in Vitis vinifera—A Review
by Daniel Marusig and Sergio Tombesi
Int. J. Mol. Sci. 2020, 21(22), 8648; https://doi.org/10.3390/ijms21228648 - 17 Nov 2020
Cited by 50 | Viewed by 5230
Abstract
The foreseen increase in evaporative demand and reduction in rainfall occurrence are expected to stress the abiotic constrains of drought and salt concentration in soil. The intensification of abiotic stresses coupled with the progressive depletion in water pools is a major concern especially [...] Read more.
The foreseen increase in evaporative demand and reduction in rainfall occurrence are expected to stress the abiotic constrains of drought and salt concentration in soil. The intensification of abiotic stresses coupled with the progressive depletion in water pools is a major concern especially in viticulture, as most vineyards rely on water provided by rainfall. Because its economical relevance and its use as a model species for the study of abiotic stress effect on perennial plants, a significant amount of literature has focused on Vitis vinifera, assessing the physiological mechanisms occurring under stress. Despite the complexity of the stress-resistance strategy of grapevine, the ensemble of phenomena involved seems to be regulated by the key hormone abscisic acid (ABA). This review aims at summarizing our knowledge on the role of ABA in mediating mechanisms whereby grapevine copes with abiotic stresses and to highlight aspects that deserve more attention in future research. Full article
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<p>Abscisic acid biosynthesis and metabolism. In the first step, β-carotene is di-hydroxylated by β-carotene hydroxilase (BCH) proteins to produce the transisomer zeaxanthin. Hence, zeaxanthin is epoxidated by zeaxanthin oxidase (ZEP) to antheraxanthin and, then, to violaxanthin. ZEP-mediated reactions can be reversed by violaxanthin de-epoxidase (VDE). Violaxanthin can be transformed into neoxanthin by neoxanthin synthase (NSY) and both violaxanthin and neoxanthin are converted in the respective 9-cis-isomer by isomerase catalysts. The 15-carbons apocarotenoid sesquiterpenoid xanthoxin is then produced by cis-xanthophylls cleavage, whose reaction is catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED). Subsequently, xanthoxin is oxidized to abscisic aldehyde by short-chain alcohol dehydrogenase (SCAD), and finally abscisic acid (ABA) is produced by oxidation of abscisic aldehyde through the combined action of ABA-aldehyde oxidase (AAO) and a molybdenum cofactor sulfurase (MOCOSU). ABA can be inactivated by oxidation or by conjugation with monosaccharides. In the first way, ABA is oxidized (Ox) at first to phaseic acid and then to 4′-dihydrophaseic acid. In the second one, ABA is conjugated with glucose to produce ABA-β-D-glucose ester. Based on [<a href="#B9-ijms-21-08648" class="html-bibr">9</a>].</p>
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<p>Abscisic acid (ABA) signaling in guard cells. (<b>a</b>) ABA inducing stomatal closure. ABA binds to PYR/PYL/RCAR receptors on the guard cells membrane and triggers the accumulation in cytosol of Ca<sup>2+</sup> by activation of Ca<sup>2+</sup> channels (I<sub>Ca</sub>). Under elevated Ca<sup>2+</sup> concentration ([Ca<sup>2+</sup>]), the cell-efflux of Cl<sup>−</sup> is enhanced. This efflux is mediated by rapid transient (R-type) and slow-activating sustained (S-type) Cl<sup>−</sup> channels, and it causes plasma membrane depolarization. Thence, the K<sup>+</sup> uptake is downregulated by inward-rectifying K<sup>+</sup> channels (I<sub>k-in</sub>) activity, while the K<sup>+</sup> efflux is promoted through outward-rectifying K<sup>+</sup> channels (I<sub>k-out</sub>). The overall reduction in ions content triggers water efflux through aquaporins by osmosis, causing loss of turgor in guard cells and stomatal closure. (<b>b</b>) Stomatal reopening by ABA inactivation. As ABA does not bind further to PYR/PYL/RCAR receptors, Ca<sup>2+</sup> accumulation ceases. The osmotic balance is restored by K<sup>+</sup> uptake through I<sub>k-in</sub>, promoting water uptake and reacquiring turgidity. Based on [<a href="#B56-ijms-21-08648" class="html-bibr">56</a>].</p>
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<p>Number of publications dealing with iso-/anisohydry from 1990 to 2019, for all plant species (white bars) and <span class="html-italic">Vitis vinifera</span> (black bars) only. Data collected from the Scopus database (<a href="https://www.scopus.com/" target="_blank">https://www.scopus.com/</a>), according to the method of [<a href="#B60-ijms-21-08648" class="html-bibr">60</a>].</p>
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<p>Modelled dynamic of stomatal conductance (g<sub>s</sub>) and foliar ABA concentration in a near-isohydric cv (Montepulciano) and an anisohydric cv (Sangiovese). Elaboration on data by [<a href="#B19-ijms-21-08648" class="html-bibr">19</a>].</p>
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11 pages, 883 KiB  
Review
Mediator Complex: A Pivotal Regulator of ABA Signaling Pathway and Abiotic Stress Response in Plants
by Leelyn Chong, Pengcheng Guo and Yingfang Zhu
Int. J. Mol. Sci. 2020, 21(20), 7755; https://doi.org/10.3390/ijms21207755 - 20 Oct 2020
Cited by 36 | Viewed by 5988
Abstract
As an evolutionarily conserved multi-protein complex, the Mediator complex modulates the association between transcription factors and RNA polymerase II to precisely regulate gene transcription. Although numerous studies have shown the diverse functions of Mediator complex in plant development, flowering, hormone signaling, and biotic [...] Read more.
As an evolutionarily conserved multi-protein complex, the Mediator complex modulates the association between transcription factors and RNA polymerase II to precisely regulate gene transcription. Although numerous studies have shown the diverse functions of Mediator complex in plant development, flowering, hormone signaling, and biotic stress response, its roles in the Abscisic acid (ABA) signaling pathway and abiotic stress response remain largely unclear. It has been recognized that the phytohormone, ABA, plays a predominant role in regulating plant adaption to various abiotic stresses as ABA can trigger extensive changes in the transcriptome to help the plants respond to environmental stimuli. Over the past decade, the Mediator complex has been revealed to play key roles in not only regulating the ABA signaling transduction but also in the abiotic stress responses. In this review, we will summarize current knowledge of the Mediator complex in regulating the plants’ response to ABA as well as to the abiotic stresses of cold, drought and high salinity. We will particularly emphasize the involvement of multi-functional subunits of MED25, MED18, MED16, and CDK8 in response to ABA and environmental perturbation. Additionally, we will discuss potential research directions available for further deciphering the role of Mediator complex in regulating ABA and other abiotic stress responses. Full article
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<p>The pivotal role of Mediator complex in the ABA signaling pathway. ABA is perceived by its receptors PYL/RCARs, which promotes the interaction between PP2Cs (negative regulators of the ABA signaling pathway) and PYLs, hence releasing the positive regulators SnRK2s to activate ABA downstream signaling events. Additionally, RAFs can directly phosphorylate SnRK2s for the activation of SnRK2s, which subsequently interact with and phosphorylate several downstream TFs including ABFs, ABI5 and RAP2.6 to transduce the ABA signals. Mediator subunits of CDK8, MED25, and MED18 relay the signals from TFs RAP2.6, ABI5, and ABI4, respectively, and help recruit the RNA Pol II to the TFs-targeted promoters of ABA-responsive genes, thereby promoting the transcription of ABA-responsive genes.</p>
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<p>The simplified network of TFs and Mediator complex in regulating the abiotic stress responses. In response to cold stress, CBF1 activates the expression of <span class="html-italic">COR</span> genes through MED2, MED14, and MED16, which are required for the recruitment of RNA Pol II to the promoters of <span class="html-italic">COR</span> genes; In response to drought stress, CDK8 physically interact with WIN1 and RAP2.6 to positively regulate the cuticle wax biosynthesis and expression of stress-responsive genes; in contrast, MED25 negatively regulates the transcriptional activity of DREB2A and the expression of stress-responsive genes, thereby negatively contributing to the drought tolerance; in response to salt stress, ZFHD1 and MYB-like interact with MED25 to positively regulate the salt response.</p>
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