GRUPO: Abscisic Acid Signaling

Abscisic acid (ABA) plays a crucial role to integrate plant response to abiotic stress (particularly drought and salinity) into the regulation of plant growth and development. An increase in ABA levels and the subsequent plant response to the hormone are key components of the adaptive mechanism to resist/avoid those forms of abiotic stress. Accordingly, the characterization of ABA signaling offers a high biotechnological potential to improve plant tolerance to drought and salinity.

Because of its essential function in plant stress physiology, elucidating the abscisic acid (ABA) signaling pathway holds enormous promise for application in agriculture. A breakthrough in ABA signaling occurred in 2009, i.e. the discovery of the 14-member PYR/PYL/RCAR family of ABA receptors (Park et al., 2009; Ma et al., 2009; Santiago et al., 2009). Control of ABA signaling by PYR/PYL/RCAR ABA-receptors involves direct ABA-dependent inhibition of clade A phosphatases type-2C (PP2Cs), for instance, ABI1, HAB1, PP2CA, which are key negative regulators of the pathway (Saez et al., 2006; Rubio et al., 2009). Inhibition of PP2Cs leads to activation of sucrose non-fermenting 1-related subfamily 2 (SnRK2) kinases, which regulate stomatal aperture and transcriptional response to ABA. Thus, a core signaling network for ABA has emerged from these findings (Fujii et al., 2009; Cutler et al., 2010).

Agrochemicals: Crystal structures are available for ABA receptors and receptor-ABA-phosphatase complexes, which reveal key details on the mode of interaction between the receptor, the hormone, and the protein phosphatase as well as the mechanism of activation of ABA signaling. This information will be used in the structure-assisted identification of synthetic molecules able to act as agonists of ABA receptors and activate ABA signaling in plants. These molecules might have the potential to improve the yield of crop plants under drought stress or any other properties modulated by the ABA pathway in crop or ornamental plants. To this end, we will clone and produce recombinant ABA receptors in tomato, orange, grapevine, and monocots as targets for screening small molecules capable of acting as agrochemicals through the activation of ABA receptors. Additionally, through collaboration with organic chemists and protein crystallographers, direct synthesis of small molecules that fit into the ABA binding ligand pocket has been performed.  Recently, we achieved the structure-guided engineering of a receptor-agonist pair for inducible activation of the ABA adaptive response to drought, protected by the EP21382948 patent.

Molecular genetics: The ABA signaling group has played a key role in the discovery and characterization of the PYR/PYL/RCAR family of ABA receptors, and their connection with PP2Cs and SnRK2s (Rodriguez et al., 1998; Gonzalez-Guzman et al., 2002; Saez et al., 2004, 2006, 2008; Park et al., 2009; Santiago et al., 2009a, 2009b; Rubio et al., 2009; Vlad et al., 2009; Fujii et al., 2009; Cutler et al., 2010; Vlad et al., 2010; Dupeux et al., 2011a, 2011b; Antoni et al. 2012; Gonzalez-Guzman et al., 2012; Antoni et al., 2013; Merilo et al., 2013; Pizzio et al., 2013).

Some of these works are landmarks in ABA signaling and together with other results have brought about a breakthrough in our knowledge of the pathway. Later on, we provided insight into the subcellular location of ABA receptors and we discovered that C2-domain abscisic acid-related proteins mediate the interaction of PYR/PYL/RCAR receptors with the plasma membrane (Rodriguez et al., 2014; Diaz et al., 2016). Concerning ABA-induced transcriptional regulation, we have discovered a link between SWI/SNF chromatin remodeling complexes and core components of ABA signaling (Saez et al., 2008; Han et al., 2012; Peirats-Llobet et al., 2016). We have contributed several genetic strategies that enhance ABA signaling as a valuable tool for improving plant water use. Among them, the constitutive inactivation of PP2Cs (Saez et al., 2006), overexpression of monomeric ABA receptors (Santiago et al., 2009a; Gonzalez-Guzman et al., 2014), and the generation of mutated ABA receptors that enhance ABA-dependent inhibition of PP2Cs (Pizzio et al., 2013). As a result, three patents were filled to cover these findings.

We have played a pioneering role in studies that address the turnover of core ABA signaling components, particularly ABA receptors and PP2Cs (Bueso et al., 2014; Irigoyen et al., 2014; Wu et al., 2016; Belda-Palazon et al., 2016, 2018 and 2019; Fernandez et al., 2020; Coego et al., 2021). We uncovered the unique role of PYL8 in root ABA signaling, which involves a non-cell-autonomous mechanism like mobile transcription factors, ABA-induced stabilization, and predominant nuclear localization (Belda-Palazon et al., 2018). We have further studied the mechanisms that affect subcellular localization and half-life of ABA receptors. We have uncovered a novel route for endosomal degradation of ABA receptors through the ESCRT pathway (Belda-Palazón et al., 2016; Yu et al., 2016; Garcia-Leon et al., 2019) and we have identified two novel families of E3 ligases that mediate the turnover of PP2Cs (Wu et al., 2016; Belda-Palazon et al., 2019; Julian et al., 2019). Physiological studies and structural biology of crop ABA receptors are a long-lasting interest of our group (Gonzalez-Guzman et al., 2014), to elucidate the formation of receptor-ABA-phosphatase complexes (Moreno-Alvero et al., 2017) and the key role of PYL8-like receptors in crop response to abiotic stress (Garcia-Maquilon et al., 2021; Pizzio et al., 2022). International collaborations (4 publications in Nature Plants, 1 Science Advances, 1 Dev Cell) have provided key findings on the role of SnRK2s-PP2Cs-ABA receptors in root hydrotropism (Dietrich et al., 2017; Miao et al., 2021), regulation of plant growth via SnRK1 and TOR (Belda-Palazon et al., 2020), specific roles of ABA receptors in stomatal response to ABA and high CO2 (Dittrich et al., 2019) and moonlight roles of FYVE1/FREE1 in repression of ABA signaling (Li et al., 2019).

Eckardt,N.A., Avin-Wittenberg,T., Bassham,D.C., Chen,P., Chen,Q., Fang,J., Genschik,P., Ghifari,A.S., Guercio,A.M., Gibbs,D.J., Heese,M., Jarvis,R.P., Michaeli,S., Murcha,M.W., Mursalimov,S., Noir,S., Palayam,M., Peixoto,B., Rodriguez,P.L., Schaller,A., Schnittger,A., Serino,G., Shabek,N., Stintzi,A., Theodoulou,F.L., Üstün,S., van Wijk,K.J., Wei,N., Xie,Q., Yu,F. and Zhang,H. (2024). The lowdown on breakdown: Open questions in plant proteolysis.

Plant Cell 2024 Jul 9:koae193. doi: 10.1093/plcell/koae193

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Sanchez-Olvera, M., Martin-Vasquez, C., Mayordomo, C., Illescas-Miranda, J., Bono, M., Coego, A., Alonso, J., Hernández-González, M., Jiménez-Arias, D., Forment, J., Albert, A., Granell, A., Borges, A. A., Rodriguez, P. L* (2024). ABA-receptor agonist iSB09 decreases soil water consumption and increases tomato CO2 assimilation and water use efficiency under drought stress.

Environmental and Experimental Botany 225, 105847.https://doi.org/10.1016/j.envexpbot.2024.105847

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Morales-Sierra,S., Luis,J.C., Jimenez-Arias,D., Rancel-Rodriguez,N.M., Coego,A., Rodriguez,P.L., Cueto,M. and Borges,A.A. (2023). Biostimulant activity of Galaxaura rugosa seaweed extracts against water deficit stress in tomato seedlings involves activation of ABA signaling.

Front Plant Sci. 14, 1251442. doi: 10.3389/fpls.2023.1251442

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Jiménez-Arias D, Morales-Sierra S, Suárez E, Lozano-Juste J, Coego A, Estevez JC, Borges AA, Rodriguez PL* (2023). Abscisic acid mimic-fluorine derivative 4 (AMF4) alleviates water deficit stress by regulating ABA-responsive genes, proline accumulation, CO2 assimilation, water use efficiency and better nutrient uptake in tomato plants.

Front Plant Sci. 14:1191967. doi: 10.3389/fpls.2023.1191967

Yang C, Li X, Chen S, Liu C, Yang L, Li K, Liao J, Zheng X, Li H, Li Y, Zeng S, Zhuang X, Rodriguez PL, Luo M, Wang Y, Gao C. (2023).ABI5-FLZ13 Module Transcriptionally Represses Growth-related Genes to Delay Seed Germination in Response to ABA.

Plant Commun. 2023 Jun 9:100636. doi: 10.1016/j.xplc.2023.100636.

Lozano-Juste J, Infantes L, Garcia-Maquilon I, Ruiz-Partida R, Merilo E, Benavente JL, Velazquez-Campoy A, Coego A, Bono M, Forment J, Pampín B, Destito P, Monteiro A, Rodríguez R, Cruces J, Rodriguez PL*, Albert A* (2023). Structure-guided engineering of a receptor-agonist pair for inducible activation of the ABA adaptive response to drought.

Sci Adv. 2023 Mar 10;9(10):eade9948. doi: 10.1126/sciadv.ade9948.

Hirt H, Al-Babili S, Almeida-Trapp M, Martin A, Aranda M, Bartels D, Bennett M, Blilou I, Boer D, Boulouis A, Bowler C, Brunel-Muguet S, Chardon F, Colcombet J, Colot V, Daszkowska-Golec A, Dinneny JR, Field B, Froehlich K, Gardener CH, Gojon A, Gomès E, Gomez-Alvarez EM, Gutierrez C, Havaux M, Hayes S, Heard E, Hodges M, Alghamdi AK, Laplaze L, Lauersen KJ, Leonhardt N, Johnson X, Jones J, Kollist H, Kopriva S, Krapp A, Masson ML, McCabe MF, Merendino L, Molina A, Moreno Ramirez JL, Mueller-Roeber B, Nicolas M, Nir I, Orduna IO, Pardo JM, Reichheld JP, Rodriguez PL, Rouached H, Saad MM, Schlögelhofer P, Singh KA, De Smet I, Stanschewski C, Stra A, Tester M, Walsh C, Weber APM, Weigel D, Wigge P, Wrzaczek M, Wulff BBH, Young IM (2023). PlantACT! – how to tackle the climate crisis.

Trends Plant Sci. 2023 28(5):537-543. doi: 10.1016/j.tplants.2023.01.005.

Chen W, Zhou H, Xu F, Yu M, Coego A, Rodriguez L, Lu Y, Xie Q, Fu Q, Chen J, Xu G, Wu D, Li X, Li X, Jaillais Y, Rodriguez PL, Zhu S, Yu F. (2023). CAR modulates plasma membrane nano-organization and immune signaling downstream of RALF1-FERONIA signaling pathway 

New Phytol. 2023 Mar;237(6):2148-2162. doi: 10.1111/nph.18687

Pizzio GA, Rodriguez PL* (2022). Dual regulation of SnRK2 signaling by Raf-like MAPKKKs

Mol Plant. 2022 Aug 1;15(8):1260-1262. doi: 10.1016/j.molp.2022.07.002.

Infantes L, Rivera-Moreno M, Daniel-Mozo M, Benavente JL, Ocaña-Cuesta J, Coego A, Lozano-Juste J, Rodriguez PL, Albert A (2022). Structure-Based Modulation of the Ligand Sensitivity of a Tomato Dimeric Abscisic Acid Receptor Through a Glu to Asp Mutation in the Latch Loop.

Front Plant Sci. 2022 Jun 6;13:884029. doi: 10.3389/fpls.2022.884029. eCollection 2022.

Franco-Aragón D, García-Maquilón I, Manicardi A, Rodríguez PL, Lozano-Juste J (2022). Evaluation of the Anti-transpirant Activity of ABA Receptor Agonists in Monocot and Eudicot Plants.

Methods Mol Biol. 2022;2494:229-238. doi: 10.1007/978-1-0716-2297-1_16.

Garcia-Maquilon I, Lozano-Juste J, Alrefaei AF, Rodriguez PL* (2022). Hydrotropism: Analysis of the Root Response to a Moisture Gradient.

Methods Mol Biol. 2022;2494:17-24. doi: 10.1007/978-1-0716-2297-1_2.

Pizzio GA, Mayordomo C, Lozano-Juste J, Garcia-Carpintero V, Vazquez-Vilar M, Nebauer SG, Kaminski KP, Ivanov NV, Estevez JC, Rivera-Moreno M, Albert A, Orzaez D, Rodriguez PL* (2022). PYL1- and PYL8-like ABA Receptors of Nicotiana benthamiana Play a Key Role in ABA Response in Seed and Vegetative Tissue. 

Cells. 2022 Feb 24;11(5):795. doi: 10.3390/cells11050795.

Belda-Palazón B, Rodriguez PL (2022). Microscopic Imaging of Endosomal Trafficking of ABA Receptors.

Methods Mol Biol. 2022;2462:59-69. doi: 10.1007/978-1-0716-2156-1_5

Julian J, Coego A, Alrefaei AF, Rodriguez PL* (2022). Affinity Purification of Ubiquitinated Proteins Using p62-Agarose to Assess Ubiquitination of Clade A PP2Cs.

Methods Mol Biol. 2022;2462:45-57. doi: 10.1007/978-1-0716-2156-1_4.

Miao R, Russinova E, Rodriguez PL* (2022). Tripartite hormonal regulation of plasma membrane H+-ATPase activity.

Trends Plant Sci Jun;27(6):588-600 https://doi.org/10.1016/j.tplants.2021.12.011

• García-Maquilón I, Rodriguez PL, Vaidya AS, Lozano-Juste J. (2021)
  A luciferase reporter assay to identify chemical activators of ABA signaling

  Methods Mol Biol. 2021;2213:113-121

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• Lozano-Juste J, García-Maquilón I, Brea J, Piña R, Albert A, Rodriguez PL, Loza MI. (2021)
  Identification of ABA receptor agonists using a multiplexed high-throughput chemical screening

  Methods Mol Biol. 2021;2213:99-111

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• Miao R, Yuan W, Wang Y, Garcia-Maquilon I, Dang X, Li Y, Zhang J, Zhu Y, Rodriguez PL, Xu W. (2021)
  Low ABA concentration promotes root growth and hydrotropism through relief of ABA INSENSITIVE 1-mediated inhibition of plasma membrane H+-ATPase 2

  Science Advances 7 (12): eabd4113.

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• Garcia-Maquilon,I., Coego,A., Lozano-Juste,J., Messerer,M., de Ollas,C., Julian,J., Ruiz-Partida,R., Pizzio,G., Belda-Palazon,B., Gomez-Cadenas,A., Mayer,K.F.X., Geiger,D., Alquraishi,S.A., Alrefaei,A.F., Ache,P., Hedrich,R. and Rodriguez,P.L* (2021)
  PYL8 ABA receptors of Phoenix dactylifera play a crucial role in response to abiotic stress and are stabilized by ABA.

  J.Exp.Bot. 72(2):757-774

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• Belda-Palazon,B., Adamo,M., Valerio,C., Ferreira,L.J., Confraria,A., Reis-Barata,D., Rodrigues,A., Meyer,C., Rodriguez,P.L. and Baena-Gonzalez,E. (2020)
  A dual function of SnRK2 kinases in the regulation of SnRK1 and plant growth.

  Nature Plants 6(11):1345-1353

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• Belda-Palazon, B., and Rodriguez, P. L.* (2020)
  Degradation of abscisic acid receptors through the endosomal pathway

  Methods Mol Biol. 2177:35-48

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• Lozano-Juste,J., Garcia-Maquilon,I., Ruiz-Partida,R. and Rodriguez,P.L. (2020)
  Drug Discovery for Thirsty Crops

  Trends Plant Sci. Sep;25(9):844-846.

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• Lozano-Juste,J., Alrefaei,A.F. and Rodriguez,P.L.* (2020)
  Plant Osmotic Stress Signaling: MAPKKKs Meet SnRK2s

  Trends Plant Sci. Dec;25(12):1179-1182

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• Fernandez, M. A., Belda-Palazon, B., Julian, J., Coego, A., Lozano-Juste, J., Inigo, S., Rodriguez, L., Bueso, E., Goossens, A., and Rodriguez, P. L.* (2020)
  RBR-type E3 ligases and the Ub-conjugating enzyme UBC26 regulate ABA receptor levels and signaling

  Plant Physiol. 182(4):1723-1742

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