Editorial: Ion Homeostasis in Plant Stress and Development

This paper was funded by grants PID2019-104054GB-I00 and RTC-2017-6468-2-AR from the Spanish "Agencia Estatal de Investigacion".Mulet, JM.; Campos, F.; Yenush, L. (2020). Editorial: Ion Homeostasis in Plant Stress and Development. Frontiers in Plant Science. 11:1-3. https://doi.org/10.3389/fpls.2020.618273S1311Bromham, L., Hua, X., & Cardillo, M. (2020). Macroevolutionary and macroecological approaches to understanding the evolution of stress tolerance in plants. Plant, Cell & Environment, 43(12), 2832-2846. doi:10.1111/pce.13857Colmenero-Flores, J. M., Franco-Navarro, J. D., Cubero-Font, P., Peinado-Torrubia, P., & Rosales, M. A. (2019). Chloride as a Beneficial Macronutrient in Higher Plants: New Roles and Regulation. International Journal of Molecular Sciences, 20(19), 4686. doi:10.3390/ijms20194686Dreyer, I., & Uozumi, N. (2011). Potassium channels in plant cells. FEBS Journal, 278(22), 4293-4303. doi:10.1111/j.1742-4658.2011.08371.xFlowers, T. J., & Colmer, T. D. (2015). Plant salt tolerance: adaptations in halophytes. Annals of Botany, 115(3), 327-331. doi:10.1093/aob/mcu267Galvan-Ampudia, C. S., Julkowska, M. M., Darwish, E., Gandullo, J., Korver, R. A., Brunoud, G., … Testerink, C. (2013). Halotropism Is a Response of Plant Roots to Avoid a Saline Environment. Current Biology, 23(20), 2044-2050. doi:10.1016/j.cub.2013.08.042Korver, R. A., Berg, T., Meyer, A. J., Galvan‐Ampudia, C. S., Tusscher, K. H. W. J., & Testerink, C. (2019). Halotropism requires phospholipase Dζ1‐mediated modulation of cellular polarity of auxin transport carriers. Plant, Cell & Environment, 43(1), 143-158. doi:10.1111/pce.13646Locascio, A., Marqués, M. C., García-Martínez, G., Corratgé-Faillie, C., Andrés-Colás, N., Rubio, L., … Yenush, L. (2019). BCL2-ASSOCIATED ATHANOGENE4 Regulates the KAT1 Potassium Channel and Controls Stomatal Movement. Plant Physiology, 181(3), 1277-1294. doi:10.1104/pp.19.00224Mahajan, S., & Tuteja, N. (2005). Cold, salinity and drought stresses: An overview. Archives of Biochemistry and Biophysics, 444(2), 139-158. doi:10.1016/j.abb.2005.10.018Ruyter-Spira, C., Kohlen, W., Charnikhova, T., van Zeijl, A., van Bezouwen, L., de Ruijter, N., … Bouwmeester, H. (2010). Physiological Effects of the Synthetic Strigolactone Analog GR24 on Root System Architecture in Arabidopsis: Another Belowground Role for Strigolactones?      . Plant Physiology, 155(2), 721-734. doi:10.1104/pp.110.16664

The Biostimulant, Potassium Humate Ameliorates Abiotic Stress in Arabidopsis thaliana by Increasing Starch Availability

[EN] Potassium humate is a widely used biostimulant known for its ability to enhance growth and improve tolerance to abiotic stress. However, the molecular mechanisms explaining its effects remain poorly understood. In this study, we investigated the mechanism of action of potassium humate using the model plant Arabidopsis thaliana. We demonstrated that a formulation of potassium humate effectively increased the fresh weight accumulation of Arabidopsis plants under normal conditions, salt stress (sodium or lithium chloride), and particularly under osmotic stress (mannitol). Interestingly, plants treated with potassium humate exhibited a reduced antioxidant response and lower proline accumulation, while maintaining photosynthetic activity under stress conditions. The observed sodium and osmotic tolerance induced by humate was not accompanied by increased potassium accumulation. Additionally, metabolomic analysis revealed that potassium humate increased maltose levels under control conditions but decreased levels of fructose. However, under stress, both maltose and glucose levels decreased, suggesting changes in starch utilization and an increase in glycolysis. Starch concentration measurements in leaves showed that plants treated with potassium humate accumulated less starch under control conditions, while under stress, they accumulated starch to levels similar to or higher than control plants. Taken together, our findings suggest that the molecular mechanism underlying the abiotic stress tolerance conferred by potassium humate involves its ability to alter starch content under normal growth conditions and under salt or osmotic stress.This research was funded by the CDTI program project EXP 00137666/IDI-20210456. awarded to CALDIC Ibérica S.L. and the research contract. "DESARROLLO DE FORMULADOS BIOESTIMULANTES Y BIOFERTILIZANTES INNOVADORES DE ORIGEN NATURAL (CALBIO) DESTINADOS A LA AGRICULTURA CONVENCIONAL Y ECOLÓGICA. ESTUDIO CIENTÍFICO DE EFECTOS SINÉRGICOS ENTRE BIOACTIVOS MICROBIANOS Y NO MICROBIANOS" Between CALDIC Ibérica S.L. and Universitat Politècnica de València. The APC was funded by the aforementioned research contract.Benito, P.; Bellón, J.; Porcel, R.; Yenush, L.; Mulet, JM. (2023). The Biostimulant, Potassium Humate Ameliorates Abiotic Stress in Arabidopsis thaliana by Increasing Starch Availability. International Journal of Molecular Sciences. 24(15):1-21. https://doi.org/10.3390/ijms241512140121241

Use of Yucca (Yucca schidigera) Extracts as Biostimulants to Promote Germination and Early Vigor and as Natural Fungicides

[EN] Climate change is increasing drought and salinity in many cultivated areas, therefore threatening food production. There is a great demand for novel agricultural inputs able to maintain yield under the conditions imposed by the anthropogenic global warming. Biostimulants have been proposed as a useful tool to achieve this objective. We have investigated the biostimulant effect of different yucca (Yucca schidigera) extracts on plant growth at different stages of development under different abiotic stress conditions. The extracts were tested in the model plant Arabidopsis thaliana, and in three different crops; tomato (Solanum lycopersicum var microtom), broccoli (Brassica oleracea var. italica) and lettuce (Lactuca sativa var romana). We have found that the investigated extracts are able to promote germination and early vigor under drought/osmotic and salt stress induced either by sodium chloride or lithium chloride. This effect is particularly strong in Arabidopsis thaliana and in the Brassicaceae broccoli. We have also determined using antibiograms against the model yeast Saccharomyces cerevisiae that the evaluated extracts may be used also as a natural fungicide. The results in this report show that yucca extracts may be used to enhance early vigor in some crops and as a natural fungicide, providing a new and useful tool for farmers.This research was funded by the CDTI program project EXP 00137666/IDI-20210456. awarded to CALDIC Iberica S.L. and the research contract. "DESARROLLO DE FORMULADOS BIOESTIMULANTES Y BIOFERTILIZANTES INNOVADORES DE ORIGEN NATURAL (CALBIO) DESTINADOS A LA AGRICULTURA CONVENCIONAL Y ECOLOGICA. ESTUDIO CIENTIFICO DE EFECTOS SINERGICOS ENTRE BIOACTIVOS MICROBIANOS Y NO MICROBIANOS" Between CALDIC Iberica S.L. and Universitat Politecnica de Valencia. The APC was funded by the afore mentioned research contract.Benito, P.; Ligorio, D.; Bellón, J.; Yenush, L.; Mulet, JM. (2023). Use of Yucca (Yucca schidigera) Extracts as Biostimulants to Promote Germination and Early Vigor and as Natural Fungicides. Plants. 12(2):1-12. https://doi.org/10.3390/plants1202027411212

A functional Rim101 complex is required for proper accumulation of the Ena1 Na+-ATPase protein in response to salt stress in Saccharomyces cerevisiae

[EN] The maintenance of ionic homeostasis is essential for cell viability, thus the activity of plasma membrane ion transporters must be tightly controlled. Previous studies in Saccharomyces cerevisiae revealed that the proper trafficking of several nutrient permeases requires the E3 ubiquitin ligase Rsp5 and, in many cases, the presence of specific adaptor proteins needed for Rsp5 substrate recognition. Among these adaptor proteins are nine members of the arrestin-related trafficking adaptor (ART) family. We studied the possible role of the ART family in the regulation of monovalent cation transporters. We show here that the salt sensitivity phenotype of the rim8/art9 mutant is due to severe defects in Ena1 protein accumulation, which is not attributable to transcriptional defects. Many components of the Rim pathway are required for correct Ena1 accumulation, but not for the accumulation of other nutrient permeases. Moreover, we observe that strains lacking components of the endosomal sorting complexes required for transport (ESCRT) pathway previously described to play a role in Rim complex formation present similar defects in Ena1 accumulation. Our results show that, in response to salt stress, a functional Rim complex via specific ESCRT interactions is required for the proper accumulation of the Ena1 protein, but not induction of the ENA1 gene.This work was supported by grants BFU2011-30197-C03-03 from the Spanish Ministry of Science and Innovation (Madrid, Spain) and EUI2009-04147 [Systems Biology of Microorganisms (SysMo2) European Research Area-Network (ERA-NET)]. V. L-T. was supported by a pre-doctoral fellowship from the Polytechnic University of Valencia.Marqués, MC.; Zamarbide-Forés, S.; Pedelini, L.; Llopis Torregrosa, V.; Yenush, L. (2015). 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Saccharomyces cerevisiae as a Tool to Investigate Plant Potassium and Sodium Transporters

[EN] Sodium and potassium are two alkali cations abundant in the biosphere. Potassium is essential for plants and its concentration must be maintained at approximately 150 mM in the plant cell cytoplasm including under circumstances where its concentration is much lower in soil. On the other hand, sodium must be extruded from the plant or accumulated either in the vacuole or in specific plant structures. Maintaining a high intracellular K+/Na+ ratio under adverse environmental conditions or in the presence of salt is essential to maintain cellular homeostasis and to avoid toxicity. The baker's yeast, Saccharomyces cerevisiae, has been used to identify and characterize participants in potassium and sodium homeostasis in plants for many years. Its utility resides in the fact that the electric gradient across the membrane and the vacuoles is similar to plants. Most plant proteins can be expressed in yeast and are functional in this unicellular model system, which allows for productive structure-function studies for ion transporting proteins. Moreover, yeast can also be used as a high-throughput platform for the identification of genes that confer stress tolerance and for the study of protein-protein interactions. In this review, we summarize advances regarding potassium and sodium transport that have been discovered using the yeast model system, the state-of-the-art of the available techniques and the future directions and opportunities in this field.This work was supported by the Spanish Ministry of Economy and Competitiveness (BIO201677776-P and BIO2016-81957-REDT) and the Valencian Government (AICO/2018/300)Locascio, AAM.; Andrés-Colás, N.; Mulet, JM.; Yenush, L. (2019). Saccharomyces cerevisiae as a Tool to Investigate Plant Potassium and Sodium Transporters. 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Regulation of the Yeast Hxt6 Hexose Transporter by the Rod1 α-Arrestin, the Snf1 Protein Kinase, and the Bmh2 14-3-3 Protein

[EN] Cell viability requires adaptation to changing environmental conditions. Ubiquitin-mediated endocytosis plays a crucial role in this process, because it provides a mechanism to remove transport proteins from the membrane. Arrestin-related trafficking proteins are important regulators of the endocytic pathway in yeast, facilitating selective ubiquitylation of target proteins by the E3 ubiquitin ligase, Rsp5. Specifically, Rod1 (Art4) has been reported to regulate the endocytosis of both the Hxt1, Hxt3, and Hxt6 glucose transporters and the Jen1 lactate transporter. Also, the AMP kinase homologue, Snf1, and 14-3-3 proteins have been shown to regulate Jen1 via Rod1. Here, we further characterized the role of Rod1, Snf1, and 14-3-3 in the signal transduction route involved in the endocytic regulation of the Hxt6 high affinity glucose transporter by showing that Snf1 interacts specifically with Rod1 and Rog3 (Art7), that the interaction between the Bmh2 and several arrestin-related trafficking proteins may be modulated by carbon source, and that both the 14-3-3 protein Bmh2 and the Snf1 regulatory domain interact with the arrestin-like domain containing the N-terminal half of Rod1 (amino acids 1-395). Finally, using both co-immunoprecipitation and bimolecular fluorescence complementation, we demonstrated the interaction of Rod1 with Hxt6 and showed that the localization of the Rod1-Hxt6 complex at the plasma membrane is affected by carbon source and is reduced upon overexpression of SNF1 and BMH2.Supported by a predoctoral fellowship from the Polytechnic University of Valencia.Llopis Torregrosa, V.; Ferri-Blazquez, A.; Adam-Artigues, A.; Deffontaines, E.; Van Heusden, GPH.; Yenush, L. (2016). Regulation of the Yeast Hxt6 Hexose Transporter by the Rod1 α-Arrestin, the Snf1 Protein Kinase, and the Bmh2 14-3-3 Protein. Journal of Biological Chemistry. 291(29):14973-14985. https://doi.org/10.1074/jbc.M116.733923S149731498529129Mulet, J. M., Llopis-Torregrosa, V., Primo, C., Marqués, M. C., & Yenush, L. (2013). Endocytic regulation of alkali metal transport proteins in mammals, yeast and plants. 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Yeast Saccharomyces cerevisiae adiponectin receptor homolog Izh2 is involved in the regulation of zinc, phospholipid and pH homeostasis

[EN] The functional link between zinc homeostasis and membrane-related processes, including lipid metabolism regulation, extends from yeast to humans, and has a likely role in the pathogenesis of diabetes. The yeast Izh2 protein has been previously implicated in zinc ion homeostasis and in the regulation of lipid and phosphate metabolism, but its precise molecular function is not known. We performed a chemogenomics experiment to determine the genes conferring resistance or sensitivity to different environmental zinc concentrations. We then determined at normal, depleted and excess zinc concentrations, the genetic interactions of IZH2 at the genome-wide level and measured changes in the transcriptome caused by deletion of IZH2. We found evidence for an important cellular function of the Rim101 pathway in zinc homeostasis in neutral or acidic environments, and observed that phosphatidylinositol is a source of inositol when zinc availability is limited. Comparison of our experimental profiles with published gene expression and genetic interaction profiles revealed pleiotropic functions for Izh2. We propose that Izh2 acts as an integrator of intra- and extracellular signals in providing adequate cellular responses to maintain homeostasis under different external conditions, including but not limited to alterations in zinc concentrations. Guardar / Salir Siguiente >This work was supported by grant P1-0207 from the Slovenian Research Agency. M.M.U. was supported by the Young Investigator fellowship scheme from the Slovenian Research Agency. Work done in the group of L.Y. was funded by grant BFU2011-30197-C03-03 from the Spanish Ministry of Science and Innovation (Madrid, Spain). C.P. was supported by a pre-doctoral fellowship from the Spanish Research Council.Mattiazzi Usaj, M.; Prelec, M.; Brioznic, M.; Primo Planta, C.; Curk, T.; Scancar, J.; Yenush, L.... (2015). Yeast Saccharomyces cerevisiae adiponectin receptor homolog Izh2 is involved in the regulation of zinc, phospholipid and pH homeostasis. Metallomics. 7(9):1338-1351. https://doi.org/10.1039/c5mt00095e133813517

Identification of distinctive physiological and molecular responses to salt stress among tolerant and sensitive cultivars of broccoli (Brassica oleracea var. Italica)

[EN] Background Salt stress is one of the main constraints determining crop productivity, and therefore one of the main limitations for food production. The aim of this study was to characterize the salt stress response at the physiological and molecular level of different Broccoli (Brassica oleracea L. var. Italica Plenck) cultivars that were previously characterized in field and greenhouse trials as salt sensitive or salt tolerant. This study aimed to identify functional and molecular traits capable of predicting the ability of uncharacterized lines to cope with salt stress. For this purpose, this study measured different physiological parameters, hormones and metabolites under control and salt stress conditions. Results This study found significant differences among cultivars for stomatal conductance, transpiration, methionine, proline, threonine, abscisic acid, jasmonic acid and indolacetic acid. Salt tolerant cultivars were shown to accumulate less sodium and potassium in leaves and have a lower sodium to potassium ratio under salt stress. Analysis of primary metabolites indicated that salt tolerant cultivars have higher concentrations of several intermediates of the Krebs cycle and the substrates of some anaplerotic reactions. Conclusions This study has found that the energetic status of the plant, the sodium extrusion and the proline content are the limiting factors for broccoli tolerance to salt stress. Our results establish physiological and molecular traits useful as distinctive markers to predict salt tolerance in Broccoli or to design novel biotechnological or breeding strategies for improving broccoli tolerance to salt stress.This work was funded by Grant RTC-2017-6468-2-AR (APROXIMACIONES MOLECULARES PARA INCREMENTAR LA TOLERANCIA A SALINIDAD Y SEQUiA DEL BROCOLI) funded by MCIN/AEI/10.13039/501100011033 and by "ERDF A way of making Europe" by the European Union. S.C. is a recipient of grant FPU19/01977 from the Spanish Ministerio de Universidades. L.M. was supported by the Spanish MICINN (PTA2019-018094). L.M and A.V. activities were founded by Prometeu program (IMAGINA project, PROMETEU/2019/110). CEAM foundation is funded by Generalitat Valenciana. None of the funding bodies has participated in the design of the study or the collection, analysis, interpretation of data, nor in writing the manuscript.Chevilly-Tena, S.; Dolz-Edo, L.; Morcillo, L.; Vilagrosa, A.; López-Nicolás, JM.; Yenush, L.; Mulet, JM. (2021). Identification of distinctive physiological and molecular responses to salt stress among tolerant and sensitive cultivars of broccoli (Brassica oleracea var. Italica). BMC Plant Biology. 21(1):1-16. https://doi.org/10.1186/s12870-021-03263-4S11621

Salicylic Acid Is Involved in the Basal Resistance of Tomato Plants to Citrus Exocortis Viroid and Tomato Spotted Wilt Virus

[EN] Tomato plants expressing the NahG transgene, which prevents accumulation of endogenous salicylic acid (SA), were used to study the importance of the SA signalling pathway in basal defence against Citrus Exocortis Viroid (CEVd) or Tomato Spotted Wilt Virus (TSWV). The lack of SA accumulation in the CEVd- or TSWV-infected NahG tomato plants led to an early and dramatic disease phenotype, as compared to that observed in the corresponding parental Money Maker. Addition of acibenzolar-S-methyl, a benzothiadiazole (BTH), which activates the systemic acquired resistance pathway downstream of SA signalling, improves resistance of NahG tomato plants to CEVd and TSWV. CEVd and TSWV inoculation induced the accumulation of the hydroxycinnamic amides p-coumaroyltyramine, feruloyltyramine, caffeoylputrescine, and feruloylputrescine, and the defence related proteins PR1 and P23 in NahG plants earlier and with more intensity than in Money Maker plants, indicating that SA is not essential for the induction of these plant defence metabolites and proteins. In addition, NahG plants produced very high levels of ethylene upon CEVd or TSWV infection when compared with infected Money Maker plants, indicating that the absence of SA produced additional effects on other metabolic pathways. This is the first report to show that SA is an important component of basal resistance of tomato plants to both CEVd and TSWV, indicating that SA-dependent defence mechanisms play a key role in limiting the severity of symptoms in CEVd- and TSWV-infected NahG tomato plants.This work was supported by grant BIO2012-33419 from the Spanish Ministry of Economy and Competitiveness received by JMB. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.López-Gresa, MP.; Lisón, P.; Yenush, L.; Conejero Tomás, V.; Rodrigo Bravo, I.; Belles Albert, JM. (2016). Salicylic Acid Is Involved in the Basal Resistance of Tomato Plants to Citrus Exocortis Viroid and Tomato Spotted Wilt Virus. PLoS ONE. 11(11). https://doi.org/10.1371/journal.pone.0166938S111

FungalBraid 2.0: expanding the synthetic biology toolbox for the biotechnological exploitation of filamentous fungi

[EN] Fungal synthetic biology is a rapidly expanding field that aims to optimize the biotechnological exploitation of fungi through the generation of standard, readyto-use genetic elements, and universal syntax and rules for contributory use by the fungal research community. Recently, an increasing number of synthetic biology toolkits have been developed and applied to filamentous fungi, which highlights the relevance of these organisms in the biotechnology field. The FungalBraid (FB) modular cloning platform enables interchangeability of DNA parts with the GoldenBraid (GB) platform, which is designed for plants, and other systems that are compatible with the standard Golden Gate cloning and syntax, and uses binary pCAMBIA-derived vectors to allow Agrobacterium tumefaciensmediated transformation of a wide range of fungal species. In this study, we have expanded the original FB catalog by adding 27 new DNA parts that were functionally validated in vivo. Among these are the resistance selection markers for the antibiotics phleomycin and terbinafine, as well as the uridine-auxotrophic marker pyr4. We also used a normalized luciferase reporter system to validate several promoters, such as PpkiA,P7760,Pef1¿, and PafpB constitutive promoters, and PglaA,PamyB, and PxlnA inducible promoters. Additionally, the recently developed dCas9-regulated GB_SynP synthetic promoter collection for orthogonal CRISPR activation (CRISPRa) in plants has been adapted in fungi through the FB system. In general, the expansion of the FB catalog is of great interest to the scientific community since it increases the number of possible modular and interchangeable DNA assemblies, exponentially increasing the possibilities of studying, developing, and exploiting filamentous fungi.This work was supported by PROMETEO/2018/066 from "Conselleria d'Educacio" (Generalitat Valenciana, Comunitat Valenciana, Spain), grant PID2021-125858OB-100, and the Severo Ochoa Excellence Program CEX 2021-001189-S funded by MCIN/AEI/10.13039/501100011033 and by "ERDF A way of making Europe." EM-G was the recipient of a predoctoral grant FPU18/02019 funded by MCIN/AEI/10.13039/501100011033 and by "ESF Investing in your future." SG holds a Juan de la Cierva Incorporacion grant (IJC 2020-042749-I) funded by MCIN/AEI/10.13039/501100011033 and the European Union NextGenerationEU/PRTR.Moreno-Giménez, E.; Gandía, M.; Sáez, Z.; Manzanares, P.; Yenush, L.; Orzáez Calatayud, DV.; Marcos, JF.... (2023). FungalBraid 2.0: expanding the synthetic biology toolbox for the biotechnological exploitation of filamentous fungi. Frontiers in Bioengineering and Biotechnology. 11:1-17. https://doi.org/10.3389/fbioe.2023.12228121171