Sustainable one-pot immobilization of enzymes in/on metal-organic framework materials

peer-reviewedThe industrial use of enzymes generally necessitates their immobilization onto solid supports. The well-known high affinity of enzymes for metal-organic framework (MOF) materials, together with the great versatility of MOFs in terms of structure, composition, functionalization and synthetic approaches, has led the scientific community to develop very different strategies for the immobilization of enzymes in/on MOFs. This review focuses on one of these strategies, namely, the one-pot enzyme immobilization within sustainable MOFs, which is particularly enticing as the resultant biocomposite Enzyme@MOFs have the potential to be: (i) prepared in situ, that is, in just one step; (ii) may be synthesized under sustainable conditions: with water as the sole solvent at room temperature with moderate pHs, etc.; (iii) are able to retain high enzyme loading; (iv) have negligible protein leaching; and (v) give enzymatic activities approaching that given by the corresponding free enzymes. Moreover, this methodology seems to be near-universal, as success has been achieved with different MOFs, with different enzymes and for different applications. So far, the metal ions forming the MOF materials have been chosen according to their low price, low toxicity and, of course, their possibility for generating MOFs at room temperature in water, in order to close the cycle of economic, environmental and energy sustainability in the synthesis, application and disposal life cycle

Flower induction and development in saffron: Timing and hormone signalling pathways

[EN] The demand for saffron is expected to rise in the coming years due to its nutraceutical and medicinal properties. To cope with this, it will be necessary to develop a mechanised production of saffron. Upgrading the production methods requires accurate control of the flowering time in this species. Nevertheless, little is known about the control of flowering time in Crocus sativus L. The aim of this study is to gain insight into the floral induction regulatory networks operating in this species. A transcriptomic analysis was performed from saffron main buds in different stages of development. The identification of putative integrators of flowering time signals, like FT, as well as meristem identity genes, such as LFY and TFL1, permitted the definition of the time of flowering induction of the buds, being able to use them as molecular markers. The identification of the transcripts encoded by a DROOPING LEAF-like (DL) gene is of particular relevance because this gene might be a novel factor for carpel specification in saffron. To elucidate the hormonal signalling networks working during flower induction, transcriptomic data were used, and the content of IAA, ABA and gibberellins was determined in competent and non-competent buds to flower, during the saffron life cycle. Our results suggested that ABA might be negatively regulating corm dormancy release, but its involvement in flower induction cannot be ruled out. ABI5 and the mediator of ABA regulated dormancy gene MARD1, could be key players of this pathway. In addition, a drop in GA4 levels may also be a necessary, but insufficient, condition for floral induction and development. DELLA, TFL1 and PIF3 genes might be involved in the gibberellin pathway. Notably, IAA seems to be a positive regulator of the process, involving MP/ARF5 and ANT genes in the pathway. Taken together, these results pave the way to the unveiling of the regulatory networks controlling the vegetative-to-reproductive phase change in saffron.The activities of this study have been supported by a project funded by the "Ministerio de Ciencia, Innovacion y Universidades de Espana" [AGL2016-77078-R].Renau-Morata, B.; Nebauer, SG.; García-Carpintero, V.; Cañizares Sales, J.; Minguet, E.; De Los Mozos, M.; Molina Romero, RV. (2021). Flower induction and development in saffron: Timing and hormone signalling pathways. Industrial Crops and Products. 164:1-19. https://doi.org/10.1016/j.indcrop.2021.113370S11916

Propuesta de cambio en organizaciones actuales desde la estrategia de los recursos intangibles

Hoy se habla cada vez más de las organizaciones basadas en el conocimiento, lo cual ha hecho que las organizaciones se ocupen con gran interés de cómo crearlo, utilizarlo de manera eficaz, y compartirlo. Nace así la denominada era o sociedad de la información y del conocimiento. Teniendo en cuenta esta realidad que también se encuentra presente en nuestra región se propone este proyecto que tiene por finalidad diseñar desarrollar, modelar estratégicamente propuestas de cambio que permitan a las organizaciones actuales Gestionar sus Recursos Intangibles. Para lograr este objetivo es necesario poder vincular las herramientas de TIC con la GC y de esta manera fortalecer la gestión. La investigación aplicada que se propone permitirá una capacitación y entrenamiento para analizar, modelar, diseñar y valorar estratégicamente el cambio mediante la Gestión de los Recursos Intangibles. Toda la experiencia y los resultados que se obtengan se transferirán y difundirán mediante actividades formales y sistémicas.Eje: Base de datos y minería de datosRed de Universidades con Carreras en Informática (RedUNCI

Gender- and hydration-associated differences in the physiological response to spinning

UniveIntroduction: There is scarce and inconsistent information about gender-related differences in the hydration of sports persons, as well as about the effects of hydration on performance, especially during indoor sports. Objective: To determine the physiological differences between genders during in indoor physical exercise, with and without hydration. Methods: 21 spinning sportspeople (12 men and 9 women) participated in three controlled, randomly assigned and non-sequential hydration protocols, including no fluid intake and hydration with plain water or a sports drink (volume adjusted to each individual every 15 min), during 90 min of spinning exercise. The response variables included body mass, body temperature, heart rate and blood pressure. Results: During exercise without hydration, men and women lost ~2% of body mass, and showed higher body temperature (~0.2°C), blood pressure (~4 mmHg) and heart rate (~7 beats/min) compared to exercises with hydration. Body temperature and blood pressure were higher for men than for women during exercise without hydration, differences not observed during exercise with hydration. Between 42-99% of variance in body temperature, blood pressure and heart rate could be explained by the physical characteristics of subjects and the work done. Conclusions: During exercise with hydration (either with water or sport drink), the physiological response was similar for both genders. Exercise without hydration produced physical stress, which could be prevented with either of the fluids (plain water was sufficient). Gender differences in the physiological response to spinning (body temperature, mean blood pressure and heart rate) can be explained in part by the distinct physical characteristics of each individual. (Nutr Hosp. 2014;29:644-651) DOI:10.3305/nh.2014.29.3.701

The targeted overexpression of SlCDF4 in the fruit enhances tomato size and yield involving gibberellin signalling

[EN] Tomato is one of the most widely cultivated vegetable crops and a model for studying fruit biology. Although several genes involved in the traits of fruit quality, development and size have been identified, little is known about the regulatory genes controlling its growth. In this study, we characterized the role of the tomato SlCDF4 gene in fruit development, a cycling DOF-type transcription factor highly expressed in fruits. The targeted overexpression of SlCDF4 gene in the fruit induced an increased yield based on a higher amount of both water and dry matter accumulated in the fruits. Accordingly, transcript levels of genes involved in water transport and cell division and expansion during the fruit enlargement phase also increased. Furthermore, the larger amount of biomass partitioned to the fruit relied on the greater sink strength of the fruits induced by the increased activity of sucrose-metabolising enzymes. Additionally, our results suggest a positive role of SlCDF4 in the gibberellin-signalling pathway through the modulation of GA(4) biosynthesis. Finally, the overexpression of SlCDF4 also promoted changes in the profile of carbon and nitrogen compounds related to fruit quality. Overall, our results unveil SlCDF4 as a new key factor controlling tomato size and composition.Renau-Morata, B.; Carrillo, L.; Cebolla Cornejo, J.; Molina Romero, RV.; Martí-Renau, R.; Domínguez-Figueroa, J.; Vicente-Carbajosa, J.... (2020). The targeted overexpression of SlCDF4 in the fruit enhances tomato size and yield involving gibberellin signalling. Scientific Reports. 10(1):1-14. https://doi.org/10.1038/s41598-020-67537-x1141011FAO. Crops production database. FAOSTAT. Latest update: 04/03/2020. Food and Agriculture Organization of the United Nations. Rome https://www.fao.org/faostat (2018).Willcox, J. K., Catignani, G. L. & Lazarus, S. Tomatoes and cardiovascular health. Crit. Rev. Food Sci. Nutr. 43, 1–18. https://doi.org/10.1080/10408690390826437 (2003).Bai, Y. L. & Lindhout, P. Domestication and breeding of tomatoes: what have we gained and what can we gain in the future?. Ann. Bot. 100, 1085–1094. https://doi.org/10.1093/aob/mcm150 (2007).Gascuel, Q., Diretto, G., Monforte, A. J., Fortes, A. M. & Granell, A. Use of natural diversity and biotechnology to increase the quality and nutritional content of tomato and grape. Front. Plant Sci. https://doi.org/10.3389/fpls.2017.00652 (2017).Handa, A. K., Anwar, R. & Mattoo, A. K. in Fruit Ripening Physiology, Signaling and Genomics (eds Nath, P. et al.) 259–290 (CABI, 2014).van der Knaap, E. et al. What lies beyond the eye: the molecular mechanisms regulating tomato fruit weight and shape. Front. Plant Sci. https://doi.org/10.3389/fpls.2014.00227 (2014).Okello, R. C. O., Heuvelink, E., de Visser, P. H. B., Struik, P. C. & Marcelis, L. F. M. What drives fruit growth?. Funct. Plant Biol. 42(9), 817–827. https://doi.org/10.1071/fp15060 (2015).Bertin, N. Analysis of the tomato fruit growth response to temperature and plant fruit load in relation to cell division, cell expansion and DNA endoreduplication. Ann. Bot. 95, 439–447. https://doi.org/10.1093/aob/mci042 (2005).Smith, M. R., Rao, I. M. & Merchant, A. Source-sink relationships in crop plants and their influence on yield development and nutritional quality. Front. Plant Sci. https://doi.org/10.3389/fpls.2018.01889 (2018).Osorio, S., Ruan, Y. L. & Fernie, A. R. An update on source-to-sink carbon partitioning in tomato. Front. Plant Sci. https://doi.org/10.3389/fpls.2014.00516 (2014).Ho, L. C. The mechanism of assimilate partitioning and carbohydrate compartmentation in fruit in relation Ito the quality and yield of tomato. J. Exp. Bot. 47, 1239–1243. https://doi.org/10.1093/jxb/47.Special_Issue.1239 (1996).Koch, K. Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. Curr. Opin. Plant Biol. 7, 235–246. https://doi.org/10.1016/j.pbi.2004.03.014 (2004).Carrari, F. et al. Integrated analysis of metabolite and transcript levels reveals the metabolic shifts that underlie tomato fruit development and highlight regulatory aspects of metabolic network behavior. Plant Physiol. 142, 1380–1396. https://doi.org/10.1104/pp.106.088534 (2006).Mounet, F. et al. Gene and metabolite regulatory network analysis of early developing fruit tissues highlights new candidate genes for the control of tomato fruit composition and development. Plant Physiol. 149, 1505–1528. https://doi.org/10.1104/pp.108.133967 (2009).Ozga, J. A. & Reinecke, D. M. Hormonal interactions in fruit development. J. Plant Growth Regul. 22, 73–81. https://doi.org/10.1007/s00344-003-0024-9 (2003).Liu, S. Y. et al. Tomato AUXIN RESPONSE FACTOR 5 regulates fruit set and development via the mediation of auxin and gibberellin signaling. Sci. Rep. https://doi.org/10.1038/s41598-018-21315-y (2018).Serrani, J. C., Sanjuan, R., Ruiz-Rivero, O., Fos, M. & Garcia-Martinez, J. L. Gibberellin regulation of fruit set and growth in tomato. Plant Physiol. 145, 246–257. https://doi.org/10.1104/pp.107.098335 (2007).McAtee, P., Karim, S., Schaffer, R. & David, K. A dynamic interplay between phytohormones is required for fruit development, maturation, and ripening. Front. Plant Sci. https://doi.org/10.3389/fpls.2013.00079 (2013).Kataoka, K., Yashiro, Y., Habu, T., Sunamoto, K. & Kitajima, A. The addition of gibberellic acid to auxin solutions increases sugar accumulation and sink strength in developing auxin-induced parthenocarpic tomato fruits. Sci. Hortic. 123, 228–233. https://doi.org/10.1016/j.scienta.2009.09.001 (2009).Zhang, C. X., Tanabe, K., Tamura, F., Itai, A. & Yoshida, M. Roles of gibberellins in increasing sink demand in Japanese pear fruit during rapid fruit growth. Plant Growth Regul. 52, 161–172. https://doi.org/10.1007/s10725-007-9187-x (2007).Shinozaki, Y. et al. High-resolution spatiotemporal transcriptome mapping of tomato fruit development and ripening. Nat. Commun. https://doi.org/10.1038/s41467-017-02782-9 (2018).Ariizumi, T., Shinozaki, Y. & Ezura, H. Genes that influence yield in tomato. Breed. Sci. 63, 3–13. https://doi.org/10.1270/jsbbs.63.3 (2013).Azzi, L. et al. Fruit growth-related genes in tomato. J. Exp. Bot. 66, 1075–1086. https://doi.org/10.1093/jxb/eru527 (2015).Lemaire-Chamley, M. et al. Changes in transcriptional profiles are associated with early fruit tissue specialization in tomato. Plant Physiol. 139, 750–769. https://doi.org/10.1104/pp.105.063719 (2005).Tanksley, S. D. The genetic, developmental, and molecular bases of fruit size and shape variation in tomato. Plant Cell 16, S181–S189. https://doi.org/10.1105/tpc.018119 (2004).Allan, A. C. & Espley, R. V. MYBs drive novel consumer traits in fruits and vegetables. Trends Plant Sci. 23, 693–705. https://doi.org/10.1016/j.tplants.2018.06.001 (2018).Karlova, R. et al. Transcriptional control of fleshy fruit development and ripening. J. Exp. Bot. 65, 4527–4541. https://doi.org/10.1093/jxb/eru316 (2014).Rohrmann, J. et al. Combined transcription factor profiling, microarray analysis and metabolite profiling reveals the transcriptional control of metabolic shifts occurring during tomato fruit development. Plant J. 68, 999–1013. https://doi.org/10.1111/j.1365-313X.2011.04750.x (2011).Zhang, S. B. et al. Spatiotemporal transcriptome provides insights into early fruit development of tomato (Solanum lycopersicum). Sci. Rep. https://doi.org/10.1038/srep23173 (2016).Corrales, A. R. et al. Characterization of tomato Cycling Dof factors reveals conserved and new functions in the control of flowering time and abiotic stress responses. J. Exp. Bot. 65, 995–1012. https://doi.org/10.1093/jxb/ert451 (2014).Renau-Morata, B. et al. Ectopic Expression of CDF3 genes in tomato enhances biomass production and yield under salinity stress conditions. Front. Plant Sci. 8, 18. https://doi.org/10.3389/fpls.2017.00660 (2017).Guillet, C. et al. Regulation of the fruit-specific PEP carboxylase SlPPC2 promoter at early stages of tomato fruit development. PLoS ONE https://doi.org/10.1371/journal.pone.0036795 (2012).Bourdon, M. et al. Evidence for karyoplasmic homeostasis during endoreduplication and a ploidy-dependent increase in gene transcription during tomato fruit growth. Development 139, 3817–3826. https://doi.org/10.1242/dev.084053 (2012).de Jong, M. et al. Solanum lycopersicum AUXIN RESPONSE FACTOR 9 regulates cell division activity during early tomato fruit development. J Exp. Bot. 66, 3405–3416. https://doi.org/10.1093/jxb/erv152 (2015).Serrani, J. C., Fos, M., Atares, A. & Garcia-Martinez, J. L. Effect of gibberellin and auxin on parthenocarpic fruit growth induction in the cv micro-tom of tomato. J. Plant Growth Regul. 26, 211–221. https://doi.org/10.1007/s00344-007-9014-7 (2007).Srivastava, A. & Handa, A. K. Hormonal regulation of tomato fruit development: a molecular perspective. J. Plant Growth Regul. 24, 67–82. https://doi.org/10.1007/s00344-005-0015-0 (2005).Exposito-Rodriguez, M., Borges, A. A., Borges-Perez, A., Hernandez, M. & Perez, J. A. Cloning and biochemical characterization of ToFZY, a tomato gene encoding a flavin monooxygenase involved in a tryptophan-dependent auxin biosynthesis pathway. J. Plant Growth Regul. 26, 329–340. https://doi.org/10.1007/s00344-007-9019-2 (2007).Li, Z. M. et al. High invertase activity in tomato reproductive organs correlates with enhanced sucrose import into, and heat tolerance of, young fruit. J. Exp. Bot. 63, 1155–1166. https://doi.org/10.1093/jxb/err329 (2012).Wang, F., Sanz, A., Brenner, M. L. & Smith, A. Sucrose synthase, starch accumulation, and tomato fruit sink strength. Plant Physiol. 101, 321–327. https://doi.org/10.1104/pp.101.1.321 (1993).Pattison, R. J. et al. Comprehensive tissue-specific transcriptome analysis reveals distinct regulatory programs during early tomato fruit development. Plant Physiol. 168, 1684-U1002. https://doi.org/10.1104/pp.15.00287 (2015).Musseau, C. et al. Identification of two new mechanisms that regulate fruit growth by cell expansion in tomato. Front. Plant Sci. https://doi.org/10.3389/fpls.2017.00988 (2017).Shiota, H., Sudoh, T. & Tanaka, I. Expression analysis of genes encoding plasma membrane aquaporins during seed and fruit development in tomato. Plant Sci. 171, 277–285. https://doi.org/10.1016/j.plantsci.2006.03.021 (2006).Wang, L. et al. Ectopically expressing MdPIP1;3, an aquaporin gene, increased fruit size and enhanced drought tolerance of transgenic tomatoes. BMC Plant Biol. https://doi.org/10.1186/s12870-017-1212-2 (2017).Long, S. P., Zhu, X. G., Naidu, S. L. & Ort, D. R. Can improvement in photosynthesis increase crop yields?. Plant Cell Environ. 29, 315–330. https://doi.org/10.1111/j.1365-3040.2005.01493.x (2006).D’Aoust, M. A., Yelle, S. & Nguyen-Quoc, B. Antisense inhibition of tomato fruit sucrose synthase decreases fruit setting and the sucrose unloading capacity of young fruit. Plant Cell 11, 2407–2418. https://doi.org/10.1105/tpc.11.12.2407 (1999).Liu, T., Hu, Y. Q. & Li, X. X. Characterization of a chestnut FLORICAULA/LEAFY homologous gene. Afr. J. Biotechnol. 10, 3978–3985 (2011).Fridman, E., Carrari, F., Liu, Y. S., Fernie, A. R. & Zamir, D. Zooming in on a quantitative trait for tomato yield using interspecific introgressions. Science 305, 1786–1789. https://doi.org/10.1126/science.1101666 (2004).Ikeda, H. et al. Dynamic metabolic regulation by a chromosome segment from a wild relative during fruit development in a tomato introgression line, IL8-3. Plant Cell Physiol. 57, 1257–1270. https://doi.org/10.1093/pcp/pcw075 (2016).Ho, L. C. Partitioning of assimilates in fruiting tomato plants. Plant Growth Regul. 2, 277–285. https://doi.org/10.1007/bf00027287 (1984).Beauvoit, B. et al. Putting primary metabolism into perspective to obtain better fruits. Ann. Bot. 122, 1–21. https://doi.org/10.1093/aob/mcy057 (2018).Corrales, A. R. et al. Multifaceted role of cycling DOF factor 3 (CDF3) in the regulation of flowering time and abiotic stress responses in Arabidopsis. Plant Cell Environ. 40, 748–764. https://doi.org/10.1111/pce.12894 (2017).Carrari, F. & Fernie, A. R. Metabolic regulation underlying tomato fruit development. J. Exp. Bot. 57, 1883–1897. https://doi.org/10.1093/jxb/erj020 (2006).Osorio, S. et al. Alteration of the interconversion of pyruvate and malate in the plastid or cytosol of ripening tomato fruit invokes diverse consequences on sugar but similar effects on cellular organic acid, metabolism, and transitory starch accumulation. Plant Physiol. 161, 628–643. https://doi.org/10.1104/pp.112.211094 (2013).Gillaspy, G., Bendavid, H. & Gruissem, W. Fruits—a developmental perspective. Plant Cell 5, 1439–1451. https://doi.org/10.1105/tpc.5.10.1439 (1993).Carrera, E., Ruiz-Rivero, O., Peres, L. E. P., Atares, A. & Garcia-Martinez, J. L. Characterization of the procera tomato mutant shows novel functions of the SlDELLA protein in the control of flower morphology, cell division and expansion, and the auxin-signaling pathway during fruit-set and development. Plant Physiol. 160, 1581–1596. https://doi.org/10.1104/pp.112.204552 (2012).Chen, S. et al. Identification and characterization of tomato gibberellin 2-oxidases (GA2oxs) and effects of fruit-specific SlGA2ox1 overexpression on fruit and seed growth and development. Hortic. Res. https://doi.org/10.1038/hortres.2016.59 (2016).Mignolli, F., Vidoz, M. L., Picciarelli, P. & Mariotti, L. Gibberellins modulate auxin responses during tomato (Solanum lycopersicum L.) fruit development. Physiol. Plant. 165, 768–779. https://doi.org/10.1111/ppl.12770 (2019).De Jong, M., Wolters-Arts, M., Feron, R., Mariani, C. & Vriezen, W. H. The Solanum lycopersicum auxin response factor 7 (SlARF7) regulates auxin signaling during tomato fruit set and development. Plant J. 57, 160–170. https://doi.org/10.1111/j.1365-313X.2008.03671.x (2009).Ellul, P. et al. The ploidy level of transgenic plants in Agrobacterium-mediated transformation of tomato cotyledons (Lycopersicon esculentum L. Mill.) is genotype and procedure dependent. Theor. Appl. Genet. 106, 231–238. https://doi.org/10.1007/s00122-002-0928-y (2003).Renau-Morata, R. et al. The use of corms produced under storage at low temperatures as a source of explants for the in vitro propagation of saffron reduces contamination levels and increases multiplication rates. Ind. Crops Prod. 46, 97–104. https://doi.org/10.1016/j.indcrop.2013.01.013 (2013).Cebolla-Cornejo, J., Valcarcel, M., Herrero-Martinez, J. M., Rosello, S. & Nuez, F. High efficiency joint CZE determination of sugars and acids in vegetables and fruits. Electrophoresis 33, 2416–2423. https://doi.org/10.1002/elps.201100640 (2012).Nebauer, S. G. et al. Influence of crop load on the expression patterns of starch metabolism genes in alternate-bearing citrus trees. Plant Physiol. Biochem. 80, 105–113. https://doi.org/10.1016/j.plaphy.2014.03.032 (2014).Hoffman, N. E., Ko, K., Milkowski, D. & Pichersky, E. Isolation and characterization of tomato cDNA and genomic clones encoding the ubiquitin gene UBI3. Plant Mol. Biol. 17, 1189–1201. https://doi.org/10.1007/bf00028735 (1991).Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(−Delta Delta C) method. Methods 25, 402–408. https://doi.org/10.1006/meth.2001.1262 (2001).Miedes, E. & Lorences, E. P. Changes in cell wall pectin and pectinase activity in apple and tomato fruits during Penicillium expansum infection. J. Sci. Food Agric. 86, 1359–1364 (2006)

Ongoing Evolution in the Genus Crocus: Diversityof Flowering Strategies on the Way to Hysteranthy

[EN] Species of the genus Crocus are found over a wide range of climatic areas. In natural habitats, these geophytes diverge in the flowering strategies. This variability was assessed by analyzing the flowering traits of the Spanish collection of wild crocuses, preserved in the Bank of Plant Germplasm of Cuenca. Plants of the seven Spanish species were analyzed both in their natural environments (58 native populations) and in common garden experiments (112 accessions). Differences among species observed in the native habitats were maintained under uniform environmental conditions, suggesting a genetic basis for flowering mechanisms. Two eco-morphological types, autumn- and spring-flowering species, share similar patterns of floral induction and differentiation period in summer. The optimal temperature for this process was 23 degrees C for both types. Unlike Irano-Turanian crocuses, spring-flowering Spanish species do not require low winter temperatures for flower elongation. Hysteranthous crocuses flower in autumn prior to leaf elongation. We conclude that the variability in flowering traits in crocuses is related to the genetic and environmental regulation of flower primordia differentiation and elongation prior to emergence above the soil surface. The elucidation of the physiological differences between eco-morphological types of crocuses: synanthous with cold requirements and synanthous and hysteranthous without cold requirements, unlocks a new approach to the flowering evolution of geophytes in Mediterranean regions. Crocus species can serve both as a new model in the study of the molecular basis of hysteranthy and for the purposes of developing the molecular markers for desirable flowering traits.The collection activities of plant materials included in this study were mainly supported by successive Spanish research projects funded by the "Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria" [INIA RF2004-00032-C03, INIA RF2008-00012-C03, INIA RF2011-00005-C03], co-funded by the European Regional Development Fund (ERDF-FEDER), and also by means of the European Action 018 Agri Gen Res (CrocusBank). The activities of PhD. TPF were supported by a pre-doctoral grant from the "Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria" within the framework of the project INIA RF2011-0005-C03-01. The preservation of these materials in the facilities of the Bank of Plant Germplasm of Cuenca (CIAF Albaladejito -IRIAF), as part of the Spanish Germplasm Collection of Saffron and other Crocus, is currently supported by the Spanish National Program for Conservation and Utilization of Plant Genetic Resources for Agriculture and Food (action INIA RFP2014-00012). Some activities have been also funded by the "Ministerio de Ciencia, Innovacion y Universidades de Espana" [AGL2016-77078-R].Pastor-Férriz, T.; De-Los-Mozos-Pascual, M.; Renau-Morata, B.; Nebauer, SG.; Sanchís, E.; Busconi, M.; Fernández, J.... (2021). Ongoing Evolution in the Genus Crocus: Diversityof Flowering Strategies on the Way to Hysteranthy. Plants. 10(3):1-18. https://doi.org/10.3390/plants1003047711810

Multifaceted role of cycling DOF factor 3 (CDF3) in the regulation of flowering time and abiotic stress responses in Arabidopsis

[EN] DNA-binding with one finger (DOF)-type transcription factors are involved in many fundamental processes in higher plants, from responses to light and phytohormones to flowering time and seed maturation, but their relation with abiotic stress tolerance is largely unknown. Here, we identify the roles of CDF3, an Arabidopsis DOF gene in abiotic stress responses and developmental processes like flowering time. CDF3 is highly induced by drought, extreme temperatures and abscisic acid treatment. The CDF3 T-DNA insertion mutant cdf3-1 is much more sensitive to drought and low temperature stress, whereas CDF3 overexpression enhances the tolerance of transgenic plants to drought, cold and osmotic stress and promotes late flowering. Transcriptome analysis revealed that CDF3 regulates a set of genes involved in cellular osmoprotection and oxidative stress, including the stress tolerance transcription factors CBFs, DREB2A and ZAT12, which involve both gigantea-dependent and independent pathways. Consistently, metabolite profiling disclosed that the total amount of some protective metabolites including -aminobutyric acid, proline, glutamine and sucrose were higher in CDF3-overexpressing plants. Taken together, these results indicate that CDF3 plays a multifaceted role acting on both flowering time and abiotic stress tolerance, in part by controlling the CBF/DREB2A-CRT/DRE and ZAT10/12 modules.We thank Dr Pablo Gonzalez-Melendi and Dr Jan Zouhar for technical handling of the confocal microscope and Dr Rafael Catala for the assistance with the low temperature stress assays. This work was supported by grants from Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA; projects 2009-0004-C01, 2012-0008-C01), Spanish Ministry of Science and Innovation (projects BIO2010-1487, BFU2013-49665-EXP). A.R.C. and J.D.F. were supported by INIA pre-doctoral fellowshipsCorrales, AR.; Carrillo, L.; Lasierra, P.; Nebauer, SG.; Dominguez-Figueroa, J.; Renau-Morata, B.; Pollmann, S.... (2017). Multifaceted role of cycling DOF factor 3 (CDF3) in the regulation of flowering time and abiotic stress responses in Arabidopsis. Plant Cell & Environment. 40(5):748-764. https://doi.org/10.1111/pce.12894S748764405Achard, P., Gong, F., Cheminant, S., Alioua, M., Hedden, P., & Genschik, P. (2008). The Cold-Inducible CBF1 Factor–Dependent Signaling Pathway Modulates the Accumulation of the Growth-Repressing DELLA Proteins via Its Effect on Gibberellin Metabolism. The Plant Cell, 20(8), 2117-2129. doi:10.1105/tpc.108.058941Ahuja, I., de Vos, R. C. H., Bones, A. M., & Hall, R. D. (2010). Plant molecular stress responses face climate change. Trends in Plant Science, 15(12), 664-674. doi:10.1016/j.tplants.2010.08.002Alonso, R., Oñate-Sánchez, L., Weltmeier, F., Ehlert, A., Diaz, I., Dietrich, K., … Dröge-Laser, W. (2009). A Pivotal Role of the Basic Leucine Zipper Transcription Factor bZIP53 in the Regulation of Arabidopsis Seed Maturation Gene Expression Based on Heterodimerization and Protein Complex Formation. The Plant Cell, 21(6), 1747-1761. doi:10.1105/tpc.108.062968BEUVE, N., RISPAIL, N., LAINE, P., CLIQUET, J.-B., OURRY, A., & LE DEUNFF, E. (2004). Putative role of gamma -aminobutyric acid (GABA) as a long-distance signal in up-regulation of nitrate uptake in Brassica napus L. Plant, Cell and Environment, 27(8), 1035-1046. doi:10.1111/j.1365-3040.2004.01208.xBlümel, M., Dally, N., & Jung, C. (2015). Flowering time regulation in crops — what did we learn from Arabidopsis? Current Opinion in Biotechnology, 32, 121-129. doi:10.1016/j.copbio.2014.11.023Bouche, N., Fait, A., Bouchez, D., Moller, S. G., & Fromm, H. (2003). Mitochondrial succinic-semialdehyde dehydrogenase of the  -aminobutyrate shunt is required to restrict levels of reactive oxygen intermediates in plants. Proceedings of the National Academy of Sciences, 100(11), 6843-6848. doi:10.1073/pnas.1037532100Catala, R., Medina, J., & Salinas, J. (2011). Integration of low temperature and light signaling during cold acclimation response in Arabidopsis. Proceedings of the National Academy of Sciences, 108(39), 16475-16480. doi:10.1073/pnas.1107161108Chaves, M. M., Flexas, J., & Pinheiro, C. (2008). Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Annals of Botany, 103(4), 551-560. doi:10.1093/aob/mcn125Chen, H., Hwang, J. E., Lim, C. J., Kim, D. Y., Lee, S. Y., & Lim, C. O. (2010). Arabidopsis DREB2C functions as a transcriptional activator of HsfA3 during the heat stress response. Biochemical and Biophysical Research Communications, 401(2), 238-244. doi:10.1016/j.bbrc.2010.09.038Claussen, W. (2005). Proline as a measure of stress in tomato plants. Plant Science, 168(1), 241-248. doi:10.1016/j.plantsci.2004.07.039Clough, S. J., & Bent, A. F. (1998). Floral dip: a simplified method forAgrobacterium-mediated transformation ofArabidopsis thaliana. The Plant Journal, 16(6), 735-743. doi:10.1046/j.1365-313x.1998.00343.xCorrales, A., Carrillo, L., Nebauer, S., Renau-Morata, B., Sánchez-Perales, M., Fernández-Nohales, P., … Medina, J. (2014). Salinity Assay in Arabidopsis. BIO-PROTOCOL, 4(16). doi:10.21769/bioprotoc.1216Corrales, A.-R., Nebauer, S. G., Carrillo, L., Fernández-Nohales, P., Marqués, J., Renau-Morata, B., … Medina, J. (2014). Characterization of tomato Cycling Dof Factors reveals conserved and new functions in the control of flowering time and abiotic stress responses. Journal of Experimental Botany, 65(4), 995-1012. doi:10.1093/jxb/ert451Davletova, S., Schlauch, K., Coutu, J., & Mittler, R. (2005). The Zinc-Finger Protein Zat12 Plays a Central Role in Reactive Oxygen and Abiotic Stress Signaling in Arabidopsis. Plant Physiology, 139(2), 847-856. doi:10.1104/pp.105.068254DÉJARDIN, A., SOKOLOV, L. N., & KLECZKOWSKI, L. A. (1999). Sugar/osmoticum levels modulate differential abscisic acid-independent expression of two stress-responsive sucrose synthase genes in Arabidopsis. Biochemical Journal, 344(2), 503-509. doi:10.1042/bj3440503Dubois, M., Skirycz, A., Claeys, H., Maleux, K., Dhondt, S., De Bodt, S., … Inzé, D. (2013). ETHYLENE RESPONSE FACTOR6 Acts as a Central Regulator of Leaf Growth under Water-Limiting Conditions in Arabidopsis. Plant Physiology, 162(1), 319-332. doi:10.1104/pp.113.216341Farrant, J. M., & Moore, J. P. (2011). Programming desiccation-tolerance: from plants to seeds to resurrection plants. Current Opinion in Plant Biology, 14(3), 340-345. doi:10.1016/j.pbi.2011.03.018Fornara, F., Montaigu, A., Sánchez‐Villarreal, A., Takahashi, Y., Ver Loren van Themaat, E., Huettel, B., … Coupland, G. (2015). The GI – CDF module of Arabidopsis affects freezing tolerance and growth as well as flowering. The Plant Journal, 81(5), 695-706. doi:10.1111/tpj.12759Fornara, F., Panigrahi, K. C. S., Gissot, L., Sauerbrunn, N., Rühl, M., Jarillo, J. A., & Coupland, G. (2009). Arabidopsis DOF Transcription Factors Act Redundantly to Reduce CONSTANS Expression and Are Essential for a Photoperiodic Flowering Response. Developmental Cell, 17(1), 75-86. doi:10.1016/j.devcel.2009.06.015Fowler, S. (1999). GIGANTEA: a circadian clock-controlled gene that regulates photoperiodic flowering in Arabidopsis and encodes a protein with several possible membrane-spanning domains. The EMBO Journal, 18(17), 4679-4688. doi:10.1093/emboj/18.17.4679Galmés, J., Medrano, H., & Flexas, J. (2007). Photosynthetic limitations in response to water stress and recovery in Mediterranean plants with different growth forms. New Phytologist, 175(1), 81-93. doi:10.1111/j.1469-8137.2007.02087.xGill, S. S., & Tuteja, N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry, 48(12), 909-930. doi:10.1016/j.plaphy.2010.08.016Gilmour, S. J., Fowler, S. G., & Thomashow, M. F. (2004). Arabidopsis Transcriptional Activators CBF1, CBF2, and CBF3 have Matching Functional Activities. Plant Molecular Biology, 54(5), 767-781. doi:10.1023/b:plan.0000040902.06881.d4Gong, P., Zhang, J., Li, H., Yang, C., Zhang, C., Zhang, X., … Ye, Z. (2010). Transcriptional profiles of drought-responsive genes in modulating transcription signal transduction, and biochemical pathways in tomato. Journal of Experimental Botany, 61(13), 3563-3575. doi:10.1093/jxb/erq167Gould, P. D., Locke, J. C. W., Larue, C., Southern, M. M., Davis, S. J., Hanano, S., … Hall, A. (2006). The Molecular Basis of Temperature Compensation in the Arabidopsis Circadian Clock. The Plant Cell, 18(5), 1177-1187. doi:10.1105/tpc.105.039990Han, Q., Kang, G., & Guo, T. (2013). Proteomic analysis of spring freeze-stress responsive proteins in leaves of bread wheat (Triticum aestivum L.). Plant Physiology and Biochemistry, 63, 236-244. doi:10.1016/j.plaphy.2012.12.002Hernando-Amado, S., González-Calle, V., Carbonero, P., & Barrero-Sicilia, C. (2012). The family of DOF transcription factors in Brachypodium distachyon: phylogenetic comparison with rice and barley DOFs and expression profiling. BMC Plant Biology, 12(1), 202. doi:10.1186/1471-2229-12-202Zou, H.-F., Zhang, Y.-Q., Wei, W., Chen, H.-W., Song, Q.-X., Liu, Y.-F., … Chen, S.-Y. (2012). The transcription factor AtDOF4.2 regulates shoot branching and seed coat formation in Arabidopsis. Biochemical Journal, 449(2), 373-388. doi:10.1042/bj20110060Hussain, S. S., Kayani, M. A., & Amjad, M. (2011). Transcription factors as tools to engineer enhanced drought stress tolerance in plants. Biotechnology Progress, 27(2), 297-306. doi:10.1002/btpr.514Imaizumi, T. (2005). FKF1 F-Box Protein Mediates Cyclic Degradation of a Repressor of CONSTANS in Arabidopsis. Science, 309(5732), 293-297. doi:10.1126/science.1110586Ingram, J., & Bartels, D. (1996). THE MOLECULAR BASIS OF DEHYDRATION TOLERANCE IN PLANTS. Annual Review of Plant Physiology and Plant Molecular Biology, 47(1), 377-403. doi:10.1146/annurev.arplant.47.1.377Jarillo, J. A., Del Olmo, I., Gómez-Zambrano, A., Lázaro, A., López-González, L., Miguel, E., … Piñeiro, M. (2008). Photoperiodic control of flowering time: a review. Spanish Journal of Agricultural Research, 6(S1), 221. doi:10.5424/sjar/200806s1-391Izaurralde, E. (1997). The asymmetric distribution of the constituents of the Ran system is essential for transport into and out of the nucleus. The EMBO Journal, 16(21), 6535-6547. doi:10.1093/emboj/16.21.6535Karimi, M., Depicker, A., & Hilson, P. (2007). Recombinational Cloning with Plant Gateway Vectors. Plant Physiology, 145(4), 1144-1154. doi:10.1104/pp.107.106989Kim, S. Y., & Nam, K. H. (2010). Physiological roles of ERD10 in abiotic stresses and seed germination of Arabidopsis. Plant Cell Reports, 29(2), 203-209. doi:10.1007/s00299-009-0813-0Kiyosue, T., Yamaguchi-Shinozaki, K., & Shinozaki, K. (1994). Cloning of cDNAs for genes that are early-responsive to dehydration stress (ERDs) inArabidopsis thaliana L.: identification of three ERDs as HSP cognate genes. Plant Molecular Biology, 25(5), 791-798. doi:10.1007/bf00028874Kurai, T., Wakayama, M., Abiko, T., Yanagisawa, S., Aoki, N., & Ohsugi, R. (2011). Introduction of the ZmDof1 gene into rice enhances carbon and nitrogen assimilation under low-nitrogen conditions. Plant Biotechnology Journal, 9(8), 826-837. doi:10.1111/j.1467-7652.2011.00592.xLiu, Q., Kasuga, M., Sakuma, Y., Abe, H., Miura, S., Yamaguchi-Shinozaki, K., & Shinozaki, K. (1998). Two Transcription Factors, DREB1 and DREB2, with an EREBP/AP2 DNA Binding Domain Separate Two Cellular Signal Transduction Pathways in Drought- and Low-Temperature-Responsive Gene Expression, Respectively, in Arabidopsis. The Plant Cell, 10(8), 1391. doi:10.2307/3870648Matsui, A., Ishida, J., Morosawa, T., Mochizuki, Y., Kaminuma, E., Endo, T. A., … Seki, M. (2008). Arabidopsis Transcriptome Analysis under Drought, Cold, High-Salinity and ABA Treatment Conditions using a Tiling Array. Plant and Cell Physiology, 49(8), 1135-1149. doi:10.1093/pcp/pcn101Medina, J., Bargues, M., Terol, J., Pérez-Alonso, M., & Salinas, J. (1999). The Arabidopsis CBF Gene Family Is Composed of Three Genes Encoding AP2 Domain-Containing Proteins Whose Expression Is Regulated by Low Temperature but Not by Abscisic Acid or Dehydration. Plant Physiology, 119(2), 463-470. doi:10.1104/pp.119.2.463Messerli, G., Partovi Nia, V., Trevisan, M., Kolbe, A., Schauer, N., Geigenberger, P., … Zeeman, S. C. (2007). Rapid Classification of Phenotypic Mutants of Arabidopsis via Metabolite Fingerprinting. Plant Physiology, 143(4), 1484-1492. doi:10.1104/pp.106.090795Mittler, R. (2006). Abiotic stress, the field environment and stress combination. Trends in Plant Science, 11(1), 15-19. doi:10.1016/j.tplants.2005.11.002Mizoguchi, T., Wright, L., Fujiwara, S., Cremer, F., Lee, K., Onouchi, H., … Coupland, G. (2005). Distinct Roles of GIGANTEA in Promoting Flowering and Regulating Circadian Rhythms in Arabidopsis. The Plant Cell, 17(8), 2255-2270. doi:10.1105/tpc.105.033464Munns, R., & Tester, M. (2008). Mechanisms of Salinity Tolerance. Annual Review of Plant Biology, 59(1), 651-681. doi:10.1146/annurev.arplant.59.032607.092911Murashige, T., & Skoog, F. (1962). A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum, 15(3), 473-497. doi:10.1111/j.1399-3054.1962.tb08052.xNishiyama, R., Le, D. T., Watanabe, Y., Matsui, A., Tanaka, M., Seki, M., … Tran, L.-S. P. (2012). Transcriptome Analyses of a Salt-Tolerant Cytokinin-Deficient Mutant Reveal Differential Regulation of Salt Stress Response by Cytokinin Deficiency. PLoS ONE, 7(2), e32124. doi:10.1371/journal.pone.0032124Noguero, M., Atif, R. M., Ochatt, S., & Thompson, R. D. (2013). The role of the DNA-binding One Zinc Finger (DOF) transcription factor family in plants. Plant Science, 209, 32-45. doi:10.1016/j.plantsci.2013.03.016Novillo, F., Medina, J., & Salinas, J. (2007). Arabidopsis CBF1 and CBF3 have a different function than CBF2 in cold acclimation and define different gene classes in the CBF regulon. Proceedings of the National Academy of Sciences, 104(52), 21002-21007. doi:10.1073/pnas.0705639105OliverosJ.C.(2007)Venny an interactive tool for comparing lists with Venn's diagrams.http://bioinfogp.cnb.csic.es/tools/venny/index.html.Oñate-Sánchez, L., & Vicente-Carbajosa, J. (2008). DNA-free RNA isolation protocols for Arabidopsis thaliana, including seeds and siliques. BMC Research Notes, 1(1), 93. doi:10.1186/1756-0500-1-93Osakabe, Y., Kajita, S., & Osakabe, K. (2011). Genetic engineering of woody plants: current and future targets in a stressful environment. Physiologia Plantarum, 142(2), 105-117. doi:10.1111/j.1399-3054.2011.01451.xPark, D. H. (1999). Control of Circadian Rhythms and Photoperiodic Flowering by the Arabidopsis GIGANTEA Gene. Science, 285(5433), 1579-1582. doi:10.1126/science.285.5433.1579Rajasekaran, L. R., Aspinall, D., & Paleg, L. G. (2000). Physiological mechanism of tolerance of Lycopersicon spp. exposed to salt stress. Canadian Journal of Plant Science, 80(1), 151-159. doi:10.4141/p99-003Rizhsky, L., Liang, H., Shuman, J., Shulaev, V., Davletova, S., & Mittler, R. (2004). When Defense Pathways Collide. The Response of Arabidopsis to a Combination of Drought and Heat Stress. Plant Physiology, 134(4), 1683-1696. doi:10.1104/pp.103.033431Rosso, M. G., Li, Y., Strizhov, N., Reiss, B., Dekker, K., & Weisshaar, B. (2003). An Arabidopsis thaliana T-DNA mutagenized population (GABI-Kat) for flanking sequence tag-based reverse genetics. Plant Molecular Biology, 53(1/2), 247-259. doi:10.1023/b:plan.0000009297.37235.4aRueda-López, M., Crespillo, R., Cánovas, F. M., & Ávila, C. (2008). Differential regulation of two glutamine synthetase genes by a single Dof transcription factor. The Plant Journal, 56(1), 73-85. doi:10.1111/j.1365-313x.2008.03573.xSakuma, Y., Maruyama, K., Osakabe, Y., Qin, F., Seki, M., Shinozaki, K., & Yamaguchi-Shinozaki, K. (2006). Functional Analysis of an Arabidopsis Transcription Factor, DREB2A, Involved in Drought-Responsive Gene Expression. The Plant Cell, 18(5), 1292-1309. doi:10.1105/tpc.105.035881Sato, Y., & Yokoya, S. (2007). Enhanced tolerance to drought stress in transgenic rice plants overexpressing a small heat-shock protein, sHSP17.7. Plant Cell Reports, 27(2), 329-334. doi:10.1007/s00299-007-0470-0Sawa, M., Nusinow, D. A., Kay, S. A., & Imaizumi, T. (2007). FKF1 and GIGANTEA Complex Formation Is Required for Day-Length Measurement in Arabidopsis. Science, 318(5848), 261-265. doi:10.1126/science.1146994Scarpeci, T. E., Zanor, M. I., Mueller-Roeber, B., & Valle, E. M. (2013). Overexpression of AtWRKY30 enhances abiotic stress tolerance during early growth stages in Arabidopsis thaliana. Plant Molecular Biology, 83(3), 265-277. doi:10.1007/s11103-013-0090-8Seki, M., Kamei, A., Yamaguchi-Shinozaki, K., & Shinozaki, K. (2003). Molecular responses to drought, salinity and frost: common and different paths for plant protection. Current Opinion in Biotechnology, 14(2), 194-199. doi:10.1016/s0958-1669(03)00030-2Shelp, B. J., Mullen, R. T., & Waller, J. C. (2012). Compartmentation of GABA metabolism raises intriguing questions. Trends in Plant Science, 17(2), 57-59. doi:10.1016/j.tplants.2011.12.006Shi, H., & Chan, Z. (2014). The cysteine2/histidine2-type transcription factor ZINC FINGER OF ARABIDOPSIS THALIANA 6 -activated C-REPEAT-BINDING FACTOR pathway is essential for melatonin-mediated freezing stress resistance in Arabidopsis. Journal of Pineal Research, 57(2), 185-191. doi:10.1111/jpi.12155Shinozaki, K., & Yamaguchi-Shinozaki, K. (2006). Gene networks involved in drought stress response and tolerance. Journal of Experimental Botany, 58(2), 221-227. doi:10.1093/jxb/erl164Shinozaki, K., Yamaguchi-Shinozaki, K., & Seki, M. (2003). Regulatory network of gene expression in the drought and cold stress responses. Current Opinion in Plant Biology, 6(5), 410-417. doi:10.1016/s1369-5266(03)00092-xSkirycz, A., & Inzé, D. (2010). More from less: plant growth under limited water. Current Opinion in Biotechnology, 21(2), 197-203. doi:10.1016/j.copbio.2010.03.002Snedden, W. A., Arazi, T., Fromm, H., & Shelp, B. J. (1995). Calcium/Calmodulin Activation of Soybean Glutamate Decarboxylase. Plant Physiology, 108(2), 543-549. doi:10.1104/pp.108.2.543STITT, M., & KRAPP, A. (1999). The interaction between elevated carbon dioxide and nitrogen nutrition: the physiological and molecular background. Plant, Cell and Environment, 22(6), 583-621. doi:10.1046/j.1365-3040.1999.00386.xStudart-Guimarães, C., Fait, A., Nunes-Nesi, A., Carrari, F., Usadel, B., & Fernie, A. R. (2007). Reduced Expression of Succinyl-Coenzyme A Ligase Can Be Compensated for by Up-Regulation of the γ-Aminobutyrate Shunt in Illuminated Tomato Leaves. Plant Physiology, 145(3), 626-639. doi:10.1104/pp.107.103101Supek, F., Bošnjak, M., Škunca, N., & Šmuc, T. (2011). REVIGO Summarizes and Visualizes Long Lists of Gene Ontology Terms. PLoS ONE, 6(7), e21800. doi:10.1371/journal.pone.0021800Suzuki, M., Kao, C.-Y., Cocciolone, S., & McCarty, D. R. (2002). Maize VP1 complements Arabidopsisabi3 and confers a novel ABA/auxin interaction in roots. The Plant Journal, 28(4), 409-418. doi:10.1046/j.1365-313x.2001.01165.xTaji, T., Ohsumi, C., Iuchi, S., Seki, M., Kasuga, M., Kobayashi, M., … Shinozaki, K. (2002). Important roles of drought- and cold-inducible genes for galactinol synthase in stress tolerance in Arabidopsis thaliana. The Plant Journal, 29(4), 417-426. doi:10.1046/j.0960-7412.2001.01227.xThomashow, M. F. (2010). Molecular Basis of Plant Cold Acclimation: Insights Gained from Studying the CBF Cold Response Pathway: Figure 1. Plant Physiology, 154(2), 571-577. doi:10.1104/pp.110.161794Toufighi, K., Brady, S. M., Austin, R., Ly, E., & Provart, N. J. (2005). The Botany Array Resource: e-Northerns, Expression Angling, and promoter analyses. The Plant Journal, 43(1), 153-163. doi:10.1111/j.1365-313x.2005.02437.xVogel, J. T., Zarka, D. G., Van Buskirk, H. A., Fowler, S. G., & Thomashow, M. F. (2004). Roles of the CBF2 and ZAT12 transcription factors in configuring the low temperature transcriptome of Arabidopsis. The Plant Journal, 41(2), 195-211. doi:10.1111/j.1365-313x.2004.02288.xWang, W., Vinocur, B., Shoseyov, O., & Altman, A. (2004). Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends in Plant Science, 9(5), 244-252. doi:10.1016/j.tplants.2004.03.006Yamaguchi-Shinozaki, K., & Shinozaki, K. (2006). TRANSCRIPTIONAL REGULATORY NETWORKS IN CELLULAR RESPONSES AND TOLERANCE TO DEHYDRATION AND COLD STRESSES. Annual Review of Plant Biology, 57(1), 781-803. doi:10.1146/annurev.arplant.57.032905.105444Yanagisawa, S. (2001). The Transcriptional Activation Domain of the Plant-Specific Dof1 Factor Functions in Plant, Animal, and Yeast Cells. Plant and Cell Physiology, 42(8), 813-822. doi:10.1093/pcp/pce105Yanagisawa, S. (2002). The Dof family of plant transcription factors. Trends in Plant Science, 7(12), 555-560. doi:10.1016/s1360-1385(02)02362-2Yanagisawa, S., Akiyama, A., Kisaka, H., Uchimiya, H., & Miwa, T. (2004). Metabolic engineering with Dof1 transcription factor in plants: Improved nitrogen assimilation and growth under low-nitrogen conditions. Proceedings of the National Academy of Sciences, 101(20), 7833-7838. doi:10.1073/pnas.0402267101Yanagisawa, S., & Schmidt, R. J. (1999). Diversity and similarity among recognition sequences of Dof transcription factors. The Plant Journal, 17(2), 209-214. doi:10.1046/j.1365-313x.1999.00363.xYanagisawa, S., & Sheen, J. (1998). Involvement of Maize Dof Zinc Finger Proteins in Tissue-Specific and Light-Regulated Gene Expression. The Plant Cell, 10(1), 75-89. doi:10.1105/tpc.10.1.75Yang, X., Srivastava, R., Howell, S. H., & Bassham, D. C. (2015). Activation of autophagy by unfolded proteins d

Arbuscular mycorrhizal symbiosis modulates the apocarotenoid biosynthetic pathway in saffron

[EN] Crocus sativus L. (saffron) has been propagated for millennia to produce the precious spice saffron from the red stigmas. The inebriant organoleptic and bioactive properties mainly depend on the content of crocins (dyeing capacity), picrocrocin (flavor), and safranal (aroma), apocarotenoids deriving from zeaxanthin. In this study, an integrated biochemical and molecular analysis was carried out on fresh saffron stigmas to investigate the in-fluence exerted by the arbuscular mycorrhizal fungus (AMF) Rhizophagus intraradices on the production of the main saffron apocarotenoids responsible for the properties of the spice. Since mineral enrichment due to AM symbiosis has been related to changes in the secondary metabolism of plants, the mineral content of saffron corms at flowering was also analyzed. Rare arbuscules (AMF trade structures) were found in mycorrhized plants. However, the expression of D27, CCD7, and NCED involved in the synthesis of strigolactones (SLs) and abscisic acid (ABA), which promote AM symbiosis, did not change in the stigmas. The transcription of beta-LYC and CCD4a/ b was not affected by AMF, whereas that of CCD2, which encodes the key enzyme producing major apocar-otenoids, was upregulated. The crocin content was reduced in treated plants even if the expression of ALDH, UGT74AD1, and UGT91P3, involved in crocin synthesis, did not change. Conversely, UGT709G1, implicated in picrocrocin synthesis, was overexpressed in the inoculated plants, thus the safranal content was increased in the spice.This research was funded by the program Interreg V-A Francia Italia Alcotra (Grant No. 1139 "ANTEA - Attivita innovative per lo sviluppo della filiera transfrontaliera del fiore edule"; and grant no. 8336 "ANTES-Fiori eduli e piante aromatiche: attivita capitalizzazione dei progetti ANTEA ed ESSICA").Stelluti, S.; Grasso, G.; Nebauer, SG.; Alonso, GL.; Renau-Morata, B.; Caser, M.; Demasi, S.... (2024). Arbuscular mycorrhizal symbiosis modulates the apocarotenoid biosynthetic pathway in saffron. Scientia Horticulturae. 323. https://doi.org/10.1016/j.scienta.2023.11244132

New Insights about How to Make an Intervention in Children and Adolescents with Metabolic Syndrome: Diet, Exercise vs. Changes in Body Composition. A Systematic Review of RCT

Objective: To record which interventions produce the greatest variations in body composition in patients ≤19 years old with metabolic syndrome (MS). Method: search dates between 2005 and 2017 in peer reviewed journals, following the PRISMA method (Preferred Reporting Items for Systematic reviews and Meta-Analyses). The selection criteria were: diagnostic for MS or at least a criterion for diagnosis; randomized clinical trials, ≤19 years of age; intervention programs that use diet and/or exercise as a tool (interventions showing an interest in body composition). Results: 1781 clinical trials were identified under these criteria but only 0.51% were included. The most frequent characteristics of the selected clinical trials were that they used multidisciplinary interventions and were carried out in America. The most utilized parameters were BMI (body mass index) in kg/m2 and BW (body weight) in kg. Conclusions: Most of the clinical trials included had been diagnosed through at least 2 diagnostic criteria for MS. Multidisciplinary interventions obtained greater changes in body composition in patients with MS. This change was especially prevalent in the combinations of dietary interventions and physical exercise. It is proposed to follow the guidelines proposed for patients who are overweight, obese, or have diabetes type 2, and extrapolate these strategies as recommendations for future clinical trials designed for patients with MS.This research is partially supported by Generalitat Valenciana, Grant GV/2017/177

Xylem anatomical study in diverse Capsicum sp. accessions, implication to drought tolerance

Drought is a limiting factor for plant survival and food production. Plants have developed various strategies in response to this stress. The immediate response to drought is the closure of the stomata. However, it prevents the transpiration and the photosynthesis. From the point of view of crops, it is interesting the ability of the plants to keep photosynthesizing under drought. Leaf water content is affected by the difference among leaf and soil water potentials (Ψleaf-Ψh) and by the xylem-specific hydraulic conductivity (Ks). It has been hypothesized that the number and diameter of the xylem vessels would affect the drought tolerance. In this experiment the influence of the xylem anatomy in the tolerance to water stress of three Capsicum sp accessions: Bol 58, Piquillo, Chimayo was studied. Plants were grown in plastic pots filled with soil:sand mixture in a glasshouse at UPV, during spring season. Two different treatments were used in the assay: normal irrigation and restricted irrigation (50% less irrigated than the control). Three and a half months after transplantation plants were harvested; roots, stems and fruits were weighted. Portions of the stem bases were fixed in FPA and rehydrated for the analysis of the xylem total area. Material was sectioned with a freezing microtome and dyed with phloroglucinol. Vessel element area was calculated with Autocad software. There were significant differences of biomass among genotypes and treatments. Piquillo showed lowest reduction of biomass under stress. The genotypes studied showed differences in the xylem structure. In addition, there were changes in the vessel diameter pattern from control to drought conditions. Generally, plants grown under water deficit reduce the number of bigger vessels (>7500µm2). Chimayo was the accession with bigger vessels, in normal and stress conditions, whereas Piquillo had smaller vessels. Therefore results showed a negative correlation between vessel size and production under stress conditions. Big vessels are prone to cavitation and a high proportion of small vessel may contribute to maintain Ks without collapse risk.Guijarro Real, C.; Molina Romero, RV.; Pérez Domingo, T.; Ribes Moya, AM.; Rodríguez Burruezo, A.; Fita, A. (2014). Xylem anatomical study in diverse Capsicum sp. accessions, implication to drought tolerance. Bulletin of University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca : Horticulture. 71(2):256-260. doi:10.15835/buasvmcn-hort:1067625626071