Gibberellins Regulate Ovule Integument Development by Interfering with the Transcription Factor ATS

[EN] Gibberellins (GAs) are plant hormones that regulate most plant life cycle aspects, including flowering and fruit development. Here, we demonstrate the implication of GAs in ovule development. DELLA proteins, negative GA response regulators, act as positive factors for ovule integument development in a mechanism that involves transcription factor ABERRANT TESTA SHAPE (ATS). The seeds of the della global mutant, a complete loss-of-function of DELLA, and the ats-1 mutant are remarkably similar, with a round shape, a disorganized testa, and viviparism. These defects are the result of an alteration in integuments that fail to fully develop and are shorter than in wild-type plants. ats-1 also shows some GA-related phenotypes, for example, higher germination rates and early flowering. In fact, ats-1 has elevated GA levels due to the activation of GA biosynthesis genes, which indicates that ATS inhibits GA biosynthesis. Moreover, DELLAs and ATS proteins interact, which suggests the formation of a transcriptional complex that regulates the expression of genes involved in integument growth. Therefore, the repression of GA biosynthesis by ATS would result in the stabilization of DELLAs to ensure correct ATS-DELLA complex formation. The requirement of both activities to coordinate proper ovule development strongly argues that the ATS-DELLA complex acts as a key molecular factor. This work provides the first evidence for a role of GAs in ovule and seed development.This work was supported by grants BIO2011-26302 and BIO2014-55946 from the Spanish Ministry of Science and Innovation and the Spanish Ministry of Economy and Competitiveness, respectively, and ACOMP/2013/048 and ACOMP/2014/106 from the Generalitat Valenciana to M.A.P.-A. R.S. received a PhD fellowship from the Spanish Ministry of Science and Innovation.Gómez Jiménez, MD.; Ventimilla-Llora, D.; Sacristán Tarrazó, R.; Perez Amador, MA. (2016). Gibberellins Regulate Ovule Integument Development by Interfering with the Transcription Factor ATS. Plant Physiology. 172(4):2403-2415. doi:10.1104/pp.16.01231S24032415172

RGL2 controls flower development, ovule number and fertility in Arabidopsis

[EN] DELLA proteins are a group of plant specific GRAS proteins of transcriptional regulators that have a key role in gibberellin (GA) signaling. In Arabidopsis, the DELLA family is formed by five members. The complexity of this gene family raises the question on whether single DELLA proteins have specific or overlapping functions in the control of several GA-dependent developmental processes. To better understand the roles played by RGL2, one of the DELLA proteins in Arabidopsis, two transgenic lines that express fusion proteins of Venus-RGL2 and a dominant version of RGL2, YPet-rgl2A17, were generated by recombineering strategy using a genomic clone that contained the RGL2 gene. The dominant YPet-rg12 Delta 17 protein is not degraded by GAs, and therefore it blocks the RGL2-dependent GA signaling and hence RGL2-dependent development. The RGL2 role in seed germination was further confirmed using these genetic tools, while new functions of RGL2 in plant development were uncovered. RGL2 has a clear function in the regulation of flower development, particularly stamen growth and anther dehiscence, which has a great impact in fertility. Moreover, the increased ovule number in the YPet-rg12 Delta 17 line points out the role of RGL2 in the determination of ovule number.We wish to thank Ms. J. Yun,M.A. Argomániz for technical assistance, and the IBMCP microscopy facility. Edit Syndicate (http://www.editsyndicate.com/) provided proofreading of the manuscript. This work was supported by grants from the Spanish Ministry of Economy and Competitiveness-FEDER [BI02011-26302 and BI02014-55946] and Generalitat Valenciana [ACOMP/2013/048 and ACOMP/2014/106] to M.A.P-A. and National Science Foundation [MCB-0923727] to J.M.A. MAP-A. received a fellowship of the 'Salvador de Madariaga' program from Spanish Ministry of Science and Innovation.Gómez Jiménez, MD.; Fuster Almunia, C.; Ocaña-Cuesta, J.; Alonso, J.; Perez Amador, MA. (2019). RGL2 controls flower development, ovule number and fertility in Arabidopsis. Plant Science. 281:82-92. https://doi.org/10.1016/j.plantsci.2019.01.014S829228

Molecular program of senescence in dry and fleshy fruit

[EN] Fruits of angiosperms can be divided into dry and fleshy fruits, depending on their dispersal strategies. Despite their apparently different developmental programmes, researchers have attempted to compare dry and fleshy fruits to establish analogies of the distinct biochemical and physiological processes that occur. But what are the common and specific phenomena in both biological strategies? Is valve dehiscence and senescence of dry fruits comparable to final ripening of fleshy fruits, when seeds become mature and fruits are competent for seed dispersal, or to over-ripening when advanced senescence occurs? We briefly review current knowledge on dry and fleshy fruit development, which has been extensively reported recently, and is the topic of this special issue. We compare the processes taking place in Arabidopsis (dry) and tomato (fleshy) fruit during final development steps using transcriptome data to establish possible analogies. Interestingly, the transcriptomic programme of Arabidopsis silique shares little similarity in gene number to tomato fruit ripening or over-ripening. In contrast, the biological processes carried out by these common genes from ripening and over-ripening programmes are similar, as most biological processes are shared during both programmes. On the other hand, several biological terms are specific of Arabidopsis and tomato ripening, including senescence, but little or no specific processes occur during Arabidopsis and tomato over-ripening. These suggest a closer analogy between silique senescence and ripening than over-ripening, but a major common biological programme between Arabidopsis silique senescence and the last steps of tomato development, irrespective of its distinction between ripening and over-ripening.We wish to thank Dr J. Carbonell for critically reading the manuscript. We also thank Clara Pons for the preliminary tomato microarray data mining. Our work has been supported by grants BIO2008-01039 and BIO2011-26302 from the Spanish Ministry of Science and Innovation and ACOMP/2010/079, ACOMP/2011/287, and ACOMP/2013/048 from the Generalitat Valenciana.Gómez Jiménez, MD.; Vera Sirera, FJ.; Perez Amador, MA. (2014). Molecular program of senescence in dry and fleshy fruit. Journal of Experimental Botany. 65(16):4515-4526. https://doi.org/10.1093/jxb/eru093S45154526651

Role of the gibberellin receptors GID1 during fruit-set in Arabidopsis

[EN] Gibberellins (GAs) play a critical role in fruit-set and fruit growth. Gibberellin is perceived by its nuclear receptors GA INSENSITIVE DWARF1s (GID1s), which then trigger degradation of downstream repressors DELLAs. To understand the role of the three GA receptor genes (GID1A, GID1B and GID1C) in Arabidopsis during fruit initiation, we have examined their temporal and spatial localization, in combination with analysis of mutant phenotypes. Distinct expression patterns are revealed for each GID1: GID1A is expressed throughout the whole pistil, while GID1B is expressed in ovules, and GID1C is expressed in valves. Functional study of gid1 mutant combinations confirms that GID1A plays a major role during fruit-set and growth, whereas GID1B and GID1C have specific roles in seed development and pod elongation, respectively. Therefore, in ovules, GA perception is mediated by GID1A and GID1B, while GID1A and GID1C are involved in GA perception in valves. To identify tissue-specific interactions between GID1s and DELLAs, we analyzed spatial expression patterns of four DELLA genes that have a role in fruit initiation (GAI, RGA, RGL1 and RGL2). Our data suggest that GID1A can interact with RGA and GAI in all tissues, whereas GID1C-RGL1 and GID1B-RGL2 interactions only occur in valves and ovules, respectively. These results uncover specific functions of each GID1-DELLA in the different GA-dependent processes that occur upon fruit-set. In addition, the distribution of GA receptors in valves along with lack of expression of GA biosynthesis genes in this tissue, strongly suggests transport of GAs from the developing seeds to promote fruit growth.We wish to thank Dr Masatoshi Nakajima (University of Tokyo, Japan) for providing the pGID1:GID1-GUS lines, and Dr Peter Hedden (Rothamsted Research, UK) for the pGA20ox:GA20ox-GUS lines. We also thank Ms C. Fuster and M. A. Argomaniz for technical assistance. This work has been supported by grants BIO2008-01039 and BIO2011-26302 from the Spanish Ministry of Science and Innovation and ACOMP/2010/079 and ACOMP/2011/287 from the Generalitat Valenciana for M. A. P.-A. and USDA grants 2010-65116-20460 and 2014-67013-21548 for T. P. S. C. G.-G. received a JAE PhD fellowship from the Spanish Council for Scientific Research (CSIC).Gallego Giraldo, C.; Hu, J.; Urbez Lagunas, C.; Gómez Jiménez, MD.; Sun, TP.; Perez Amador, MA. (2014). Role of the gibberellin receptors GID1 during fruit-set in Arabidopsis. Plant Journal. 79(6):1020-1032. doi:10.1111/tpj.12603S1020103279

PsPMEP, a pollen specific pectin methylesterase of pea (Pisum sativum L.)

[EN] Pectin methylesterases (PMEs) are a family of enzymes involved in plant reproductive processes such as pollen development and pollen tube growth. We have isolated and characterized PsPMEP, a pea (Pisum sativum L.) pollen-specific gene that encodes a protein with homology to PMEs. Sequence analysis showed that PsPMEP belongs to group 2 PMEs, which are characterized by the presence of a processable amino-terminal PME inhibitor domain followed by the catalytic PME domain. Moreover, PsPMEP contains several motifs highly conserved among PMEs with the essential amino acid residues involved in enzyme substrate binding and catalysis. Northern blot and in situ hybridization analyses showed that PsPMEP is expressed in pollen grains from 4 days before anthesis till anther dehiscence and in pollinated carpels. In the PsPMEP promoter region, we have identified several conserved cis-regulatory elements that have been associated with gene pollen-specific expression. Expression analysis of PsPMEP promoter fused to the uidA reporter gene in Arabidopsis thaliana plants showed a similar expression pattern when compared with pea, indicating that this promoter is also functional in a non-leguminous plant. GUS expression was detected in mature pollen grains, during pollen germination, during pollen tube elongation along the transmitting tract, and when the pollen tube reaches the embryo sac in the ovule.This work was funded by grants BIO2006-09374 and BIO2009-08134 from the Spanish Ministry of Science and Innovation (MICINN). The collaboration and assistance of Julia Marin-Navarro in the catalytic activity assays of PsPMEP in yeast and Rafael Martinez-Pardo in the greenhouse is gratefully acknowledged. We would like to thank the HAPRECI consortium (COST Action FA0903) to bring us the opportunity to collaborate with other European research groups working in the field of Plant Reproduction and to select our manuscript to be published in this special issue.Gómez Jiménez, MD.; Renau Morata, B.; Roque Mesa, EM.; Polaina, J.; Beltran Porter, JP.; Cañas Clemente, LA. (2013). PsPMEP, a pollen specific pectin methylesterase of pea (Pisum sativum L.). Plant Reproduction. 26(3):245-254. https://doi.org/10.1007/s00497-013-0220-0S245254263Bate N, Twell D (1998) Functional architecture of a late pollen promoter: pollen specific transcription is developmentally regulated by multiple stage-specific and co-dependent activator elements. Plant Mol Biol 37:859–869Bechtold N, Ellis J, Pelletier G (1993) In planta Agrobacterium-mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. C R Acad Sci 316:1194–1199Bosch M, Hepler PK (2005) Pectin methylesterases and pectin dynamics in pollen tubes. Plant Cell 17:3219–3226Bosch M, Hepler PK (2006) Silencing of the tobacco pollen pectin methylesterase NtPPME1 results in retarded in vivo pollen tube growth. Planta 233:736–745Camardella L (2000) Kiwi protein inhibitor of pectin methylesterase–amino-acid sequence and structural importance of two disulfide bridges. Eur J Biochem 267:4561–4565Carpenter JL, Ploense SE, Snustad DP, Silflow CD (1992) Preferential expression of an alpha-tubulin gene of Arabidopsis in pollen. Plant Cell 4:557–571Combet C, Blanchet C, Geourjon C, Deléage G (2000) NPS@: network protein sequence analysis. Trend Biochem Sci 25:147–150De Almeida Engler J, Van Montagu M, Engler G (1998) Whole-mount in situ hybridization in plants. Methods Mol Biol (Clifton, NJ) 82:373–384De Caroli M, Lenucci MS, Di Sansebastiano GP, Dalessandro G, De Lorenzo G, Piro G (2011) Protein trafficking to the cell wall occurs through mechanisms distinguishable from default sorting in tobacco. Plant J 65(2):295–308Francis KE, Lam SY, Copenhaver GP (2006) Separation of Arabidopsis pollen tetrads is regulated by QUARTET1, a pectin methylesterase gene. Plant Physiol 142:1004–1013Futamura N, Mori H, Kouchi H, Shinohara K (2000) Male flower-specific expression of genes for polygalacturonase, pectin methylesterase and β-1,3-glucanase in a dioecious willow (Salix gilgliana Seemen). Plant Cell Physiol 41:16–26García-Sogo B, Pineda B, Castelblanque L, Antón T, Medina M, Roque E, Torresi C, Beltrán JP, Moreno V, Cañas LA (2010) Efficient transformation of Kalanchoe blossfeldiana and production of male sterile plants by engineered anther ablation. Plant Cell Rep 29:61–77García-Sogo B, Pineda B, Roque E, Antón T, Atarés A, Borja M, Beltrán JP, Moreno V, Cañas LA (2012) Production of engineered long-life and male sterile Pelargonium plants. BMC Plant Biol 12:156–162Gómez MD, Beltrán JP, Cañas LA (2004) The pea END1 promoter drives anther-specific gene expression in different plant species. Planta 219:967–981Gummadi SN, Manoj N, Kumar DS (2007) Structure and biochemical properties of pectinases. In: Polaina J, MacCabe AP (eds) Industrial enzymes: structure, function and applications. Springer, Berlin, pp 99–115. ISBN 9781402053764Gurgu L, Lafraya A, Polaina J, Marín-Navarro J (2011) Fermentation of cellobiose to ethanol by industrial Saccharomyces strains carrying the β-glucosidase gene (BGL1) from Saccharomycopsis fibuligera. Bioresour Technol 102:5229–5236Hamilton DA, Schwarz YH, Mascarenhas JP (1998) A monocot pollen-specific promoter contains separable pollen-specific and quantitative elements. 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Causas de incumplimiento de los pacientes que acuden a una farmacia comunitaria de Granada

Este trabajo fue presentado como comunicación- póster en el II Congreso Nacional de la SEFaC en A Coruña el 17-19 de noviembre de 2006.Objetivo: Determinar si los pacientes que acuden a una oficina de farmacia de Granada incumplen el tratamiento terapéutico e identificar su causa. Diseño: Estudio descriptivo, observacional, transversal. Emplazamiento: Farmacia comunitaria de Granada. Participantes: Pacientes mayores de 12 años y sin ninguna dificultad cognitiva, que acudieron a la farmacia en septiembre de 2005. Mediciones principales: Se midió el cumplimiento farmacoterapéutico mediante el cuestionario de Morisky-Green1. Se pasó un segundo cuestionario a los pacientes incumplidores preparado ad hoc para determinar las posibles causas de incumplimiento. Resultados: Participaron y concluyeron el estudio 139 pacientes. De ellos, 108 (77,7%) incumplían el tratamiento; el 62% eran mujeres y el resto, varones, con una mayoría en el rango de edad de 19-45 años (76%) y el 64,8% tenía un nivel cultural medio

Hormonal regulation of temperature-induced growth in Arabidopsis

[EN] Successful plant survival depends upon the proper integration of information from the environment with endogenous cues to regulate growth and development. We have investigated the interplay between ambient temperature and hormone action during the regulation of hypocotyl elongation, and we have found that gibberellins (GAs) and auxin are quickly and independently recruited by temperature to modulate growth rate, whereas activity of brassinosteroids (BRs) seems to be required later on. Impairment of GA biosynthesis blocked the increased elongation caused at higher temperatures, but hypocotyls of pentuple DELLA knockout mutants still reduced their response to higher temperatures when BR synthesis or auxin polar transport were blocked. The expression of several key genes involved in the biosynthesis of GAs and auxin was regulated by temperature, which indirectly resulted in coherent variations in the levels of accumulation of nuclear GFP-RGA (repressor of GA1) and in the activity of the DR5 reporter. DNA microarray and genetic analyses allowed the identification of the transcription factor PIF4 (phytochrome-interacting factor 4) as a major target in the promotion of growth at higher temperature. These results suggest that temperature regulates hypocotyl growth by individually impinging on several elements of a pre-existing network of signaling pathways involving auxin, BRs, GAs, and PIF4.We thank G. Choi (KAIST, Daejeon, South Korea), C. Fankhauser (University of Lausanne, Lausanne, Switzerland), T. Guilfoyle (Department of Biochemistry, University of Missouri, MO, USA), N. P. Harberd (Department of Plant Sciences, University of Oxford, Oxford, UK), E. Huq (University of Texas, Austin, TX, USA), T-p Sun (Department of Biology, Duke University, Durham, USA), S. G. Thomas (Rothamsted Research, Hertfordshire, UK), G. Vert (Institut de Biologie Integrative des Plantes, Montpellier, France), Z. Y. Wang (Department of Plant Biology, Carnegie Institution, Stanford, USA), Y. Yin (Plant Science Institute, Iowa State University, Ames, IA, USA), and the Arabidopsis Biological Resource Center for seeds; and X. W. Deng (Yale University, New Haven, CT, USA) for antibodies against RPT5. We also thank Dr Jorge Casal (Universidad de Buenos Aires, Buenos Aires, Argentina) for helpful suggestions on this work. Work in the authors' laboratories is funded by grant BIO2007-60923 from the Spanish Ministry of Science and Innovation and by grant 167890/110 from the Norwegian Research Council. JG-B was supported by a JAE pre-doctoral fellowship from CSIC.Stavang, JA.; Gallego-Bartolomé, J.; Gómez Jiménez, MD.; Yoshida, S.; Asami, T.; Olsen, JE.; García-Martínez, JL.... (2009). Hormonal regulation of temperature-induced growth in Arabidopsis. The Plant Journal. 60(4):589-601. https://doi.org/10.1111/j.1365-313X.2009.03983.x58960160

Regulation of ovule initiation by gibberellins and brassinosteroids in tomato and Arabidopsis: two plant species, two molecular mechanisms

This is the peer reviewed version of the following article: Barro¿Trastoy, D., Carrera, E., Baños, J., Palau-Rodríguez, J., Ruiz-Rivero, O., Tornero, P., Alonso, J.M., López-Díaz, I., Gómez, M.D. and Pérez-Amador, M.A. (2020), Regulation of ovule initiation by gibberellins and brassinosteroids in tomato and Arabidopsis: two plant species, two molecular mechanisms. Plant J, 102: 1026-1041, which has been published in final form at https://doi.org/10.1111/tpj.14684. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.[EN] Ovule primordia formation is a complex developmental process with a strong impact on the production of seeds. In Arabidopsis this process is controlled by a gene network, including components of the signalling pathways of auxin, brassinosteroids (BRs) and cytokinins. Recently, we have shown that gibberellins (GAs) also play an important role in ovule primordia initiation, inhibiting ovule formation in both Arabidopsis and tomato. Here we reveal that BRs also participate in the control of ovule initiation in tomato, by promoting an increase on ovule primordia formation. Moreover, molecular and genetic analyses of the co-regulation by GAs and BRs of the control of ovule initiation indicate that two different mechanisms occur in tomato and Arabidopsis. In tomato, GAs act downstream of BRs. BRs regulate ovule number through the downregulation of GA biosynthesis, which provokes stabilization of DELLA proteins that will finally promote ovule primordia initiation. In contrast, in Arabidopsis both GAs and BRs regulate ovule number independently of the activity levels of the other hormone. Taken together, our data strongly suggest that different molecular mechanisms could operate in different plant species to regulate identical developmental processes even, as for ovule primordia initiation, if the same set of hormones trigger similar responses, adding a new level of complexity.We wish to thank B. Janssen (Horticulture and Food Research Institute, New Zealand) for the pBJ60 shuttle vector, C. Ferrandiz and M. Colombo (IBMCP, CSIC-UPV, Valencia, Spain) for their help in the generation of 35S:ANT lines and L.E.P. Peres (Universidade de Sao Paulo, Brazil) for the tomato mutant lines. Our thanks also go to C. Fuster for technical assistance. This work was supported by grants from the Spanish Ministry of Economy and Competitiveness-FEDER (BIO2017-83138R) to MAPA and from NSF (DBI-0820755, MCB-1158181, and IOS-1444561) to JMA.Barro-Trastoy, D.; Carrera, E.; Baños, J.; Palau-Rodríguez, J.; Ruiz-Rivero, O.; Tornero Feliciano, P.; Alonso, JM.... (2020). 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The EPICTER score: a bedside and easy tool to predict mortality at 6 months in acute heart failure

Aims: Estimating the prognosis in heart failure (HF) is important to decide when to refer to palliative care (PC). Our objective was to develop a tool to identify the probability of death within 6 months in patients admitted with acute HF. Methods and results: A total of 2848 patients admitted with HF in 74 Spanish hospitals were prospectively included and followed for 6 months. Each factor independently associated with death in the derivation cohort (60% of the sample) was assigned a prognostic weight, and a risk score was calculated. The accuracy of the score was verified in the validation cohort. The characteristics of the population were as follows: advanced age (mean 78 years), equal representation of men and women, significant comorbidity, and predominance of HF with preserved ejection fraction. During follow-up, 753 patients (26%) died. Seven independent predictors of mortality were identified: age, chronic obstructive pulmonary disease, cognitive impairment, New York Heart Association class III-IV, chronic kidney disease, estimated survival of the patient less than 6 months, and acceptance of a palliative approach by the family or the patient. The area under the ROC curve for 6 month death was 0.74 for the derivation and 0.68 for the validation cohort. The model showed good calibration (Hosmer and Lemeshow test, P value 0.11). The 6 month death rates in the score groups ranged from 6% (low risk) to 54% (very high risk). Conclusions: The EPICTER score, developed from a prospective and unselected cohort, is a bedside and easy-to-use tool that could help to identify high-risk patients requiring PC

Development and Validation of an Early Mortality Risk Score for Older Patients Treated with Chemotherapy for Cancer

Background: Estimation of life expectancy in older patients is relevant to select the best treatment strategy. We aimed to develop and validate a score to predict early mortality in older patients with cancer. Patients and Methods: A total of 749 patients over 70 years starting new chemotherapy regimens were prospectively included. A prechemotherapy assessment that included sociodemographic variables, tumor/treatment variables, and geriatric assessment variables was performed. Association between these factors and early death was examined using multivariable logistic regression. Score points were assigned to each risk factor. External validation was performed on an independent cohort. Results: In the training cohort, the independent predictors of 6-month mortality were metastatic stage (OR 4.8, 95% CI [2.4-9.6]), ECOG-PS 2 (OR 2.3, 95% CI [1.1-5.2]), ADL ≤ 5 (OR 1.7, 95% CI [1.1-3.5]), serum albumin levels ≤ 3.5 g/dL (OR 3.4, 95% CI [1.7-6.6]), BMI < 23 kg/m2 (OR 2.5, 95% CI [1.3-4.9]), and hemoglobin levels < 11 g/dL (OR 2.4, 95% CI (1.2-4.7)). With these results, we built a prognostic score. The area under the ROC curve was 0.78 (95% CI, 0.73 to 0.84), and in the validation set, it was 0.73 (95% CI: 0.67-0.79). Conclusions: This simple and highly accurate tool can help physicians making decisions in elderly patients with cancer who are planned to initiate chemotherapy treatment