Aceites esenciales: productos antimicrobianos y antioxidantes naturales en la industria agroalimentaria

[ES] Los consumidores son conscientes del peligro derivado del uso de antioxidantes y antimicrobianos sintéticos en la industria agroalimentaria, demandando alternativas más seguras y ecológicas. En este estudio, se ha determinado la actividad antioxidante de aceites esenciales comerciales mediante el método DPPH y su efecto antimicrobiano frente a la bacteria Pseudomonas syringae y el hongo fitopatógeno Fusarium oxysporum a través del empleo del método estandarizado de disco. Los aceites esenciales de clavo, ajedrea, canela y orégano, así como carvacrol, mostraron la máxima actividad antioxidante, comparable a antioxidantes establecidos. El aceite esencial de gaulteria fue el más potente inhibidor del crecimiento de P. syringae en las dosis más altas (20 y 10 µL) ensayadas. El aceite esencial de orégano, así como su componente principal carvacrol, detuvieron el crecimiento de la bacteria incluso a la dosis más baja ensayada (1 µL). Los aceites esenciales de canela, orégano y menta inhibieron el desarrollo de F. oxysporum en todas las dosis (20, 10 y 5 µL) aplicadas. En general, la mayoría de aceites esenciales mostraron más actividad antifúngica que antibacteriana y antioxidante.[EN] Consumers are aware of the dangers arising from the use of synthetic antioxidants and antimicrobials in the agrifood industry, demanding safer and "greener" alternatives. In this study, the antioxidant activity of commercial essential oils through DPPH method, their antimicrobial effects against the bacterium Pseudomonas syringae and the phytopathogenic fungus Fusarium oxysporum by means of the standardized disk method were determined. Clove along with winter savory, cinnamon and oregano essential oils as well as carvacrol showed the highest antioxidant activity comparable to reference standards. Wintergreen essential oil was the most potent inhibitor against P. syringae growth at the highest doses (20 and 10 µL). Oregano essential oil and its main component carvacrol were able to stop the bacterium growth even at the lowest treatment (1 µL). Cinnamon, oregano and peppermint essential oils inhibited F. oxysporum development at all doses (20, 10 and 5 µL) assayed. In general, most of the essential oils displayed more antifungal than antibacterial and antioxidant activities.Ibáñez, MD.; López-Gresa, MP.; Lisón, P.; Rodrigo Bravo, I.; Belles Albert, JM.; González-Mas, MC.; Blázquez, MA. (2020). Essential oils as natural antimicrobial and antioxidant products in the Agrifood Industry. Nereis. Revista Iberoamericana Interdisciplinar de Métodos, Modelización y Simulación. (12):55-69. https://doi.org/10.46583/nereis_2020.12.585S55691

Evidences towards deciphering the mode of action of dimethylpyrazole-based nitrification inhibitors in soil and pure cultures of Nitrosomonas europaea

Background: Agriculture relies on the intensive use of synthetic nitrogen (N) fertilizers to maximize crop yields, which has led to the transformation of agricultural soils into high-nitrifying environments. Nevertheless, nitrification inhibitors (Nis) have been developed to suppress soil-nitrifier activity and decrease N losses. The Nis 3,4-dimethylpyrazole phosphate (DMPP) and 2-(3,4-dimethyl-1H-pyrazol-1-yl) succinic acid isomeric mixture (DMPSA) are able to reduce N2O emissions and maintain soil NH4+ for a longer time. Although both Nls have been proven to be effective to inhibit soil nitrification, their exact mode of action has not been confirmed. We aimed to provide novel insights to further understand the mode of action of DMP-based Nis. We evaluated the performance of DMPP and DMPSA in soil and pure cultures of nitrifying bacteria Nitrosomonas europaea. Results: DMPSA did not inhibit nitrification in pure cultures of N. europaea. In the soil, we evidenced that DMPSA needs to be broken into DMP to achieve the inhibition of nitrification, which is mediated by a soil biological process that remains to be identified. Moreover, both DMPP and DMPSA are thought to inhibit nitrification due to their ability to chelate the Cu2+ cations that the ammonia monooxygenase enzyme (AMO) needs to carry on the first step of NH4+ oxidation. However, the efficiency of DMPP was not altered regardless the Cu2+ concentration in the medium. In addition, we also showed that DMPP targets AMO but not hydroxylamine oxidoreductase enzyme (HAO). Conclusions: The inability of DMPSA to inhibit nitrification in pure cultures together with the high efficiency of DMPP to inhibit nitrification even in presence of toxic Cu2+ concentration in the medium, suggest that the mode of action of DMP-based Nis does not rely on their capacity as metal chelators.This project was funded by the Spanish Government (RTI2018-094623-B-C21 MCIU/AEI/FEDER, UE), by the Basque Government (IT-932-16), and by EuroChem Agro Iberia S.L.U. Dr. Adrian Bozal-Leorri held a grant from the Basque Government (PRE-2020-2-0142)

A Multi-Species Analysis Defines Anaplerotic Enzymes and Amides as Metabolic Markers for Ammonium Nutrition

Nitrate and ammonium are the main nitrogen sources in agricultural soils. In the last decade, ammonium (NH4+), a double-sided metabolite, has attracted considerable attention by researchers. Its ubiquitous presence in plant metabolism and its metabolic energy economy for being assimilated contrast with its toxicity when present in high amounts in the external medium. Plant species can adopt different strategies to maintain NH4+ homeostasis, as the maximization of its compartmentalization and assimilation in organic compounds, primarily as amino acids and proteins. In the present study, we report an integrative metabolic response to ammonium nutrition of seven plant species, belonging to four different families: Gramineae (ryegrass, wheat, Brachypodium distachyon), Leguminosae (clover), Solanaceae (tomato), and Brassicaceae (oilseed rape, Arabidopsis thaliana). We use principal component analysis (PCA) and correlations among metabolic and biochemical data from 40 experimental conditions to understand the whole-plant response. The nature of main amino acids is analyzed among species, under the hypothesis that those Asn-accumulating species will show a better response to ammonium nutrition. Given the provision of carbon (C) skeletons is crucial for promotion of the nitrogen assimilation, the role of different anaplerotic enzymes is discussed in relation to ammonium nutrition at a whole-plant level. Among these enzymes, isocitrate dehydrogenase (ICDH) shows to be a good candidate to increase nitrogen assimilation in plants. Overall, metabolic adaptation of different carbon anaplerotic activities is linked with the preference to synthesize Asn or Gln in their organs. Lastly, glutamate dehydrogenase (GDH) reveals as an important enzyme to surpass C limitation during ammonium assimilation in roots, with a disparate collaboration of glutamine synthetase (GS).The design of the study, analysis, and interpretation of data and writing of the manuscript was supported by the Basque Government [IT932-16] or GIC15/179, the Spanish Ministry of Economy and Competitiveness [AGL2015-64582-C3-2-R] and [BIO2017-84035-R]. IVM held a postdoctoral grant from the Basque Government (conv. 2018) and MDLP held a PhD grant by COLCIENCIAS (conv. 672)

Biochar reduces the efficiency of nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) mitigating N2O emissions

Among strategies suggested to decrease agricultural soil N2O losses, the use of nitrification inhibitors such as DMPP (3,4-dimethylpyrazole phosphate) has been proposed. However, the efficiency of DMPP might be affected by soil amendments, such as biochar, which has been shown to reduce N2O emissions. This study evaluated the synergic effect of a woody biochar applied with DMPP on soil N2O emissions. A incubation study was conducted with a silt loam soil and a biochar obtained from Pinus taeda at 500 degrees C. Two biochar rates (0 and 2% (w/w)) and three different nitrogen treatments (unfertilized, fertilized and fertilized + DMPP) were assayed under two contrasting soil water content levels (40% and 80% of water filled pore space (WFPS)) over a 163 day incubation period. Results showed that DMPP reduced N2O emissions by reducing ammonia-oxidizing bacteria (AOB) populations and promoting the last step of denitrification (measured by the ratio nosZI + nosZII/nirS + nirK genes). Biochar mitigated N2O emissions only at 40% WFPS due to a reduction in AOB population. However, when DMPP was applied to the biochar amended soil, a counteracting effect was observed, since the N2O mitigation induced by DMPP was lower than in control soil, demonstrating that this biochar diminishes the efficiency of the DMPP both at low and high soil water contents.This work was funded by the Spanish Government (AGL2015-64582-C3-2-R MINECO/FEDER), by the Basque Government (IT-932-16) and by the European Union (FACCE-CSA no 276610/MIT04-DESIGN-UPVASC, FACCE-CSA no 2814ERA01A and 2814ERA02A). This work is also supported by the USDA/NIFA Interagency Climate Change Grant Proposal number 2014-02114 [Project number 6657-12130-002-08I, Accession number 1003011] under the Multi-Partner Call on Agricultural Greenhouse Gas Research of the FACCE-Joint Program Initiative. Any opinions, findings, or recommendation expressed in this publication are those of the authors and do not necessarily reflect the view of the USDA. MLC was supported by a Ramon y Cajal contract from the Spanish Ministry of Economy and Competitiveness and thanks Fundacion Seneca for financing the project 19281/PI/14

Comparative Analysis of the Volatile Fraction of Fruit Juice from Different Citrus Species

The volatile composition of fruit from four Citrus varieties (Powell Navel orange, Clemenules mandarine, and Fortune mandarine and Chandler pummelo) covering four different species has been studied. Over one hundred compounds were profiled after HS-SPME-GC-MS analysis, including 27 esters, 23 aldehydes, 21 alcohols, 13 monoterpene hydrocarbons, 10 ketones, 5 sesquiterpene hydrocarbons, 4 monoterpene cyclic ethers, 4 furans, and 2 aromatic hydrocarbons, which were all confirmed with standards. The differences in the volatile profile among juices of these varieties were essentially quantitative and only a few compounds were found exclusively in a single variety, mainly in Chandler. The volatile profile however was able to differentiate all four varieties and revealed complex interactions between them including the participation in the same biosynthetic pathway. Some compounds (6 esters, 2 ketones, 1 furan and 2 aromatic hydrocarbons) had never been reported earlier in Citrus juices. This volatile profiling platform for Citrus juice by HS-SPME-GC-MS and the interrelationship detected among the volatiles can be used as a roadmap for future breeding or biotechnological applications

Tumor xenograft modeling identifies an association between TCF4 loss and breast cancer chemoresistance

Understanding the mechanisms of cancer therapeutic resistance is fundamental to improving cancer care. There is clear benefit from chemotherapy in different breast cancer settings; however, knowledge of the mutations and genes that mediate resistance is incomplete. In this study, by modeling chemoresistance in patientderived xenografts (PDXs), we show that adaptation to therapy is genetically complex and identify that loss of transcription factor 4 (TCF4; also known as ITF2) is associated with this process. A triple-negative BRCA1-mutaied PDX was used to study the genetics of chemoresistance. The PDX was treated in parallel with four chemotherapies for five iterative cycles. Exome sequencing identified few genes with de novo or enriched mutations in common among the different therapies, whereas many common depleted mutations/ genes were observed. Analysis of somatic mutations from The Cancer Genome Atlas (TCGA) supported the prognostic relevance of the identified genes. A mutation in TCF4 was found de novo in all treatments, and analysis of drug sensitivity profiles across cancer cell lines supported the link to chemoresistance. Loss of TCF4 conferred chemoresistance in breast cancer cell models, possibly by altering cell cycle regulation. Targeted sequencing in chemoresistant tumors identified an intronic variant of TCF4 that may represent an expression quantitative trait locus associated with relapse outcome in TCGA. Immunohistochemical studies suggest a common loss of nuclear TCF4 expression post-chemotherapy. Together, these results from tumor xenograft modeling depict a link between altered TCF4 expression and breast cancer chemoresistance

Homenaje a Elena Romero

Edición a cargo de Aitor García MorenoEste volumen no quiere ser sino, desde el punto de vista del contenido, representación del sefardismo en la actualidad en sus múltiples facetas, con estudios que den muestra de su admirable variedad como campo de estudios, muestra asimismo de la increíble experiencia y peripecia vital de un grupo cultural como el de los judeoespañoles.Este volumen es un resultado más del proyecto «Sefarad, siglo XXI (2009-2011): Edición y estudio filológico de textos sefardíes» del Plan Nacional de I+D+I (ref. FFI2009-10672).Peer reviewe

Fruit load modulates flowering-related gene expression in buds of alternate-bearing 'Moncada' mandarin

Background and Aims Gene determination of flowering is the result of complex interactions involving both promoters and inhibitors. In this study, the expression of flowering-related genes at the meristem level in alternate-bearing citrus trees is analysed, together with the interplay between buds and leaves in the determination of flowering. Methods First defruiting experiments were performed to manipulate blossoming intensity in `Moncada¿ mandarin, Citrus clementina. Further defoliation was performed to elucidate the role leaves play in the flowering process. In both cases, the activity of flowering-related genes was investigated at the flower induction (November) and differentiation (February) stages. Key Results Study of the expression pattern of flowering-genes in buds from on (fully loaded) and off (without fruits) trees revealed that homologues of FLOWERING LOCUS T (CiFT), TWIN SISTER OF FT (TSF), APETALA1 (CsAP1) and LEAFY (CsLFY) were negatively affected by fruit load. CiFT and TSF activities showed a marked increase in buds from off trees through the study period (ten-fold in November). By contrast, expression of the homologues of the flowering inhibitors of TERMINAL FLOWER 1 (CsTFL), TERMINAL FLOWER 2 (TFL2) and FLOWERING LOCUS C (FLC) was generally lower in off trees. Regarding floral identity genes, the increase in CsAP1 expression in off trees was much greater in buds than in leaves, and significant variations in CsLFY expression (approx. 20 %) were found only in February. Defoliation experiments further revealed that the absence of leaves completely abolished blossoming and severely affected the expression of most of the flowering-related genes, particularly decreasing the activity of floral promoters and of CsAP1 at the induction stage. Conclusions These results suggest that the presence of fruit affects flowering by greatly altering gene-expression not only at the leaf but also at the meristem level. Although leaves are required for flowering to occur, their absence strongly affects the activity of floral promoters and identity genes.This work was supported by a grant from the Instituto Nacional Investigaciones Agrarias, Spain (RTA2009-00147). M. C. Gonzalez was the recipient of a contract by the Fundacion Agroalimed (Conselleria d'Agricultura, Pesca i Alimentacio, Generalitat Valenciana).Muñoz Fambuena, N.; Mesejo Conejos, C.; Gonzalez Más, MC.; Primo-Millo, E.; Agustí Fonfría, M.; Iglesias, DJ. (2012). Fruit load modulates flowering-related gene expression in buds of alternate-bearing 'Moncada' mandarin. Annals of Botany. 110(6):1109-1118. doi:10.1093/aob/mcs190S110911181106Abe, M. (2005). FD, a bZIP Protein Mediating Signals from the Floral Pathway Integrator FT at the Shoot Apex. Science, 309(5737), 1052-1056. doi:10.1126/science.1115983Bustin, S. (2002). Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. 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The Journal of Horticultural Science and Biotechnology, 84(2), 149-154. doi:10.1080/14620316.2009.11512496Esumi, T., Kitamura, Y., Hagihara, C., Yamane, H., & Tao, R. (2010). Identification of a TFL1 ortholog in Japanese apricot (Prunus mume Sieb. et Zucc.). Scientia Horticulturae, 125(4), 608-616. doi:10.1016/j.scienta.2010.05.016Giakountis, A., & Coupland, G. (2008). Phloem transport of flowering signals. Current Opinion in Plant Biology, 11(6), 687-694. doi:10.1016/j.pbi.2008.10.003Hashimoto, J. G., Beadles-Bohling, A. S., & Wiren, K. M. (2004). Comparison of RiboGreen®and 18S rRNA quantitation for normalizing real-time RT-PCR expression analysis. BioTechniques, 36(1), 54-60. doi:10.2144/04361bm06Jaeger, K. E., Graf, A., & Wigge, P. A. (2006). The control of flowering in time and space. Journal of Experimental Botany, 57(13), 3415-3418. doi:10.1093/jxb/erl159Jang, S., Torti, S., & Coupland, G. (2009). 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Plant and Cell Physiology, 51(4), 561-575. doi:10.1093/pcp/pcq021Li, D., Liu, C., Shen, L., Wu, Y., Chen, H., Robertson, M., … Yu, H. (2008). A Repressor Complex Governs the Integration of Flowering Signals in Arabidopsis. Developmental Cell, 15(1), 110-120. doi:10.1016/j.devcel.2008.05.002Lord, E. M., & Eckard, K. J. (1985). Shoot Development in Citrus sinensis L. (Washington Navel Orange). I. Floral and Inflorescence Ontogeny. Botanical Gazette, 146(3), 320-326. doi:10.1086/337531Mathieu, J., Warthmann, N., Küttner, F., & Schmid, M. (2007). Export of FT Protein from Phloem Companion Cells Is Sufficient for Floral Induction in Arabidopsis. Current Biology, 17(12), 1055-1060. doi:10.1016/j.cub.2007.05.009Matsuda, N., Ikeda, K., Kurosaka, M., Takashina, T., Isuzugawa, K., Endo, T., & Omura, M. (2009). Early Flowering Phenotype in Transgenic Pears (Pyrus communis L.) Expressing the CiFT Gene. Journal of the Japanese Society for Horticultural Science, 78(4), 410-416. doi:10.2503/jjshs1.78.410Michaels, S. D., Himelblau, E., Kim, S. Y., Schomburg, F. M., & Amasino, R. M. (2004). Integration of Flowering Signals in Winter-Annual Arabidopsis. Plant Physiology, 137(1), 149-156. doi:10.1104/pp.104.052811Moss, G. I. (1971). Effect of fruit on flowering in relation to biennial bearing in sweet orange(Citrus sinensis). Journal of Horticultural Science, 46(2), 177-184. doi:10.1080/00221589.1971.11514396Muñoz-Fambuena, N., Mesejo, C., Carmen González-Mas, M., Primo-Millo, E., Agustí, M., & Iglesias, D. J. (2011). Fruit regulates seasonal expression of flowering genes in alternate-bearing ‘Moncada’ mandarin. Annals of Botany, 108(3), 511-519. doi:10.1093/aob/mcr164Muñoz-Fambuena, N., Mesejo, C., González-Mas, M. C., Iglesias, D. J., Primo-Millo, E., & Agustí, M. (2012). Gibberellic Acid Reduces Flowering Intensity in Sweet Orange [Citrus sinensis (L.) Osbeck] by Repressing CiFT Gene Expression. Journal of Plant Growth Regulation, 31(4), 529-536. doi:10.1007/s00344-012-9263-yNishikawa, F., Endo, T., Shimada, T., Fujii, H., Shimizu, T., Omura, M., & Ikoma, Y. (2007). Increased CiFT abundance in the stem correlates with floral induction by low temperature in Satsuma mandarin (Citrus unshiu Marc.). Journal of Experimental Botany, 58(14), 3915-3927. doi:10.1093/jxb/erm246Nishikawa, F., Endo, T., Shimada, T., Fujii, H., Shimizu, T., Kobayashi, Y., … Omura, M. (2010). Transcriptional changes in CiFT-introduced transgenic trifoliate orange (Poncirus trifoliata L. Raf.). Tree Physiology, 30(3), 431-439. doi:10.1093/treephys/tpp122Notaguchi, M., Abe, M., Kimura, T., Daimon, Y., Kobayashi, T., Yamaguchi, A., … Araki, T. (2008). Long-Distance, Graft-Transmissible Action of Arabidopsis FLOWERING LOCUS T Protein to Promote Flowering. 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Diverse effects of overexpression of LEAFY and PTLF, a poplar (Populus) homolog of LEAFY/FLORICAULA, in transgenic poplar and Arabidopsis. The Plant Journal, 22(3), 235-245. doi:10.1046/j.1365-313x.2000.00734.xSherman, W. B., & Beckman, T. G. (2003). CLIMATIC ADAPTATION IN FRUIT CROPS. Acta Horticulturae, (622), 411-428. doi:10.17660/actahortic.2003.622.43Southerton, S. G., Strauss, S. H., Olive, M. R., Harcourt, R. L., Decroocq, V., Zhu, X., … Dennis, E. S. (1998). Plant Molecular Biology, 37(6), 897-910. doi:10.1023/a:1006056014079Sreekantan, L., & Thomas, M. R. (2006). VvFT and VvMADS8, the grapevine homologues of the floral integrators FT and SOC1, have unique expression patterns in grapevine and hasten flowering in Arabidopsis. Functional Plant Biology, 33(12), 1129. doi:10.1071/fp06144Takada, S., & Goto, K. (2003). TERMINAL FLOWER2, an Arabidopsis Homolog of HETEROCHROMATIN PROTEIN1, Counteracts the Activation of FLOWERING LOCUS T by CONSTANS in the Vascular Tissues of Leaves to Regulate Flowering Time. The Plant Cell, 15(12), 2856-2865. doi:10.1105/tpc.016345Tan, F.-C., & Swain, S. M. (2007). Functional characterization of AP3, SOC1 and WUS homologues from citrus (Citrus sinensis). Physiologia Plantarum, 131(3), 481-495. doi:10.1111/j.1399-3054.2007.00971.xTränkner, C., Lehmann, S., Hoenicka, H., Hanke, M.-V., Fladung, M., Lenhardt, D., … Flachowsky, H. (2010). Over-expression of an FT-homologous gene of apple induces early flowering in annual and perennial plants. Planta, 232(6), 1309-1324. doi:10.1007/s00425-010-1254-2Vemmos, S. N. (1999). Carbohydrate content of inflorescent buds of defruited and fruiting pistachio (Pistacia vera L) branches in relation to biennial bearing. The Journal of Horticultural Science and Biotechnology, 74(1), 94-100. doi:10.1080/14620316.1999.11511079Wada, M., Cao, Q., Kotoda, N., Soejima, J., & Masuda, T. (2002). Plant Molecular Biology, 49(6), 567-577. doi:10.1023/a:1015544207121Wigge, P. A. (2005). Integration of Spatial and Temporal Information During Floral Induction in Arabidopsis. Science, 309(5737), 1056-1059. doi:10.1126/science.1114358Yahata, D., Matsumoto, K., & Ushijima, K. (2004). Relationship between Flower-bud Differentiation and Carbohydrate Contents in Spring Shoots of Very-early, Early and Late Maturing Cultivars of Satsuma Mandarin. Engei Gakkai zasshi, 73(5), 405-410. doi:10.2503/jjshs.73.405Yamaguchi, A., Kobayashi, Y., Goto, K., Abe, M., & Araki, T. (2005). TWIN SISTER OF FT (TSF) Acts as a Floral Pathway Integrator Redundantly with FT. Plant and Cell Physiology, 46(8), 1175-1189. doi:10.1093/pcp/pci151Yan, J., Yuan, F., Long, G., Qin, L., & Deng, Z. (2011). 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The management of acute venous thromboembolism in clinical practice. Results from the European PREFER in VTE Registry

Venous thromboembolism (VTE) is a significant cause of morbidity and mortality in Europe. Data from real-world registries are necessary, as clinical trials do not represent the full spectrum of VTE patients seen in clinical practice. We aimed to document the epidemiology, management and outcomes of VTE using data from a large, observational database. PREFER in VTE was an international, non-interventional disease registry conducted between January 2013 and July 2015 in primary and secondary care across seven European countries. Consecutive patients with acute VTE were documented and followed up over 12 months. PREFER in VTE included 3,455 patients with a mean age of 60.8 ± 17.0 years. Overall, 53.0 % were male. The majority of patients were assessed in the hospital setting as inpatients or outpatients (78.5 %). The diagnosis was deep-vein thrombosis (DVT) in 59.5 % and pulmonary embolism (PE) in 40.5 %. The most common comorbidities were the various types of cardiovascular disease (excluding hypertension; 45.5 %), hypertension (42.3 %) and dyslipidaemia (21.1 %). Following the index VTE, a large proportion of patients received initial therapy with heparin (73.2 %), almost half received a vitamin K antagonist (48.7 %) and nearly a quarter received a DOAC (24.5 %). Almost a quarter of all presentations were for recurrent VTE, with >80 % of previous episodes having occurred more than 12 months prior to baseline. In conclusion, PREFER in VTE has provided contemporary insights into VTE patients and their real-world management, including their baseline characteristics, risk factors, disease history, symptoms and signs, initial therapy and outcomes

Fruit regulates seasonal expression of flowering genes in alternate-bearing 'Moncada' mandarin

Background and Aims The presence of fruit has been widely reported to act as an inhibitor of flowering in fruit trees. This study is an investigation into the effect of fruit load on flowering of `Moncada¿ mandarin and on the expression of putative orthologues of genes involved in flowering pathways to provide insight into the molecular mechanisms underlying alternate bearing in citrus. Methods The relationship between fruit load and flowering intensity was examined first. Defruiting experiments were further conducted to demonstrate the causal effect of fruit removal upon flowering. Finally, the activity of flowering-related genes was investigated to determine the extent to which their seasonal expression is affected by fruit yield. Key Results First observations and defruiting experiments indicated a significant inverse relationship between preceding fruit load and flowering intensity. Moreover, data indicated that when fruit remained on the tree from November onwards, a dramatic inhibition of flowering occurred the following spring. The study of the expression pattern of flowering-genes of on (fully loaded) and off (without fruits) trees revealed that homologues of FLOWERING LOCUS T (FT), SUPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1), APETALA1 (AP1) and LEAFY (LFY) were negatively affected by fruit load. Thus, CiFT expression showed a progressive increase in leaves from off trees through the study period, the highest differences found from December onwards (10-fold). Whereas differences in the relative expression of SOC1 only reached significance from September to mid-December, CsAP1 expression was constantly higher in those trees through the whole study period. Significant variations in CsLFY expression only were found in late February (close to 20 %). On the other hand, the expression of the homologues of TERMINAL FLOWER 1 (TFL1) and FLOWERING LOCUS C (FLC) did not appear to be related to fruit load. Conclusions These results suggest for the first time that fruit inhibits flowering by repressing CiFT and SOC1 expression in leaves of alternate-bearing citrus. Fruit also reduces CsAP1 expression in leaves, and the significant increase in leaf CsLFY expression from off trees in late February was associated with the onset of floral differentiation.We thank Dr D. Westall for her help in editing the manuscript. M. C. Gonzalez was recipient of a contract by the Fundacion Agroalimed (Conselleria d'Agricultura, Pesca i Alimentacio, Generalitat Valenciana). This work was supported by a grant from the Instituto Nacional Investigaciones Agrarias, Spain (RTA2009-00147).Muñoz Fambuena, N.; Mesejo Conejos, C.; Gonzalez Más, MC.; Primo-Millo, E.; Agustí Fonfría, M.; Iglesias, DJ. (2011). Fruit regulates seasonal expression of flowering genes in alternate-bearing 'Moncada' mandarin. Annals of Botany. 108:511-519. doi:10.1093/aob/mcr164S51151910