Tackling functional redundancy of Arabidopsis fatty acid elongase complexes

Very-long-chain fatty acids (VLCFA) are precursors for various lipids playing important physiological and structural roles in plants. Throughout plant tissues, VLCFA are present in multiple lipid classes essential for membrane homeostasis, and also stored in triacylglycerols. VLCFA and their derivatives are also highly abundant in lipid barriers, such as cuticular waxes in aerial epidermal cells and suberin monomers in roots. VLCFA are produced by the fatty acid elongase (FAE), which is an integral endoplasmic reticulum membrane multi-enzymatic complex consisting of four core enzymes. The 3-ketoacyl-CoA synthase (KCS) catalyzes the first reaction of the elongation and determines the chain-length substrate specificity of each elongation cycle, whereas the other three enzymes have broad substrate specificities and are shared by all FAE complexes. Consistent with the co-existence of multiple FAE complexes, performing sequential and/or parallel reactions to produce the broad chain-length-range of VLCFA found in plants, twenty-one KCS genes have been identified in the genome of Arabidopsis thaliana. Using CRISPR-Cas9 technology, we established an expression platform to reconstitute the different Arabidopsis FAE complexes in yeast. The VLCFA produced in these yeast strains were analyzed in detail to characterize the substrate specificity of all KCS candidates. Additionally, Arabidopsis candidate proteins were transiently expressed in Nicotiana benthamiana leaves to explore their activity and localization in planta. This work sheds light on the genetic and biochemical redundancy of fatty acid elongation in plants

Screening of Iberian Coinage in the 2(th)-1(th) BCE Period Using the Voltammetry of Immobilized Particles

This is the peer reviewed version of the following article: A. Doménech-Carbó, M. T. Doménech-Carbó, C. Álvarez-Romero, T. Pasíes, M. Buendía, Electroanalysis 2019, 31, 1164, which has been published in final form at https://doi.org/10.1002/elan.201900090. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.[EN] The voltammetry of immobilized particles (VIMP) was applied for grouping a series of 86 Iberian coins nominally minted in the cities of Iltirta, Castulo and Obulco in the 2(th)-1(th) BCE period for which there are no chronological data. Using characteristic signatures for the reduction of cuprite, tenorite and lead corrosion products in the patina of the coins, voltammetric grouping of coins was proposed. Voltammetric data were found to be consistent with textural and compositional properties of the surface and subsurface of selected coins using FIB-FESEM-EDX. The obtained data confirmed a clear separation between the productions of Iltirta on one side, and those of Castulo and Obulco on the other side, indicating the possibility to establish a rough chronology for these productions.Project CTQ2017-85317-C2-1-P, supported with Ministerio de Economia, Industria y Competitividad (MINECO), Fondo Europeo de Desarrollo Regional (ERDF) and Agencia Estatal de Investigacion (AEI), is gratefully acknowledged. The authors wish also to thank Mr. Manuel Planes and Dr. Jose Luis Moya, technical supervisors of the Electron Microscopy Service of the Universitat Politecnica de Valencia. Thanks to Manuel Gozalbes for his technical assistance in the numismatic domain and Gonzalo Cores and the Museu de Prehistrica de Valencia for facilitating the access to its collections.Doménech-Carbó, A.; Domenech Carbo, MT.; Álvarez-Romero, C.; Pasíes, T.; Buendía, M. (2019). Screening of Iberian Coinage in the 2(th)-1(th) BCE Period Using the Voltammetry of Immobilized Particles. Electroanalysis. 31(6):1164-1173. https://doi.org/10.1002/elan.201900090S11641173316P. P. Ripollès V. Heuchert A. Burnett Coinage and identity in the Roman provinces Oxford University Press London 79 93M. Gozalbes Circulación y uso de los denarios ibéricos in M. Campo Ús i circulació de la moneda a la Hispania Citerior XIII Curs d'història monetària d'Hispania Museu de Prehistòria de València València 83 103Constantinides, I., Gritsch, M., Adriaens, A., Hutter, H., & Adams, F. (2001). Microstructural characterisation of five simulated archaeological copper alloys using light microscopy, scanning electron microscopy, energy dispersive X-ray microanalysis and secondary ion mass spectrometry. Analytica Chimica Acta, 440(2), 189-198. doi:10.1016/s0003-2670(01)01061-3Linke, R., & Schreiner, M. (2000). Energy Dispersive X-Ray Fluorescence Analysis and X-Ray Microanalysis of Medieval Silver Coins. Microchimica Acta, 133(1-4), 165-170. doi:10.1007/s006040070087Dowsett, M., & Adriaens, A. (2004). The role of SIMS in cultural heritage studies. Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms, 226(1-2), 38-52. doi:10.1016/j.nimb.2003.12.086Shalev, S., Shilstein, S. S., & Yekutieli, Y. (2006). XRF study of archaeological and metallurgical material from an ancient copper-smelting site near Ein-Yahav, Israel☆. Talanta, 70(5), 909-913. doi:10.1016/j.talanta.2006.05.052Gaudiuso, R., Dell’Aglio, M., De Pascale, O., Loperfido, S., Mangone, A., & De Giacomo, A. (2014). Laser-induced breakdown spectroscopy of archaeological findings with calibration-free inverse method: Comparison with classical laser-induced breakdown spectroscopy and conventional techniques. Analytica Chimica Acta, 813, 15-24. doi:10.1016/j.aca.2014.01.020Del Hoyo-Meléndez, J. M., Świt, P., Matosz, M., Woźniak, M., Klisińska-Kopacz, A., & Bratasz, Ł. (2015). Micro-XRF analysis of silver coins from medieval Poland. Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms, 349, 6-16. doi:10.1016/j.nimb.2015.02.018Tomassetti, M., Marini, F., Bucci, R., & Campanella, L. (2016). A survey on innovative dating methods in archaeometry with focus on fossil bones. TrAC Trends in Analytical Chemistry, 79, 371-379. doi:10.1016/j.trac.2015.11.017BUDD, P., GALE, D., POLLARD, A. M., THOMAS, R. G., & WILLIAMS, P. A. (1993). EVALUATING LEAD ISOTOPE DATA: FURTHER OBSERVATIONS. Archaeometry, 35(2), 241-247. doi:10.1111/j.1475-4754.1993.tb01038.xAttanasio, D., Bultrini, G., & Ingo, G. M. (2001). The Possibility of Provenancing A Series of Bronze Punic Coins Found At Tharros (Western Sardinia) Using the Literature Lead Isotope Database. Archaeometry, 43(4), 529-547. doi:10.1111/1475-4754.00035Scott, D. A. (1994). An Examination of the Patina and Corrosion Morphology of Some Roman Bronzes. Journal of the American Institute for Conservation, 33(1), 1. doi:10.2307/3179666Constantinides, I., Adriaens, A., & Adams, F. (2002). Surface characterization of artificial corrosion layers on copper alloy reference materials. Applied Surface Science, 189(1-2), 90-101. doi:10.1016/s0169-4332(02)00005-3Robbiola, L., & Portier, R. (2006). A global approach to the authentication of ancient bronzes based on the characterization of the alloy–patina–environment system. Journal of Cultural Heritage, 7(1), 1-12. doi:10.1016/j.culher.2005.11.001Robbiola, L., Blengino, J.-M., & Fiaud, C. (1998). Morphology and mechanisms of formation of natural patinas on archaeological Cu–Sn alloys. Corrosion Science, 40(12), 2083-2111. doi:10.1016/s0010-938x(98)00096-1L. Robbiola L.-P. Hurtel Standard nature of the passive layers of buried archaeological bronze – The example of two Roman half-length portraits in I. MacLeod S. Pennec L. Robbiola 1997 109 117F. Scholz B. Meyer 1998 1 86Doménech-Carbó, A., Labuda, J., & Scholz, F. (2012). Electroanalytical chemistry for the analysis of solids: Characterization and classification (IUPAC Technical Report). Pure and Applied Chemistry, 85(3), 609-631. doi:10.1351/pac-rep-11-11-13Doménech-Carbó, A., Doménech-Carbó, M. T., & Costa, V. (Eds.). (2009). Electrochemical Methods in Archaeometry, Conservation and Restoration. Monographs in Electrochemistry. doi:10.1007/978-3-540-92868-3Doménech, A. (2011). Tracing, authenticating and dating archaeological metal using the voltammetry of microparticles. Analytical Methods, 3(10), 2181. doi:10.1039/c1ay05416cDoménech-Carbó, A., & Doménech-Carbó, M. T. (2018). Electroanalytical techniques in archaeological and art conservation. Pure and Applied Chemistry, 90(3), 447-461. doi:10.1515/pac-2017-0508Costa, V., Leyssens, K., Adriaens, A., Richard, N., & Scholz, F. (2009). Electrochemistry reveals archaeological materials. Journal of Solid State Electrochemistry, 14(3), 449-451. doi:10.1007/s10008-009-0864-8Arjmand, F., & Adriaens, A. (2011). Electrochemical quantification of copper-based alloys using voltammetry of microparticles: optimization of the experimental conditions. Journal of Solid State Electrochemistry, 16(2), 535-543. doi:10.1007/s10008-011-1365-0Souissi, N., Bousselmi, L., Khosrof, S., & Triki, E. (2004). Voltammetric behaviour of an archeaological bronze alloy in aqueous chloride media. Materials and Corrosion, 55(4), 284-292. doi:10.1002/maco.200303719Ottenwelter, E., & Costa, V. (2014). Evidence of Metallic Plating on Archaeological Artefacts by Voltammetry of Microparticles. Archaeometry, 57(3), 497-504. doi:10.1111/arcm.12091Doménech-Carbó, A., Doménech-Carbó, M., & Martínez-Lázaro, I. (2007). Electrochemical identification of bronze corrosion products in archaeological artefacts. A case study. Microchimica Acta, 162(3-4), 351-359. doi:10.1007/s00604-007-0839-3Šatović, D., Martinez, S., & Bobrowski, A. (2010). Electrochemical identification of corrosion products on historical and archaeological bronzes using the voltammetry of micro-particles attached to a carbon paste electrode. Talanta, 81(4-5), 1760-1765. doi:10.1016/j.talanta.2010.03.037Doménech-Carbó, A., Doménech-Carbó, M. T., Redondo-Marugán, J., Osete-Cortina, L., & Vivancos-Ramón, M. V. (2015). Electrochemical Characterization of Corrosion Products in Leaded Bronze Sculptures Considering Ohmic Drop Effects on Tafel Analysis. Electroanalysis, 28(4), 833-845. doi:10.1002/elan.201500613Blum, D., Leyffer, W., & Holze, R. (1996). Pencil-Leads as new electrodes for abrasive stripping voltammetry. Electroanalysis, 8(3), 296-297. doi:10.1002/elan.1140080317Doménech-Carbó, A., Doménech-Carbó, M. T., & Peiró-Ronda, Mªa. (2011). ‘One-Touch’ Voltammetry of Microparticles for the Identification of Corrosion Products in Archaeological Lead. Electroanalysis, 23(6), 1391-1400. doi:10.1002/elan.201000739Doménech, A., Lastras, M., Rodríguez, F., & Osete, L. (2013). Mapping of corrosion products of highly altered archeological iron using voltammetry of microparticles. Microchemical Journal, 106, 41-50. doi:10.1016/j.microc.2012.05.002Doménech, A., Doménech-Carbó, M. T., & Martínez-Lázaro, I. (2010). Layer-by-layer identification of copper alteration products in metallic works of art using the voltammetry of microparticles. Analytica Chimica Acta, 680(1-2), 1-9. doi:10.1016/j.aca.2010.09.002DOMÉNECH-CARBÓ, A., DOMÉNECH-CARBÓ, M. T., PEIRÓ-RONDA, M. A., & OSETE-CORTINA, L. (2011). ELECTROCHEMISTRY AND AUTHENTICATION OF ARCHAEOLOGICAL LEAD USING VOLTAMMETRY OF MICROPARTICLES: APPLICATION TO THE TOSSAL DE SANT MIQUEL IBERIAN PLATE. Archaeometry, 53(6), 1193-1211. doi:10.1111/j.1475-4754.2011.00608.xDoménech-Carbó, A., Doménech-Carbó, M. T., Pérez, M. L., & Herrero-Cortell, M. (2015). Detection of archaeological forgeries of Iberian lead plates using nanoelectrochemical techniques. The lot of fake plates from Bugarra (Spain). Forensic Science International, 247, 79-88. doi:10.1016/j.forsciint.2014.12.001Doménech-Carbó, A., Doménech-Carbó, M. T., & Peiró-Ronda, M. A. (2011). Dating Archeological Lead Artifacts from Measurement of the Corrosion Content Using the Voltammetry of Microparticles. Analytical Chemistry, 83(14), 5639-5644. doi:10.1021/ac200731qDoménech-Carbó, A., Doménech-Carbó, M. T., Capelo, S., Pasíes, T., & Martínez-Lázaro, I. (2014). Dating Archaeological Copper/Bronze Artifacts by Using the Voltammetry of Microparticles. Angewandte Chemie International Edition, 53(35), 9262-9266. doi:10.1002/anie.201404522Doménech-Carbó, A., Scholz, F., Doménech-Carbó, M. T., Piquero-Cilla, J., Montoya, N., Pasíes-Oviedo, T., … Oliver, A. (2018). Dating of Archaeological Gold by Means of Solid State Electrochemistry. ChemElectroChem, 5(15), 2113-2117. doi:10.1002/celc.201800435Doménech-Carbó, A., Doménech-Carbó, M. T., Redondo-Marugán, J., Osete-Cortina, L., Barrio, J., Fuentes, A., … Pasíes, T. (2017). Electrochemical Characterization and Dating of Archaeological Leaded Bronze Objects Using the Voltammetry of Immobilized Particles. Archaeometry, 60(2), 308-324. doi:10.1111/arcm.12308Di Turo, F., Montoya, N., Piquero-Cilla, J., De Vito, C., Coletti, F., Favero, G., … Doménech-Carbó, A. (2017). Dating Archaeological Strata in theMagna MaterTemple Using Solid-state Voltammetric Analysis of Leaded Bronze Coins. Electroanalysis, 30(2), 361-370. doi:10.1002/elan.201700724Doménech-Carbó, A., Doménech-Carbó, M. T., Álvarez-Romero, C., Montoya, N., Pasíes-Oviedo, T., & Buendía, M. (2017). Electrochemical Characterization of Coinage Techniques the 17thCentury: ThemaravedísCase. Electroanalysis, 29(9), 2008-2018. doi:10.1002/elan.201700326Pavlov, D., Monakhov, B., Maja, M., & Penazzi, N. (1989). Mechanism of Action of Sn on the Passivation Phenomena in the Lead‐Acid Battery Positive Plate (Sn‐Free Effect). Journal of The Electrochemical Society, 136(1), 27-33. doi:10.1149/1.2096603Cai, W.-B., Wan, Y.-Q., Liu, H.-T., & Zhou, W.-F. (1995). A study of the reduction process of anodic PbO2 film on Pb in sulfuric acid solution. Journal of Electroanalytical Chemistry, 387(1-2), 95-100. doi:10.1016/0022-0728(94)03866-2MEEKS, N. D. (1986). TIN-RICH SURFACES ON BRONZE?SOME EXPERIMENTAL AND ARCHAEOLOGICAL CONSIDERATIONS. Archaeometry, 28(2), 133-162. doi:10.1111/j.1475-4754.1986.tb00383.xSerghini-Idrissi, M., Bernard, M. C., Harrif, F. Z., Joiret, S., Rahmouni, K., Srhiri, A., … Ziani, M. (2005). Electrochemical and spectroscopic characterizations of patinas formed on an archaeological bronze coin. Electrochimica Acta, 50(24), 4699-4709. doi:10.1016/j.electacta.2005.01.050Bongiorno, V., Campodonico, S., Caffara, R., Piccardo, P., & Carnasciali, M. M. (2012). Micro‐Raman spectroscopy for the characterization of artistic patinas produced on copper‐based alloys. Journal of Raman Spectroscopy, 43(11), 1617-1622. doi:10.1002/jrs.4167Basso, E., Invernizzi, C., Malagodi, M., La Russa, M. F., Bersani, D., & Lottici, P. P. (2014). Characterization of colorants and opacifiers in roman glass mosaictesseraethrough spectroscopic and spectrometric techniques. Journal of Raman Spectroscopy, 45(3), 238-245. doi:10.1002/jrs.4449Ingo, G. M., Plescia, P., Angelini, E., Riccucci, C., & de Caro, T. (2006). Bronze roman mirrors: the secret of brightness. Applied Physics A, 83(4), 611-615. doi:10.1007/s00339-006-3535-

The Arabidopsis cer26 mutant, like the cer2 mutant, is specifically affected in the very long chain fatty acid elongation process

Plant aerial organs are covered by cuticular waxes, which form a hydrophobic crystal layer that mainly serves as a waterproof barrier. Cuticular wax is a complex mixture of very long chain lipids deriving from fatty acids, predominantly of chain lengths from 26 to 34 carbons, which result from acyl-CoA elongase activity. The biochemical mechanism of elongation is well characterized; however, little is known about the specific proteins involved in the elongation of compounds with more than 26 carbons available as precursors of wax synthesis. In this context, we characterized the three Arabidopsis genes of the CER2-like family: CER2, CER26 and CER26-like . Expression pattern analysis showed that the three genes are differentially expressed in an organ- and tissue-specific manner. Using individual TDNA insertion mutants, together with a cer2 cer26 double mutant, we characterized the specific impact of the inactivation of the different genes on cuticular waxes. In particular, whereas the cer2 mutation impaired the production of wax components longer than 28 carbons, the cer26 mutant was found to be affected in the production of wax components longer than 30 carbons. The analysis of the acyl-CoA pool in the respective transgenic lines confirmed that inactivation of both genes specifically affects the fatty acid elongation process beyond 26 carbons. Furthermore, ectopic expression of CER26 in transgenic plants demonstrates that CER26 facilitates the elongation of the very long chain fatty acids of 30 carbons or more, with high tissular and substrate specificity

Suspensions cellulaires embryogènes de bananiers et bananiers plantain

This guideline presents two protocols to produce embryogenic cell suspensions by using scalps or immature male flowers. The protocols have been developed by the Laboratory of Tropical Crop Improvement of Katholieke Universiteit Leuven (KULeuven) and the cellular biology laboratory of the Centre de coopération internationale en recherche agronomique pour le développement (Cirad)

Suspensiones de células embriogénicas de banano y plátano

This guideline presents two protocols to produce embryogenic cell suspensions by using scalps or immature male flowers. The protocols have been developed by the Laboratory of Tropical Crop Improvement of Katholieke Universiteit Leuven (KULeuven) and the cellular biology laboratory of the Centre de coopération internationale en recherche agronomique pour le développement (Cirad)

Banana and plantain embryogenic cell suspensions

This guideline presents two protocols to produce embryogenic cell suspensions by using scalps or immature male flowers. The protocols have been developed by the Laboratory of Tropical Crop Improvement of Katholieke Universiteit Leuven (KULeuven) and the cellular biology laboratory of the Centre de coopération internationale en recherche agronomique pour le développement (Cirad)

The BET bromodomain inhibitor I-BET-151 induces structural and functional alterations of the heart mitochondria in healthy male mice and rats

The bromodomain and extra-terminal domain family inhibitors (BETi) are a promising new class of anticancer agents. Since numerous anticancer drugs have been correlated to cardiomyopathy, and since BETi can affect non-cancerous tissues, we aimed to investigate in healthy animals any ultrastructural BETi-induced alterations of the heart as compared to skeletal muscle. Male Wistar rats were either treated during 3 weeks with I-BET-151 (2 or 10 mg/kg/day) (W3) or treated for 3 weeks then allowed to recover for another 3 weeks (W6) (3-weeks drug washout). Male C57Bl/6J mice were only treated during 5 days (50 mg/kg/day). We demonstrated the occurrence of ultrastructural alterations and progressive destruction of cardiomyocyte mitochondria after I-BET-151 exposure. Those mitochondrial alterations were cardiac muscle-specific, since the skeletal muscles of exposed animals were similar in ultrastructure presentation to the non-exposed animals. I-BET-151 decreased the respiration rate of heart mitochondria in a dose-dependent manner. At the higher dose, it also decreased mitochondrial mass, as evidenced by reduced right ventricular citrate synthase content. I-BET-151 reduced the right and left ventricular fractional shortening. The concomitant decrease in the velocity-time-integral in both the aorta and the pulmonary artery is also suggestive of an impaired heart function. The possible context-dependent cardiac side effects of these drugs have to be appreciated. Future studies should focus on the basic mechanisms of potential cardiovascular toxicities induced by BETi and strategies to minimize these unexpected complications

Contribution of CgPDR1-Regulated Genes in Enhanced Virulence of Azole-Resistant Candida glabrata

In Candida glabrata, the transcription factor CgPdr1 is involved in resistance to azole antifungals via upregulation of ATP binding cassette (ABC)-transporter genes including at least CgCDR1, CgCDR2 and CgSNQ2. A high diversity of GOF (gain-of-function) mutations in CgPDR1 exists for the upregulation of ABC-transporters. These mutations enhance C. glabrata virulence in animal models, thus indicating that CgPDR1 might regulate the expression of yet unidentified virulence factors. We hypothesized that CgPdr1-dependent virulence factor(s) should be commonly regulated by all GOF mutations in CgPDR1. As deduced from transcript profiling with microarrays, a high number of genes (up to 385) were differentially regulated by a selected number (7) of GOF mutations expressed in the same genetic background. Surprisingly, the transcriptional profiles resulting from expression of GOF mutations showed minimal overlap in co-regulated genes. Only two genes, CgCDR1 and PUP1 (for PDR1 upregulated and encoding a mitochondrial protein), were commonly upregulated by all tested GOFs. While both genes mediated azole resistance, although to different extents, their deletions in an azole-resistant isolate led to a reduction of virulence and decreased tissue burden as compared to clinical parents. As expected from their role in C. glabrata virulence, the two genes were expressed as well in vitro and in vivo. The individual overexpression of these two genes in a CgPDR1-independent manner could partially restore phenotypes obtained in clinical isolates. These data therefore demonstrate that at least these two CgPDR1-dependent and -upregulated genes contribute to the enhanced virulence of C. glabrata that acquired azole resistance