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1. Combined sensory, volatilome and transcriptome analyses identify a limonene terpene synthase as a major contributor to the characteristic aroma of a Coffea arabica L. specialty coffee

Author

Marie, Lison, Breitler, Jean-Christophe, Bamogo, Pingdwende Kader Aziz, Bordeaux, Mélanie, Lacombe, Séverine, Rios, Maëlle, Lebrun, Marc, Boulanger, Renaud, Lefort, Eveline, Nakamura, Sunao, Motoyoshi, Yudai, Mieulet, Delphine, Campa, Claudine, Legendre, Laurent, and Bertrand, Benoît

Published
2024
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11. Discovery and functional characterization of two diterpene synthases for sclareol biosynthesis in Salvia sclarea(L.) and their relevance for perfume manufacture

Author

Caniard, Anne, Zerbe, Philipp, Legrand, Sylvain, Cohade, Allison, Valot, Nadine, Magnard, Jean-Louis, Bohlmann, Jörg, and Legendre, Laurent

Subjects
Plant Biology, Biological Sciences, Alkyl and Aryl Transferases, Amino Acid Sequence, Chromatography, Liquid, DNA, Complementary, Diterpenes, Gas Chromatography-Mass Spectrometry, Gene Expression Regulation, Plant, Genetic Engineering, Molecular Sequence Data, Perfume, Phylogeny, Protein Transport, Reference Standards, Saccharomyces cerevisiae, Salvia, Sequence Alignment, Subcellular Fractions, Transcriptome, Diterpene, Sage, Salvia sclarea, Sclareol, Terpene synthase, Microbiology, Crop and Pasture Production, Plant Biology & Botany, Crop and pasture production, Plant biology
Abstract

BackgroundSclareol is a diterpene natural product of high value for the fragrance industry. Its labdane carbon skeleton and its two hydroxyl groups also make it a valued starting material for semisynthesis of numerous commercial substances, including production of Ambrox® and related ambergris substitutes used in the formulation of high end perfumes. Most of the commercially-produced sclareol is derived from cultivated clary sage (Salvia sclarea) and extraction of the plant material. In clary sage, sclareol mainly accumulates in essential oil-producing trichomes that densely cover flower calices. Manool also is a minor diterpene of this species and the main diterpene of related Salvia species.ResultsBased on previous general knowledge of diterpene biosynthesis in angiosperms, and based on mining of our recently published transcriptome database obtained by deep 454-sequencing of cDNA from clary sage calices, we cloned and functionally characterized two new diterpene synthase (diTPS) enzymes for the complete biosynthesis of sclareol in clary sage. A class II diTPS (SsLPPS) produced labda-13-en-8-ol diphosphate as major product from geranylgeranyl diphosphate (GGPP) with some minor quantities of its non-hydroxylated analogue, (9 S, 10 S)-copalyl diphosphate. A class I diTPS (SsSS) then transformed these intermediates into sclareol and manool, respectively. The production of sclareol was reconstructed in vitro by combining the two recombinant diTPS enzymes with the GGPP starting substrate and in vivo by co-expression of the two proteins in yeast (Saccharomyces cerevisiae). Tobacco-based transient expression assays of green fluorescent protein-fusion constructs revealed that both enzymes possess an N-terminal signal sequence that actively targets SsLPPS and SsSS to the chloroplast, a major site of GGPP and diterpene production in plants.ConclusionsSsLPPS and SsSS are two monofunctional diTPSs which, together, produce the diterpenoid specialized metabolite sclareol in a two-step process. They represent two of the first characterized hydroxylating diTPSs in angiosperms and generate the dihydroxylated labdane sclareol without requirement for additional enzymatic oxidation by activities such as cytochrome P450 monoxygenases. Yeast-based production of sclareol by co-expresssion of SsLPPS and SsSS was efficient enough to warrant the development and use of such technology for the biotechnological production of scareol and other oxygenated diterpenes.

Published
2012

13. Discovery and functional characterization of two diterpene synthases for sclareol biosynthesis in Salvia sclarea (L.) and their relevance for perfume manufacture.

Author

Caniard, Anne, Zerbe, Philipp, Legrand, Sylvain, Cohade, Allison, Valot, Nadine, Magnard, Jean-Louis, Bohlmann, Jörg, and Legendre, Laurent

Subjects
Subcellular Fractions, Saccharomyces cerevisiae, Salvia, Diterpenes, Alkyl and Aryl Transferases, DNA, Complementary, Perfume, Chromatography, Liquid, Genetic Engineering, Sequence Alignment, Phylogeny, Gene Expression Regulation, Plant, Amino Acid Sequence, Protein Transport, Reference Standards, Molecular Sequence Data, Gas Chromatography-Mass Spectrometry, Transcriptome, Diterpene, Sage, Salvia sclarea, Sclareol, Terpene synthase, DNA, Complementary, Chromatography, Liquid, Gene Expression Regulation, Plant, Microbiology, Plant Biology, Crop and Pasture Production, Plant Biology & Botany
Abstract

BackgroundSclareol is a diterpene natural product of high value for the fragrance industry. Its labdane carbon skeleton and its two hydroxyl groups also make it a valued starting material for semisynthesis of numerous commercial substances, including production of Ambrox® and related ambergris substitutes used in the formulation of high end perfumes. Most of the commercially-produced sclareol is derived from cultivated clary sage (Salvia sclarea) and extraction of the plant material. In clary sage, sclareol mainly accumulates in essential oil-producing trichomes that densely cover flower calices. Manool also is a minor diterpene of this species and the main diterpene of related Salvia species.ResultsBased on previous general knowledge of diterpene biosynthesis in angiosperms, and based on mining of our recently published transcriptome database obtained by deep 454-sequencing of cDNA from clary sage calices, we cloned and functionally characterized two new diterpene synthase (diTPS) enzymes for the complete biosynthesis of sclareol in clary sage. A class II diTPS (SsLPPS) produced labda-13-en-8-ol diphosphate as major product from geranylgeranyl diphosphate (GGPP) with some minor quantities of its non-hydroxylated analogue, (9 S, 10 S)-copalyl diphosphate. A class I diTPS (SsSS) then transformed these intermediates into sclareol and manool, respectively. The production of sclareol was reconstructed in vitro by combining the two recombinant diTPS enzymes with the GGPP starting substrate and in vivo by co-expression of the two proteins in yeast (Saccharomyces cerevisiae). Tobacco-based transient expression assays of green fluorescent protein-fusion constructs revealed that both enzymes possess an N-terminal signal sequence that actively targets SsLPPS and SsSS to the chloroplast, a major site of GGPP and diterpene production in plants.ConclusionsSsLPPS and SsSS are two monofunctional diTPSs which, together, produce the diterpenoid specialized metabolite sclareol in a two-step process. They represent two of the first characterized hydroxylating diTPSs in angiosperms and generate the dihydroxylated labdane sclareol without requirement for additional enzymatic oxidation by activities such as cytochrome P450 monoxygenases. Yeast-based production of sclareol by co-expresssion of SsLPPS and SsSS was efficient enough to warrant the development and use of such technology for the biotechnological production of scareol and other oxygenated diterpenes.

Published
2012

16. First record of a freshwater cave sponge (Porifera, unknown gen. and sp.) in a cave inhabited by Astyanax cavefish in the Sierra de El Abra, San Luis Potosí, Mexico

Author

Legendre, Laurent, Espinasa, Luis, Lacaille-Múzquiz, Jean-Louis, Alaniz-Garfía, Gabriel, Ornelas-García, Patricia, and Rétaux, Sylvie

Subjects
Vertebrata, Gnathostomata, Actinopterygii, Osteichthyes, Characidae, Astyanax, Animalia, subterranean, Characiformes, karst, Chordata, Biota, Porifera
Abstract

The karstic cave, la Cueva de Los Sabinos, located in the Sierra de El Abra in the state of San Luis Potosí, Mexico, is mostly known for hosting a population of blind, depigmented Astyanax mexicanus cavefish. Herein, we report the discovery of a non-pigmented sponge (Porifera) in the final sump of this cave. No genus or species name could be attributed because we did not collect any specimen. Up to now, the sponge distribution seems restricted to a single pool in la Cueva de Los Sabinos, but further careful exploration of other pools of the cave as well as closely related cavities is warranted. To our knowledge, this observation constitutes the fourth report of a freshwater, white, cave-adapted sponge in the world and the first for Mexico and North America. It is also the eleventh troglobite species encountered in Los Sabinos. Our discovery confirms the exceptionally rich biodiversity of this cave ecosystem.

Published
2023

18. Genetic identification and reiterated captures suggests that theAstyanax mexicanusEl Pachón cavefish population is closed and declining

Author

Legendre, Laurent, primary, Rode, Julie, additional, Germon, Isabelle, additional, Pavie, Marie, additional, Quiviger, Carla, additional, Policarpo, Maxime, additional, Leclercq, Julien, additional, Père, Stéphane, additional, Fumey, Julien, additional, Hyacinthe, Carole, additional, Ornelas-García, Patricia, additional, Espinasa, Luis, additional, Rétaux, Sylvie, additional, and Casane, Didier, additional

Published
2022
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20. Genetic identification and reiterated captures suggest that the Astyanax mexicanus El Pachón cavefish population is closed and declining.

Author

Legendre, Laurent, Rode, Julie, Germon, Isabelle, Pavie, Marie, Quiviger, Carla, Policarpo, Maxime, Leclercq, Julien, Père, Stéphane, Fumey, Julien, Hyacinthe, Carole, Ornelas-García, Patricia, Espinasa, Luis, Rétaux, Sylvie, and Casane, Didier

Subjects
ASTYANAX, IDENTIFICATION, DEMOGRAPHIC change, CAVES, CENSUS
Abstract

The sizes of Astyanax mexicanus blind cavefish populations of North-East Mexico are demographic parameters of great importance for investigating a variety of ecological, evolutionary, and conservation issues. However, few estimates have been obtained. For these mobile animals living in an environment difficult to explore as a whole, methods based on capture-mark-recapture are appropriate, but their feasibility and interpretation of results depend on several assumptions that must be carefully examined. Here, we provide evidence that minimally invasive genetic identification from captures at different time intervals (three days and three years) can give insights into cavefish population size dynamics as well as other important demographic parameters of interest. We also provide tools to calibrate sampling and genotyping efforts necessary to reach a given level of precision. Our results suggest that the El Pachón cave population is currently very small, of an order of magnitude of a few hundreds of individuals, and is distributed in a relatively isolated area. The probable decline in population size in the El Pachón cave since the last census in 1971 raises serious conservation issues. [ABSTRACT FROM AUTHOR]

Published
2023
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22. Additional file 1 of Coevolution of the olfactory organ and its receptor repertoire in ray-finned fishes

Author

Policarpo, Maxime, Bemis, Katherine E., Laurenti, Patrick, Legendre, Laurent, Sandoz, Jean-Christophe, Rétaux, Sylvie, and Casane, Didier

Abstract

Additional file 1: Figure S1. Diversity of the olfactory receptor gene repertoire in ray-finned fishes. Figure S2. Distribution of ORA7 and ORA8 subfamilies in ray-finned fishes. Figure S3. Distribution of birth and death rates of OR, TAAR, OlfC and ORA genes in ray-finned fishes. Figure S4. Correlation of the number of gene losses (or gene gains) between gene families, estimated using the 368 branches of the phylogenetic tree. Figure S5. Correlation between the number of OR, TAAR and OlfC pseudogenes in 185 ray-finned fishes. Figure S6. Narial tubes of four species of ray-finned fishes with complex olfactory organs and large gene repertoires. Figure S7. Takifugu rubripes, USNM 57620, 290 mm TL.

Published
2022
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23. Additional file 4 of Coevolution of the olfactory organ and its receptor repertoire in ray-finned fishes

Author

Policarpo, Maxime, Bemis, Katherine E., Laurenti, Patrick, Legendre, Laurent, Sandoz, Jean-Christophe, Rétaux, Sylvie, and Casane, Didier

Abstract

Additional file 4. Supplementary Data 3. (A) Phylogeny of OR genes from 44 species representing 44 orders of ray-finned fishes sampled in this study. Branches are colored according to the gene subfamily classification. (B) Phylogeny of all TAAR genes retrieved from 185 ray-finned fishes. Branches are colored according to gene family classification. Outgroup sequences (nonTAAR GPCRs) are colored in black. (C) Phylogeny of all OlfC genes retrieved from 185 ray-finned fish. Branches are colored according to the gene subfamily classification. Outgroup sequences (CasR and V2R2 genes) are colored in black. (D) Phylogeny of all ORA genes retrieved from 185 ray-finned fishes. Branches are colored according to the gene subfamily classification. Outgroup sequences (T2R genes) are colored in black.

Published
2022
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27. Eye morphogenesis in the blind Mexican cavefish

Author

Devos, Lucie, primary, Agnès, François, additional, Edouard, Joanne, additional, Simon, Victor, additional, Legendre, Laurent, additional, El Khallouki, Naima, additional, Barbachou, Sosthène, additional, Sohm, Frédéric, additional, and Rétaux, Sylvie, additional

Published
2021
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29. Unravelling the metabolic and hormonal machinery during key steps of somatic embryogenesis: A case study in coffee

Author

Awada, Rayan, Campa, Claudine, Gibault, Estelle, Déchamp, Eveline, Georget, Frédéric, Lepelley, Maud, Abdallah, Cécile, Erban, Alexender, Martinez-Seidel, Frederico, Kopka, Joachim, LEGENDRE, Laurent, Léran, Sophie, Conejero, Geneviève, Verdeil, Jean-Luc, Crouzillat, Dominique, Breton, David, Bertrand, Benoit, Etienne, Hervé, UMR - Interactions Plantes Microorganismes Environnement (UMR IPME), Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Institut de Recherche pour le Développement (IRD [Bolivie]), Université de Montpellier (UM), Diversité, adaptation, développement des plantes (UMR DIADE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Max-Planck-Institut für Molekulare Pflanzenphysiologie (MPI-MP), Max-Planck-Gesellschaft, Laboratoire d'Ecologie Microbienne - UMR 5557 (LEM), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Ecole Nationale Vétérinaire de Lyon (ENVL), Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Plateforme d'histocytologie et d'imagerie cellulaire végétale (PHIV), Biochimie et Physiologie Moléculaire des Plantes (BPMP), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Institut National de la Recherche Agronomique (INRA)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Institut National de la Recherche Agronomique (INRA)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro), French National Research Agency (ANR) : ANR-10-INBS-04, ANR-10-INBS-0004,France-BioImaging,Développment d'une infrastructure française distribuée coordonnée(2010), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut de Recherche pour le Développement (IRD [France-Sud]), Université de Lyon-Université de Lyon-Ecole Nationale Vétérinaire de Lyon (ENVL)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro)-Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro)-Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro), Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro), Plant Science Unit, Institute for Agricultural and Fisheries Research, Max Planck Institute for Molecular Plant Physiology, Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Lyon (ENVL)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Université de Montpellier (UM)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Plateforme RIO Imaging (PHIV MRI), and Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro)-Institut National de la Recherche Agronomique (INRA)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)

Subjects
Plant Somatic Embryogenesis Techniques, coffee, Coffea, Article, F30 - Génétique et amélioration des plantes, lcsh:Chemistry, histology, histologie, cellule totipotente, Plant Growth Regulators, cell imaging, Expression des gènes, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, totipotency, Marqueur génétique, hormone content, Microscopie, lcsh:QH301-705.5, Différenciation cellulaire, imagerie cellulaire, cell fate, Vegetal Biology, somatic embryogenesis, embryogénèse somatique, metabolomics, Plant Leaves, physiologie hormonale, lcsh:Biology (General), lcsh:QD1-999, Microscopie immunofluorescente, U30 - Méthodes de recherche, Biologie végétale
Abstract

RATIONALE - Somatic embryogenesis (SE) is one of the most promising processes for large-scale dissemination of elite varieties. However, whatever the species considered, SE research still remains essentially empirical resulting in many drawbacks to fulfil market demands, especially due to an overall slow technical progress over the last 20 years. Knowledge about the molecular events involved in the key steps of the SE process is urgently needed to pilot the optimization of SE protocols. In this study, we took advantage of the latest metabolomics technologies and applied them to one of the most advanced and reliable large-scale SE processes, the one developed for coffee. METHODS - Sampling covered 15 key developmental stages. Five independent leaf introductions were carried out with more than 4,000 leaf explants and a total of 25 independent cell lines. All obtained cell lines were high-yielding and time-synchronized during embryo regeneration enabling a successful sampling. Primary metabolites, secondary metabolites and phytohormones were quantified using GC-MS, HPLC and UPLC- MS/MS respectively. A robust statistical method was used to identify metabolic pathway changes associated with the main developmental phases and phase switches. Histological analysis and cell imaging were also required to characterize developmental stages and associate metabolic profiles with cell structure organization. Lastly, comparing Arabica embryogenic and non-embryogenic calli enabled the identification of metabolic markers of the embryogenic capacity. RESULTS - Statistical analysis performed on 104 metabolites revealed that massive re-configuration of metabolic pathways induced SE. During initial dedifferentiation, a sharp decrease in phenolic compounds and caffeine levels was observed while auxins, cytokinins and ethylene levels were at their highest. Totipotency reached its highest expression during the callus stages when a shut-off in hormonal and metabolic pathways related to sugar and energetic substance hydrolysis was evidenced. Abscisic acid, leucine, maltotriose, myo-inositol, proline, tricarboxylic acid cycle metabolites and zeatin appeared as key metabolic markers of the embryogenic capacity. Combining metabolomics with multiphoton microscopy led to the identification of chlorogenic acids as markers of embryo redifferentiation. CONCLUSIONS & PERSPECTIVES - The present analysis shows that metabolite fingerprints are signatures of cell fate and represent a starting point for optimizing SE protocols in a rational way. These findings should be informative and useful to a wide range of plant species, offering unprecedented perspectives in plant micropropagation.

Published
2021
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30. 5-CQA and mangiferin, two leaf biomarkers of adaptation to full sun or shade conditions in Coffea arabica L.

Author

Duangsodsri, Teerarat, Villain, Luc, Vestalys, Ialy Rojo, Michalet, Serge, Abdallah, Cécile, Breitler, Jean-Christophe, Bordeaux, Mélanie, Villegas, Andres Mauricio, Raherimandimby, Marson, Legendre, Laurent, Etienne, Hervé, Bertrand, Benoît, Campa, Claudine, Duangsodsri, Teerarat, Villain, Luc, Vestalys, Ialy Rojo, Michalet, Serge, Abdallah, Cécile, Breitler, Jean-Christophe, Bordeaux, Mélanie, Villegas, Andres Mauricio, Raherimandimby, Marson, Legendre, Laurent, Etienne, Hervé, Bertrand, Benoît, and Campa, Claudine

Abstract

Phenolic compounds are involved in plant response to environmental conditions and are highly present in leaves of Coffea arabica L., originally an understory shrub. To increase knowledge of C. arabica leaf phenolic compounds and their patterns in adaptation to light intensity, mature leaves of Ethiopian wild accessions, American pure lines and their relative F1 hybrids were sampled in full sun or under 50% shade field plots in Mexico and at two contrasting elevations in Nicaragua and Colombia. Twenty-one phenolic compounds were identified by LC-DAD-MS2 and sixteen were quantified by HPLC-DAD. Four of them appeared to be involved in C. arabica response to light intensity. They were consistently more accumulated in full sun, presenting a stable ratio of leaf content in the sun vs. shade for all the studied genotypes: 1.6 for 5-CQA, F-dihex and mangiferin and 2.8 for rutin. Moreover, 5-CQA and mangiferin contents, in full sun and shade, allowed for differentiating the two genetic groups of Ethiopian wild accessions (higher contents) vs. cultivated American pure lines. They appear, therefore, to be potential biomarkers of adaptation of C. arabica to light intensity for breeding programs. We hypothesize that low 5-CQA and mangiferin leaf contents should be searched for adaptation to full-sun cropping systems and high contents used for agroforestry systems.

Published
2020

31. Phylogenetic analysis of Pinguicula (Lentibulariaceae): chloroplast DNA sequences and morphology support several geographically distinct radiations

Author

Cieslak, Thomas, Polepalli, Jai Santosh, White, Adam, Muller, Kai, Borsch, Thomas, Barthlott, Wilhelm, Steiger, Juerg, Marchant, Adam, and Legendre, Laurent

Subjects
Lamiales -- Genetic aspects, Biogeography -- Research, Botany -- Morphology, Botany -- Research, Biological sciences
Abstract

The genus Pinguicula is one of the three genera of the carnivorous Lentibulariaceae, comprising approximately 80 species. Phylogeny inference using nucleotide sequences of the chloroplast gone matK and the trnK group II intron, as well as a set of 32 morphological characters revealed five well-supported, major lineages within the genus. These lineages largely reflect radiations in clearly defined geographic regions, whereas most previously recognized sections of the genus are shown to be para-or polyphyletic. A species-rich Mexican-Central American-Caribbean clade has the Eurasian P. alpina and an East Asian clade as successive sisters. All three are characterized by a production of flower buds on winter-resting plants, a specific corolla hair structure and a very large corolla lower central lobe. Another diverse clade is composed of species with primarily European distribution including the widespread type species P. vulgaris. For this clade, vegetative reproduction during dormancy is synapomorphic. Species native to SE North America and the South American Andes and a group of Mediterranean and NE Atlantic coast species together appear in a fifth well- supported clade, that is characterized by a tropical-type growth habit. It is the only clade that has reached temperate zones of the southern hemisphere. Key words: Lamiales; Lentibulariaceae; matK/trnK; morphology; northern hemisphere biogeography; Pinguicula.

Published
2005

33. 5-CQA and Mangiferin, Two Leaf Biomarkers of Adaptation to Full Sun or Shade Conditions in Coffea arabica L.

Author

Duangsodsri, Teerarat, primary, Villain, Luc, additional, Vestalys, Ialy Rojo, additional, Michalet, Serge, additional, Abdallah, Cécile, additional, Breitler, Jean-Christophe, additional, Bordeaux, Mélanie, additional, Villegas, Andres Mauricio, additional, Raherimandimby, Marson, additional, Legendre, Laurent, additional, Etienne, Hervé, additional, Bertrand, Benoît, additional, and Campa, Claudine, additional

Published
2020
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38. Culture of Adult Ascidians and Ascidian Genetics

Author

Hendrickson, Carolyn, primary, Christiaen, Lionel, additional, Deschet, Karine, additional, Jiang, Di, additional, Joly, Jean-Stéphane, additional, Legendre, Laurent, additional, Nakatani, Yuki, additional, Tresser, Jason, additional, and Smith, William C., additional

Published
2004
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39. Morphogenetic and patterning defects explain the coloboma phenotype of the eye in the Mexican cavefish

Author

Devos, Lucie, Klee, Florent, Edouard, Joanne, Simon, Victor, Legendre, Laurent, Khallouki, Naima El, Blin, Maryline, Barbachou, Sosthène, Sohm, Frédéric, Rétaux, Sylvie, Institut des Neurosciences Paris-Saclay (NeuroPSI), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Animaux Modèles Aquatiques : ingéniérie GENétique (AMAGEN), and Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, genetic structures, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, sense organs, eye diseases
Abstract

The morphogenesis of the vertebrate eye consists of a complex choreography of cell movements, tightly coupled to axial regionalization and cell type specification processes. Any disturbance in these events can lead to developmental defects and blindness. Here we have deciphered the sequence of defective events leading to coloboma phenotype in the embryonic eye of the blind cavefish of the species Astyanax mexicanus . Using comparative live imaging on targeted enhancer-trap Zic1:hsp70:GFP reporter lines of both the normal, river-dwelling morph and the cave morph of the species, we identified major defects in initial optic vesicle size and optic cup invagination in cavefish. Combining these results with gene expression analyses, we also discovered defects in axial patterning affecting mainly the temporal retina, in optic stalk tissue specification, and in the spreading processes involving the retinal pigmented epithelium cells. Based on these results, we propose a developmental scenario to explain the cavefish phenotype and discuss developmental constraints to morphological evolution. The cavefish eye appears as an outstanding natural mutant model to study molecular and cellular processes involved in optic region morphogenesis.

Published
2019
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40. Unravelling the metabolic and hormonal machinery during key steps of somatic embryogenesis: A case study in coffee

Author

Awada, Rayan, Campa, Claudine, Gibault, Estelle, Dechamp, Eveline, Georget, Frederic, Lepelley, Maud, Abdallah, Cécile, Erban, Alexander, Martinez-Seidel, Federico, Kopka, Joachim, Legendre, Laurent, Leran, Sophie, Conejero, Geneviève, Verdeil, Jean-Luc, Crouzillat, Dominique, Breton, David, Bertrand, Benoît, Etienne, Hervé, Awada, Rayan, Campa, Claudine, Gibault, Estelle, Dechamp, Eveline, Georget, Frederic, Lepelley, Maud, Abdallah, Cécile, Erban, Alexander, Martinez-Seidel, Federico, Kopka, Joachim, Legendre, Laurent, Leran, Sophie, Conejero, Geneviève, Verdeil, Jean-Luc, Crouzillat, Dominique, Breton, David, Bertrand, Benoît, and Etienne, Hervé

Abstract

Somatic embryogenesis (SE) is one of the most promising processes for large-scale dissemination of elite varieties. However, for many plant species, optimizing SE protocols still relies on a trial-and-error approach. Using coffee as a model plant, we report here the first global analysis of metabolome and hormone dynamics aiming to unravel mechanisms regulating cell fate and totipotency. Sampling from leaf explant dedifferentiation until embryo development covered 15 key stages. An in-depth statistical analysis performed on 104 metabolites revealed that massive re-configuration of metabolic pathways induced SE. During initial dedifferentiation, a sharp decrease in phenolic compounds and caffeine levels was also observed while auxins, cytokinins and ethylene levels were at their highest. Totipotency reached its highest expression during the callus stages when a shut-off in hormonal and metabolic pathways related to sugar and energetic substance hydrolysis was evidenced. Abscisic acid, leucine, maltotriose, myo-inositol, proline, tricarboxylic acid cycle metabolites and zeatin appeared as key metabolic markers of the embryogenic capacity. Combining metabolomics with multiphoton microscopy led to the identification of chlorogenic acids as markers of embryo redifferentiation. The present analysis shows that metabolite fingerprints are signatures of cell fate and represent a starting point for optimizing SE protocols in a rational way.

Published
2019

41. Kavalactones and flavokavins profiles contribute to quality assessment of kava (Piper methysticum G. Forst.), the Traditional beverage of the Pacific

Author

Lebot, Vincent, Michalet, Serge, Legendre, Laurent, Lebot, Vincent, Michalet, Serge, and Legendre, Laurent

Abstract

Kava (Piper methysticum) is increasingly traded internationally and there is need for a rapid method to analyze kava raw material before export. The objectives of the present study were: (i) to develop a simple and robust protocol for high throughput simultaneous quantification of kavalactones (KLs) and flavokavins (FKs) in kava and (ii) to assess its potential for quality control. Methysticin; dihydromethysticin; kavain; desmethoxyyangonin; dihydrokavain; yangonin; and flavokavin A, B and C were quantified using HPTLC in acetonic extracts of 174 kava varieties. UHPLC analysis was conducted on a subset of six varieties representing the genetic variation of the species. The genetically distinct groups of nobles, two-day and wichmannii varieties were clearly differentiated and multivariate analyses of UHPLC and HPTLC data were congruent. Noble varieties have significantly low FKs/KLs (0.13) and high kavain/flavokavin B (K/FKB = 7.31). Two-day and wichmannii varieties are characterized by high FKs/KLs (0.36, 0.21) and low K/FKB (1.5, 1.7). A high-throughput HPTLC protocol was developed with a total analytical time of 50 min for 20 samples and only 10 mL of mobile phase. The use of acetone, sonication and two different detection wavelengths improves the accuracy compared to previous HPLC studies and confirms that kava varieties exhibit distinct chemotypes clearly differentiated by their FKs/KLs profiles. These results will strengthen the use of Codex Alimentarius regional standards.

Published
2019

43. Eye morphogenesis in the blind Mexican cavefish

Author

Devos, Lucie, primary, Agnès, François, additional, Edouard, Joanne, additional, Simon, Victor, additional, Legendre, Laurent, additional, Khallouki, Naima El, additional, Barbachou, Sosthène, additional, Sohm, Frédéric, additional, and Rétaux, Sylvie, additional

Published
2019
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46. Comparing growth in surface and cave morphs of the species Astyanax mexicanus: insights from scales

Author

Simon, Victor, Elleboode, Romain, Mahé, Kélig, Legendre, Laurent, Ornelas-Garcia, Patricia, Espinasa, Luis, Rétaux, Sylvie, Institut des Neurosciences Paris-Saclay (NeuroPSI), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), IFREMER, Fisheries Laboratory, Sclerochronology Centre, Laboratoire Ressources halieutiques, Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Animaux Modèles Aquatiques : ingéniérie GENétique (AMAGEN), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Department of Zoology, Universidad Nacional Autónoma de Mexico, School of Science, Marist College, Marist College, and Institut des Neurosciences de Paris-Saclay (Neuro-PSI)

Subjects
genetic structures, [SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, Research, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, lcsh:Evolution, Cave, Scales, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, social sciences, Comparative biology, Growth, Life history traits, musculoskeletal system, humanities, Age, Fish, Size, Food, lcsh:QH359-425, sense organs
Abstract

Background Life in the darkness of caves is accompanied, throughout phyla, by striking phenotypic changes including the loss or severe reduction in eyes and pigmentation. On the other hand, cave animals have undergone constructive changes, thought to be adaptive, to survive in this extreme environment. The present study addresses the question of the evolution of growth in caves, taking advantage of the comparison between the river-dwelling and the cave-dwelling morphs of the Mexican tetra, Astyanax mexicanus. Results A sclerochronology approach was undertaken to document the growth of the species in these two very distinct habitats. Scales from 158 wild Astyanax mexicanus specimens were analyzed from three caves (Pachón, Tinaja and Subterráneo) and two rivers (Rio Gallinas and Arroyo Lagarto) in San Luis Potosi and Tamaulipas, Mexico. A 10–13% reduction in scales size was observed in the cave morphs compared to the surface morphs. Age could be reliably inferred from annual growth increments on the scales from the two morphs of the species. Further comparisons with growth curves in laboratory conditions, obtained using the von Bertalanffy growth model, were also performed. In the wild and in the laboratory, cavefish originating from the Pachón cave reached smaller sizes than surface fish from three different locations: Rio Gallinas and Arroyo Lagarto (wild sampling) and Texas (laboratory population), respectively. Wild Pachón cavefish also seemed to grow to smaller sizes than the two other wild cavefish populations studied, Tinaja and Subterráneo. Finally, growth in the laboratory was faster than in the wild, particularly in the two first years of life. Conclusions These data suggest that cavefish originating from the Pachón cave are subjected to an intrinsic limitation of their final size, which is at least in part independent from energy/food availability. This growth limitation may be an advantageous way of limiting energy expenditure and food needs in the cave environment. Moreover, growth regulation evolved differently in independently evolved cave populations. These results are discussed with regard to the sources of energy or general ecological conditions present in caves, and to the differences in behavior or feeding skills known in cavefish.

Published
2017
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47. Lens apoptosis in the Astyanax blind cavefish is not triggered by its small size or defects in morphogenesis

Author

Hinaux, Hélène, Recher, Gaëlle, Alié, Alexandre, Legendre, Laurent, Blin, Maryline, Rétaux, Sylvie, Institut des Neurosciences de Paris-Saclay (Neuro-PSI), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), BioEmergences, Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Animaux Modèles Aquatiques : ingéniérie GENétique (AMAGEN), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Institut des Neurosciences Paris-Saclay (NeuroPSI), Agence Nationale pour la Recherche (ANR), Centre National de la Recherche Scientifique (CNRS), Retina France, and ANR

Subjects
Embryology, Ocular Anatomy, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Equipment, lcsh:Medicine, Apoptosis, Blindness, Retina, Ocular System, evolution, expression, Lens, Crystalline, Medicine and Health Sciences, Morphogenesis, Animals, gene, lcsh:Science, Lens (Anatomy), eye degeneration, fish, mechanisms, Cell Death, [SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, Characidae, Lasers, Embryos, lcsh:R, Biology and Life Sciences, Gene Expression Regulation, Developmental, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, Cell Differentiation, Cell Biology, zebrafish, Optical Lenses, Biological Evolution, alpha-a-crystallin, anterior segment dysgenesis, mexicanus, Optical Equipment, Cell Processes, Engineering and Technology, Eyes, lcsh:Q, Anatomy, Head, Research Article, Developmental Biology
Abstract

International audience; Blindness is a convergent trait in many cave animals of various phyla. Astyanax mexicanus cavefish is one of the best studied cave animals; however the mechanisms underlying eye degeneration in this species are not yet completely understood. The lens seems to play a central role, but only relatively late differentiation defects have been implicated in the cavefish lens apoptosis phenotype so far. Here, we used genetic crosses between Astyanax cavefish and surface fish to confirm that during development, lens size is independent of retina size. We then investigated whether the small size of the cavefish lens could directly cause cell death. Laser ablation experiments of lens placode cells in surface fish embryos showed that a small lens size is not sufficient to trigger lens apoptosis. We further examined potential lens morphogenesis defects through classical histology and live-imaging microscopy. From lens placode to lens ball, we found that lens invagination and formation of the lens epithelium and fiber cells occur normally in cavefish. We conclude that the main and deleterious defect in the Astyanax cavefish lens must concern the molecular control of lens cell function.

Published
2017
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