@article{
 title = {Multimodal assessment of flax dew retting and its functional impact on fibers and natural fiber composites},
 type = {article},
 year = {2020},
 keywords = {Composites,Enzymes,Fibers,Metabarcoding,Retting,Tensile},
 pages = {112255},
 volume = {148},
 websites = {https://doi.org/10.1016/j.indcrop.2020.112255,https://linkinghub.elsevier.com/retrieve/pii/S0926669020301710},
 month = {6},
 publisher = {Elsevier},
 id = {0d95f633-bb91-36bd-a598-4ecf564d5b13},
 created = {2020-03-09T16:53:09.513Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.853Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Chabbert2020},
 private_publication = {false},
 abstract = {Flax (Linum usitatissinum L.) is an economically important fiber crop species as it produces long cellulosic fibers with high tensile strength. The first step in flax fiber extraction occurs after harvest via a process known as dew retting in which pulled (uprooted) plants are placed on the soil. During this process microorganisms from the soil and the phyllosphere develop on the plants, leading to partial decomposition of the stem tissues and facilitated mechanical extraction and improved fiber quality. Despite its importance, the management of dew retting is mainly based on empirical knowledge that makes the process difficult to control. In this study the dynamics of flax dew retting were investigated for the first time by combining targeted-metagenomics (metabarcoding), enzymatic activities and chemical and microscopic characterization of the stem. This multimodal approach indicated that intensive microbial colonization and major chemical changes in the stem cell wall composition occurred during the first weeks of retting leading to progressive fiber bundle decohesion in the outer stem tissues. The main changes could be explained by the degradation of pectin-rich and non-lignified thin walls of both the parenchyma cells surrounding fiber bundles, and the differentiating xylem and cambial cells. Field emission scanning electron microscopy suggested that the ultrastructure of the fiber secondary wall is weakly impacted by dew-retting. Analyses indicated that the mechanical properties of extracted technical fibers were improved during the last retting stages. Overall data on thermoplastic composites produced by a twin-screw extrusion and injection molding processes suggested that the retting degree had a small but significant effect on tensile properties.},
 bibtype = {article},
 author = {Chabbert, Brigitte and Padovani, Justine and Djemiel, Christophe and Ossemond, Jordane and Lemaître, Alain and Yoshinaga, Arata and Hawkins, Simon and Grec, Sébastien and Beaugrand, Johnny and Kurek, Bernard},
 doi = {10.1016/j.indcrop.2020.112255},
 journal = {Industrial Crops and Products},
 number = {January}
}

Flax (Linum usitatissinum L.) is an economically important fiber crop species as it produces long cellulosic fibers with high tensile strength. The first step in flax fiber extraction occurs after harvest via a process known as dew retting in which pulled (uprooted) plants are placed on the soil. During this process microorganisms from the soil and the phyllosphere develop on the plants, leading to partial decomposition of the stem tissues and facilitated mechanical extraction and improved fiber quality. Despite its importance, the management of dew retting is mainly based on empirical knowledge that makes the process difficult to control. In this study the dynamics of flax dew retting were investigated for the first time by combining targeted-metagenomics (metabarcoding), enzymatic activities and chemical and microscopic characterization of the stem. This multimodal approach indicated that intensive microbial colonization and major chemical changes in the stem cell wall composition occurred during the first weeks of retting leading to progressive fiber bundle decohesion in the outer stem tissues. The main changes could be explained by the degradation of pectin-rich and non-lignified thin walls of both the parenchyma cells surrounding fiber bundles, and the differentiating xylem and cambial cells. Field emission scanning electron microscopy suggested that the ultrastructure of the fiber secondary wall is weakly impacted by dew-retting. Analyses indicated that the mechanical properties of extracted technical fibers were improved during the last retting stages. Overall data on thermoplastic composites produced by a twin-screw extrusion and injection molding processes suggested that the retting degree had a small but significant effect on tensile properties.

@article{
 title = {Targeted Metagenomics of Retting in Flax: The Beginning of the Quest to Harness the Secret Powers of the Microbiota},
 type = {article},
 year = {2020},
 keywords = {cell wall,flax,holobiont,meta-omics,microbiota,natural fibers,retting,soil},
 pages = {1-22},
 volume = {11},
 id = {559054be-ca91-3406-a2c4-9a1efa7ee213},
 created = {2020-11-30T08:53:49.389Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.355Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Djemiel2020},
 private_publication = {false},
 abstract = {The mechanical and chemical properties of natural plant fibers are determined by many different factors, both intrinsic and extrinsic to the plant, during growth but also after harvest. A better understanding of how all these factors exert their effect and how they interact is necessary to be able to optimize fiber quality for use in different industries. One important factor is the post-harvest process known as retting, representing the first step in the extraction of bast fibers from the stem of species such as flax and hemp. During this process microorganisms colonize the stem and produce hydrolytic enzymes that target cell wall polymers thereby facilitating the progressive destruction of the stem and fiber bundles. Recent advances in sequencing technology have allowed researchers to implement targeted metagenomics leading to a much better characterization of the microbial communities involved in retting, as well as an improved understanding of microbial dynamics. In this paper we review how our current knowledge of the microbiology of retting has been improved by targeted metagenomics and discuss how related ‘-omics’ approaches might be used to fully characterize the functional capability of the retting microbiome.},
 bibtype = {article},
 author = {Djemiel, Christophe and Goulas, Estelle and Badalato, Nelly and Chabbert, Brigitte and Hawkins, Simon and Grec, Sébastien},
 doi = {10.3389/fgene.2020.581664},
 journal = {Frontiers in Genetics},
 number = {October}
}

The mechanical and chemical properties of natural plant fibers are determined by many different factors, both intrinsic and extrinsic to the plant, during growth but also after harvest. A better understanding of how all these factors exert their effect and how they interact is necessary to be able to optimize fiber quality for use in different industries. One important factor is the post-harvest process known as retting, representing the first step in the extraction of bast fibers from the stem of species such as flax and hemp. During this process microorganisms colonize the stem and produce hydrolytic enzymes that target cell wall polymers thereby facilitating the progressive destruction of the stem and fiber bundles. Recent advances in sequencing technology have allowed researchers to implement targeted metagenomics leading to a much better characterization of the microbial communities involved in retting, as well as an improved understanding of microbial dynamics. In this paper we review how our current knowledge of the microbiology of retting has been improved by targeted metagenomics and discuss how related ‘-omics’ approaches might be used to fully characterize the functional capability of the retting microbiome.

@inbook{
 type = {inbook},
 year = {2019},
 pages = {145-165},
 websites = {http://link.springer.com/10.1007/978-3-030-23964-0_10},
 id = {b1d2b513-b1a8-31c3-b339-e52546323c7f},
 created = {2019-09-11T16:11:30.736Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.426Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Grec2019},
 private_publication = {false},
 abstract = {The assembled genome sequence has been augmented by other molecular resources including the characterization of both coding and small RNA molecules. The transcriptomic analysis of the flax seed development will further the improvement of oil production and health-modulating phytochemicals of flax, which, along with the bast fiber, are arguably the most economically important traits in flax. The assembled genome sequence has also facilitated the theoretical identification of small RNA molecules as well as their physical identification. Although there is more variation in the wild flax progenitor germplasm than in the cultivated varieties, chemical mutagenesis-derived mutant populations have been successfully used to identify important genes involved in cell wall formation among others. Flax is also amenable to transformation, and the development of the floral dip procedure for this species may facilitate other approaches to mutant production and increasing available variation. However, the issues with the transgenic flax, Triffid, and its unregulated escape into the commercial seed provide a cautionary tale for the development of genetically modified flax. Some flax varieties have a particular characteristic, which is not known in other plant species, namely, the rapid modulation of its genome under certain growth conditions. The variation, with the generation of altered stable lines termed genotrophs, has been shown to occur within a specific defined subset of the genome. Understanding the mechanisms and characteristics of this genome compartment that appears to have the function of modifying the phenotype without the deleterious effects associated with a random mutagenesis makes it an interesting evolutionary mechanism. The plethora of molecular tools available allows this phenomenon to be understood at the molecular level, and potentially capable of manipulation. The molecular resources available, the commercially important, and the health-related characteristics make flax an accessible interesting model system for understanding complex pathways while also providing the basis for improving the commercial attractiveness of the crop. The development of the molecular resources for flax is directly attributable to the investment in the TUFGEN program and to the collaborations that were fostered, and continue to thrive, through the program.},
 bibtype = {inbook},
 author = {Grec, Sébastien and Dalmais, Marion and Chatterjee, Manash and Bendahmane, Abdelhafid and Hawkins, Simon},
 doi = {10.1007/978-3-030-23964-0_10},
 chapter = {Development of Flax (Linum usitatissimum L.) Mutant Populations for Forward and Reverse Genetics},
 title = {Plant Genetics and Genomics: Crops and Models}
}

The assembled genome sequence has been augmented by other molecular resources including the characterization of both coding and small RNA molecules. The transcriptomic analysis of the flax seed development will further the improvement of oil production and health-modulating phytochemicals of flax, which, along with the bast fiber, are arguably the most economically important traits in flax. The assembled genome sequence has also facilitated the theoretical identification of small RNA molecules as well as their physical identification. Although there is more variation in the wild flax progenitor germplasm than in the cultivated varieties, chemical mutagenesis-derived mutant populations have been successfully used to identify important genes involved in cell wall formation among others. Flax is also amenable to transformation, and the development of the floral dip procedure for this species may facilitate other approaches to mutant production and increasing available variation. However, the issues with the transgenic flax, Triffid, and its unregulated escape into the commercial seed provide a cautionary tale for the development of genetically modified flax. Some flax varieties have a particular characteristic, which is not known in other plant species, namely, the rapid modulation of its genome under certain growth conditions. The variation, with the generation of altered stable lines termed genotrophs, has been shown to occur within a specific defined subset of the genome. Understanding the mechanisms and characteristics of this genome compartment that appears to have the function of modifying the phenotype without the deleterious effects associated with a random mutagenesis makes it an interesting evolutionary mechanism. The plethora of molecular tools available allows this phenomenon to be understood at the molecular level, and potentially capable of manipulation. The molecular resources available, the commercially important, and the health-related characteristics make flax an accessible interesting model system for understanding complex pathways while also providing the basis for improving the commercial attractiveness of the crop. The development of the molecular resources for flax is directly attributable to the investment in the TUFGEN program and to the collaborations that were fostered, and continue to thrive, through the program.

@article{
 title = {Characterization of bacterial and fungal community dynamics by high-throughput sequencing (HTS) metabarcoding during flax dew-retting},
 type = {article},
 year = {2017},
 keywords = {16s rrna and its,amplicons,bacterial and fungal microbiota,cazyme predictions,dynamics,flax dew-retting,flax dew-retting, bacterial and fungal microbiota,frontiers in microbiology,frontiersin,high-throughput sequencing,hts,metabarcoding,org,trophic modes,www},
 pages = {1-16},
 volume = {8},
 id = {a8562b9b-f204-3071-8666-9992e32a339e},
 created = {2017-12-04T15:26:42.704Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.169Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Djemiel2017},
 private_publication = {false},
 bibtype = {article},
 author = {Djemiel, Christophe and Grec, Sebastien and Hawkins, Simon},
 doi = {10.3389/fmicb.2017.02052},
 journal = {Frontiers in Microbiology},
 number = {October}
}

@article{
 title = {Ectopic lignification in the flax lignified bast fiber1 mutant stem is associated with tissue-specific modifications in gene expression and cell wall composition.},
 type = {article},
 year = {2014},
 keywords = {Alcohol Oxidoreductases,Alcohol Oxidoreductases: genetics,Alcohol Oxidoreductases: metabolism,Aldehyde Oxidoreductases,Aldehyde Oxidoreductases: genetics,Aldehyde Oxidoreductases: metabolism,Cell Wall,Cell Wall: chemistry,Cell Wall: ultrastructure,Computational Biology,Flax,Flax: chemistry,Flax: enzymology,Flax: genetics,Flax: ultrastructure,Gene Expression Profiling,Gene Expression Regulation, Plant,Hydrogen Peroxide,Hydrogen Peroxide: metabolism,Lignin,Lignin: chemistry,Lignin: metabolism,Methyltransferases,Methyltransferases: genetics,Methyltransferases: metabolism,Mutation,Oligonucleotide Array Sequence Analysis,Organ Specificity,Phylogeny,Plant Proteins,Plant Proteins: genetics,Plant Proteins: metabolism,Plant Stems,Plant Stems: chemistry,Plant Stems: enzymology,Plant Stems: genetics,Plant Stems: ultrastructure,Plants, Genetically Modified,Transcriptome,Xylem,Xylem: chemistry,Xylem: enzymology,Xylem: genetics,Xylem: ultrastructure},
 pages = {4462-82},
 volume = {26},
 websites = {http://www.plantcell.org/content/26/11/4462.long},
 id = {3e870d19-df57-3aa8-a13e-2da64571ffae},
 created = {2017-06-08T11:48:17.195Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.464Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Chantreau2014},
 private_publication = {false},
 abstract = {Histochemical screening of a flax ethyl methanesulfonate population led to the identification of 93 independent M2 mutant families showing ectopic lignification in the secondary cell wall of stem bast fibers. We named this core collection the Linum usitatissimum (flax) lbf mutants for lignified bast fibers and believe that this population represents a novel biological resource for investigating how bast fiber plants regulate lignin biosynthesis. As a proof of concept, we characterized the lbf1 mutant and showed that the lignin content increased by 350% in outer stem tissues containing bast fibers but was unchanged in inner stem tissues containing xylem. Chemical and NMR analyses indicated that bast fiber ectopic lignin was highly condensed and rich in G-units. Liquid chromatography-mass spectrometry profiling showed large modifications in the oligolignol pool of lbf1 inner- and outer-stem tissues that could be related to ectopic lignification. Immunological and chemical analyses revealed that lbf1 mutants also showed changes to other cell wall polymers. Whole-genome transcriptomics suggested that ectopic lignification of flax bast fibers could be caused by increased transcript accumulation of (1) the cinnamoyl-CoA reductase, cinnamyl alcohol dehydrogenase, and caffeic acid O-methyltransferase monolignol biosynthesis genes, (2) several lignin-associated peroxidase genes, and (3) genes coding for respiratory burst oxidase homolog NADPH-oxidases necessary to increase H2O2 supply.},
 bibtype = {article},
 author = {Chantreau, Maxime and Portelette, Antoine and Dauwe, Rebecca and Kiyoto, Shingo and Crônier, David and Morreel, Kris and Arribat, Sandrine and Neutelings, Godfrey and Chabi, Malika and Boerjan, Wout and Yoshinaga, Arata and Mesnard, François and Grec, Sebastien and Chabbert, Brigitte and Hawkins, Simon},
 doi = {10.1105/tpc.114.130443},
 journal = {The Plant cell},
 number = {11}
}

Histochemical screening of a flax ethyl methanesulfonate population led to the identification of 93 independent M2 mutant families showing ectopic lignification in the secondary cell wall of stem bast fibers. We named this core collection the Linum usitatissimum (flax) lbf mutants for lignified bast fibers and believe that this population represents a novel biological resource for investigating how bast fiber plants regulate lignin biosynthesis. As a proof of concept, we characterized the lbf1 mutant and showed that the lignin content increased by 350% in outer stem tissues containing bast fibers but was unchanged in inner stem tissues containing xylem. Chemical and NMR analyses indicated that bast fiber ectopic lignin was highly condensed and rich in G-units. Liquid chromatography-mass spectrometry profiling showed large modifications in the oligolignol pool of lbf1 inner- and outer-stem tissues that could be related to ectopic lignification. Immunological and chemical analyses revealed that lbf1 mutants also showed changes to other cell wall polymers. Whole-genome transcriptomics suggested that ectopic lignification of flax bast fibers could be caused by increased transcript accumulation of (1) the cinnamoyl-CoA reductase, cinnamyl alcohol dehydrogenase, and caffeic acid O-methyltransferase monolignol biosynthesis genes, (2) several lignin-associated peroxidase genes, and (3) genes coding for respiratory burst oxidase homolog NADPH-oxidases necessary to increase H2O2 supply.

@article{
 title = {Dual function of the CYP76 family from Arabidopsis thaliana in the metabolism of monoterpenols and phenylurea herbicides.},
 type = {article},
 year = {2014},
 pages = {1149-1161},
 volume = {166},
 websites = {http://www.ncbi.nlm.nih.gov/pubmed/25082892},
 id = {48bfa6c2-884a-3dff-8a5b-1400b2c6288a},
 created = {2017-06-08T11:48:17.227Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.592Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Hofer2014},
 private_publication = {false},
 abstract = {Comparative genomics analysis unravels lineage-specific bursts of gene duplications related to the emergence of specialized pathways. The CYP76C subfamily of cytochrome P450 enzymes is specific to Brassicaceae. Two of its members were recently associated with monoterpenol metabolism. This prompted us to investigate the CYP76C subfamily genetic and functional diversification. Our study revealed high rates of CYP76C gene duplication and loss in Brassicaceae, suggesting the association of the CYP76C subfamily with species-specific adaptive functions. Gene differential expression and enzyme functional specialization in Arabidopsis thaliana, including metabolism of different monoterpenols and formation of different products, support this hypothesis. In addition to linalool metabolism, CYP76C1, CYP76C2 and CYP76C4 metabolized herbicides belonging to the class of phenylurea. Their ectopic expression in the whole plant conferred herbicide tolerance. CYP76Cs from A. thaliana thus provide a first example of promiscuous P450 enzymes endowing effective metabolism of both natural and xenobiotic compounds. Our data also suggest that the CYP76C gene family provides a suitable genetic background for a quick evolution of herbicide resistance.},
 bibtype = {article},
 author = {Höfer, René and Boachon, Benoit and Renault, Hugues and Gavira, Carole and Miesch, Laurence and Iglesias, Juliana and Ginglinger, Jean-François and Allouche, Lionel and Miesch, Michel and Grec, Sebastien and Larbat, Romain and Werck, Daniele},
 doi = {10.1104/pp.114.244814},
 journal = {Plant physiology},
 number = {November}
}

Comparative genomics analysis unravels lineage-specific bursts of gene duplications related to the emergence of specialized pathways. The CYP76C subfamily of cytochrome P450 enzymes is specific to Brassicaceae. Two of its members were recently associated with monoterpenol metabolism. This prompted us to investigate the CYP76C subfamily genetic and functional diversification. Our study revealed high rates of CYP76C gene duplication and loss in Brassicaceae, suggesting the association of the CYP76C subfamily with species-specific adaptive functions. Gene differential expression and enzyme functional specialization in Arabidopsis thaliana, including metabolism of different monoterpenols and formation of different products, support this hypothesis. In addition to linalool metabolism, CYP76C1, CYP76C2 and CYP76C4 metabolized herbicides belonging to the class of phenylurea. Their ectopic expression in the whole plant conferred herbicide tolerance. CYP76Cs from A. thaliana thus provide a first example of promiscuous P450 enzymes endowing effective metabolism of both natural and xenobiotic compounds. Our data also suggest that the CYP76C gene family provides a suitable genetic background for a quick evolution of herbicide resistance.

@article{
 title = {Geraniol hydroxylase and hydroxygeraniol oxidase activities of the CYP76 family of cytochrome P450 enzymes and potential for engineering the early steps of the (seco)iridoid pathway},
 type = {article},
 year = {2013},
 keywords = {Catharanthus roseus,Cytochrome P450,Geraniol oxidation,In planta enzyme assay,Multifunctional enzyme,Pathway reconstitution},
 pages = {221-232},
 volume = {20},
 publisher = {Academic Press Inc.},
 id = {6728d664-1af6-3ce0-b6bb-62386eacdce9},
 created = {2017-06-08T11:48:17.352Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.106Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Hofer2013},
 private_publication = {false},
 abstract = {The geraniol-derived (seco)iridoid skeleton is a precursor for a large group of bioactive compounds with diverse therapeutic applications, including the widely used anticancer molecule vinblastine. Despite of this economic prospect, the pathway leading to iridoid biosynthesis from geraniol is still unclear. The first geraniol hydroxylation step has been reported to be catalyzed by cytochrome P450 enzymes such as CYP76B6 from Catharanthus roseus and CYP76C1 from Arabidopsis thaliana. In the present study, an extended functional analysis of CYP76 family members was carried-out to identify the most effective enzyme to be used for pathway reconstruction. This disproved CYP76C1 activity and led to the characterization of CYP76C4 from A. thaliana as a geraniol 9- or 8-hydroxylase. CYP76B6 emerged as a highly specialized multifunctional enzyme catalyzing two sequential oxidation steps leading to the formation of 8-oxogeraniol from geraniol. This dual function was confirmed in planta using a leaf-disc assay. The first step, geraniol hydroxylation, was very efficient and fast enough to outcompete geraniol conjugation in plant tissues. When the enzyme was expressed in leaf tissues, 8-oxogeraniol was converted into further oxidized and/or reduced compounds in the absence of the next enzyme of the iridoid pathway. © 2013 International Metabolic Engineering Society.},
 bibtype = {article},
 author = {Höfer, René and Dong, Lemeng and André, François and Ginglinger, Jean François and Lugan, Raphael and Gavira, Carole and Grec, Sebastien and Lang, Gerhard and Memelink, Johan and Van Der Krol, Sander and Bouwmeester, Harro and Werck-Reichhart, Danièle},
 journal = {Metabolic Engineering}
}

The geraniol-derived (seco)iridoid skeleton is a precursor for a large group of bioactive compounds with diverse therapeutic applications, including the widely used anticancer molecule vinblastine. Despite of this economic prospect, the pathway leading to iridoid biosynthesis from geraniol is still unclear. The first geraniol hydroxylation step has been reported to be catalyzed by cytochrome P450 enzymes such as CYP76B6 from Catharanthus roseus and CYP76C1 from Arabidopsis thaliana. In the present study, an extended functional analysis of CYP76 family members was carried-out to identify the most effective enzyme to be used for pathway reconstruction. This disproved CYP76C1 activity and led to the characterization of CYP76C4 from A. thaliana as a geraniol 9- or 8-hydroxylase. CYP76B6 emerged as a highly specialized multifunctional enzyme catalyzing two sequential oxidation steps leading to the formation of 8-oxogeraniol from geraniol. This dual function was confirmed in planta using a leaf-disc assay. The first step, geraniol hydroxylation, was very efficient and fast enough to outcompete geraniol conjugation in plant tissues. When the enzyme was expressed in leaf tissues, 8-oxogeraniol was converted into further oxidized and/or reduced compounds in the absence of the next enzyme of the iridoid pathway. © 2013 International Metabolic Engineering Society.

@article{
 title = {PT-Flax (phenotyping and TILLinG of flax): development of a flax (Linum usitatissimum L.) mutant population and TILLinG platform for forward and reverse genetics.},
 type = {article},
 year = {2013},
 keywords = {Base Pairing,Base Pairing: genetics,Ethyl Methanesulfonate,Flax,Flax: genetics,Flowers,Flowers: genetics,Genes, Plant,Genes, Plant: genetics,Genome, Plant,Genome, Plant: genetics,Genotype,Lignin,Lignin: genetics,Mutagenesis,Mutagenesis: genetics,Mutation,Mutation Rate,Mutation: genetics,Nucleotide Motifs,Nucleotide Motifs: genetics,Phenotype,Phylogeny,Reverse Genetics,Reverse Genetics: methods,Seeds,Seeds: genetics},
 pages = {159},
 volume = {13},
 websites = {http://bmcplantbiol.biomedcentral.com/articles/10.1186/1471-2229-13-159},
 publisher = {BioMed Central},
 id = {d5992a68-27b5-35c7-8297-4f3bcad2561b},
 created = {2017-06-08T11:48:17.489Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:00.817Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Chantreau2013},
 private_publication = {false},
 abstract = {BACKGROUND: Flax (Linum usitatissimum L.) is an economically important fiber and oil crop that has been grown for thousands of years. The genome has been recently sequenced and transcriptomics are providing information on candidate genes potentially related to agronomically-important traits. In order to accelerate functional characterization of these genes we have generated a flax EMS mutant population that can be used as a TILLinG (Targeting Induced Local Lesions in Genomes) platform for forward and reverse genetics.\n\nRESULTS: A population of 4,894 M2 mutant seed families was generated using 3 different EMS concentrations (0.3%, 0.6% and 0.75%) and used to produce M2 plants for subsequent phenotyping and DNA extraction. 10,839 viable M2 plants (4,033 families) were obtained and 1,552 families (38.5%) showed a visual developmental phenotype (stem size and diameter, plant architecture, flower-related). The majority of these families showed more than one phenotype. Mutant phenotype data are organised in a database and can be accessed and searched at UTILLdb (http://urgv.evry.inra.fr/UTILLdb). Preliminary screens were also performed for atypical fiber and seed phenotypes. Genomic DNA was extracted from 3,515 M2 families and eight-fold pooled for subsequent mutant detection by ENDO1 nuclease mis-match cleavage. In order to validate the collection for reverse genetics, DNA pools were screened for two genes coding enzymes of the lignin biosynthesis pathway: Coumarate-3-Hydroxylase (C3H) and Cinnamyl Alcohol Dehydrogenase (CAD). We identified 79 and 76 mutations in the C3H and CAD genes, respectively. The average mutation rate was calculated as 1/41 Kb giving rise to approximately 9,000 mutations per genome. Thirty-five out of the 52 flax cad mutant families containing missense or codon stop mutations showed the typical orange-brown xylem phenotype observed in CAD down-regulated/mutant plants in other species.\n\nCONCLUSIONS: We have developed a flax mutant population that can be used as an efficient forward and reverse genetics tool. The collection has an extremely high mutation rate that enables the detection of large numbers of independant mutant families by screening a comparatively low number of M2 families. The population will prove to be a valuable resource for both fundamental research and the identification of agronomically-important genes for crop improvement in flax.},
 bibtype = {article},
 author = {Chantreau, Maxime and Grec, Sébastien and Gutierrez, Laurent and Dalmais, Marion and Pineau, Christophe and Demailly, Hervé and Paysant-Leroux, Christine and Tavernier, Reynald and Trouvé, Jean-Paul and Chatterjee, Manash and Guillot, Xavier and Brunaud, Véronique and Chabbert, Brigitte and van Wuytswinkel, Olivier and Bendahmane, Abdelhafid and Thomasset, Brigitte and Hawkins, Simon},
 doi = {10.1186/1471-2229-13-159},
 journal = {BMC plant biology},
 number = {1}
}

BACKGROUND: Flax (Linum usitatissimum L.) is an economically important fiber and oil crop that has been grown for thousands of years. The genome has been recently sequenced and transcriptomics are providing information on candidate genes potentially related to agronomically-important traits. In order to accelerate functional characterization of these genes we have generated a flax EMS mutant population that can be used as a TILLinG (Targeting Induced Local Lesions in Genomes) platform for forward and reverse genetics.\n\nRESULTS: A population of 4,894 M2 mutant seed families was generated using 3 different EMS concentrations (0.3%, 0.6% and 0.75%) and used to produce M2 plants for subsequent phenotyping and DNA extraction. 10,839 viable M2 plants (4,033 families) were obtained and 1,552 families (38.5%) showed a visual developmental phenotype (stem size and diameter, plant architecture, flower-related). The majority of these families showed more than one phenotype. Mutant phenotype data are organised in a database and can be accessed and searched at UTILLdb (http://urgv.evry.inra.fr/UTILLdb). Preliminary screens were also performed for atypical fiber and seed phenotypes. Genomic DNA was extracted from 3,515 M2 families and eight-fold pooled for subsequent mutant detection by ENDO1 nuclease mis-match cleavage. In order to validate the collection for reverse genetics, DNA pools were screened for two genes coding enzymes of the lignin biosynthesis pathway: Coumarate-3-Hydroxylase (C3H) and Cinnamyl Alcohol Dehydrogenase (CAD). We identified 79 and 76 mutations in the C3H and CAD genes, respectively. The average mutation rate was calculated as 1/41 Kb giving rise to approximately 9,000 mutations per genome. Thirty-five out of the 52 flax cad mutant families containing missense or codon stop mutations showed the typical orange-brown xylem phenotype observed in CAD down-regulated/mutant plants in other species.\n\nCONCLUSIONS: We have developed a flax mutant population that can be used as an efficient forward and reverse genetics tool. The collection has an extremely high mutation rate that enables the detection of large numbers of independant mutant families by screening a comparatively low number of M2 families. The population will prove to be a valuable resource for both fundamental research and the identification of agronomically-important genes for crop improvement in flax.

@article{
 title = {A "novel" protocol for the analysis of hydroxycinnamic acids in leaf tissue of chicory (<i>Cichorium intybus</i> L., Asteraceae).},
 type = {article},
 year = {2012},
 keywords = {Asteraceae,Chemistry,Chicory,Chromatography,Coumaric Acids,Coumaric Acids: analysis,Coumaric Acids: chemistry,High Pressure Liquid,High Pressure Liquid: methods,Liquid,Liquid: methods,Mass Spectrometry,Mass Spectrometry: methods,Pharmaceutical,Pharmaceutical: methods,Pharmaceutical: trends,Plant Leaves},
 pages = {142983},
 volume = {2012},
 websites = {http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3523586&tool=pmcentrez&rendertype=abstract},
 id = {fbf9d9a0-ada7-31a3-8873-3b8ab0926db5},
 created = {2017-06-08T11:48:17.175Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.516Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Bahri2012},
 private_publication = {false},
 abstract = {A "novel" protocol is presented for easy and reliable estimation of soluble hydroxycinnamate levels in Cichorium intybus L. leaf tissue in large-scale experiments. Samples were standardized by punching 6 discs per leaf, and hydroxycinnamates were extracted by submerging the discs in 80% ethanol with 5% acetic acid for at least 48 h in the darkness at 4°C. Residual dry mass of the discs was used for a posteriori correction of compound levels. Chlorophyll was eliminated by chloroform, and the aqueous phases were transferred to microplates, dried, and dissolved in 50% methanol for HPLC analysis and storage. An HPLC program of 8 min was developed for the analysis of the extracts. Comparisons with extractions of liquid nitrogen powders indicated that the novel extraction method was reliable. No degradation of the major hydroxycinnamates-caftaric, chlorogenic, and chicoric acids-was observed, during maceration at ambient temperatures, or after storage for 1 year.},
 bibtype = {article},
 author = {Bahri, Meriem and Hance, Philippe and Grec, Sébastien and Quillet, Marie-Christine and Trotin, Francis and Hilbert, Jean-Louis and Hendriks, Theo},
 doi = {10.1100/2012/142983},
 journal = {The Scientific World Journal}
}

A "novel" protocol is presented for easy and reliable estimation of soluble hydroxycinnamate levels in Cichorium intybus L. leaf tissue in large-scale experiments. Samples were standardized by punching 6 discs per leaf, and hydroxycinnamates were extracted by submerging the discs in 80% ethanol with 5% acetic acid for at least 48 h in the darkness at 4°C. Residual dry mass of the discs was used for a posteriori correction of compound levels. Chlorophyll was eliminated by chloroform, and the aqueous phases were transferred to microplates, dried, and dissolved in 50% methanol for HPLC analysis and storage. An HPLC program of 8 min was developed for the analysis of the extracts. Comparisons with extractions of liquid nitrogen powders indicated that the novel extraction method was reliable. No degradation of the major hydroxycinnamates-caftaric, chlorogenic, and chicoric acids-was observed, during maceration at ambient temperatures, or after storage for 1 year.

@article{
 title = {Natural Hypolignification Is Associated with Extensive Oligolignol Accumulation in Flax Stems},
 type = {article},
 year = {2012},
 pages = {1893-1915},
 volume = {158},
 id = {c3ba7ffa-1744-38d5-b53b-2f3fe757216d},
 created = {2017-06-08T11:48:17.551Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.170Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Huis2012},
 private_publication = {false},
 abstract = {Flax (Linum usitatissimum) stems contain cells showing contrasting cell wall structure: lignified in inner stem xylem tissue and hypolignified in outer stem bast fibers. We hypothesized that stem hypolignification should be associated with extensive phenolic accumulation and used metabolomics and transcriptomics to characterize these two tissues. (1)H nuclear magnetic resonance clearly distinguished inner and outer stem tissues and identified different primary and secondary metabolites, including coniferin and p-coumaryl alcohol glucoside. Ultrahigh-performance liquid chromatography-Fourier transform ion cyclotron resonance-mass spectrometry aromatic profiling (lignomics) identified 81 phenolic compounds, of which 65 were identified, to our knowledge, for the first time in flax and 11 for the first time in higher plants. Both aglycone forms and glycosides of monolignols, lignin oligomers, and (neo)lignans were identified in both inner and outer stem tissues, with a preponderance of glycosides in the hypolignified outer stem, indicating the existence of a complex monolignol metabolism. The presence of coniferin-containing secondary metabolites suggested that coniferyl alcohol, in addition to being used in lignin and (neo)lignan formation, was also utilized in a third, partially uncharacterized metabolic pathway. Hypolignification of bast fibers in outer stem tissues was correlated with the low transcript abundance of monolignol biosynthetic genes, laccase genes, and certain peroxidase genes, suggesting that flax hypolignification is transcriptionally regulated. Transcripts of the key lignan genes Pinoresinol-Lariciresinol Reductase and Phenylcoumaran Benzylic Ether Reductase were also highly abundant in flax inner stem tissues. Expression profiling allowed the identification of NAC (NAM, ATAF1/2, CUC2) and MYB transcription factors that are likely involved in regulating both monolignol production and polymerization as well as (neo)lignan production.},
 bibtype = {article},
 author = {Huis, R. and Morreel, K. and Fliniaux, O. and Lucau-Danila, a. and Fenart, S. and Grec, S. and Neutelings, G. and Chabbert, B. and Mesnard, F. and Boerjan, W. and Hawkins, S.},
 doi = {10.1104/pp.111.192328},
 journal = {Plant Physiology},
 number = {4}
}

Flax (Linum usitatissimum) stems contain cells showing contrasting cell wall structure: lignified in inner stem xylem tissue and hypolignified in outer stem bast fibers. We hypothesized that stem hypolignification should be associated with extensive phenolic accumulation and used metabolomics and transcriptomics to characterize these two tissues. (1)H nuclear magnetic resonance clearly distinguished inner and outer stem tissues and identified different primary and secondary metabolites, including coniferin and p-coumaryl alcohol glucoside. Ultrahigh-performance liquid chromatography-Fourier transform ion cyclotron resonance-mass spectrometry aromatic profiling (lignomics) identified 81 phenolic compounds, of which 65 were identified, to our knowledge, for the first time in flax and 11 for the first time in higher plants. Both aglycone forms and glycosides of monolignols, lignin oligomers, and (neo)lignans were identified in both inner and outer stem tissues, with a preponderance of glycosides in the hypolignified outer stem, indicating the existence of a complex monolignol metabolism. The presence of coniferin-containing secondary metabolites suggested that coniferyl alcohol, in addition to being used in lignin and (neo)lignan formation, was also utilized in a third, partially uncharacterized metabolic pathway. Hypolignification of bast fibers in outer stem tissues was correlated with the low transcript abundance of monolignol biosynthetic genes, laccase genes, and certain peroxidase genes, suggesting that flax hypolignification is transcriptionally regulated. Transcripts of the key lignan genes Pinoresinol-Lariciresinol Reductase and Phenylcoumaran Benzylic Ether Reductase were also highly abundant in flax inner stem tissues. Expression profiling allowed the identification of NAC (NAM, ATAF1/2, CUC2) and MYB transcription factors that are likely involved in regulating both monolignol production and polymerization as well as (neo)lignan production.

@article{
 title = {Phenolic profiles and antioxidative effects of hawthorn cell suspensions, fresh fruits, and medicinal dried parts},
 type = {article},
 year = {2009},
 keywords = {Anthocyanin,Antioxidant,Cell culture suspension,Crataegus monogyna,Hawthorn,Idaein,Phenolic content},
 pages = {897-903},
 volume = {115},
 id = {0471299e-78a7-3ec2-ba14-45f4c17a0c89},
 created = {2017-06-08T11:48:17.223Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.486Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Froehlicher2009},
 private_publication = {false},
 abstract = {The polyphenolic content of two cell suspension lines (red and yellow) initiated from the ovarian wall of Crataegus monogyna flower and their antioxidative potencies against ABTSradical dot+, DPPHradical dot, and human LDL oxidation were compared to those of red fresh and dry fruits, flower buds and flowering tops. Maximal phenolics and proanthocyanidins contents were found in red suspension extracts displaying high antioxidative effects. In contrast, yellow cell extracts were always the poorest in both phenolics and activity. Flower buds and flowering tops have significant phenolic yields and effects. Both fresh and dried fruits are less active. The amounts in some major phenolic compounds were determined in all tested samples: again, the most antioxidant samples were richer, the red cell line showing particularly high amounts in epicatechin and chlorogenic acid, whilst dried flower buds contained mainly hyperoside and chlorogenic acid. (-)-Epicatechin was confirmed to be more efficient as an antioxidant compound than hyperoside and chlorogenic acid in all assays and more generally, proanthocyanidins were found to be more clearly related to antioxidant activity than other classes of phenolics. The major anthocyanin characterising the red cells of C. monogyna was isolated and identified as cyanidin-3-O-galactoside. © 2009 Elsevier Ltd. All rights reserved.},
 bibtype = {article},
 author = {Froehlicher, Thomas and Hennebelle, Thierry and Martin-Nizard, Françoise and Cleenewerck, Patricia and Hilbert, Jean Louis and Trotin, Francis and Grec, Sébastien},
 journal = {Food Chemistry},
 number = {3}
}

The polyphenolic content of two cell suspension lines (red and yellow) initiated from the ovarian wall of Crataegus monogyna flower and their antioxidative potencies against ABTSradical dot+, DPPHradical dot, and human LDL oxidation were compared to those of red fresh and dry fruits, flower buds and flowering tops. Maximal phenolics and proanthocyanidins contents were found in red suspension extracts displaying high antioxidative effects. In contrast, yellow cell extracts were always the poorest in both phenolics and activity. Flower buds and flowering tops have significant phenolic yields and effects. Both fresh and dried fruits are less active. The amounts in some major phenolic compounds were determined in all tested samples: again, the most antioxidant samples were richer, the red cell line showing particularly high amounts in epicatechin and chlorogenic acid, whilst dried flower buds contained mainly hyperoside and chlorogenic acid. (-)-Epicatechin was confirmed to be more efficient as an antioxidant compound than hyperoside and chlorogenic acid in all assays and more generally, proanthocyanidins were found to be more clearly related to antioxidant activity than other classes of phenolics. The major anthocyanin characterising the red cells of C. monogyna was isolated and identified as cyanidin-3-O-galactoside. © 2009 Elsevier Ltd. All rights reserved.

@article{
 title = {Caffeoyl coenzyme A O-methyltransferase down-regulation is associated with modifications in lignin and cell-wall architecture in flax secondary xylem},
 type = {article},
 year = {2009},
 keywords = {CCoAOMT,Cell wall,Flax,Irregular xylem,Lignin,Monolignol},
 pages = {9-19},
 volume = {47},
 id = {39c5d951-73cd-34a1-82db-5c4535c88c90},
 created = {2017-06-08T11:48:17.300Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.749Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Day2009},
 private_publication = {false},
 abstract = {Caffeoyl coenzyme A O-methyltransferase (CCoAOMT, EC 2.1.1.104) down-regulated-flax (Linum usitatissimum) plants were generated using an antisense strategy and functionally characterized. Chemical analyses (acetyl bromide and thioacidolysis) revealed that the lignin quantity was reduced and that the Syringyl/Gua??acyl (S/G) lignin monomer ratio was modified in the non-condensed lignin fraction of two independent down-regulated lines. These modifications were associated with altered xylem organization (both lines), reduced cell-wall thickness (one line) and the appearance of an irregular xylem (irx) phenotype (both lines). In addition UV microspectroscopy also indicated that CCoAOMT down-regulation induced changes in xylem cell-wall structure and the lignin fractions. Microscopic examination also suggested that CCoAOMT down-regulation could influence individual xylem cell size and identity. As a first step towards investigating the cellular mechanisms responsible for the unusual structure of flax lignin (G-rich, condensed), recombinant flax CCoAOMT protein was produced and its affinity for different potential substrates evaluated. Results indicated that the preferred substrate was caffeoyl coenzyme A, followed by 5-hydroxyconiferaldehyde suggesting that flax CCoAOMT possesses a small, but probably significant 5??? methylating activity, in addition to a more usual 3??? methylating activity. ?? 2008 Elsevier Masson SAS. All rights reserved.},
 bibtype = {article},
 author = {Day, Arnaud and Neutelings, Godfrey and Nolin, Fr??d??rique and Grec, S??bastien and Habrant, Anouk and Cr??nier, David and Maher, Bouchra and Rolando, Christian and David, H??l??ne and Chabbert, Brigitte and Hawkins, Simon},
 doi = {10.1016/j.plaphy.2008.09.011},
 journal = {Plant Physiology and Biochemistry},
 number = {1}
}

Caffeoyl coenzyme A O-methyltransferase (CCoAOMT, EC 2.1.1.104) down-regulated-flax (Linum usitatissimum) plants were generated using an antisense strategy and functionally characterized. Chemical analyses (acetyl bromide and thioacidolysis) revealed that the lignin quantity was reduced and that the Syringyl/Gua??acyl (S/G) lignin monomer ratio was modified in the non-condensed lignin fraction of two independent down-regulated lines. These modifications were associated with altered xylem organization (both lines), reduced cell-wall thickness (one line) and the appearance of an irregular xylem (irx) phenotype (both lines). In addition UV microspectroscopy also indicated that CCoAOMT down-regulation induced changes in xylem cell-wall structure and the lignin fractions. Microscopic examination also suggested that CCoAOMT down-regulation could influence individual xylem cell size and identity. As a first step towards investigating the cellular mechanisms responsible for the unusual structure of flax lignin (G-rich, condensed), recombinant flax CCoAOMT protein was produced and its affinity for different potential substrates evaluated. Results indicated that the preferred substrate was caffeoyl coenzyme A, followed by 5-hydroxyconiferaldehyde suggesting that flax CCoAOMT possesses a small, but probably significant 5??? methylating activity, in addition to a more usual 3??? methylating activity. ?? 2008 Elsevier Masson SAS. All rights reserved.

@article{
 title = {Polyphenol contents and antioxidant activities of extracts from flowers of two Crataegus azarolus L. varieties},
 type = {article},
 year = {2009},
 keywords = {Crataegus azarolus L.,DPPH free radical scavenger,Organic extract,Phenolic contents},
 pages = {660-668},
 volume = {12},
 websites = {http://www.ncbi.nlm.nih.gov/pubmed/19634468},
 month = {5},
 day = {1},
 id = {c626ab90-b447-38c1-a628-ea7f2f66d00e},
 created = {2017-06-08T11:48:17.415Z},
 accessed = {2016-07-11},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.756Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Bahri-Sahloul2009a},
 private_publication = {false},
 abstract = {Phenolic contents of the ethyl acetate extracts prepared from floral buds and opened flowers harvested on Crataegus azarolus trees native in two localities were performed. The antioxidant activity was measured by DPPH' (2,2-diphenyl-picrylhydrazyl), ABTS+ (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radicals scavenging using spectrophotometric method. The C. azarolus var. aronia (Willd.) Batt., producing yellow fruits, was richer in total phenols (1638.7 +/- 89.9 mg acid gallic/100 g dry weight) according to C. azarolus var. eu-azarolus Maire (1415.5 +/- 23.8 mg acid gallic/100 g dry weight), producing red ones. Ethyl acetate extract from opened flowers has less content in total phenols, proanthocyanidins and flavonoids compared to this from floral buds. Floral buds from the two C. azarolus varieties occurring in Siliana-Djebel Serdj showed the highest radical scavenging activities (2431.8 +/- 32.7 and 2267.7 +/- 22.7 micromol Trolox/100 g dry weight). Hawthorn from Tunisia contains eight antioxidants of phenolic type (chlorogenic acid, hyperoside, rutin, spiraeoside, isoquercitrin, quercetin, (-)-epicatechin and the dimer procyanidin B2). These compounds identified specially in floral bud extracts presented a strong radical-scavenging activity.},
 bibtype = {article},
 author = {Bahri-Sahloul, R. and Ammar, S. and Fredj, R. B. and Saguem, S. and Grec, S. and Trotin, F. and Skhiri, F. H.},
 doi = {10.3923/pjbs.2009.660.668},
 journal = {Pakistan Journal of Biological Sciences},
 number = {9}
}

Phenolic contents of the ethyl acetate extracts prepared from floral buds and opened flowers harvested on Crataegus azarolus trees native in two localities were performed. The antioxidant activity was measured by DPPH' (2,2-diphenyl-picrylhydrazyl), ABTS+ (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radicals scavenging using spectrophotometric method. The C. azarolus var. aronia (Willd.) Batt., producing yellow fruits, was richer in total phenols (1638.7 +/- 89.9 mg acid gallic/100 g dry weight) according to C. azarolus var. eu-azarolus Maire (1415.5 +/- 23.8 mg acid gallic/100 g dry weight), producing red ones. Ethyl acetate extract from opened flowers has less content in total phenols, proanthocyanidins and flavonoids compared to this from floral buds. Floral buds from the two C. azarolus varieties occurring in Siliana-Djebel Serdj showed the highest radical scavenging activities (2431.8 +/- 32.7 and 2267.7 +/- 22.7 micromol Trolox/100 g dry weight). Hawthorn from Tunisia contains eight antioxidants of phenolic type (chlorogenic acid, hyperoside, rutin, spiraeoside, isoquercitrin, quercetin, (-)-epicatechin and the dimer procyanidin B2). These compounds identified specially in floral bud extracts presented a strong radical-scavenging activity.

@article{
 title = {Chemical characterisation of Crataegus azarolus L. fruit from 14 genotypes found in Tunisia},
 type = {article},
 year = {2009},
 pages = {23-28},
 volume = {84},
 id = {eb8095a9-0d85-3547-9fc4-57ef506d5e5e},
 created = {2019-03-05T15:23:14.968Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.744Z},
 read = {false},
 starred = {false},
 authored = {false},
 confirmed = {true},
 hidden = {false},
 citation_key = {Bahri-Sahloul2009},
 private_publication = {false},
 bibtype = {article},
 author = {Bahri-Sahloul, R and Ammar, S and Grec, S and Harzallah-Skhiri, F},
 journal = {The Journal of Horticultural Science and Biotechnology},
 number = {1}
}

@article{
 title = {NpPDR1, a pleiotropic drug resistance-type ATP-binding cassette transporter from \emphNicotiana plumbaginifolia, plays a major role in plant pathogen defense.},
 type = {article},
 year = {2005},
 pages = {341-352},
 volume = {139},
 websites = {http://www.plantphysiol.org/cgi/content/abstract/139/1/341},
 id = {0b07f10c-863d-35da-aaad-8cd0864a5844},
 created = {2017-06-08T11:48:17.176Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.565Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Stukkens2005},
 private_publication = {false},
 abstract = {Nicotiana plumbaginifolia NpPDR1, a plasma membrane pleiotropic drug resistance-type ATP-binding cassette transporter formerly named NpABC1, has been suggested to transport the diterpene sclareol, an antifungal compound. However, direct evidence for a role of pleiotropic drug resistance transporters in the plant defense is still lacking. In situ immunolocalization and histochemical analysis using the gusA reporter gene showed that NpPDR1 was constitutively expressed in the whole root, in the leaf glandular trichomes, and in the flower petals. However, NpPDR1 expression was induced in the whole leaf following infection with the fungus Botrytis cinerea, and the bacteria Pseudomonas syringae pv tabaci, Pseudomonas fluorescens, and Pseudomonas marginalis pv marginalis, which do not induce a hypersensitive response in N. plumbaginifolia, whereas a weaker response was observed using P. syringae pv syringae, which does induce a hypersensitive response. Induced NpPDR1 expression was more associated with the jasmonic acid than the salicylic acid signaling pathway. These data suggest that NpPDR1 is involved in both constitutive and jasmonic acid-dependent induced defense. Transgenic plants in which NpPDR1 expression was prevented by RNA interference showed increased sensitivity to sclareol and reduced resistance to B. cinerea. These data show that NpPDR1 is involved in pathogen resistance and thus demonstrate a new role for the ATP-binding cassette transporter family.},
 bibtype = {article},
 author = {Stukkens, Yvan and Bultreys, Alain and Trombik, Tomasz and Vanham, Delphine and Boutry, Marc},
 doi = {10.1104/pp.105.062372.1},
 journal = {Plant Physiology},
 number = {September}
}

Nicotiana plumbaginifolia NpPDR1, a plasma membrane pleiotropic drug resistance-type ATP-binding cassette transporter formerly named NpABC1, has been suggested to transport the diterpene sclareol, an antifungal compound. However, direct evidence for a role of pleiotropic drug resistance transporters in the plant defense is still lacking. In situ immunolocalization and histochemical analysis using the gusA reporter gene showed that NpPDR1 was constitutively expressed in the whole root, in the leaf glandular trichomes, and in the flower petals. However, NpPDR1 expression was induced in the whole leaf following infection with the fungus Botrytis cinerea, and the bacteria Pseudomonas syringae pv tabaci, Pseudomonas fluorescens, and Pseudomonas marginalis pv marginalis, which do not induce a hypersensitive response in N. plumbaginifolia, whereas a weaker response was observed using P. syringae pv syringae, which does induce a hypersensitive response. Induced NpPDR1 expression was more associated with the jasmonic acid than the salicylic acid signaling pathway. These data suggest that NpPDR1 is involved in both constitutive and jasmonic acid-dependent induced defense. Transgenic plants in which NpPDR1 expression was prevented by RNA interference showed increased sensitivity to sclareol and reduced resistance to B. cinerea. These data show that NpPDR1 is involved in pathogen resistance and thus demonstrate a new role for the ATP-binding cassette transporter family.

@article{
 title = {Identification of regulatory sequence elements within the transcription promoter region of NpABC1, a gene encoding a plant ABC transporter induced by diterpenes},
 type = {article},
 year = {2003},
 keywords = {ABC transporter,Gene regulation,Methyl jasmonate,Promoter,Sclareol,Secondary metabolites,Transcription},
 pages = {237-250},
 volume = {35},
 id = {cc3bec93-532d-3ab9-bb18-850c09c05b04},
 created = {2017-06-08T11:48:17.226Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:00.829Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Grec2003},
 private_publication = {false},
 abstract = {Expression of NpABC1, a gene encoding a plasma membrane ATP binding cassette (ABC) transporter in Nicotiana plumbaginifolia, is induced by sclareol, an antifungal diterpene produced at the leaf surface, as well as by sclareolide, a close analog. A genomic fragment including the 1282-bp region upstream of the NpABC1 transcription start was fused to the reporter beta-glucuronidase (gus) gene and introduced into N. tabacum BY2 cells for stable transformation. A 25-fold increase in gus expression was observed when cells were treated with sclareolide and some other terpenes. The combined use of 5'-deletion promoter analysis, gel mobility shift assays, DNase I footprinting, and site-directed mutagenesis allowed us to identify three cis-elements (sclareol box 1 (SB1), SB2, and SB3) located, respectively, within nucleotides -827 to -802, -278 to -243, and -216 to -190 upstream of the NpABC1 transcription start. In vivo evaluation of these elements on sclareolide-induced expression showed that mutation of SB1 reduced expression by twofold, while that of SB2 had no effect. On the other hand, SB3 had a marked effect as it completely abolished sclareolide-mediated expression. NpABC1-gus expression was not induced by the stress signals, salicylic acid and ethylene, but was mediated, to some extent, by methyl jasmonate, which is known to promote diterpene synthesis.},
 bibtype = {article},
 author = {Grec, Sébastien and Vanham, Delphine and De Ribaucourt, Jeoffrey Christyn and Purnelle, Bénédicte and Boutry, Marc},
 doi = {10.1046/j.1365-313X.2003.01792.x},
 journal = {Plant Journal},
 number = {2}
}

Expression of NpABC1, a gene encoding a plasma membrane ATP binding cassette (ABC) transporter in Nicotiana plumbaginifolia, is induced by sclareol, an antifungal diterpene produced at the leaf surface, as well as by sclareolide, a close analog. A genomic fragment including the 1282-bp region upstream of the NpABC1 transcription start was fused to the reporter beta-glucuronidase (gus) gene and introduced into N. tabacum BY2 cells for stable transformation. A 25-fold increase in gus expression was observed when cells were treated with sclareolide and some other terpenes. The combined use of 5'-deletion promoter analysis, gel mobility shift assays, DNase I footprinting, and site-directed mutagenesis allowed us to identify three cis-elements (sclareol box 1 (SB1), SB2, and SB3) located, respectively, within nucleotides -827 to -802, -278 to -243, and -216 to -190 upstream of the NpABC1 transcription start. In vivo evaluation of these elements on sclareolide-induced expression showed that mutation of SB1 reduced expression by twofold, while that of SB2 had no effect. On the other hand, SB3 had a marked effect as it completely abolished sclareolide-mediated expression. NpABC1-gus expression was not induced by the stress signals, salicylic acid and ethylene, but was mediated, to some extent, by methyl jasmonate, which is known to promote diterpene synthesis.

@article{
 title = {Cryptic polyadenylation sites within the coding sequence of three yeast genes expressed in tobacco},
 type = {article},
 year = {2000},
 keywords = {ABC transporter,DNA polymerase,Plant,Transgenic},
 pages = {87-95},
 volume = {242},
 id = {30cf5a13-2e3c-306d-9b4c-6e5ddf986a07},
 created = {2017-06-08T11:48:17.396Z},
 file_attached = {false},
 profile_id = {bae88dda-ac8b-323a-8293-ff3f78f068fe},
 group_id = {7766420b-c45a-3034-bfaf-9f92b0bbbda7},
 last_modified = {2021-04-09T15:21:01.651Z},
 read = {false},
 starred = {false},
 authored = {true},
 confirmed = {true},
 hidden = {false},
 citation_key = {Grec2000},
 private_publication = {false},
 abstract = {Three yeast genes, MIP (mitochondrial DNA polymerase) and two genes, YCF1 (yeast cadmium factor 1) and PDR5 (pleiotropic drug resistance 5), conferring multidrug resistance, were provided with the cauliflower mosaic virus 35S transcription promoter and introduced into tobacco using an Agrobacterium tumefaciens T-DNA-derived vector. Transcripts of each gene much shorter than those expected were found in the transgenic plants. RT-PCR and S1 nuclease mapping of the PDR5 and MIP transcripts demonstrated the presence of one (PDR5), or several close (MIP), cryptic polyadenylation site(s) within the coding sequence of these yeast genes. Possible sequences involved in polyadenylation are discussed. (C) 2000 Elsevier Science B.V. All rights reserved.},
 bibtype = {article},
 author = {Grec, Sébastien and Wang, Yingchun and Le Guen, Laurence and Negrouk, Valentine and Boutry, Marc},
 doi = {10.1016/S0378-1119(99)00544-2},
 journal = {Gene},
 number = {1-2}
}

Three yeast genes, MIP (mitochondrial DNA polymerase) and two genes, YCF1 (yeast cadmium factor 1) and PDR5 (pleiotropic drug resistance 5), conferring multidrug resistance, were provided with the cauliflower mosaic virus 35S transcription promoter and introduced into tobacco using an Agrobacterium tumefaciens T-DNA-derived vector. Transcripts of each gene much shorter than those expected were found in the transgenic plants. RT-PCR and S1 nuclease mapping of the PDR5 and MIP transcripts demonstrated the presence of one (PDR5), or several close (MIP), cryptic polyadenylation site(s) within the coding sequence of these yeast genes. Possible sequences involved in polyadenylation are discussed. (C) 2000 Elsevier Science B.V. All rights reserved.