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Dauchot N.,University of Namur | Raulier P.,Catholic University of Louvain | Maudoux O.,Chicoline Cosucra | Notte C.,Chicoline Cosucra | And 2 more authors.
Frontiers in Plant Science | Year: 2015

Key Message: The loss of mini-exon 2 in the 1-FEH IIb glycosyl-hydrolase results in a putative non-functional allele. This loss of function has a strong impact on the susceptibility to post-harvest inulin depolymerization. Significant variation of copy number was identified in its close paralog 1-FEH IIa, but no quantitative effect of copy number on carbohydrates-related phenotypes was detected. Inulin polyfructan is the second most abundant storage carbohydrate in flowering plants. After harvest, it is depolymerized by fructan exohydrolases (FEHs) as an adaptive response to end-season cold temperatures. In chicory, the intensity of this depolymerization differs between cultivars but also between individuals within a cultivar. Regarding this phenotypic variability, we recently identified statistically significant associations between inulin degradation and genetic polymorphisms located in three FEHs. We present here new results of a systematic analysis of copy number variation (CNV) in five key members of the chicory (Cichorium intybus) GH32 multigenic family, including three FEH genes and the two inulin biosynthesis genes: 1 -SST and 1 -FFT. qPCR analysis identified a significant variability of relative copy number only in the 1-FEH IIa gene. However, this CNV had no quantitative effect. Instead, cloning of the full length gDNA of a close paralogous sequence (1-FEH IIb) identified a 1028 bp deletion in lines less susceptible to post-harvest inulin depolymerization. This region comprises a 9 bp mini-exon containing one of the three conserved residues of the active site. This results in a putative non-functional 1-FEH IIb allele and an observed lower inulin depolymerization. Extensive genotyping confirmed that the loss of mini-exon 2 in 1-FEH IIb and the previously identified 47 bp duplication located in the 3'UTR of 1-FEH IIa belong to a single haplotype, both being statistically associated with reduced susceptibility to post-harvest inulin depolymerization. Emergence of these haplotypes is discussed. © 2015 Dauchot, Raulier, Maudoux, Notté, Draye and Van Cutsem. Source


Raulier P.,Catholic University of Louvain | Maudoux O.,Chicoline Cosucra | Notte C.,Chicoline Cosucra | Draye X.,Catholic University of Louvain | Bertin P.,Catholic University of Louvain
Genetic Resources and Crop Evolution | Year: 2016

The present study used 15 simple sequence repeat loci to characterize the genetic diversity of the germplasm that originated the current industrial chicory and to establish the relationships between and inside Cichorium intybus L. and Cichorium endivia L. Initially we analyzed 19 cultivated C. endivia accessions, 27 wild and 155 cultivated C. intybus accessions distributed among three groups: 83 root chicories, 42 Witloof and 30 leaf chicories. The leaf chicories comprised cultivars corresponding to the Radicchio, Sugarloaf and Catalogne subgroups. The latter has not been previously included in any genetic diversity study. Subsequently, 1297 individuals from the 15 modern root chicory cultivars at the origin of the breeding of the current industrial root chicory cultivars were analyzed. Although the accessions of C. endivia and C. intybus were clearly separated from each other, seven wild C. intybus individuals were genetically closer to C. endivia than to C. intybus, revealing complex genetic interrelationships between these species. The differentiation of C. intybus into three cultivar groups (Witloof, root chicory and leaf chicory) was confirmed. The leaf chicory individuals were divided into three genetic subgroups, corresponding to the Radicchio, Sugarloaf and Catalogne cultivars, thus attesting to the validity of the classification based on morphological factors. Clear differentiation was observed among the Belgian, Polish and Austrian modern industrial root cultivars, but not among the French industrial modern root cultivars. The high phenotypic and genetic variability of the modern industrial root cultivars indicates that this germplasm constitutes a useful gene pool for cultivar improvement and selection. © Springer Science+Business Media Dordrecht 2015. Source


Dauchot N.,University of Namur | Raulier P.,Catholic University of Louvain | Maudoux O.,Chicoline Cosucra | Notte C.,Chicoline Cosucra | And 3 more authors.
Theoretical and Applied Genetics | Year: 2014

Key message: Nucleotidic polymorphisms were identified in fructan exohydrolases genes which are statistically associated with enhanced susceptibility to post-harvest inulin depolymerization. Industrial chicory (Cichorium intybus L.) root is the main commercial source of inulin, a linear fructose polymer used as dietary fiber. Post-harvest, inulin is depolymerized into fructose which drastically increases processing cost. To identify genetic variations associated with enhanced susceptibility to post-harvest inulin depolymerization and related free sugars content increase, we used a candidate-gene approach focused on inulin and sucrose synthesis and degradation genes, all members of the family 32 of glycoside hydrolases (GH32). Polymorphism in these genes was first investigated by carrying out EcoTILLING on two groups of chicory breeding lines exhibiting contrasted response to post-harvest inulin depolymerization. This allowed the identification of polymorphisms significantly associated with depolymerization in three fructan exohydrolase genes (FEH). This association was confirmed on a wider panel of 116 unrelated families in which the FEH polymorphism explained 35 % of the post-harvest variance for inulin content, 36 % of variance for sucrose content, 18 % for inulin degree of polymerization, 23 % for free fructose content and 22 % for free glucose content. These polymorphisms were associated with significant post-harvest changes of inulin content, inulin chain length and free sugars content. © 2013 Springer-Verlag Berlin Heidelberg. Source

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