Transcriptome driven characterization of curly-and smooth-leafed endives reveals molecular differences in the sesquiterpenoid pathway

Endives(Cichorium endivia L.)are popular vegetables,diversified into curly/frisée-and smooth/broad-leafed(escaroles)cultivar types(cultigroups),and consumed as fresh and bagged salads.They are rich in sesquiterpene lactones(STL)that exert proven function on bitter taste and human health.The assembly of a reference transcriptome of 77,022 unigenes and RNA-sequencing experiments were carried out to characterize the differences between endives and escaroles at the gene structural and expression levels.A set of 3177 SNPs distinguished smooth from curly cultivars,and an SNP-supported phylogenetic tree separated the cultigroups into two distinct clades,consistently with the botanical varieties of origin(crispum and latifolium,respectively).A pool of 699 genes maintained differential expression pattern(core-DEGs)in pairwise comparisons between curly vs smooth cultivars grown in the same environment.Accurate annotation allowed the identification of 26 genes in the sesquiterpenoid biosynthesis pathway,which included several germacrene A synthase,germacrene A oxidase and costunolide synthase members(GAS/GAO/COS module),required for the synthesis of costunolide,a key precursor of lactucopicrin-and lactucin-like sesquiterpene lactones.The core-DEGs contained a GAS gene(contig83192)that was positively correlated with STL levels and recurrently more expressed in curly than smooth endives,suggesting a cultigroup-specific behavior.The significant positive correlation of GAS/GAO/COS transcription and STL abundance(2.4-fold higher in frisée endives)suggested that sesquiterpenoid pathway control occurs at the transcriptional level.Based on correlation analyses,five transcription factors(MYB,MYB-related and WRKY)were inferred to act on contig83192/GAS and specific STL,suggesting the occurrence of two distinct routes in STL biosynthesis.

Leaf nutrient content and transcriptomic analyses of endive (Cichorium endivia) stressed by downpour-induced waterlog reveal a gene network regulating kestose and inulin contents

Endive(Cichorium endivia L.),a vegetable consumed as fresh or packaged salads,is mostly cultivated outdoors and known to be sensitive to waterlogging in terms of yield and quality.Phenotypic,metabolic and transcriptomic analyses were used to study variations in curly-(‘Domari’,‘Myrna’)and smooth-leafed(‘Flester’,‘Confiance’)cultivars grown in short-term waterlog due to rainfall excess before harvest.After recording loss of head weights in all cultivars(6-35%),which was minimal in‘Flester’,NMR untargeted profiling revealed variations as influenced by genotype,environment and interactions,and included drop of total carbohydrates(6–50%)and polyols(3–37%),gain of organic acids(2–30%)and phenylpropanoids(98–560%),and cultivar-specific fluctuations of amino acids(−37 to+15%).The analysis of differentially expressed genes showed GO term enrichment consistent with waterlog stress and included the carbohydrate metabolic process.The loss of sucrose,kestose and inulin recurred in all cultivars and the sucrose-inulin route was investigated by covering over 50 genes of sucrose branch and key inulin synthesis(fructosyltransferases)and catabolism(fructan exohydrolases)genes.The lowered expression of a sucrose gene subset together with that of SUCROSE:SUCROSE-1-FRUCTOSYLTRANSFERASE(1-SST)may have accounted for sucrose and kestose contents drop in the leaves of waterlogged plants.Two anti-correlated modules harbouring candidate hub-genes,including 1-SST,were identified by weighted gene correlation network analysis,and proposed to control positively and negatively kestose levels.In silico analysis further pointed at transcription factors of GATA,DOF,WRKY types as putative regulators of 1-SST.