The Common Cichory (Cichorium intybus L.) as a Source of Extracts with Health-Promoting Properties-A Review.

Natural products are gaining more interest recently, much of which focuses on those derived from medicinal plants. The common chicory (Cichorium intybus L.), of the Astraceae family, is a prime example of this trend. It has been proven to be a feasible source of biologically relevant elements (K, Fe, Ca), vitamins (A, B1, B2, C) as well as bioactive compounds (inulin, sesquiterpene lactones, coumarin derivatives, cichoric acid, phenolic acids), which exert potent pro-health effects on the human organism. It displays choleretic and digestion-promoting, as well as appetite-increasing, anti-inflammatory and antibacterial action, all owing to its varied phytochemical composition. Hence, chicory is used most often to treat gastrointestinal disorders. Chicory was among the plants with potential against SARS-CoV-2, too. To this and other ends, roots, herb, flowers and leaves are used. Apart from its phytochemical applications, chicory is also used in gastronomy as a coffee substitute, food or drink additive. The aim of this paper is to present, in the light of the recent literature, the chemical composition and properties of chicory.

Rejuvenation of chicory and lettuce plants following phase change in tissue culture.

BACKGROUND: A frequent problem associated with the tissue culture of Compositae species such as chicory (Cichorium intybus L.) and lettuce (Lactuca sativa L.) is the premature bolting to in vitro flowering of regenerated plants. Plants exhibiting such phase changes have poor survival and poor seed set upon transfer from tissue culture to greenhouse conditions. This can result in the loss of valuable plant lines following applications of cell and tissue culture for genetic manipulation. RESULTS: This study demonstrates that chicory and lettuce plants exhibiting stable in vitro flowering can be rejuvenated by a further cycle of adventitious shoot regeneration from cauline leaves. The resulting rejuvenated plants exhibit substantially improved performance following transfer to greenhouse conditions, with increased frequency of plant survival, a doubling of the frequency of plants that flowered, and substantially increased seed production. CONCLUSION: As soon as in vitro flowering is observed in unique highly-valued chicory and lettuce lines, a further cycle of adventitious shoot regeneration from cauline leaves should be implemented to induce rejuvenation. This re-establishes a juvenile phase accompanied by in vitro rosette formation, resulting in substantially improved survival, flowering and seed set in a greenhouse, thereby ensuring the recovery of future generations from lines genetically manipulated in cell and tissue culture.

An in vivo whole-plant experimental system for the analysis of gene expression in extraradical mycorrhizal mycelium.

Arbuscular mycorrhizal fungi (AMF) establish beneficial mutualistic symbioses with land plants, receiving carbon in exchange for mineral nutrients absorbed by the extraradical mycelium (ERM). With the aim of obtaining in vivo produced ERM for gene expression analyses, a whole-plant bi-dimensional experimental system was devised and tested with three host plants and three fungal symbionts. In such a system, Funneliformis mosseae in symbiosis with Cichorium intybus var. foliosum, Lactuca sativa, and Medicago sativa produced ERM whose lengths ranged from 9.8 ± 0.8 to 20.8 ± 1.2 m per plant. Since ERM produced in symbiosis with C. intybus showed the highest values for the different structural parameters assessed, this host was used to test the whole-plant system with F. mosseae, Rhizoglomus irregulare, and Funneliformis coronatus. The whole-plant system yielded 1-7 mg of ERM fresh biomass per plant per harvest, and continued producing new ERM for 6 months. Variable amounts of high-quality and intact total RNA, ranging from 15 to 65 µg RNA/mg ERM fresh weight, were extracted from the ERM of the three AMF isolates. Ammonium transporter gene expression was successfully determined in the cDNAs obtained from ERM of the three fungal symbionts by RT-qPCR using gene-specific primers designed on available (R. irregulare) and new (F. mosseae and F. coronatus) ammonium transporter gene sequences. The whole-plant experimental system represents a useful research tool for large production and easy collection of ERM for morphological, physiological, and biochemical analyses, suitable for a wide variety of AMF species, for a virtually limitless range of host plants and for studies involving diverse symbiotic interactions.

A root chicory MADS box sequence and the Arabidopsis flowering repressor FLC share common features that suggest conserved function in vernalization and de-vernalization responses.

Root chicory (Cichorium intybus var. sativum) is a biennial crop, but is harvested to obtain root inulin at the end of the first growing season before flowering. However, cold temperatures may vernalize seeds or plantlets, leading to incidental early flowering, and hence understanding the molecular basis of vernalization is important. A MADS box sequence was isolated by RT-PCR and named FLC-LIKE1 (CiFL1) because of its phylogenetic positioning within the same clade as the floral repressor Arabidopsis FLOWERING LOCUS C (AtFLC). Moreover, over-expression of CiFL1 in Arabidopsis caused late flowering and prevented up-regulation of the AtFLC target FLOWERING LOCUS T by photoperiod, suggesting functional conservation between root chicory and Arabidopsis. Like AtFLC in Arabidopsis, CiFL1 was repressed during vernalization of seeds or plantlets of chicory, but repression of CiFL1 was unstable when the post-vernalization temperature was favorable to flowering and when it de-vernalized the plants. This instability of CiFL1 repression may be linked to the bienniality of root chicory compared with the annual lifecycle of Arabidopsis. However, re-activation of AtFLC was also observed in Arabidopsis when a high temperature treatment was used straight after seed vernalization, eliminating the promotive effect of cold on flowering. Cold-induced down-regulation of a MADS box floral repressor and its re-activation by high temperature thus appear to be conserved features of the vernalization and de-vernalization responses in distant species.

Water stress drastically reduces root growth and inulin yield in Cichorium intybus (var. sativum) independently of photosynthesis.

Root chicory (Cichorium intybus var. sativum) is a cash crop cultivated for inulin production in Western Europe. This plant can be exposed to severe water stress during the last 3 months of its 6-month growing period. The aim of this study was to quantify the effect of a progressive decline in water availability on plant growth, photosynthesis, and sugar metabolism and to determine its impact on inulin production. Water stress drastically decreased fresh and dry root weight, leaf number, total leaf area, and stomatal conductance. Stressed plants, however, increased their water-use efficiency and leaf soluble sugar concentration, decreased the shoot-to-root ratio and lowered their osmotic potential. Despite a decrease in photosynthetic pigments, the photosynthesis light phase remained unaffected under water stress. Water stress increased sucrose phosphate synthase activity in the leaves but not in the roots. Water stress inhibited sucrose:sucrose 1-fructosyltransferase and fructan:fructan 1 fructosyltransferase after 19 weeks of culture and slightly increased fructan 1-exohydrolase activity. The root inulin concentration, expressed on a dry-weight basis, and the mean degree of polymerization of the inulin chain remained unaffected by water stress. Root chicory displayed resistance to water stress, but that resistance was obtained at the expense of growth, which in turn led to a significant decrease in inulin production.

Impact of vernalization and heat on flowering induction, development and fertility in root chicory (Cichorium intybus L. var. sativum).

Root chicory (Cichorium intybus var. sativum) is a biennial plant that requires vernalization for flowering initiation. However, we previously showed that heat can induce root chicory flowering independently of vernalization. To deepen our understanding of the temperature control of flowering in this species, we investigated the impact of heat, vernalization and their interaction on flowering induction and reproductive development. Heat increased the flowering percentage of non-vernalized plants by 25% but decreased that of vernalized plants by 65%. After bolting, heat negatively affected inflorescence development, decreasing the proportion of sessile capitula on the floral stem by 40% and the floral stem dry weight by 42% compared to control conditions, although it did not affect the number of flowers per capitulum. Heat also decreased flower fertility: pollen production, pollen viability and stigma receptivity were respectively 25%, 3% and 82% lower in heat-treated plants than in untreated control plants. To investigate the genetic control of flowering by temperature in root chicory, we studied the expression of the FLC-LIKE1 (CiFL1) gene in response to heat; CiFL1 was previously shown to be repressed by vernalization in chicory and to repress flowering when over-expressed in Arabidopsis. Heat treatment increased CiFL1 expression, as well as the percentage of bolting and flowering shoot apices. Heat thus has a dual impact on flowering initiation in root chicory since it appears to both induce flowering and counteract vernalization. However, after floral transition, heat has a primarily negative impact on root chicory reproduction.

Proteomic analysis of chicory root identifies proteins typically involved in cold acclimation.

Chicory (Cichorium intybus) roots contain high amounts of inulin, a fructose polymer used as a storage carbohydrate by the plant and as a human dietary and prebiotic compound. We performed 2-D electrophoretic analysis of proteins from root material before the first freezing period. The proteins were digested with trypsin and the peptides analyzed by MS (MALDI-TOF/TOF). From the 881 protein spots analyzed, 714 proteins corresponded to a database accession, 619 of which were classified into functional categories. Besides expected proteins (e.g. related to metabolism, energy, protein synthesis, or cell structure), other well-represented categories were proteins related to folding and stability (49 spots), proteolysis (49 spots), and the stress response (67 spots). The importance of abiotic stress response was confirmed by the observation that 7 of the 21 most intense protein spots are known to be involved in cold acclimation. These results suggest a major effect of the low temperature period that preceded root harvesting.

Arsenic effects on some photophysical parameters of Cichorium intybus under different radiation and water irrigation regimes.

The presence of arsenic (As) in groundwater is a major problem in several parts of Latin America. In the present work, non-destructive approaches to monitor the effects of As on plants of Cichorium intybus, an herbaceous Asteraceae, were explored. In this sense, the effects of As at different levels of water and radiation were evaluated on these crops. Plants were grown in a greenhouse, watered daily with As solutions and exposed to different water and/or light conditions for four months, using a three-factor (As, water, radiation) and two-level resource (As vs non As, field capacity vs half-field capacity condition, light vs shade condition) factorial design. The parameters most affected by this treatment were the area under the first derivative of the reflectance spectrum in the blue region, chlorophyll concentration, the Fred/Ffar-red fluorescence ratio and the quantum yield for the photophysical decay. These changes indicated the ability of this plant species to be a biomonitor for the presence of arsenic in irrigation water. Interestingly, it was further proved in this work that the biomonitoring capacity was enhanced in the presence of sunlight.

Does particulate matter along roadsides interfere with plant reproduction? A comparison of effects of different road types on Cichorium intybus pollen deposition and germination.

The roadside habitat can be challenging for plants, which must maintain normal biological processes despite an influx of airborne pollutants. While the effects of many gases on plants have been quantified, the impacts of particulate pollutants have been relatively less studied. This is especially true of field experiments where particle dispersion may be influenced by meteorology and roadway use. We examined chicory (Cichorium intybus L.) along roadsides in the Cincinnati, Ohio metropolitan area to assess particulate influence on plant pollination through stigmatic clogging. We compared flowers collected from plants situated along interstates, U.S. highways, state highways, and county roads as these different road-types vary in motor vehicle usage and thus should have varying levels of particulate deposition on flowers. We examined floral stigmas for total particulates, total pollen, and percentage of pollen tube germination to determine whether particulates may interfere with early reproductive processes. Our results suggest that there was minimal variation of particulate matter found on chicory stigmas among road-types. Furthermore, the deposition of particulates on stigmas based on road-type did not show a strong link to variation in pollen deposition and pollen germination. There was also no significant relationship between total particulate levels and pollen germination rates across all road types. Future studies should investigate other plant species that may be more sensitive to roadside pollution, such as economically important crops. Locations in which vehicle use is increasing and where pollutants are not regulated strictly should also be examined as the effects of airborne particulates in early plant reproduction would be expected to be more substantial in these areas.

[Effects of Eucalyptus grandis leaf litter at its early stage of decomposition on the growth and photosynthetic characteristics of Cichorium intybus].

From March to May, 2010, a pot experiment was conducted to investigate the effects of Eucalyptus grandis leaf litter at its early stage of decomposition on the growth and photosynthetic characteristics of Cichorium intybus. Four treatments with different application rate of the leaf litter, i.e., 0 g x pot(-1) (CK), 30 g x pot(-1) (A1), 60 g x pot(-1) (A2), and 90 g x pot(-1) (A3), were installed. Each pot contained 12 kg soil mixed with the leaf litter, and then, C. intybus was sown. The growth indicators of the C. intybus were measured at the 30, 45, 60, and 75 d after sowing, and the photosynthetic characteristics of the C. intybus in treatment A3 were studied after the seedlings third leaf fully expanded. At each measured time, the biomass accumulation and leaf area growth of C. intybus in treatments A1, A2, and A3 were inhibited significantly. At the early stage of the leaf litter decomposition, the synthesis of photosynthetic pigments of the C. intybus seedlings was inhibited significantly, and the inhibition effect was getting stronger with the increasing amount of the leaf litter addition. The diurnal change of the seedlings photosynthetic rate in all treatments showed a bimodal curve with midday depression, the stomatal conductance and water use efficiency had the same variation trend with the net photosynthetic rate, and the total diurnal photosynthesis decreased in the order of CK > A1 > A2 > A3. The GC-MS analysis showed there were 33 kinds of small molecule compounds released gradually with the decomposition of the leaf litter, among which, allelopathic substance terpenoid dominated.

[Effects of Eucalyptus grandis leaf litter decomposition on the growth and photosynthetic characteristics of Cichorium intybus].

A pot experiment was conducted to study the effects of Eucalyptus grandis leaf litter during its early stage decomposition on the growth and the photosynthesis of Cichorium intybus. Each pot contained 12 kg soil mixed with different amounts of E. grandis leaf litter (30 g x pot(-1), A1; 60 g x pot(-1), A2; 90 g x pot(-1), A3; and 0 g x pot(-1), CK), and sowed with C. intybus. The growth indicators and the photosynthetic characteristics of C. intybus were measured after the third leaf of C. intybus seedlings fully expanded in treatment A3. At the early stage of leaf litter decomposition, the C. intybus biomass accumulation, leaf area growth, and synthesis of photosynthetic pigments were inhibited significantly, and the inhibition effect was getting stronger with the increasing amount of the leaf litter addition. The intercellular CO2 concentration of C. intybus was increased by litter addition, while the net photosynthetic rate, stomatal conductance, and transpiration rate were significantly lower than those of the control. With the increase of leaf litter addition, all the parameters of C. intybus light response and CO2 response except CO2 compensation point showed an obvious downward trend, and there existed significant differences between the treatments of litter additions and the control. It was suggested that during the decomposition of E. grandis leaf litter, its allelopathic substances released gradually and acted on receptor plants, inhibited the synthesis of photosynthetic pigments and the photosynthesis of the receptors, decreased the receptors environmental adaptation ability, and accordingly, inhibited the growth of C. intybus.

Models analyses for allelopathic effects of chicory at equivalent coupling of nitrogen supply and pH level on F. arundinacea, T. repens and M. sativa.

Alllelopathic potential of chicory was investigated by evaluating its effect on seed germination, soluble sugar, malondialdehyde (MDA) and the chlorophyll content of three target plants species (Festuca arundinacea, Trifolium repens and Medicago sativa). The secretion of allelochemicals was regulated by keeping the donor plant (chicory) separate from the three target plant species and using different pH and nitrogen levels. Leachates from donor pots with different pH levels and nitrogen concentrations continuously irrigated the target pots containing the seedlings. The allelopathic effects of the chicory at equivalent coupling of nitrogen supply and pH level on the three target plants species were explored via models analyses. The results suggested a positive effect of nitrogen supply and pH level on allelochemical secretion from chicory plants. The nitrogen supply and pH level were located at a rectangular area defined by 149 to 168 mg/l nitrogen supply combining 4.95 to 7.0 pH value and point located at nitrogen supply 177 mg/l, pH 6.33 when they were in equivalent coupling effects; whereas the inhibitory effects of equivalent coupling nitrogen supply and pH level were located at rectangular area defined by 125 to 131 mg/l nitrogen supply combining 6.71 to 6.88 pH value and two points respectively located at nitrogen supply 180 mg/l with pH 6.38 and nitrogen supply 166 mg/l with pH 7.59. Aqueous extracts of chicory fleshy roots and leaves accompanied by treatment at different sand pH values and nitrogen concentrations influenced germination, seedling growth, soluble sugar, MDA and chlorophyll of F. arundinacea, T. repens and M. sativa. Additionally, we determined the phenolics contents of root and leaf aqueous extracts, which were 0.104% and 0.044% on average, respectively.

Endophytic fungi occurring in fennel, lettuce, chicory, and celery--commercial crops in southern Italy.

The occurrence of endophytic fungi in fennel, lettuce, chicory, and celery crops was investigated in southern Italy. A total of 186 symptomless plants was randomly collected and sampled at the stage of commercial ripeness. Fungal species of Acremonium, Alternaria, Fusarium, and Plectosporium were detected in all four crops; Plectosporium tabacinum was the most common in all crop species and surveyed sites. The effect of eight endophytic isolates (five belonging to Plectosporium tabacinum and three to three species of Acremonium) inoculated on lettuce plants grown in gnotobiosis was assessed by recording plant height, root length and dry weight, collar diameter, root necrosis, and leaf yellowing. P. tabacinum and three species of Acremonium, inoculated on gnotobiotically grown lettuce plants, showed pathogenic activity that varied with the fungal isolate. Lettuce plants inoculated with the isolates Ak of Acremonium kiliense, Ac of Acremonium cucurbitacearum, and P35 of P. tabacinum showed an increased root growth, compared to the non-inoculated control. The high frequency of P. tabacinum isolation recorded in lettuce plants collected in Bari and Metaponto, and in fennel plants from Foggia agricultural districts, suggests a relationship not only between a crop species and P. tabacinum, but also between the occurrence of the endophyte and the crop rotation history of the soil.

Growth, fructan yield, and quality of chicory (Cichorium intybus L.) as related to photosynthetic capacity, harvest time, and water regime.

Fructans are polymers that are widely used in several industrial applications. In the last few years they have received increasing interest because of their positive effects on health. At present, fructans are mostly supplied by chicory, which is only grown and processed in The Netherlands, France, and Belgium. It would therefore be an attractive concept to expand its cultivation to the southern European countries, although water shortage and high temperatures may hinder its growth and yield. So far, few experiments have been carried out on the effects of water, so the present research was focused on the course of growth and fructan quality in rainfed (W(0)) and well-watered (W(1)) conditions. The positive effects of water restoration mostly concerned the above-ground dry weight (ADW), whereas the root dry weight (RDW) was less influenced. No significant differences on RDW were found in 1999, whereas it was 14% higher (P <0.01) in W(1) in 2000. The effect of water was very clear on assimilate allocation: the overall priority at the whole plant scale seemed to be root structures, then storage reserves, and finally ADW. Therefore, the fructan content was higher in W(0), and insignificant differences between W(0) and W(1) were found on fructan yield at the final harvests. The only significant effect of the water regime on fructans was to speed up their storage. The leaf photosynthetic capacity (A) was poorly affected by water availability, whereas it appeared consistently modulated by leaf temperature and leaf nitrogen content. Stomatal conductance appeared to be mostly affected by the soil water content and it was mostly related to A up to about 300 mmol m(-2) s(-1). The fructan chain length (DP) was not affected by water regime. Besides, DP classes showed a normal statistical distribution; skewness and kurtosis significantly changed only when the harvest was very late. Equally, a very late harvest time significantly lowered DP.

Expression analysis of a chicory fructan 1-exohydrolase gene reveals complex regulation by cold.

The gene for a recently identified cDNA, 1-FEH IIa, encoding a fructan 1-exohydrolase was isolated and cloned from Cichorium intybus and a 1149 bp promoter fragment was characterized. An analysis of the genomic 1-FEH IIa sequence indicated that the gene (FEHIIa) consists of six introns and seven exons, which is similar to plant invertase genes. Like invertase genes, FEHIIa also contains the 9 nt mini-exon encoding the tripeptide DPN. A database search for cis-acting response elements within its promoter identified multiple elements that appear to have relevance to cold-induced expression of the gene in field-grown roots. Promoter analysis by transient expression assay demonstrated that the FEHIIa gene promoter is highly expressed in etiolated Cichorium leaves and cold-stored roots, which correlated well with the high level expression detected by RNA blot analysis. Cold also enhanced FEHIIa reporter gene expression in green leaves, however, the reporter gene activity was much lower compared with similar induction experiments in etiolated leaves. Promoter deletion analysis demonstrated the presence of potential cold-responsive ABRE and/or CRT/DRE elements in the -22 to -172 region, while regions -933 to -717 and -493 to -278 contain elements that can down-regulate expression at the conditions used. Characterization of the FEHIIa promoter may provide tools to study cold-induced expression and to increase freezing tolerance in agricultural crops.

Comparative analysis of NBS domain sequences of NBS-LRR disease resistance genes from sunflower, lettuce, and chicory.

Plant resistance to many types of pathogens and pests can be achieved by the presence of disease resistance (R) genes. The nucleotide binding site-leucine rich repeat (NBS-LRR) class of R-genes is the most commonly isolated class of R-genes and makes up a super-family, which is often arranged in the genome as large multi-gene clusters. The NBS domain of these genes can be targeted by polymerase chain reaction (PCR) amplification using degenerate primers. Previous studies have used PCR derived NBS sequences to investigate both ancient R-gene evolution and recent evolution within specific plant families. However, comparative studies with the Asteraceae family have largely been ignored. In this study, we address recent evolution of NBS sequences within the Asteraceae and extend the comparison to the Arabidopsis thaliana genome. Using multiple sets of primers, NBS fragments were amplified from genomic DNA of three species from the family Asteraceae: Helianthus annuus (sunflower), Lactuca sativa (lettuce), and Cichorium intybus (chicory). Analysis suggests that Asteraceae species share distinct families of R-genes, composed of genes related to both coiled-coil (CC) and toll-interleukin-receptor homology (TIR) domain containing NBS-LRR R-genes. Between the most closely related species, (lettuce and chicory) a striking similarity of CC subfamily composition was identified, while sunflower showed less similarity in structure. These sequences were also compared to the A. thaliana genome. Asteraceae NBS gene subfamilies appear to be distinct from Arabidopsis gene clades. These data suggest that NBS families in the Asteraceae family are ancient, but also that gene duplication and gene loss events occur and change the composition of these gene subfamilies over time.