Seasonal variation in bioactive properties and phenolic composition of cardoon (Cynara cardunculus var. altilis) bracts.

Cardoon (Cynara cardunculus L.) bracts were collected at different maturation stages to investigate seasonal changes in the phenolic compounds profile and in vitro bioactivities. Among the 12 phenolic compounds tentatively identified, 3,5-O-dicaffeoylquinic acid (21.83 mg/g extract) and apigenin-7-O-glucuronide (10.6 mg/g extract) were the most abundant. Immature bracts (C1: principal growth stage (PGS) 5) had the highest phenolic compounds content, and anti-inflammatory (IC50 = 72 µg/mL) and cytotoxic (GI50 of 30-79 µg/mL) activities. Moreover, extract C1 inhibited efficiently the formation of thiobarbituric acid reactive substances (TBARS; IC50 = 26.8 µg/mL), while extract C8 (PGS 8/9) was more effective against oxidative haemolysis (IC50 38 and 75 µg/mL). The highest antibacterial and antifungal activities were attributed to samples C1 and C6 (PGS 7/8) and samples C2 (PGS 5/6) and C4 (PGS 6/7), respectively. Overall, the obtained results suggest the seasonal changes of polyphenolic composition and bioactivity of cardoon bracts of variable maturity.

Combined application of antagonistic Wickerhamomyces anomalus BS91 strain and Cynara cardunculus L. leaf extracts for the control of postharvest decay of citrus fruit.

Combined use of biocontrol agents and plant extracts can be considered a viable and promising strategy for protecting plant tissues with different synergistic mechanisms of action that improve the antimicrobial activity of the mixtures. Treatments of citrus fruits with Wickerhamomyces anomalus BS91 have been previously reported as effective measures to reduce the incidence of green mold caused by Penicillium digitatum. On the opposite, the knowledge of the antifungal activity of cultivated cardoon (Cynara cardunculus L. var. altilis DC.) leaf extract, vegetable widespread in some Mediterranean areas, is still very limited. In this study, experimental trials were conducted to evaluate the effectiveness in vitro of leaf aqueous, methanolic and ethanolic extracts of C. cardunculus against seven fungal pathogens responsible for considerable food losses in the postharvest stage. In addition, biocontrol yeast W. anomalus BS91 and the three C. cardunculus extracts were tested in vivo both as a single treatment and in mixture, against Penicillium digitatum on 'Tarocco' oranges and 'Femminello' lemons. The combination of W. anomalus BS91 and leaf ethanolic extract reduced with the highest efficacy the incidence and severity of green mold on orange and lemon fruits with respect to the control, and was more effective than treatment with antagonistic yeast or leaf extracts applied alone. Incidence and severity of citrus decay were more consistently reduced when mixtures were applied 24 h before the inoculation of the pathogen, thus suggesting the relevance of preventive treatments. The mixtures of antagonistic W. anomalus BS91 and ethanolic leaf extract were more effective in controlling green mold decay on oranges than on lemons. These results indicate that biocontrol agents and leaf extracts, used in appropriate combination, can provide a stronger protection than when used singularly. However, compatibility between microbial antagonist and antimicrobial extract should be preliminary verified.

Influence of Genotype and Harvest Time on the Cynara cardunculus L. Sesquiterpene Lactone Profile.

The excessive and inappropriate application of herbicides has caused environmental pollution. The use of allelochemicals as bioherbicides could provide a solution to this problem. The allelopathic activity of Cynara cardunculus L. has been studied previously, and sesquiterpene lactones (STLs) were identified as the most relevant allelochemicals. The goal of the study reported here was to investigate the effect of six genotypes and three harvest times on the qualitative and quantitative composition of STLs in C. cardunculus leaves through a new ultra-high-performance liquid chromatography-tandem mass spectrometry analysis method and, thus, the effect on phytotoxicity. Overall, wild cardoon contained the highest levels of STLs of the three botanical varieties studied. Nevertheless, climatic conditions had a marked influence on the presence of STLs among the six genotypes, which was higher in the April harvest. Cynaropicrin was the most abundant STL detected. A close relationship was found between the STL profiles and the allelopathic activity, expressed as inhibition of wheat coleoptile elongation. The data provide a new and important contribution to our understanding of C. cardunculus allelopathy.

An assessment of vegetation management practices and burrow fumigation with aluminum phosphide as tools for managing voles within perennial crop fields in California, USA.

Voles (Cricetidae) cause extensive damage to a variety of crops throughout much of the Northern Hemisphere. The removal of vegetation from crop fields at the end of the growing season, combined with a subsequent burrow fumigant application of aluminum phosphide, has the potential to substantially curtail vole activity but has not been thoroughly examined. We set up a study to test the impact of these management tools in perennial globe artichoke (Cynara cardunculus var. scolymus) fields in Monterey County, CA, during 2010 and 2011, to determine their potential utility as part of an integrated pest management (IPM) program for managing California voles (Microtus californicus). We used both chewing indices and mortality estimates derived via radiotelemetry to assess the efficacy of aboveground vegetation removal and aluminum phosphide applications on vole abundance. We determined the impact of plowing artichoke fields on vole activity as well. Both removal of vegetation and applications of aluminum phosphide substantially reduced vole presence within treated fields. Plowing also reduced vole abundance to the point of little residual activity following treatment. These management practices appear to be effective at eliminating voles from crop fields. Combining these tools with management practices designed to slow down reinvasion by neighboring vole populations (e.g., barriers, repellents, traps) has the potential to substantially reduce farmer reliance on rodenticides for vole management, although rodenticides will still be needed to curtail populations that reestablish within crop fields. Such an IPM approach should substantially benefit both farmers and agro-ecosystems.

Influence of the ripening stage and the lyophilization of wild cardoon flowers on their chemical composition, enzymatic activities of extracts and technological properties of cheese curds.

The selection of an appropriate Cynara cardunculus flowers batch is a preliminary step to produce rennet with better clotting properties. For this reason, we proposed to study the influence of the ripening stage and the lyophilization of cardoon flowers on their chemical composition, enzymatic activities of extracts, and technological properties of cheese curds. Results of flowers composition have shown that lyophilized flowers harvested at the middle of ripening stage (A) could be employed to produce mainly proteins or milk-clotting proteases. To confirm this, enzymatic activities of extracts and technological properties of curds were assessed. The experimental findings revealed that flowers lyophilization seems to be an efficient way to produce rennet with better clotting properties, leading to higher yield, moisture, and texture parameters of curd. These findings allowed us to select lyophilized flowers (A) for further cheese making process.

The potential of native species as bioenergy crops on trace-element contaminated Mediterranean lands.

The establishment of energy crops could be an option for the management of degraded and contaminated lands, where they would not compete with food production for land use. Here, we aimed to explore the potential of certain native Mediterranean species for the revegetation of contaminated lands for energy production purposes. A field survey was conducted in a trace-element (TE) contaminated area from SW Spain, where the patterns of biomass production, TE accumulation and the calorific value of some thistle species were analyzed along a soil contamination gradient. In a greenhouse experiment the response of two thistle species (Cynara cardunculus and Silybum marianum) and the shrub Dittrichia viscosa to soil contamination was assessed, as well as the effects of these species on some soil microbial parameters involved in nutrient cycling (enzyme activities and arbuscular mycorrhizal colonization in roots). Silybum marianum was able to colonize highly contaminated soils. Its aboveground biomass accumulated Cd and had a relatively high calorific value; this value was similar in biomass obtained from both heavily and moderately contaminated soils. Greenhouse experiment confirmed that S. marianum biomass production and calorific value is scarcely affected by soil contamination. In addition, some soil enzyme activities were clearly enhanced in the S. marianum rhizosphere. Dittrichia viscosa is another promising species, given its capacity to produce a high biomass with appreciable calorific value in acid contaminated soils. Germination of both species was hampered in the acid contaminated soil, and therefore soil pH correction would have to be accomplished before establishing these species on extremely acid soils. Further assessment of the risk of transfer of Cd and other TE to the food chain would be needed to confirm the suitability of these species for the revegetation of contaminated lands with energy production purposes.

Optimizing the combined application of amendments to allow plant growth in a multielement-contaminated soil.

This study was aimed to 1) properly understand the dynamics of toxic elements (Al, Fe, Mn, Cu, Pb, Zn and As) in a sulphide-mine soil after combined application of compost from urban sewage sludge (SVC) and bottom ashes from biomass combustion (BA) and to 2) optimize the combination of both amendments for vegetation growth. Soil was amended following a D-optimal design and the mixtures (15 in total) were incubated during 30 d. At the end of the incubation, the effects of amendments on the assessed variables as well as the process modelling were evaluated by Response Surface Methodology (RSM). The process modelling confirmed that quadratic models were adequate to explain the behaviour of the assessed variables (R(2) ≥ 0.94 and Q(2) ≥ 0.75). Both amendments significantly increased pH and electrical conductivity, while reduced metal extractability. A different behaviour of As respect to metals was observed and high doses of BA sharply increased its extractability. The optimization process indicated that adequate conditions for vegetation growth would be reached adding the soil with 6.8% of SVC and 3.1% of BA (dry weight). After amendments application the germination and root elongation of three energy crops were significantly increased while lipid peroxidation was decreased. Therefore, the combined application of SVC and BA to a contaminated soil could improve soil conditions and might be expected to have an advantage during plant growth. Moreover, the RSM could be a powerful technique for the assessment of combined amendment effects on soil properties and their effective application in multielement-contaminated soils.

Effects of saline stress on mineral composition, phenolic acids and flavonoids in leaves of artichoke and cardoon genotypes grown in floating system.

BACKGROUND: Cynara cardunculus is a species native to the Mediterranean basin. It includes globe artichoke and cultivated cardoon as well as their progenitor wild cardoon. The species is a source of biophenols, and its leaf extracts have been widely used in herbal medicine as hepatoprotectors and choleretics since ancient times. The aim of this study was to determine the effect of increasing the level of salinity in the nutrient solution (1 or 30 mmol L⁻¹ NaCl) on biomass production, mineral composition, radical-scavenging activity, caffeoylquinic acids and flavonoids in three artichoke ('Romolo', 'Violetto di Provenza' and 'Violetto di Romagna') and three cultivated cardoon ('Bianco Avorio', 'Bianco Gigante Inerme' and 'Gigante di Romagna') cultivars grown in a floating system. RESULTS: Increased salinity in the nutrient solution decreased the leaf dry biomass and leaf number of artichoke and cultivated cardoon cultivars. Salinity reduced macro- and microelement accumulation in leaves (e.g. N, K, Ca, Mg, Fe, Mn and B) but improved their antioxidant activity, total polyphenols, chlorogenic acid, cynarin and luteolin. The cultivated cardoons, especially 'Bianco Avorio' and 'Gigante di Romagna', showed higher biomass and leaf number than those observed in artichoke genotypes. 'Violetto di Provenza' exhibited the highest content of chlorogenic acid, closely followed by 'Violetto di Romagna', whereas for cynarin content the highest values were recorded in 'Violetto di Provenza', 'Bianco Avorio' and 'Gigante di Romagna'. The highest content of luteolin was recorded in 'Gigante di Romagna' and 'Bianco Avorio', while the highest content of apigenin was observed in 'Gigante di Romagna'. CONCLUSION: The results showed that the floating system could be considered an effective tool to improve quality aspects through proper management of the salt concentration in the nutrient solution. They also suggest that specific cultivars should be selected to obtain the desired profile of bioactive compounds.

Micropropagation of globe artichoke (Cynara cardunculus L. var. scolymus).

The globe artichoke (Cynara cardunculus L. var. scolymus) is a perennial plant cultivated in the Mediterranean region and the Americas for its edible young flower heads. Although vegetative propagation by offshoots or by "ovoli" (underground dormant axillary buds) has been the primary method of propagation, the potential for the diffusion of diseases and the phenotypic variability can be very high. The propagation of this species by axillary shoot proliferation from in vitro-cultured meristems produces systemic pathogen-free plants and a higher multiplication rate as compared to that obtained by conventional agamic multiplication. Axillary shoot proliferation can be induced from excised shoot apices cultured on Murashige and Skoog agar solidified medium supplemented with various concentrations of cytokinins and auxins, depending on genotype. For the production of virus-free plants, meristems, 0.3-0.8 mm long are excised from shoot apices and surface sterilized. The transfer of artichoke microshoots to a medium lacking cytokinins or with low cytokinin concentration is critical for rooting. Adventitious roots develop within 3-5 weeks after transfer to root induction MS medium containing NAA or IAA at various concentrations. However, in vitro rooting frequency rate is dependent on the genotype and the protocol used. Acclimatization of in vitro microshoots having 3-4 roots is successfully accomplished; plantlets develop new roots in ex vitro conditions and continue to grow.

Genetic mapping and identification of QTL for earliness in the globe artichoke/cultivated cardoon complex.

BACKGROUND: The Asteraceae species Cynara cardunculus (2n = 2x = 34) includes the two fully cross-compatible domesticated taxa globe artichoke (var. scolymus L.) and cultivated cardoon (var. altilis DC). As both are out-pollinators and suffer from marked inbreeding depression, linkage analysis has focussed on the use of a two way pseudo-test cross approach. RESULTS: A set of 172 microsatellite (SSR) loci derived from expressed sequence tag DNA sequence were integrated into the reference C. cardunculus genetic maps, based on segregation among the F1 progeny of a cross between a globe artichoke and a cultivated cardoon. The resulting maps each detected 17 major linkage groups, corresponding to the species' haploid chromosome number. A consensus map based on 66 co-dominant shared loci (64 SSRs and two SNPs) assembled 694 loci, with a mean inter-marker spacing of 2.5 cM. When the maps were used to elucidate the pattern of inheritance of head production earliness, a key commercial trait, seven regions were shown to harbour relevant quantitative trait loci (QTL). Together, these QTL accounted for up to 74% of the overall phenotypic variance. CONCLUSION: The newly developed consensus as well as the parental genetic maps can accelerate the process of tagging and eventually isolating the genes underlying earliness in both the domesticated C. cardunculus forms. The largest single effect mapped to the same linkage group in each parental maps, and explained about one half of the phenotypic variance, thus representing a good candidate for marker assisted selection.

Responses of Cynara cardunculus L. to single and combined cadmium and nickel treatment conditions.

A greenhouse pot experiment was carried out to study the responses of Cynara cardunculus L. (cardoon) to cadmium and nickel. Three groups of 12 pots each were planted with cardoon plants and spiked with single and combined cadmium and nickel aqueous solutions. The bioavailable metal concentrations, measured in soil, ranged widely and were up to 246.7 mg kg⁻¹ for Cd and 61.1 mg kg⁻¹ for Ni. Under Cd treatment, cardoon growth remained unaffected, while increased Ni soil concentrations inhibited plant growth and were lethal to the highly treated plants. In the combined Cd and Ni treatments, an antagonistic effect was observed between the two metals. Cadmium and nickel concentrations in cardoon tissues rose with increasing metal concentrations in the soil. Cadmium and nickel contents in shoots reached 169.3 and 342.3 mg kg⁻¹ in the single treatments while, under the combined Cd and Ni treatments, they were up to 235.0 and 440.7 mg kg⁻¹, respectively. Generally, mean contents of both metals in the shoots were higher than in the roots and the translocation factor was greater than 1. A possible enhancing effect of nickel on cadmium uptake was observed. Cardoon showed characteristics of a Cd accumulator.

Allocation of proteolytic activity in the seedling of Cynara cardunculus L. in the initial growth stages.

Cynara cardunculus L. seeds were germinated in vitro under environmentally controlled conditions. Seeds showed a 60% germination rate, and three growth stages were established based on the seedling mean relative growth rate (RGR). Root, stem and cotyledons were compared in these stages with respect to the emergence of total proteases and cardosin activity and its allocation in the seedling. In growth stage I (1st-5th post-germinative days), seedlings grew very slowly. Total proteases and cardosins were already active at the onset of seedlings in the stem. Total soluble protein remained constant in cardoon seedlings during stage I, and the content of all free amino acids (aa) but proline (Pro) was equally allocated on the 1st post-germinative day. In growth stage II (5th-10th post-germinative days), seedlings grew intensively and exhibited fully developed cotyledons. A pronounced increase in the content of all free aa up to the middle of growth stage II in both stems and roots was observed. In addition, the allocation of the total proteolytic activity and cardosins followed a gradient from the root to the seedling shoot. However, the whole seedling soluble protein remained constant up to the 7th day in and tended to peak on the 10th post-germinative day, being allocated mainly to the seedling stem. In growth stage III (10th-15th post-germinative days), cardoon seedlings exhibited the lowest mean RGR and the highest R/S growth ratio. An intensive degradation of total soluble protein present in the whole seedling except for cotyledons (ca. 5-fold) was observed. Nevertheless, in growth stage III, both the gradients exhibited by total proteases and cardosins activities between the root and the seedling shoot were enhanced, as were contents of all aa except Pro, exhibiting the highest levels in cotyledons on the 15th post-germinative day.