Comparing annual and biennial crop cycle on the growth, yield and quality of saffron using three corm dimensions

Saffron (Crocus sativus L.) is a geophyte plant belonging to the Iridaceae family and it is appreciated for its red dried stigmas used as cooking spice and flavouring agent. Effects of crop cycle length and mother corm dimension, as well as their interaction, have been evaluated on the flowering (morphological traits of flowers, days to flowering, flowering interval and flower production), quantitative traits (stigma and daughter corm yield), vegetative development (leaf and daughter corm traits) and qualitative characteristics (coloring, bittering and aromatic powers) of spice. A two-year field study (2017–2019) was conducted to compare annual and biennial crop cycle of saffron using three corm dimensional classes (D1: 2.0–2.5 cm, D2: 2.6–3.5 cm and D3: 3.6–4.5 cm) according a split-plot design with 3 replications. The results showed that the corms of D3 class, planted in annual crop cycle, produced flowers with the highest stigma length (42.2 mm), and dry weight of stigmas (7.4 mg), stamens (11.4 mg) and tepals (40.7 mg). The highest number of flowers per m2 (311.8) and stigma yield (20.7 kg ha 1) were found when corms belonging to D2 class were planted in biennial crop cycle, meanwhile the highest daughter corms production (35.9 t ha 1) was obtained when corms belonged to D3 class were planted in annual crop cycle. Number of daughter corms per m2 with a horizontal diameter from 3.1 to 4.5 cm and weight from 10.1 to > 25 g decreased as increasing the crop cycle length. In biennial crop cycle, corms of D1 class produced more daughter corms belonged to 3.1–3.5 cm diameter class and to 15.1–20 g weight class compared to D2 and D3 classes. Regarding to the spice quality, coloring and bittering powers were positively influenced by biennial crop cycle. According to International Standardization Organization (ISO 3632) references, the maximum values of color (306.3 A1 % 1 cm 440 nm) and taste (116.2 A1 % 1 cm 257 nm) were reached in spice obtained from “biennial crop cycle x D2 class” interaction. No significant effect of all experimental factors on aromatic power was found. It was concluded that the evaluation of combination between crop cycle length and corm dimension is ecessary in the saffron management in order to achieve the optimum yield of stigmas and corms, to improve the qualitative traits of spice and to enhance the by-products as corms of D1 class

The Influence of Soil Physical and Chemical Properties on Saffron (Crocus sativus L.) Growth,Yield and Quality

Soil physical and chemical properties play a central role in plant growth, influencing the availability of air, nutrients, and water. The aim of this two-year study was to evaluate the effect of soil texture and chemical properties (pH, electrical conductivity, organic carbon, organic matter, total, and active lime) on saffron (Crocus sativus L.) growth, yield, and quality. Corms were planted in pots filled with seven different soil textures obtained mixing an increasing quantity (33% and 66%) of sand to a clay soil (S1) and to a clay loam soil (S2) compared to a full (100%) sandy soil as a control (S7). A randomized complete block design comprising of seven pots with different types of soil (S1, S2, S3, S4, S5, S6, and S7) replicated three times was used. The results showed that the highest flower number (320.3 nm2), stigma yield (2.0 gm2), daughter corm production (7.9 kgm2), and horizontal diameter (3.1 cm) were derived from S3 and S4 soils. These were characterized by a loam and sandy-loam texture, not very calcareous, with a sub-alkaline and neutral pH, low electrical conductivity, a content of organic matter between 5.46 and 8.67 g kg1, and a content of active lime between 21.25 and 26.25 g kg1. According to the International Organization for Standardization (ISO) references, although all spice samples belonged to the first qualitative category, S1, S3, and S2 soils recorded the highest value for coloring power (290.5, 289.1, and 287.6 A1m 440 nm, respectively). The highest values of bittering (109.2 A1m 257 nm) and aromatic (26.6 A1m 330 nm) power were reached by S3 soil. Positive correlations were found both between color with clay and organic matter, and aroma with total calcium carbonate. In conclusion, the assessment of soil conditions is particularly important to obtain the best saffron performance in terms of stigma and daughter corms yield as well as spice qualitative traits

Crocus sativus L. Ecotypes from Mediterranean Countries: Phenological, Morpho-Productive, Qualitative and Genetic Traits

The characterization of C. sativus ecotypes is of great interest for preserving them from a possible genetic erosion due to the decrease of European cultivation surface. In this study, we evaluated four ecotypes from Italy (Sardinia and Abruzzo), Spain (Castilla-La Mancha), and Greece (Kozani) in order to detect the existence of variability and promote the biodiversity of this crop. Thirty-one traits related to saffron flowering, flower morphology, production of spice and daughter corms, vegetative development (leaf and corm traits), and spice quality, were evaluated. In addition, a genetic analysis through three PCR-based approaches, SSRs, RAPD, and SRAP was assessed. Results highlighted a phenotypic variation among ecotypes during two consecutive years. All the studied parameters were influenced by the ecotype except for the stamen length, color coordinates of tepals, leaf length, and leaf number per plant. Sardinia had a longer flowering interval, earlier flowering, and higher spice yield and quality than the other corm origins. The maximum values of morphological traits, such as stigma length, dry weight of stigmas, tepals, flowers and leaves, leaf area, and daughter corm weight were observed in the Abruzzo ecotype. Principal component analysis (PCA) showed a clear separation among ecotypes, in which Sardinia and Spain showed more similarities than Abruzzo and Kozani. Significant negative correlation was found between days to flower with stigma yield and quality. However, we could not find molecular markers discriminating among corm origins. In conclusion, this study suggests the importance of C. sativus ecotypes as precious source of biodiversity and bioactive compounds, and of their enhancement as fundamental prerequisite for a sustainable development strategy and as an agricultural diversification opportunity for growers

Crocus sativus (L.) Grown in Pots with High Volume Capacity: From a Case of Study to a Patent

Saffron (Crocus sativus L.) cultivation is widespread in different parts of the world, including various Mediterranean areas. The crop management techniques, requiring intensive manual labor from planting, weeding, flower picking to the collecting of stigmas, contribute greatly to the high price of the spice. Traditionally, the corms are cultivated in field soil and only stigmas are collected to obtain the spice while the flower’s remaining parts, corresponding to about 90% of the total biomass, are discarded and thrown away. In this study, in order to overcome some difficulties occurring during the whole crop cycle (pathogens, fungi, pests, weeds, etc.), as well as to ease and increase floral bioresidue recovery, an alternative planting way for Crocus sativus L. was proposed relying on the use of large pots. For this aim, corms with 3.0–3.5 cm diameter size, from two different geographical origins (Spain, Holland), were planted in plastic pots with a volume of 250 L or 350 L, placed in two different areas of the Basilicata region (Italy). The effect of this new growing condition on dry stigma yield as well as daughter corm yield and size was evaluated. Although this cultivation system is more expensive than the traditional one, it offers numerous and huge advantages. Among them, it allows us to maintain a more correct posture and to preserve flower integrity during harvesting. The structural integrity of the tepals is a very important factor to obtain innovative dried flowers in their original tridimensional shape (3D). Consequently, the proposed cultivation system facilitates the achievement of a real “niche product” with high added value (absence of pollen grains). Moreover, the qualitative analysis of the spice, performed according to the International Standardization Organization Normative 3632 (ISO 3632-2/1:2010/2011), classified all investigated saffron samples in the first qualitative category. The results of the first three trial years are very exciting and promising as they are similar to those from the literature carried out in ground soil. However, corms from Spain gave the best results. Further investigations are in progress in order to optimize this alternative cultivation system

Antioxidant Activity, Total Phenolic Content and Morphological Traits in 1 to 4-Year-Old <i>Muscari comosum</i> Bulbs Cultivated in Three Growing Environments

Muscari comosum, commonly called “Lampascione”, is an edible wild plant that grows predominantly in the Mediterranean area. In this study, qualitative characteristics (size and antioxidant properties) were investigated in “Lampascione” bulbs of different ages cultivated in three different growing environments (greenhouse, shaded greenhouse and open field). The 4-year-old bulbs grown in a greenhouse showed the best shape index. The highest total phenolic content was observed in seed bulbs for all the growing environments. The antioxidant activity expressed in terms of EC50 average value had a fluctuating trend. However, the best antioxidant activity was found in bulbs cultivated in open fields and in the 4-year-olds grown in the greenhouse. Therefore, all these bulbs have a high antioxidant activity and can be considered as a very good nutraceutical source useful to consumers, as well as in the pharmaceutical sector, who are more and more interested in having products for a healthy and natural diet

Simultaneous determination of water- and fat-soluble vitamins, lycopene and beta-carotene in tomato samples and pharmaceutical formulations: Double injection single run by reverse-phase liquid chromatography with UV detection

Vitamins and carotenes represent important dietary components essential for health, because of their potential ability to decrease the risk for heart disease, cancer and sight disorders. Established chromatographic methods are typically designed for classes of water- or fat-soluble compounds, based on their different solubility and chemical structures. The simultaneous determination of both kinds of vitamins, in addition to highly lipophilic compounds, such as carotenes, increases the complexity of an analytical method, as well as the choice of proper solubilisation solvents, which must be in tune with the chromatographic eluents. In this study, a novel LC method coupled with UV detection is described for the simultaneous determination of thiamine, riboflavin, niacin, pyridoxine, folic acid, ascorbic acid, retinol, γ-tocopherol, phylloquinone, lycopene and β-carotene. An on-line focusing step was required for column head trapping of carotenes and fatsoluble vitamins, before injecting the water-soluble vitamins. Then, two consecutive injections, combined in a single run based on a ternary gradient program, allowed an efficient separation of both lipophilic and hydrophilic compounds. The method was validated in terms of linearity, recovery and precision. Detection and quantification limits were in the range 0.6–200 μg/L and 2–700 μg/L, respectively. Finally, the potential of the proposed method was confirmed by the analysis of tomatoes and pharmaceutical formulations

MATRIX ASSISTED PULSED LASER EVAPORATION FOR LACCASE BASED BIOSENSORS

Matrix Assisted Pulsed Laser Evaporation (MAPLE) is a laser\u2010based deposition technique used to deposit films of soft material [1]. In MAPLE, a pulsed laser beam is focused inside a vacuum chamber and impinges on the surface of a rotating target. The target consists of a frozen solution of the material of interest that is diluted in an appropriate solvent. Thus, when the laser beam impacts the target, the laser pulsed energy is mainly absorbed by the solvent and converted to thermal energy, allowing the solvent to vaporize while the material of interest is deposited as a thin film [1]. Therefore, MAPLE is exploitable as an alternative strategy for the immobilization of enzymes [2]. In this study we focused our attention on Laccase, since it is an enzyme widely used as biological recognition component in biosensors for detecting polyphenols that are important compounds in foodstuffs [3] because of their recognized nutritional value. Laccase has been deposited by MAPLE onto suitable substrates from water and benzene solutions, and the effectiveness of the process has been evaluated in terms of characteristic of chemical bonds (Fourier Transform InfraRed), morphology (Atomic Force Microscopy), enzyme loading (quartz crystal microbalance), and enzyme activity (colorimetric assay). The deposition process has been also performed onto screen printed carbon electrodes, and the obtained biosensor has been characterized in terms of linearity, LOD, LOQ, repeatability and stability of response. Moreover, the potential of the Laccase biosensor has been also tested by the determination of total polyphenols content in vegetable ethanolic extracts and the results have been compared with those obtained by the Folin\u2010Ciocalteau method

Amperometric biosensor based on Laccase immobilized onto a screen-printed electrode by Matrix Assisted Pulsed Laser Evaporation

A Laccase-based biosensor for the determination of phenolic compounds was developed by using Matrix Assisted Pulsed Laser Evaporation as an innovative enzyme immobilization technique. and the deriving biosensor was characterized and applied for the first time. Laccase was immobilized onto different substrates including screen printed carbon electrodes and spectroscopic, morphologic and electrochemical characterizations were carried out. A linear range from 1 to 60 \u3bcM was achieved working at 5.5 pH and 120.2 V detection potential vs Ag pseudoreference. The limits of detection and quantification were found to be 1 and 5 \u3bcM, respectively. A good fabrication reproducibility, stability of response and selectivity toward interferents were also found The potential of the developed biosensor was tested in the determination of total polyphenol content in real matrices (tea infusion, ethanolic extract from Muscari comosum bulbs and aqueous solution of a food supplement from black radish root and artichoke leaves) and the results were compared with those obtained by using the Folin\u2013Ciocalteu method

Laccase thin films produced by MAPLE and ESI techniques: Effect of some solvents

Matrix Assisted Pulsed Laser Evaporation (MAPLE) and ElectroSpray Ionization (ESI) can represent useful techniques for producing enzyme thin films [1-2] exploitable in biosensors. Previous studies were carried out to deposit Laccase thin films by MAPLE using water or benzene as solvents [3-4]. Laccase was chosen since it is a redox enzyme widely used as a biological recognition component in biosensors for the detection of polyphenols. In this work, Laccase films deposited by MAPLE using water or benzene were compared and Laccase films deposited by ESI using acetonitrile or ethanol, both at 10% concentration, were also investigated. The efficiency of the deposition processes as well as the preservation of the biomolecular structure and functionality strongly depend on the solvent. The Laccase film funzionality in terms of enzymatic activity, was determined by spectrophotometric analysis by using syringaldazine as the enzyme substrate. These analysis highlighted that MAPLE using benzene allows obtaining more active films in a shorter time with respect to those obtained with water. Also ESI using ethanol allows obtaining more active Laccase films than those obtained by ESI using acenonitrile. Therefore a successful deposition of Laccase by MAPLE or ESI requires a careful choice of the solvent