Acorn squash are a conveniently sized, inexpensive, nutritious vegetable, but the fruit quality of traditional cultivars is often unsatisfactory. New, sweet-tasting acorn squash were developed recently, but data on their fruit development and storability are limited. Our objective was to monitor the physical, carbohydrate, and antioxidant contents of 'Table Confection' sweet acorn squash fruits over the course of their development and after cold storage. Fruits were harvested at intervals spanning 15 to 60 days past anthesis (dpa) and assessed for various quality components. Maximum weight and dry matter content were attained at 40 dpa but 60-dpa fruits were firmer, with higher total soluble solids (TSS) (60 dpa: 18%; 40 dpa: 9%), sucrose, and antioxidant contents, and a more intense fruit-flesh colour. After long-term cold storage at either 10 degrees C-RH 70% or 15 degrees C-RH 95%, the fruits that had been harvested at 40 dpa had greatly increased TSS (17%), sucrose content, and flesh-colour intensity, but lost as much as 13% of their weight. The results indicate that if premature harvesting is forced by production or marketing considerations, sweet acorn squash can achieve high quality during cold storage, but at the expense of weight loss in accordance with the duration of storage.
Acorn squash (Cucurbita pepo) have been a familiar item at produce stands for decades in the United States and Canada, but little known or appreciated elsewhere. Following the breeding and development of sweet-fleshed acorn squash in Israel and its commercial introduction in 2007, acorn squash became a high-priced, popular produce item there. Scarcity of supply in winter has led to attempts to fill the consumer demand by using available protected cultivation infrastructure in the relatively mild area of southwestern Israel for production. Such production has proven feasible but it was not determined whether it would be preferable to allow the plants to simply grow sprawling on the ground or vertically, using trellises to train the plants to grow erect. Two sweet acorn squash hybrid cultivars differing in fruit size were compared, growing on the ground or on trellises, for yield, quality, and storability of the fruits. The hybrids bore fully ripe fruits from December through February, producing 56% higher yields when trellised rather than when allowed to grow on the ground. The fruits of trellised plants of both hybrids were more uniformly black-green and firmer than those of ground-grown plants. Their dry matter content at harvest and after 78 days of storage was very high, averaging 28% and 25%, respectively. Total soluble solids content of the fruit flesh from trellised plants was 19% at harvest and an extraordinary 20% after storage at 10°C, 70% RH. The fruit flesh of trellised plants was also more highly colored and had higher carotenoid, ascorbate, and anti-oxidant contents. Overall, trellising of sweet acorn squash during the winter under protected cultivation resulted in significantly higher yields and the finest fruit quality.
Marketing melons (Cucumis melo) as convenient fresh-cut products is popular nowadays. However, damage inflicted by fresh-cut processing results in fast quality degradation and food safety risks. The life of fresh-cut produce can be extended by a modified atmosphere (MA), either generated in a package by tissue respiration (a passive MA) or injected by gas flushing (an active MA). This work investigated the effect of passive and active MA formed in packages of different perforation levels on the quality of fresh-cut melons of two genetic groups: C. melo var. cantalupensis, characterized by climacteric fruit behavior, and non-climacteric C. melo inodorus. The best product preservation was achieved in passive MA packages: non-perforated for inodorus melons and micro-perforated for cantalupensis ones. The optimal packages allowed for the preservation of both genotypes for 14 days at 6–8 °C. The major factors limiting the shelf life of fresh-cut melons were microbial spoilage, translucency disorder and hypoxic fermentation associated with cantalupensis melons with enhanced ethyl acetate accumulation. Inodorus melons were found to be preferable for fresh-cut processing since they were less prone to fermented off-flavor development.
Herein, we report bioderived cross-linkers to create biopolymer-based hydrogels with tunable properties. Nucleosides (inosine and uridine) and ribose (pentose sugar lucking the nitrogenous base) were partially oxidized to yield inosine dialdehyde (IdA), uridine dialdehyde (UdA), and ribose dialdehyde (RdA). The dialdehydes were further used as cross-linkers with polysaccharide chitosan to form hydrogels. Depending on the cross-linker type and concentration, the hydrogels showed tunable rheological, mechanical, and liquid holding properties allowing the preparation of injectable, soft, and moldable hydrogels. Computational modeling and molecular dynamics simulations shed light on hydrogel formation and revealed that, in addition to covalent bonding, noncovalent interactions (π-π stacking, cation-π, and H-bonding) also significantly contributed to the cross-linking process. To demonstrate various application possibilities, the prepared hydrogels were used as a growth platform for plant cells, as injectable inks for layer-by-layer 3D printing applications, and as moldable hydrogels for soft lithography to replicate the microstructure of the plant. These findings suggest that the obtained tunable biocompatible hydrogels have the potential to be good candidates for various biotechnological applications.
The pericarp is the predominant tissue determining the structural characteristics of most fruits. However, the molecular and genetic mechanisms controlling pericarp development remain only partially understood. Previous studies have identified that CLASS-II KNOX genes regulate fruit size, shape, and maturation in Arabidopsis thaliana and Solanum lycopersicum. Here we characterized the roles of the S. lycopersicum CLASS-II KNOX (TKN-II) genes in pericarp development via a detailed histological, anatomical, and karyotypical analysis of TKN-II gene clade mRNA-knockdown (35S:amiR-TKN-II) fruits. We identify that 35S:amiR-TKN-II pericarps contain more cells around their equatorial perimeter and fewer cell layers than the control. In addition, the cell sizes but not the ploidy levels of these pericarps were dramatically reduced. Further, we demonstrate that fruit shape and pericarp layer number phenotypes of the 35S:amiR-TKN-II fruits can be overridden by the procera mutant, known to induce a constitutive response to the plant hormone gibberellin. However, neither the procera mutation nor exogenous gibberellin application can fully rescue the reduced pericarp width and cell size phenotype of 35S:amiR-TKN-II pericarps. Our findings establish that TKN-II genes regulate tomato fruit anatomy, acting via gibberellin to control fruit shape but utilizing a gibberellin-independent pathway to control the size of pericarp cells. Tomato CLASS-II KNOXgenes regulate fruit and pericarp anatomy via both gibberellin-dependent and independent pathways.
Herein, we report biocompatible hydrogel for wound healing that was prepared using nature-sourced building blocks. For the first time, OCS was employed as a building macromolecule to form bulk hydrogels along with the nature-sourced nucleoside derivative (inosine dialdehyde, IdA) as the cross-linker. A strong correlation was obtained between the mechanical properties and stability of the prepared hydrogels with a cross-linker concentration. The Cryo-SEM images of IdA/OCS hydrogels showed an interconnected spongy-like porous structure. Alexa 555 labeled bovine serum albumin was incorporated into the hydrogels matrix. The release kinetics studies under physiological conditions indicated that cross-linker concentration could also control the release rate. The potential of hydrogels in wound healing applications was tested in vitro and ex vivo on human skin. Topical application of the hydrogel was excellently tolerated by the skin with no impairment of epidermal viability or irritation, determined by MTT and IL-1α assays, respectively. The hydrogels were used to load and deliver epidermal growth factor (EGF), showing an increase in its ameliorating action, effectively enhancing wound closure inflicted by punch biopsy. Furthermore, BrdU incorporation assay performed in both fibroblast and keratinocyte cells revealed an increased proliferation in hydrogel-treated cells and an enhancement of EGF impact in keratinocytes.
This study reports significant steps toward developing anti-biofilm surfaces based on superhydrophobic properties that meet the complex demands of today's food and medical regulations. It presents inverse Pickering emulsions of water in dimethyl carbonate (DMC) stabilized by hydrophobic silica (R202) as a possible food-grade coating formulation and describes its significant passive anti-biofilm properties. The final coatings are formed by applying the emulsions on the target surface, followed by evaporation to form a rough layer. Analysis shows that the final coatings exhibited a Contact Angle (CA) of up to 155° and a Roll-off Angle (RA) lower than 1° on the polypropylene (PP) surface, along with a relatively high light transition. Dissolving polycaprolactone (PCL) into the continuous phase enhanced the average CA and coating uniformity but hindered the anti-biofilm activity and light transmission. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed a uniform coating by a "Swiss-cheese" like structure with high nanoscale and microscale roughness. Biofilm experiments confirm the coating's anti-biofilm abilities that led to the reduction in survival rates of S.aureus and E.coli, by 90-95% respectively, compared to uncoated PP surfaces.
This work addresses the challenge of delivering bioactive molecules by designing biocompatible nanogel particles (NGPs) utilizing rationally modified nature-sourced building blocks: capryl-oligochitosan and oxidized inosine. Capryl substituents endowed the resultant NGPs with membrane-penetration capabilities, while purine-containing inosine allowed H-bond/π-π/π-cation interactions. The prepared NGPs were complexed with carboxyfluorescein-labeled single-stranded oligonucleotide (FAM-oligo) and DsRed-encoding plasmid DNA. The successful delivery of FAM-oligo to the cell cytoplasm of the Nicotiana benthamiana plant was observed. Alexa 555-labeled bovine serum albumin (Alexa 555-BSA) was also efficiently encapsulated and delivered to the plant. In addition to delivering FAM-oligo and Alexa 555-BSA separately, NGPs also successfully co-delivered both biomolecules to the plant. Finally, NGPs successfully encapsulated the drug amphotericin B and reduced its toxicity while maintaining its efficacy. The presented findings suggest that NGPs may become a promising platform for the advanced delivery of bioactive molecules in various applications.
Improved postharvest storage is a major target for pepper-crop production. The three main components of postharvest improvement of pepper fruit are reducing water-loss rate, reducing chilling susceptibility, and increasing resistance to pathogens. To date, a small number of Quantitative Trait Locus (QTL) studies have been reported for reduced water loss and enhanced tolerance to chilling and anthracnose. More effort is needed to screen germplasm collections for accessions with improved postharvest traits. Molecular studies have enabled the identification of candidate genes conferring reduced susceptibility to chilling injury and pathogen infection in pepper fruit, and in related crops such as tomato ??? which may be implemented in pepper. Manipulation of the activity of these genes by genome editing can improve postharvest pepper quality.
BACKGROUND Terpene, eugenol and polyphenolic contents of basil are major determinants of quality, which is affected by genetics, weather, growing practices, pests and diseases. Here, we aimed to develop a simple predictive analytical method for determining the polyphenol, eugenol and terpene content of the leaves of major Israeli sweet basil cultivars grown hydroponically, as a function of harvest time, through the use of near-infrared (NIR) spectroscopy, liquid/gas chromatography, and chemometric methods. We also wanted to identify the harvest time associated with the highest terpene, eugenol and polyphenol content. RESULTS Six different cultivars and four different harvest times were analyzed. Partial least square regression (PLS-R) analysis yielded an accurate, predictive model that explained more than 93% of the population variance for all of the analyzed compounds. The model yielded good/excellent prediction (R-2 > 0.90, R-cv(2) and R-pre(2) > 0.80) and very good residual predictive deviation (RPD > 2) for all of the analyzed compounds. Concentrations of rosmarinic acid, eugenol and terpenes increased steadily over the first 3 weeks, peaking in the fourth week in most of the cultivars. Our PLS-discriminant analysis (PLS-DA) model provided accurate harvest classification and prediction as compared to cultivar classification. The sensitivity, specificity and accuracy of harvest classification were larger than 0.82 for all harvest time points, whereas the cultivar classification, resulted in sensitivity values lower than 0.8 in three cultivars. CONCLUSION The PLS-R model provided good predictions of rosmarinic acid, eugenol and terpene content. Our NIR coupled with a PLS-DA demonstrated reasonable solution for harvest and cultivar classification. (c) 2021 Society of Chemical Industry.
Cannabis based products are widely used for various medicinal conditions. Currently, the most common method to identify and quantify cannabinoids are liquid chromatographic methods coupled to UV-VIS or mass spectrometric detectors. As cannabinoids are fluorescent compounds, fluorescent-based detection methods are gaining increasing interest. Consequently, a comprehensive qualitative and quantitative determination of fluorescence characteristics of major cannabinoids is warranted. In the present study we have determined the fluorescent excitation-emission-spectrum of 13 major cannabinoid standards and calculated their molar extinction coefficient (epsilon) and quantum yield (Phi) values. Furthermore, we examined the correlation between epsilon and Phi to available physico-chemical substance descriptors, namely, octanol/water partitioning coefficient, molar weight and boiling point. The cannabinoids displayed a characteristic fluorescence excitation (225-400 nm)-emission (300-525 nm) signal. Neutral and acid cannabinoids could be distinguished based on their characteristic excitation-emission spectrum, namely neutral cannabinoids exhibited maximal intensities at 280 nm excitation and 300 nm emission, while acidic cannabinoids displayed maximal intensity at 300 nm excitation and 400 nm emission. Moreover, cannabinoids with overlapping UV spectrum (such as Delta 9-trans-tetrahydrocannabinolic acid, cannabidiolic acid and Cannabigerolic acid) were found to have overlapping fluorescence excitation-emission spectrum, due to the same number of conjugated double bonds and the presence or absence of a carboxylic group. The calculated epsilon and Phi values were found to negatively correlate with each other. The spectral data provided hereby could assist to develop tools for initial screening of extracted samples for the presence of cannabinoids.
Widely available enzymes, alpha-amylase from Aspergillus oryzae and pectinase from Aspergillus aculeatus, have been examined for their ability to perform nonspecific hydrolysis of polysaccharides chitosan, pectin, alginate, carboxymethyl cellulose (CMC), hydroxypropyl methylcellulose (HPMC) and methyl cellulose (MC) to form the corresponding oligosaccharides. The good yields of oligo-CMC (50-60%), oligo-pectin (40-60%) and oligo-alginate (25-54%) have been achieved. The hydrolysis of chitosan yielded 26-10% oligosaccharides, while HPMC and MC did not undergo significant hydrolysis process, pointing out the importance of polar groups, preferably carboxyl, for this reaction. Among the received oligomers, oligo-CMC is the least explored. Therefore, its bioactivity was examined, demonstrating a notable elicitation potential increasing the content of phenolic compounds and glucosinolates in broccoli sprouts. Furthermore, oligo-CMC was combined with H2O2 & nbsp;and found to diminish drawbacks and enhance advantages of this elicitor.& nbsp;As oligosaccharides find numerous applications in food, health, beauty and agriculture fields, sustainable and cost-effective method for their mass production is of importance. Moreover, the eliciting potential of oligo-CMC may allow its use in "super treatment " formulations that prevent damages, accelerate growth and increase content of beneficial nutrients in plants.
The aim of this review is to summarize our recently reported findings on the use of preharvest treatments (shade nets), applied either directly or in combination with other techniques (grafting) in order to minimize physiological disorders and maximize and maintain the phytochemical content of vegetables. The use of coloured nets for shading vegetables to protect against stress (intense solar radiation, heat stress, drought, drying winds and hailstorms) during the summer months is an effective and inexpensive method, and it provides plant protection and altered microclimate and modified intensity and quality of light. Moreover, the use of coloured nets supports a more intensive vegetative growth, longer vegetation and increased yield, and it reduces a number of physiological disorders while improving the morphological and nutritional quality of vegetables. Under colour nets, tomato plants provided fruits with thicker pericarp, firmness, a higher content of lycopene, a lower percent of physiological disorders and better tolerance to transport and storage. Shade-grown plants generally have higher total chlorophyll and carotenoid contents, an increase in the total yield and a decrease in physiological disorders accompanied with an increase in the content of total phenolic compounds and flavonoids. Grafting can increase yield and fruit size and improve or reduce external and/or internal fruit quality and retained better postharvest quality compared to the fruits from non-grafted plants. Further investigations using shade nets alone or in combination with grafting are needed to ensure the use of adequate strategies for managing plant growth of different plant species with limited physiological disorders for increased marketable yield and for maintaining quality during storage.
Fruits can be divided into dry and fleshy types. Dry fruits mature through senescence and fleshy fruits through ripening. Previous studies have indicated that partially common molecular networks could govern fruit maturation in these different fruit types. However, the nature of such networks remains obscure. CLASS-II KNOX genes were shown to regulate the senescence of the Arabidopsis (Arabidopsis thaliana) dry fruits, the siliques, but their roles in fleshy-fruit development are unknown. Here, we investigated the roles of the tomato (Solanum lycopersicum) CLASS-II KNOX (TKN-II) genes in fleshy fruit ripening using knockout alleles of individual genes and an artificial microRNA line (35S:amiR-TKN-II) simultaneously targeting all genes. 35S:amiR-TKN-II plants, as well as a subset of tkn-II single and double mutants, have smaller fruits. Strikingly, the 35S:amiR-TKN-II and tknII3 tknII7/+ fruits showed early ripening of the locular domain while their pericarp ripening was stalled. Further examination of the ripening marker-gene RIPENING INHIBITOR (RIN) expression and 35S:amiR-TKN-II rin-1 mutant fruits suggested that TKN-II genes arrest RIN activity at the locular domain and promote it in the pericarp. These findings imply that CLASS-II KNOX genes redundantly coordinate maturation in both dry and fleshy fruits. In tomato, these genes also control spatial patterns of fruit ripening, utilizing differential regulation of RIN activity at different fruit domains.
In this work, we present biocompatible nanocarriers based on modified polysaccharides capable of transporting insulin macromolecules through human skin without any auxiliary techniques. N-Alkylamidated carboxymethyl cellulose (CMC) derivatives CMC-6 and CMC-12 were synthesized and characterized using attenuated total reflectance Fourier transform infrared (ATR-FTIR) and nuclear magnetic resonance (NMR) spectroscopy, gel permeation chromatography and thermogravimetric, calorimetric and microscopic techniques. The prepared modified polysaccharides spontaneously assemble into soft nanoaggregates capable of adjusting to both aqueous and lipid environments. Due to this remarkable self-adjustment ability, CMC-6 and CMC-12 were examined for transdermal delivery of insulin. First, a significant increase in the amount of insulin present in lipid media upon encapsulation in CMC-12 was observed in vitro. Then, ex vivo studies on human skin were conducted. Those studies revealed that the CMC-12 carrier led to an enhancement of transdermal insulin delivery, showing a remarkable 85% insulin permeation. Finally, toxicity studies revealed no alteration in epidermal viability upon treatment and the absence of any skin irritation or amplified cytokine release, verifying the safety of the prepared carriers. Three-dimensional (3D) molecular modeling and conformational dynamics of CMC-6 and CMC-12 polymer chains explained their binding capacities and the ability to transport insulin macromolecules. The presented carriers have the potential to become a biocompatible, safe and feasible platform for the design of effective systems for transdermal delivery of bioactive macromolecules in medicine and cosmetics. In addition, transdermal insulin delivery reduces the pain and infection risk in comparison to injections, which may increase the compliance and glycemic control of diabetic patients.
Cannabis sativa L. is used to treat a wide variety of medical conditions, in light of its beneficial pharmacological properties of its cannabinoids and terpenes. At present, the quantitative chemical analysis of these active compounds is achieved through the use of laborious, expensive, and time-consuming technologies, such as high-pressure liquid-chromatography- photodiode arrays, mass spectrometer detectors (HPLC-PDA or MS), or gas chromatography-mass spectroscopy (GC-MS). Hence, we aimed to develop a simple, accurate, fast, and cheap technique for the quantification of major cannabinoids and terpenes using Fourier transform near infra-red spectroscopy (FT-NIRS). FT-NIRS was coupled with multivariate classification and regression models, namely partial least square-discriminant analysis (PLS-DA) and partial least squares regression (PLS-R) models. The PLS-DA model yielded an absolute major class separation (high-THC, high-CBD, hybrid, and high-CBG) and perfect class prediction. Using only three latent variables (LVs), the cross-validation and prediction model errors indicated a low probability of over-fitting the data. In addition, the PLS-DA model enabled the classification of chemovars with genetic-chemical similarities. The classification of high-THCA chemovars was more sensitive and more specific than the classifications of the remaining chemovars. The prediction of cannabinoid and terpene concentrations by PLS-R yielded 11 robust models with high predictive capabilities (R2CV and R2pred > 0.8, RPD >2.5 and RPIQ >3, RMSECV/RMSEC ratio <1.2) and additional 15 models whose performance was acceptable for initial screening purposes (R2CV > 0.7 and R2pred < 0.8, RPD >2 and RPIQ <3, 1.2 < RMSECV/RMSEC ratio <2). Our results confirm that there is sufficient information in the FT-NIRS to develop cannabinoid and terpene prediction models and major-cultivar classification models.
Cannabis is used to treat various medical conditions, and lines are commonly classified according to their total concentrations of Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD). Based on their ratio of total THC to total CBD, cannabis cultivars are commonly classified into high-THC, high-CBD, and hybrid classes. While cultivars from the same class have similar compositions of major cannabinoids, their levels of other cannabinoids and their terpene compositions may differ substantially. Therefore, a more comprehensive and accurate classification of medicinal cannabis cultivars, based on a large number of cannabinoids and terpenes is needed. For this purpose, three different chemometric-based classification models were constructed using three sets of chemical profiles. We examined those models to determine which provides the most accurate "chemovar" classification. This was done by analyzing profiles of cannabinoids, terpenes, and the combination of these substances using the partial least square-discriminant analysis multivariate (PLS-DA) technique. The chemical profiles were selected from the three major classes of medicinal cannabis that are most commonly prescribed to patients in Israel: high-THC, high-cannabigerol (CBG), and hybrid. We studied the correlations between cannabinoids and terpenes to identify major bio-indicators representing the plant's terpene and cannabinoid content. All three PLS-DA models provided highly accurate classifications, utilizing six to nine latent variables with an overall accuracy ranging from 2 to 11% CV. The PLS-DA model applied to the combined cannabinoid-and-terpene profile did the best job of differentiating between the chemovars in terms of misclassification error, sensitivity, specificity, and accuracy. The combined cannabinoid-and-terpene PLS-DA profile had cross-validation and prediction misclassification errors of 4% and 0%, respectively. This is the first study to demonstrate the highly accurate classification of samples of medicinal cannabis based on their cannabinoid and terpene profiles, as compared to cannabinoid profiles alone. Furthermore, our correlation analysis indicated that 11 cannabinoids and terpenes might serve as bio-indicators for 32 different active compounds. These findings suggest that the use of multivariate statistics could assist in breeding studies and serve as a tool for minimizing the mislabeling of cannabis inflorescences.
A series of quaternary dimethyl-(alkyl)-ammonium chitosan derivatives (QACs) was synthesized and studied for physicochemical properties and bioactivity. The QACs tended to spontaneously self-assembly into nanoaggregates. Antimicrobial activity was examined in vitro on Gram-negative Escherichia coli (E. coli) and Gram-positive Listeria innocua (L. innocua) bacteria as well as phytopathogenic fungus Botrytis cinerea. The hexyl chain-substituted QAC-6 demonstrated the highest potency causing 3.0- and 4.5-log CFU mL-1 reduction of E. coli and L. innocua, respectively. QAC-6 was tested for antimicrobial activity on stainless steel coupons and fresh spinach leaves. A traditional 'wet' application (spray) and dry Engineered Water Nanostructure (EWNS) approach were used for spinach decontamination. With both approaches, significant reduction of microbial load on the treated produce was achieved. The wet application showed a greater reduction of microbial load, while the advantages of EWNS were reaching the antimicrobial effect with miniscule dose of active agent leaving treated surface visibly dry.
This study presents anti-viral coating formulations based on Pickering emulsion. We used a versatile strategy to produce a stable Pickering emulsion by depletion interaction between an emulsion droplet and a particle using solvent-soluble polymers as a depletant. This interaction facilitates the adsorption of particles onto the droplet interface and stabilizes the emulsions. The coating formulation is based on water-in-toluene emulsions stabilized using commercial hydrophobic silica, with polycaprolactone polymer as a depletant dissolved in toluene. The structure of the emulsions and their stability were characterized by confocal microscopy, centrifugal analysis using a LUMiSizer (R), and cryogenic-scanning electron microscopy (cryo-SEM) used to confirm the depletion phenomenon. The active materials copper oxychloride and propionic acid were incorporated into the most stable water-in-toluene emulsion and applied to the polycarbonate surfaces after being oven-dried to remove the toluene while leaving the colloidosomes template with the active ingredient needed to resist viral infections. The bio-assay of the copper oxychloride and propionic add encapsulated samples were carried out on Nicotiana tabaccum var Xanthi plants. At a 6-7 leaf stage, Young plants of Xanthi were subjected to inoculation with the 0.01 M sodium-phosphate pH - 7.0 containing ToBRFV from each of the coated surfaces. The inoculated plants were dusted with carborundum to enhance virus infection, and three leaves above the two lowest leaves in each plant were rubbed. Local lesions were apparent at similar to 4 days post-inoculation, indicating virus count. The formulation encapsulated with copper oxychloride gave 87% protection, whereas the one with propionic add dissolved conferred 91% protection from infestation.