Transcriptome assessment in 'Red Globe' grapevine zygotic embryos during the cooling and warming phase of the cryopreservation procedure.

Cryopreservation has the potential for long-term germplasm storage. The metabolic pathways and gene regulation involved in cryopreservation procedures are still not well documented. Hence, the genetic expression profile was evaluated using RNA-Seq in zygotic embryos of grapevines subjected to cryopreservation by vitrification. Sequencing was performed on the Illumina NextSeq 500. The average alignment of reads was 96% against the reference genome. The expression profiles showed 229 genes differentially expressed (186 repressed and 46 induced). The main biological processes showing upregulated enrichment were related to nucleosome assembly, while downregulated processes were related to organ growth. The most highly repressed processes were associated with the organization of the cell wall and membrane components. The unnamed protein product and 17.3 kDa class II heat shock protein (HSP) were highly induced, while ATPase subunit 1 and expansin-A1 were repressed. The response to the cooling and warming process during cryopreservation probably indicates that the changes occurring in transcription may be related to epigenetics. In addition, the cell exhibits an increase in the reserve of nutrients while seeking to survive modestly using available energy and pausing the plant's development. Additionally, energy containment occurred to cope with the stress caused by the treatment where deactivation of components of the cell membrane was observed, possibly due to changes in fluidity caused by alterations in temperature.

Fruit Juices of Etcho (Pachycereus pecten-aboriginum) and Giant Cardon (Pachycereus pringlei) are Sources of Health-Promoting Ingredients with Potential Anticancer Properties.

Mexico is one of the main diversification centers of cacti in the world, with more than 500 endemic species, most of which remain nutritionally and functionally uncharacterized. The columnar cacti of the genus Pachycereus comprise five underutilized endemic Mexican species, whose nutraceutical properties have only been studied in the P. weberi species. Therefore, this study aimed to evaluate the nutritional quality and bioactive properties of etcho (P. pecten-aboriginum) and giant cardon (P. pringlei) fruit. The physical, chemical, and nutritional composition of etcho and giant cardon fruits were characterized, as well as the profile and content of bioactive compounds, antioxidant activity (ABTS•+ and DPPH•), and antiproliferative capacity in cervical (HeLa) and breast cancer (MDA-MB-231, MCF-7, and T-47D) cell lines. Our results suggest that etcho and giant cardon fruits are rich sources of essential nutrients and bioactive phytochemicals (including K, Mg, P, dietary fiber, polyphenolic compounds, vitamin C, betalains, and myo-inositol) with antioxidant and anticancer potential by inhibiting the proliferation of all evaluated cell lines with IC50 values in the range of 198 to 287 µg of gallic acid equivalents/mL. Therefore, etcho and giant cardon fruits could be used for nutraceutical purposes, and their consumption could promote health benefits.

Postharvest application of pectic-oligosaccharides on quality attributes, activities of defense-related enzymes, and anthocyanin accumulation in strawberry.

BACKGROUND: The postharvest application of pectic-oligosaccharides (POS) as an elicitor to improve the postharvest shelf-life and nutritional quality by stimulating natural defense mechanisms in strawberries was studied. Strawberries (cv. Festival) were treated with POS (at 0, 2, 5, and 9 g L-1 ) and evaluated for firmness, weight loss, color, soluble solids, titratable acidity (TA), total phenolic and anthocyanin content, antioxidant capacity, decay, and some defense-related enzyme activity during storage at 2 ± 0.5 °C for 14 days. RESULTS: Treatment with POS significantly delayed (P < 0.05) strawberry decay, and reduced the water loss and softening of fruit during storage. Strawberries treated with POS showed a significant increase in total phenolic and anthocyanin content, and antioxidant capacity when compared with controls. Interestingly, POS induced higher activity of phenylalanine ammonia-lyase (PAL), chitinase, and ß-1,3-glucanase in strawberries. Compared to the control, the activity of enzymes was markedly increased in fruit treated with all tested POS concentrations, particularly chitinase, and ß-1,3-glucanase activities, but 5 and 9 g L-1 POS were the most effective treatments for maintaining the quality attributes and improving anthocyanin accumulation and antioxidant capacity of strawberries. CONCLUSION: These findings suggest that POS treatment could potentially be applied to maintain quality attributes, reduce decay, and further enrich the anthocyanin content and antioxidant capacity of strawberries during postharvest storage. The results also suggest that the positive effects of POS on strawberries could be associated with the rapid accumulation of chitinase and ß-1,3-glucanase activities, and the increase of PAL enzyme activity leading to the synthesis and accumulation of anthocyanins. © 2019 Society of Chemical Industry.

Combination of Cymbopogon citratus and Allium cepa essential oils increased antibacterial activity in leafy vegetables.

BACKGROUND: Cymbopogon citratus and Allium cepa essential oils (EOs) are rich in terpenes and sulfur compounds respectively, both with antibacterial activity and different cell targets, supporting the idea that their combination can increase their efficacy. RESULTS: Major constituents of C. citratus were geranial and neral, while A. cepa presented dipropyl disulfide and dipropyl trisulfide. Cymbopogon citratus and A. cepa EOs inhibited the in vitro growth of Escherichia coli O157:H7 (minimal inhibitory concentrations of 2.21 and 5.13 g L-1 respectively), Salmonella Choleraesuis (3.04 and 1.28 g L-1 ), Listeria monocytogenes (1.33 and 2.56 g L-1 ) and Staphylococcus aureus (0.44 and 5.26 g L-1 ). Application of the EO combination to spinach caused a greater reduction in E. coli (2.34 log colony-forming units (CFU) g-1 ), S. Choleraesuis (2.94 log CFU g-1 ), L. monocytogenes (2.06 log CFU g-1 ) and S. aureus (1.37 log CFU g-1 ) compared with higher doses of individual EOs; a similar effect was observed for romaine lettuce. Individual and combined EOs caused a reduction in flavor acceptability level; however, no significant differences were found among odor acceptability of control vegetables and those treated with the EO combination and C. citratus EO. CONCLUSION: Leafy vegetables treated with the EO combination showed higher antibacterial protection and odor acceptability compared with individual EO treatments. © 2016 Society of Chemical Industry.

Quality and PR gene expression of table grapes treated with ozone and sulfur dioxide to control fungal decay.

BACKGROUND: Gaseous fumigants are commonly employed to control fungal decay of cold-stored grapes. So far it is not clear if these fumigants, besides the direct interaction against fungal structures, induce transcriptional responses of defensive markers. In order to contribute to understanding the mechanisms by which these fumigants exert their effect, we studied the influence of ozone (O3) and sulfur dioxide (SO2) on the decay caused by Botrytis cinerea, and the quality and expression of the defense-related genes chitinase, ß-1,3-glucanase and phenylalanine ammonia-lyase (PAL) in the table grape cultivars 'Redglobe' and 'Sugraone'. RESULTS: The application of SO2 or O3 delayed decay of both table grape cultivars caused by B. cinerea compared with the inoculated control. O3 treatments altered weight loss, firmness and shatter in both cultivars. Significant upregulation of chitinase and ß-1,3-glucanase were observed in SO2 -treated 'Redglobe' berries stored at 2 °C. O3 treatment transiently increased the expression of chitinase and PAL in 'Redglobe' and 'Sugraone' berries, respectively. CONCLUSION: Ozone and sulfur dioxide treatments can influence the expression patterns of PAL, chitinase and ß-1,3-glucanase to different extents in different grape cultivars and under different exposure conditions. The upregulation of these genes may be involved in the mechanism by which these fumigants inhibit the decay caused by pathogenic fungi.

Microtubule plus end-tracking proteins play critical roles in directional growth of hyphae by regulating the dynamics of cytoplasmic microtubules in Aspergillus nidulans.

Cytoplasmic microtubules (MTs) serve as a rate-limiting factor for hyphal tip growth in the filamentous fungus Aspergillus nidulans. We hypothesized that this function depended on the MT plus end-tracking proteins (+TIPs) including the EB1 family protein EBA that decorated the MT plus ends undergoing polymerization. The ebAΔ mutation reduced colony growth and the mutant hyphae appeared in an undulating pattern instead of exhibiting unidirectional growth in the control. These phenotypes were enhanced by a mutation in another +TIP gene clipA. EBA was required for plus end-tracking of CLIPA, the Kinesin-7 motor KipA, and the XMAP215 homologue AlpA. In addition, cytoplasmic dynein also depended on EBA to track on most polymerizing MT plus ends, but not for its conspicuous appearance at the MT ends near the hyphal apex. The loss of EBA reduced the number of cytoplasmic MTs and prolonged dwelling times for MTs after reaching the hyphal apex. Finally, we found that colonies were formed in the absence of EBA, CLIPA, and NUDA together, suggesting that they were dispensable for fundamental functions of MTs. This study provided a comprehensive delineation of the relationship among different +TIPs and their contributions to MT dynamics and unidirectional hyphal expansion in filamentous fungi.

Post-harvest control of gray mold in table grapes using volatile sulfur compounds from Allium sativum.

BACKGROUND: This study proposes the use of volatile sulfur compounds derived from garlic (Allium sativum) extracts applied via gas for the control of Botrytis cinerea, which causes post-harvest disease in table grapes. The effects of the volatile compounds emitted by garlic extract and sodium metabisulfite on conidia germination of B. cinerea were evaluated in vitro to assess their effectiveness at controlling grey mold on grapes stored at different temperatures. RESULTS: Diallyl disulfide, diallyl trisulfide and allicin were identified and quantified in a garlic extract using reversed-phase high-performance liquid chromatography. The volatile compounds emitted by the garlic extract in the form of allicin and diallyl trisulfide inhibited conidia germination of B. cinerea in vitro and significantly reduced the lesion diameters on stored grapes, which were similar to the effects of sodium metabisulfite, while the diallyl disulfide did not have any effect. The sulfhydryl groups of cysteine or reduced glutathione completely reversed the antifungal effect of these compounds. CONCLUSION: The antifungal activity that allicin and diallyl trisulfide, which are the volatile compounds emitted by a garlic extract, exerted on conidia germination of B. cinerea may be considered as an alternative for the control of gray mold in table grapes after harvest.

Pectin-derived oligosaccharides increase color and anthocyanin content in Flame Seedless grapes.

BACKGROUND: Grapes grown in warm weather do not develop a desirable red color and require the use of products to enhance berry color. Pectin-derived oligosaccharides (PDOs) have been shown to have a role in various responses including plant defense, growth and development. In this work a mixture of PDOs with 3-20 degrees of polymerization was applied to Vitis vinifera cv. Flame Seedless grapes under field conditions and compared to the effects of ethephon (an ethylene-releasing compound). The effect of treatments on grape color, anthocyanin content and phenylalanine ammonia lyase (PAL) mRNA levels was evaluated. RESULTS: PDOs treatment increased berry color measured by the Color Index of Red Grapes (CIRG) and anthocyanin content, compared to ethephon and untreated berries (control); 1.5, 1 and 0.5 mg mL⁻¹ PDOs increased berry color by 30%, 27% and 26%, respectively, when compared to control berries. Levels of PAL mRNA accumulating in berries treated with PDOs were elevated within the first 24 h of treatment. CONCLUSIONS: PDOs enhanced the color and anthocyanin content of Flame Seedless grape berries possibly due by the induction of PAL mRNA expression. The results demonstrated that PDOs can be used to improve fruit quality aspects such as berry skin color.

Are Bacteriocins a Feasible Solution for Current Diverse Global Problems?

The development of effective technologies to cope with persistent and progressive global problems in human health and sustainable development has become an imperative worldwide challenge. The search for natural alternatives has led to the discovery of bacteriocins, which are potent protein antimicrobial compounds produced by most bacteria. The relevance of these molecules is evidenced by more than 4,500 papers published in the last decade in Scopus indexed journals highlighting their versatility and potential to impact various aspects of daily life, including the food industry, medicine, and agriculture. Bacteriocins have demonstrated antibacterial, antifungal, antiviral, and anticancer activities, and they also act as microbiota regulators and plant growth promoters. This mini-review aims to provide insights into the current state and emerging roles of bacteriocins, as well as their potential and limitations as feasible solutions against current diverse global problems.

Oligosaccharins as Elicitors of Defense Responses in Wheat.

Wheat is a highly relevant crop worldwide, and like other massive crops, it is susceptible to foliar diseases, which can cause devastating losses. The current strategies to counteract wheat diseases include global monitoring of pathogens, developing resistant genetic varieties, and agrochemical applications upon diseases' appearance. However, the suitability of these strategies is far from permanent, so other alternatives based on the stimulation of the plants' systemic responses are being explored. Plants' defense mechanisms can be elicited in response to the perception of molecules mimicking the signals triggered upon the attack of phytopathogens, such as the release of plant and fungal cell wall-derived oligomers, including pectin and chitin derivatives, respectively. Among the most studied cell wall-derived bioelicitors, oligogalacturonides and oligochitosans have received considerable attention in recent years due to their ability to trigger defense responses and enhance the synthesis of antipathogenic compounds in plants. Particularly, in wheat, the application of bioelicitors induces lignification and accumulation of polyphenolic compounds and increases the gene expression of pathogenesis-related proteins, which together reduce the severity of fungal infections. Therefore, exploring the use of cell wall-derived elicitors, known as oligosaccharins, stands as an attractive option for the management of crop diseases by improving plant readiness for responding promptly to potential infections. This review explores the potential of plant- and fungal-derived oligosaccharins as a practical means to be implemented in wheat crops.

Physicochemical and transport properties of biodegradable agar films impregnated with natural semiochemical based-on hydroalcoholic garlic extract.

Biodegradable films based on agar with glycerol (GLY) as a plasticizer were developed by incorporating hydroalcoholic garlic extract (HGE) on the film surface. The effect of GLY content (0, 15, or 30 wt%) and different concentrations of HGE (0, 0.5, 1, or 1.5 µg/mL) on the physicochemical and transport properties of the films was evaluated. The optical (color and transparency), mechanical (tensile test), transport (diffusion and water vapor transmission rate), thermal (thermogravimetric analysis) structural (infrared spectroscopy and X-ray diffraction), and morphological (scanning electron microscope) properties were analyzed. The impregnation of HGE increased the transparency values and decreased the luminosity, tensile strength, elastic modulus, and crystallinity of the agar films. The formulation of 30 wt% GLY with 1.5 µg/mL HGE, identified as 30 GLY [1.5], showed a similar thermal stability that of a neat agar film. The agar films with 30 wt% GLY showed the lowest diffusion coefficient and water vapor transmission rate, indicating that volatile compounds are slowly released. From the results the formulation 30 GLY [1.5] could be used as a film to transport and to release HGE which is supported by a biodegradable matrix and this system has a potential use as insect semiochemical for plague control.

Inhibition of Glucosyltransferase Activity and Glucan Production as an Antibiofilm Mechanism of Lemongrass Essential Oil against Escherichia coli O157:H7.

The resistance of Escherichia coli O157:H7 to disinfection is associated with its ability to form biofilms, mainly constituted by glucans produced by glucosyltransferases. Citral and geraniol, terpenes found in the essential oil of Cymbopogon citratus (EO), have proven antibacterial activity against planktonic E. coli; however, no information was found about their efficacy and mode of action against E. coli biofilms. Therefore, the inhibitory effect of C. citratus EO, citral, and geraniol on glucans production and glucosyltransferase activity as anti-biofilm mechanism against E. coli was evaluated. EO, citral, and geraniol inhibited the planktonic growth of E. coli (minimal inhibitory concentration or MIC= 2.2, 1.0, and 3.0 mg/mL, respectively) and the bacterial adhesion (2.0, 2.0, and 4.0 mg/mL, respectively) on stainless steel. All compounds decreased the glucans production; citral and geraniol acted as uncompetitive inhibitors of glucosyltransferase activity (The half maximal inhibitory concentrations or IC50 were 8.5 and 6.5 µM, respectively). The evidence collected by docking analysis indicated that both terpenes could interact with the helix finger of the glucosyltransferase responsible for the polymer production. In conclusion, C. citratus EO, citral, and geraniol inhibited glucosyltransferase activity, glucans production, and the consequent biofilm formation of E. coli O157:H7.

Data on antioxidant activity in grapevine (Vitis vinifera L.) following cryopreservation by vitrification.

Cryopreservation is used for the long-term conservation of plant genetic resources. This technique very often induces lethal injury or tissue damage. In this study, we measured indicators of viability and cell damage following cryopreservation and vitrification-cryopreservation in Vitis vinifera L. axillary buds cv. "Flame seedless" stored in liquid nitrogen (LN) for: three seconds, one hour, one day, one week and one month; after LN thawed at 38 °C for three minutes. The enzymatic activity of catalase (CAT) and superoxide dismutase (SOD), as well as the amount of malondialdehyde (MDA), total protein and viability were assayed.

Fungicidal potential of methoxylated flavones from citrus for in vitro control of Colletotrichum gloeosporioides, causal agent of anthracnose disease in tropical fruits.

Four polymethoxylated flavones (3,5,6,7,3',4'-hexamethoxyflavone, 3,5,6,7,8,3',4'-heptamethoxyflavone, 5,6,7,8,4'-pentamethoxyflavone and 5,6,7,8,3',4'-hexamethoxyflavone) were isolated and characterized from cold-pressed orange oil. Their antifungal activities were evaluated against Colletotrichum gloeosporioides (Penz) Penz & Sacc, a major plant pathogen of fruits that causes significant damage to crops in tropical, sub-tropical and temperate regions. Methoxylated flavones were effective in inhibiting mycelial growth of the fungus. As flavone concentration increased, mycelial growth decreased. 5,6,7,8,3',4'-Hexamethoxyflavone completely inhibited the growth of Cgloeosporioides at a concentration of 100 microg ml(-1).

Volatiles emitted by Carya illinoinensis (Wang.) K. Koch as a prelude for semiochemical investigations to focus on Acrobasis nuxvorella Nuenzig (Lepidoptera: Pyralidae).

BACKGROUND: Plant volatiles have complex intra- and interspecific effects in the environment that include plant/herbivore interactions. Identifying the quantity and quality of volatiles produced by a plant is needed to aid the process of determining which chemicals are exerting what effects and then examining whether these effects can be manipulated to benefit society. The qualitative characterization of volatile compounds emitted by pecan, Carya illinoinensis (Wang.) K. Koch, was begun in order to establish a database for investigating how these volatiles affect Acrobasis nuxvorella Nuenzig, a monophagous pest of pecan. Headspace solid-phase microextraction combined with gas chromatography-mass spectrometry was used for the analysis of the volatile constituents of pecan during three phenological stages (dormant buds, intact new shoot growth and intact nutlets) of the Western Schley and Wichita cultivars. RESULTS: About 111 distinct compounds were identified from the two cultivars, accounting for ∼99% of the headspace volatiles. The chromatographic profiles of both varieties revealed variations in the volatile composition and proportion between cultivars, with a predominance of terpene hydrocarbons, of the sesquiterpenes class, as well as monoterpenes. CONCLUSION: The significantly higher responsiveness recorded for the larvae of A. nuxvorella to C. illinoinensis shoots indicates that the larvae may be activated by terpenes emanating from the new shoot growth. This is the first study that has examined volatiles of pecan in Mexico.

Irreversibility of chilling injury in zucchini squash (Cucurbita pepo L) could be a programmed event long before the visible symptoms are evident.

Zucchini fruits were subjected to 2.5 or 10 degrees C for 16d, followed by transfer to 20 degrees C for 24h in order to evaluate the relationship between ripening pattern, measured as CO(2) evolution and ethylene (C(2)H(4)) production, and metabolic heat production (q). Chilling injury (CI) visible symptoms were evident after 8d at 2.5 degrees C, but none were recorded on fruits kept at 10 degrees C. In fruits held at 10 degrees C, q, C(2)H(4) production, and CO(2) evolution diminished in the course of 16d, whereas in those at 2.5 degrees C CO(2) evolution showed an early burst peaking at 8d. Both C(2)H(4) production and q also showed a burst at 2.5 degrees C but they started at 4 and 8d, respectively, and peaked at 12d. The results showed that irreversibility of chilling injury in zucchini could occur long before the appearance of visible symptoms, although the metabolic activity accompanying the irreversibility process was not noticeable by isothermal calorimetry.

Effect of pectic oligomers on physiological responses of chilling injury in discs excised from zucchini (Cucurbita pepo L.).

The effect of pectic oligomers (OG) on ethylene biosynthesis, electrolyte leakage (EL), and CO(2) production was studied in discs excised from zucchini fruit (Cucurbita pepo L.) and stored at 20 or 2.5 degrees C. At 20 degrees C, OG enhanced ethylene biosynthesis and had a transient effect on decreasing EL, but showed little effect on respiratory rate; both the amount and size of the oligomer were important in changing both ethylene synthesis and EL. At 2.5 degrees C, OG increased both ethylene biosynthesis and respiratory rate with a maximum effect at 100 microg of oligomer and peaking at 6 h; shorter oligomers demonstrated an even greater effect on ethylene biosynthesis, but differences were smaller in respiratory rate. EL at 2.5 degrees C was affected most by 1 microg of OG and by monomeric galacturonic acid, with transient increases that peaked at 8 h. We suggest a signaling role for OG in the early steps of cold acclimation or chilling injury.