The influence of a simulated digestive process on some biochemical and biological aspects of strawberry grape (Vitis labrusca) was investigated. The amount of total polyphenols and anthocyanins as well as the antioxid...The influence of a simulated digestive process on some biochemical and biological aspects of strawberry grape (Vitis labrusca) was investigated. The amount of total polyphenols and anthocyanins as well as the antioxidant power were evaluated. Results evidenced that the simulated gastrointestinal transit caused a decrease of the polyphenols content and total anthocyanins;these last, however, were more resistant than polyphenols, decreasing only of 50% respect to the initial value (31.50 μg/ml of extract). The extract exhibited an excellent antioxidant power (EC50 3.8 mg/ml), which decreased of about four times after the simulated gastrointestinal transit. The antimicrobial activity of the extract, evaluated against three Escherichia coli, Pseudomonas aeruginosa and Bacillus cereus pathogen strains was enhanced by the simulated digestion, with an increase of the inhibition halo.展开更多
Torzella (Brassica oleracea acephala) is one of the most ancient kinds of cauliflower developed in the Mediterranean area. Broccolo (Brassica oleracea botrytis cimosa), var. “San Pasquale” is a leafy vegetable culti...Torzella (Brassica oleracea acephala) is one of the most ancient kinds of cauliflower developed in the Mediterranean area. Broccolo (Brassica oleracea botrytis cimosa), var. “San Pasquale” is a leafy vegetable cultivated in Campania too, mainly in the province of Naples. We evaluated the polyphenols content and the antioxidant activity of these two Brassicaceae, provided by the same experimental plant of the Campania region. Both vegetables showed high content of total polyphenols (6.37 mM GAE/g and 2.24 mM GAE/g of Torzella and Broccolo, respectively), and a remarkable antioxidant activity (EC50 1.53 mg and EC50 6.51 mg, in Torzella and Broccolo, respectively). Polyphenol composition, determined by ultra performance liquid chromatography (UPLC), allowed us to detect a certain number of compounds (gallic, ferulic, chlorogenic, p-coumaric and caffeic acids, catechin, luteolin, naringenin) common to both species. Therefore, epicatechin was found only in the extract of san Pasquale broccoli;on the contrary, rutin and apigenin were detected only in the extracts of torzella broccoli. Hence, such products can be considered therapeutic functional foods due to their extraordinary reserve of secondary metabolites and bioactive constituents that are beneficial for managing and preventing several chronic illnesses in humans.展开更多
Four types of kale-based snacks were investigated on the basis of polyphenol content and composition, glucosinolates amount, and antioxidant activity. The amount of polyphenols resulted to be higher than that of raw m...Four types of kale-based snacks were investigated on the basis of polyphenol content and composition, glucosinolates amount, and antioxidant activity. The amount of polyphenols resulted to be higher than that of raw material, fresh kale (0.57 ± 0.03 mg GAE). The antioxidant activity turned out indeed very effective, resulting sufficient just from 0.72 mg to 2.1 mg to dishearten the activity of 1 ml DPPH to 50%. Glucosinolates present in kale-based snacks ranged from 1.17 mg/gr to 1.52 mg of sinigrin equivalent/gr. Polyphenol profile, obtained by UPLC-DAD analysis allowed us to identify gallic acid, chlorogenic acid, and catechin common in all samples. Other polyphenols, that is caffeic acid, p-coumaric acid, and ferulic acid, were found in almost all samples, almost all more abundant than fresh kale, representing a precious source of beneficial metabolites. Our results suggest that such snacks could represent an effective functional dietary supplement also if compared to fresh vegetable. Manufacturing of kale-based snacks does not negatively affect the beneficial effect of the raw material;indeed, such snacks could positively meet people’s expectation, mainly children and young, well known consumers of snacks, which, also in such manner, could be addressed towards a healthier dietary regime.展开更多
The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of inter...The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of interest for improving crop management and developing new strategies for plant diseases control. The aim of this work is to investigate the components involved in this interaction and, for this purpose, a proteomic approach was used. 2-D maps of the protein extracts from the single components in various interactions between plants (potato, bean, tobacco or tomato), pathogens (Botrytis cinerea, Rhizoctonia solani or Pythium ultimum) and biocontrol fungi (Trichoderma atroviride strain P1 or Trichoderma harzianum strain T22) were obtained. The proteome of each partner was collected separately and extracted by acetone precipitation in presence of trichloroacetic acid and a reducing agent (DTT). The extracted proteins were separated by isoelectrofocusing (IEF), using IPG (Immobilized pH gradient) strips, followed by SDS-PAGE. In order to improve resolution the separations were performed both on wide than narrow pH range and on different gel lengths. Differential spots were noted in the proteome of the three-way interaction when compared to each single component. These were further characterized by mass spectrometry and in silico analysis with the aim of identifying and cloning the relative genes. During the in vitro interaction of T. harzianum strain T22 with tomato and the culture filtrate or cell walls of pathogens, the spot number was higher than in the presence of pathogen biomass. In terms of Trichoderma differential proteins displayed on 2D gels, the most important changes were obtained in the presence of P. ultimum . During the in vivo interaction with tomato, the antagonist proteome changed much more in presence of soilborne fungi R. solani and P. ultimum than with the foliar fungus B. cinerea, both in terms of total and increased or novel spots. In silico analysis of some of those spots revealed homology with intracellular enzymes (GTPases, hydrolases) and with stress-related proteins (heat shock proteins HSP70, bacteriocin cloacin). Specific proteins in the plant proteome, i.e. pathogenesis-related proteins, have been identified during the in vivo interaction of bean with R. solani and T. atroviride strain P1. This is in agreement with the demonstrated ability of these beneficial fungi to induce plant systemic disease resistance by activating expression of defence-related genes. Proteins extracted from T. atrovride strain P1 which were analysed by mass spectrometry, revealed some interesting homologies with a fungal hydrophobin of Pleurotus ostreatus and an ABC transporter of Ralstonia metallidurans. These could represent molecular factors involved in the antagonistic mechanisms of Trichoderma and play a role in the three-way interaction with the plant and other microbes.展开更多
Biochemical characteristics, antimicrobial and quorum quenching activity of the extract of the “Carciofo di Montoro”, a typical ecotype of Cynara cardunculus var. scolymus of the Campania region (Southern Italy) wer...Biochemical characteristics, antimicrobial and quorum quenching activity of the extract of the “Carciofo di Montoro”, a typical ecotype of Cynara cardunculus var. scolymus of the Campania region (Southern Italy) were studied, to consider it as potential reserve of bioactive constituents useful for food industry and beneficial for managing and preventing several chronic illnesses in humans. The extract exhibited a good polyphenol content (528 μg GAE/g) and antioxidant activity (EC50 less than 5 mg). Ultra pressure liquid chromatography (UPLC) revealed high amount of chlorogenic acid, cynarin and epicatechin. The extract showed antimicrobial activity against Escherichia coli, Staphylocccus aureus, Pseudomonas aeruginosa, Enterococcus faecalis and Bacillus cereus pathogen strains. Finally, quorum quenching activity was demonstrated. The variety Carciofo di Montoro could represent a good source of health-promoting polyphenols, encouraging a nutraceutical use of such ecotype, for several phyto-pharmaceutical applications.展开更多
Trichoderma harzianum strain T22 parasitizes and controls many phytopatogenic fungi and is applied commercially as biological control agent. The production of hydrolitic enzymes appears to be a key factor in the paras...Trichoderma harzianum strain T22 parasitizes and controls many phytopatogenic fungi and is applied commercially as biological control agent. The production of hydrolitic enzymes appears to be a key factor in the parasitic process. We tested the endo-esochitinolitic and glucanolitic activities of culture filtrates of T22 grown under carbon and nitrogen starvation or in presence of biomass or cell walls of the phytopathogenic fungi Botrytis cinerea, Rhizoctonia solani and Pythium ultimum. The highest level of enzimatic activities was found in culture where the mycoparasite interacted with a phytopathogenic fungus. Therefore we used a proteomic approach to investigate changes in the complex mixture of extracellular proteins secreted by T. harzianum strain T22 in order to identify proteins of potential biotechnology value for commercial and industrial use.Proteome technology has greatly enhanced our ability to conduct functional genomics studies. Nevertheless only a few studies have been published so far on the fungal extracellular proteome. Sample preparation remains the most critical step in analyses based on two-dimensional gel electrophoresis (2-DE), and it requires to be optimized for each specific application. In this study, our first aim was to set up the extraction protocol of the extracellular proteins secreted by T . harzianum strain T22 when it was grown in vitro .The secreted proteins were analysed by two-dimensional electrophoresis (2-DE) and substantial changes in the extracellular proteome of the mycoparasite have been observed. Comparing the 2D maps of the fungus grown in minimal medium with glycerol as carbon source (used as control condition) with those obtained in inducing conditions, a lot of novel proteins appeared. The higher number of novel and up-regulated spots was obtained in the presence of Rhizoctonia solani biomass. Other spots were specifically up-regulated by the interaction with different plant pathogens. Differentially expressed proteins were subjected to matrix assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectrometry (MS) and N-terminal Edman sequencing. The in silico analyses of some of the novel and up-regulated spots showed interesting homology to hypothetical and putative proteins from other fungal species. These include novel enzymes, such as glycosylhydrolases and metalloprotease, proteins with conserved domains involved in pathogen-host interactions, such as Ras that regulates signal transduction pathways or LRRs that is involved in host recognition, etc. Work is in progress to demonstrate the role of some of these proteins in biocontrol and ability to induce systemic resistance.展开更多
Scamorza is a pasta filata cheese produced in Southern Italy and eaten after a short ripening.The ripening phase is critical in defining the main qualitative features of the Scamorza cheese.The success of this operati...Scamorza is a pasta filata cheese produced in Southern Italy and eaten after a short ripening.The ripening phase is critical in defining the main qualitative features of the Scamorza cheese.The success of this operation is conditioned not only by the process parameters,but also by the characteristics of the ripening room in which different microclimates originate.This work intended to evaluate the influence of the different positions of cheeses within the ripening room on the evolution of their qualitative characteristics during the process of drying/ripening.For this purpose,samples of Scamorza cheese,produced in the Molise Region(Italy),were divided into two batches(C and L)and subjected to ripening for seven days in a thermo thermo-regulated room.The two batches were placed in different points of the room:the batch C in the central area and the batch L in the lateral area.During the ripening,temperature,humidity and air flow were monitored and the Scamorza cheeses were analysed to assess some qualitative characteristics.In a ripening room,the created microclimates are able to influence the quality of the product,as demonstrated by data related to temperature,humidity and air flow.In fact,from the results obtained,some appreciable differences among products from batches C and L were observed for the weight loss,the water activity and the colorimetric indexes.Differences in the behaviour of mesophilic lactic acid bacteria,pH and acidity were also found.The more rapid loss of water,characterizing the batch C,resulted in an evolution of physicochemical,physical and microbiological features,which resulted different from those observed in the samples from the batch L.Therefore,the results obtained in this study point out that,within the ripening room,the formation of different micro-environments is able to strongly influence the definition of the qualitative characteristics of the products placed in it.展开更多
文摘The influence of a simulated digestive process on some biochemical and biological aspects of strawberry grape (Vitis labrusca) was investigated. The amount of total polyphenols and anthocyanins as well as the antioxidant power were evaluated. Results evidenced that the simulated gastrointestinal transit caused a decrease of the polyphenols content and total anthocyanins;these last, however, were more resistant than polyphenols, decreasing only of 50% respect to the initial value (31.50 μg/ml of extract). The extract exhibited an excellent antioxidant power (EC50 3.8 mg/ml), which decreased of about four times after the simulated gastrointestinal transit. The antimicrobial activity of the extract, evaluated against three Escherichia coli, Pseudomonas aeruginosa and Bacillus cereus pathogen strains was enhanced by the simulated digestion, with an increase of the inhibition halo.
文摘Torzella (Brassica oleracea acephala) is one of the most ancient kinds of cauliflower developed in the Mediterranean area. Broccolo (Brassica oleracea botrytis cimosa), var. “San Pasquale” is a leafy vegetable cultivated in Campania too, mainly in the province of Naples. We evaluated the polyphenols content and the antioxidant activity of these two Brassicaceae, provided by the same experimental plant of the Campania region. Both vegetables showed high content of total polyphenols (6.37 mM GAE/g and 2.24 mM GAE/g of Torzella and Broccolo, respectively), and a remarkable antioxidant activity (EC50 1.53 mg and EC50 6.51 mg, in Torzella and Broccolo, respectively). Polyphenol composition, determined by ultra performance liquid chromatography (UPLC), allowed us to detect a certain number of compounds (gallic, ferulic, chlorogenic, p-coumaric and caffeic acids, catechin, luteolin, naringenin) common to both species. Therefore, epicatechin was found only in the extract of san Pasquale broccoli;on the contrary, rutin and apigenin were detected only in the extracts of torzella broccoli. Hence, such products can be considered therapeutic functional foods due to their extraordinary reserve of secondary metabolites and bioactive constituents that are beneficial for managing and preventing several chronic illnesses in humans.
文摘Four types of kale-based snacks were investigated on the basis of polyphenol content and composition, glucosinolates amount, and antioxidant activity. The amount of polyphenols resulted to be higher than that of raw material, fresh kale (0.57 ± 0.03 mg GAE). The antioxidant activity turned out indeed very effective, resulting sufficient just from 0.72 mg to 2.1 mg to dishearten the activity of 1 ml DPPH to 50%. Glucosinolates present in kale-based snacks ranged from 1.17 mg/gr to 1.52 mg of sinigrin equivalent/gr. Polyphenol profile, obtained by UPLC-DAD analysis allowed us to identify gallic acid, chlorogenic acid, and catechin common in all samples. Other polyphenols, that is caffeic acid, p-coumaric acid, and ferulic acid, were found in almost all samples, almost all more abundant than fresh kale, representing a precious source of beneficial metabolites. Our results suggest that such snacks could represent an effective functional dietary supplement also if compared to fresh vegetable. Manufacturing of kale-based snacks does not negatively affect the beneficial effect of the raw material;indeed, such snacks could positively meet people’s expectation, mainly children and young, well known consumers of snacks, which, also in such manner, could be addressed towards a healthier dietary regime.
文摘The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of interest for improving crop management and developing new strategies for plant diseases control. The aim of this work is to investigate the components involved in this interaction and, for this purpose, a proteomic approach was used. 2-D maps of the protein extracts from the single components in various interactions between plants (potato, bean, tobacco or tomato), pathogens (Botrytis cinerea, Rhizoctonia solani or Pythium ultimum) and biocontrol fungi (Trichoderma atroviride strain P1 or Trichoderma harzianum strain T22) were obtained. The proteome of each partner was collected separately and extracted by acetone precipitation in presence of trichloroacetic acid and a reducing agent (DTT). The extracted proteins were separated by isoelectrofocusing (IEF), using IPG (Immobilized pH gradient) strips, followed by SDS-PAGE. In order to improve resolution the separations were performed both on wide than narrow pH range and on different gel lengths. Differential spots were noted in the proteome of the three-way interaction when compared to each single component. These were further characterized by mass spectrometry and in silico analysis with the aim of identifying and cloning the relative genes. During the in vitro interaction of T. harzianum strain T22 with tomato and the culture filtrate or cell walls of pathogens, the spot number was higher than in the presence of pathogen biomass. In terms of Trichoderma differential proteins displayed on 2D gels, the most important changes were obtained in the presence of P. ultimum . During the in vivo interaction with tomato, the antagonist proteome changed much more in presence of soilborne fungi R. solani and P. ultimum than with the foliar fungus B. cinerea, both in terms of total and increased or novel spots. In silico analysis of some of those spots revealed homology with intracellular enzymes (GTPases, hydrolases) and with stress-related proteins (heat shock proteins HSP70, bacteriocin cloacin). Specific proteins in the plant proteome, i.e. pathogenesis-related proteins, have been identified during the in vivo interaction of bean with R. solani and T. atroviride strain P1. This is in agreement with the demonstrated ability of these beneficial fungi to induce plant systemic disease resistance by activating expression of defence-related genes. Proteins extracted from T. atrovride strain P1 which were analysed by mass spectrometry, revealed some interesting homologies with a fungal hydrophobin of Pleurotus ostreatus and an ABC transporter of Ralstonia metallidurans. These could represent molecular factors involved in the antagonistic mechanisms of Trichoderma and play a role in the three-way interaction with the plant and other microbes.
文摘Biochemical characteristics, antimicrobial and quorum quenching activity of the extract of the “Carciofo di Montoro”, a typical ecotype of Cynara cardunculus var. scolymus of the Campania region (Southern Italy) were studied, to consider it as potential reserve of bioactive constituents useful for food industry and beneficial for managing and preventing several chronic illnesses in humans. The extract exhibited a good polyphenol content (528 μg GAE/g) and antioxidant activity (EC50 less than 5 mg). Ultra pressure liquid chromatography (UPLC) revealed high amount of chlorogenic acid, cynarin and epicatechin. The extract showed antimicrobial activity against Escherichia coli, Staphylocccus aureus, Pseudomonas aeruginosa, Enterococcus faecalis and Bacillus cereus pathogen strains. Finally, quorum quenching activity was demonstrated. The variety Carciofo di Montoro could represent a good source of health-promoting polyphenols, encouraging a nutraceutical use of such ecotype, for several phyto-pharmaceutical applications.
文摘Trichoderma harzianum strain T22 parasitizes and controls many phytopatogenic fungi and is applied commercially as biological control agent. The production of hydrolitic enzymes appears to be a key factor in the parasitic process. We tested the endo-esochitinolitic and glucanolitic activities of culture filtrates of T22 grown under carbon and nitrogen starvation or in presence of biomass or cell walls of the phytopathogenic fungi Botrytis cinerea, Rhizoctonia solani and Pythium ultimum. The highest level of enzimatic activities was found in culture where the mycoparasite interacted with a phytopathogenic fungus. Therefore we used a proteomic approach to investigate changes in the complex mixture of extracellular proteins secreted by T. harzianum strain T22 in order to identify proteins of potential biotechnology value for commercial and industrial use.Proteome technology has greatly enhanced our ability to conduct functional genomics studies. Nevertheless only a few studies have been published so far on the fungal extracellular proteome. Sample preparation remains the most critical step in analyses based on two-dimensional gel electrophoresis (2-DE), and it requires to be optimized for each specific application. In this study, our first aim was to set up the extraction protocol of the extracellular proteins secreted by T . harzianum strain T22 when it was grown in vitro .The secreted proteins were analysed by two-dimensional electrophoresis (2-DE) and substantial changes in the extracellular proteome of the mycoparasite have been observed. Comparing the 2D maps of the fungus grown in minimal medium with glycerol as carbon source (used as control condition) with those obtained in inducing conditions, a lot of novel proteins appeared. The higher number of novel and up-regulated spots was obtained in the presence of Rhizoctonia solani biomass. Other spots were specifically up-regulated by the interaction with different plant pathogens. Differentially expressed proteins were subjected to matrix assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectrometry (MS) and N-terminal Edman sequencing. The in silico analyses of some of the novel and up-regulated spots showed interesting homology to hypothetical and putative proteins from other fungal species. These include novel enzymes, such as glycosylhydrolases and metalloprotease, proteins with conserved domains involved in pathogen-host interactions, such as Ras that regulates signal transduction pathways or LRRs that is involved in host recognition, etc. Work is in progress to demonstrate the role of some of these proteins in biocontrol and ability to induce systemic resistance.
基金Authors wish to thank Caseificio Molisano L.Barone s.n.c.for the production of cheeses.The study was performed within the project“Trasferimento di innovazione nella filiera lattiero-casearia per la valorizzazione del caciocavallo molisano e il recupero di sottoprodotti di trasformazione”,financially supported by PSR Molise 2007-2013-Misura 124.
文摘Scamorza is a pasta filata cheese produced in Southern Italy and eaten after a short ripening.The ripening phase is critical in defining the main qualitative features of the Scamorza cheese.The success of this operation is conditioned not only by the process parameters,but also by the characteristics of the ripening room in which different microclimates originate.This work intended to evaluate the influence of the different positions of cheeses within the ripening room on the evolution of their qualitative characteristics during the process of drying/ripening.For this purpose,samples of Scamorza cheese,produced in the Molise Region(Italy),were divided into two batches(C and L)and subjected to ripening for seven days in a thermo thermo-regulated room.The two batches were placed in different points of the room:the batch C in the central area and the batch L in the lateral area.During the ripening,temperature,humidity and air flow were monitored and the Scamorza cheeses were analysed to assess some qualitative characteristics.In a ripening room,the created microclimates are able to influence the quality of the product,as demonstrated by data related to temperature,humidity and air flow.In fact,from the results obtained,some appreciable differences among products from batches C and L were observed for the weight loss,the water activity and the colorimetric indexes.Differences in the behaviour of mesophilic lactic acid bacteria,pH and acidity were also found.The more rapid loss of water,characterizing the batch C,resulted in an evolution of physicochemical,physical and microbiological features,which resulted different from those observed in the samples from the batch L.Therefore,the results obtained in this study point out that,within the ripening room,the formation of different micro-environments is able to strongly influence the definition of the qualitative characteristics of the products placed in it.
