Antioxidant Compounds from Edible Mushrooms as Potential Candidates for Treating Age-Related Neurodegenerative Diseases.

The last century has seen an increase in our life expectancy. As a result, various age-related diseases, such as neurodegenerative diseases (NDs), have emerged, representing new challenges to society. Oxidative stress (OS), a condition of redox imbalance resulting from excessive production of reactive oxygen species, represents a common feature that characterizes the brains of elderly people, thus contributing to NDs. Consequently, antioxidant supplementation or dietary intake of antioxidant-containing foods could represent an effective preventive and therapeutic intervention to maintain the integrity and survival of neurons and to counteract the neurodegenerative pathologies associated with aging. Food contains numerous bioactive molecules with beneficial actions for human health. To this purpose, a wide range of edible mushrooms have been reported to produce different antioxidant compounds such as phenolics, flavonoids, polysaccharides, vitamins, carotenoids, ergothioneine, and others, which might be used for dietary supplementation to enhance antioxidant defenses and, consequently, the prevention of age-related neurological diseases. In this review, we summarized the role of oxidative stress in age-related NDs, focusing on the current knowledge of the antioxidant compounds present in edible mushrooms, and highlighting their potential to preserve healthy aging by counteracting age-associated NDs.

Antioxidant Activity of Polysaccharides from the Edible Mushroom Pleurotus eryngii.

In this study the antioxidant and neuroprotective activity of an enriched polysaccharide fraction (EPF) obtained from the fruiting body of cultivated P. eryngii was evaluated. Proximate composition (moisture, proteins, fat, carbohydrates and ash) was determined using the AOAC procedures. The EPF was extracted by using, in sequence, hot water and alkaline extractions followed by deproteinization and precipitation with cold ethanol. Total α- and ß-glucans were quantified using the Megazyme International Kit. The results showed that this procedure allows a high yield of polysaccharides with a higher content of (1-3; 1-6)-ß-D-glucans. The antioxidant activity of EPF was detected from the total reducing power, DPPH, superoxide, hydroxyl and nitric oxide radical scavenging activities. The EPF was found to scavenge DPPH, superoxide, hydroxyl and nitric oxide radicals with a IC50 values of 0.52 ± 0.02, 1.15 ± 0.09, 0.89 ± 0.04 and 2.83 ± 0.16 mg/mL, respectively. As assessed by the MTT assay, the EPF was biocompatible for DI-TNC1 cells in the range of 0.006-1 mg/mL and, at concentrations ranging from 0.05 to 0.2 mg/mL, significantly counteracted H2O2-induced reactive oxygen species production. This study demonstrated that polysaccharides extracted from P. eryngii might be used as functional food to potentiate the antioxidant defenses and to reduce oxidative stress.

Edible Mushrooms as Source of Fibrin(ogen)olytic Enzymes: Comparison between Four Cultivated Species.

Cardiovascular diseases represent the main cause of death. A common feature of cardiovascular disease is thrombosis resulting from intravascular accumulation of fibrin. In the last years, several fibrinolytic enzymes have been discovered in many medicinal or edible mushrooms as potential new antithrombotic agents. This study aimed to compare the fibrin(ogen)olytic activity of crude extracts from the fruiting bodies of four cultivated edible mushrooms: Lentinula edodes, Pleurotus ostreatus, Pleurotus eryngii, and Agrocybe aegerita. Fibrin(ogen)olytic activity was assessed by fibrin plate, spectrophotometric assay and electrophoretic analysis (SDS-PAGE and zymography). The highest activity was detected for P. ostreatus followed by P. eryngii, L. edodes and A. aegerita. Results indicated that enzymes exhibited maximum activity at pH 6-7 and 30-40 °C, respectively. Enzyme activity was inhibited by serine and metalloprotease inhibitors. We proposed a new index called the Specific Fibrin(ogen)olytic Index (SFI), which allows specification of the proportion of the total proteolytic capacity due to the fibrin(ogen)olytic activity. These data suggest that the extracts from fruiting bodies or powdered mushrooms can be used as functional ingredients for the development of new functional foods that may act as thrombolytic agents responding, at the same time, to the increasing demand for safe, healthy and sustainable food.

Severe outbreak of tomato yellow leaf curl Sardinia virus on pepper in southern Italy.

During summer and autumn 2008 a severe outbreak of pepper leaf curl disease (PLCD) was observed in pepper crops under plastic tunnels in the ionic coast of Basilicata region. Its incidence reached, in some cases, values close to 50%. The beginning of infections was recorded along the perimeter of the tunnels, where it reached a percentage of almost 100%. The infection then progressively spread towards the central areas of the greenhouses. Large populations of whiteflies, identificated as Bemisia tabaci, were observed on the infected crops. Detection assays for TYLCSV and TYLCV were performed in order to ascertain the etiologic agent: 190 symptomatic samples were collected from different fields and assayed in DAS-ELISA using a broad-spectrum reagent combination (distributed by Bioreba AG) detecting TYLCV, TYLCSV and other Begamoviruses: of these, 176 samples resulted positive. In order to discriminate between TYLCSV, TYLCV or any other Begamovirus, 15 positive samples were analyzed by PCR using a couple of synthetic oligonucleotides allowing the amplification of the whole coat protein (CP) gene. RFLP analysis performed on the PCR product, 1008 bp long, showed the presence of only TYLCSV in all assayed samples. The molecular characterization performed by phylogenetic analysis of the sequenced coat protein gene revealed that the isolate shares a similarity of about 97% with the corresponding sequence of a tomato TYLCSV isolate from Sicily (Z28390) and is almost identical with the pepper isolate CAB-It recovered in the same area in 2007 (TYLCSV was first recorded on pepper in Italy in 2007 in Policoro-MT, Fanigliulo et al., 2008. Comm. Appl. Biol. Sci, Ghent University, 73/2, 2008), indicating that there is a very low variability in TYLCSV population in the surveyed area. The further diffusion of PLCD and its hazard has to be connected with the presence of wide tomato cultivations, of weed hosts alternative to pepper (Solanum nigrum, Datura stramonium, Sonchus asper, Cardaria draba and Abutilon theophrasti) and with the strong presence of the vector B. tabaci of the B biotype, more efficient than Q biotype in the virus transmission, and able to feed and reproduce on peppers as well as on tomatoes.

Cyazofamide: a fungicide against Bremia lactucae on lettuce.

During 2007 and 2008 summers, experimentation was performed by the Bioagritest test facility, according to EPPO guidelines and Principles of Good Experimental Practice (GEP), aiming at establishing the biological efficacy of Cyazofamide 25SC (formulate MILDICUT, Belchim) and Cyazofamide 400SC + an organosilicone adjuvant containing polyalkyleneoxide modified heptamethyltrisiloxane (Ranman 400 SC, Belchim) on lettuce against Bremia lactucae. The study was performed in Nocera Inferiore (Salerno), southern Italy, in a greenhouse with polyethylene cover. Experimental design consisted in random blocks, in 3 repetitions. Two different dosages of the formulate Mildicut--3.5 lt/ha and 4.5 lt/ha--were compared with the unique dosage 0.350 lt/ha of Ranman and a commercial formulate: 2.4% metalaxyl-M + 40% copper, 4 kg/ha (Ridomil Gold R 46 WP, Syngenta Crop Protection). Four foliage applications were applied every 7 days. The intensity and diffusion of the disease were evaluated on Leaves together with eventual phytotoxic effects. The extreme climatic conditions occurred during the course of study, with rather low temperatures and high humidity in the greenhouse, accompanied by high rainfall outside, allowed for extremely serious attacks by Bremia Lactucae, as to make the 3 replications of the checks strongly compromised. Experimental results show the excellent effectiveness of Cyazofamide: both the two doses of Mildicut and Ranman, such as the standard formulate used in comparison, showed to be effective in the control of the disease, highlighting a strong biological activity against B. Lactuce. No effect of phytotoxicity was noticed on leaves.

Teppeki, selective insecticide about Bombus terrestris.

At a time when a highly controversial debate about the causes of the widespread deaths of bees is taking place all over Europe, which accused the agriculture and its practices with particular reference to the harmful effects of some insecticides, it seems important to point out as another insecticide, the Teppeki, can be selective about bumble and have a good compatibility with the activity of the apiaries. This insecticide has the active ingredient flonicamid (500 g/kg) belonging to a new chemical class, called pyridinecarboxamides: the product works systemic and is known as having a long lasting efficacy against all important aphid species. Bioagritest test facility of Pignola (PZ, Italy) has conducted in two successive production cycles an experimental trial on a tomato hydroponic cultivation within the Agricola Bonsai farm in Sibari (CS, Italy), whose objective was to measure the selectivity of flonicamid on Bombus terrestris, insects playing an important role in the pollination of certain species grown in greenhouse such as Tomato, Eggplant, Pepper and Cucumber. On the pollinated flower B. terrestris leaves some trace of its visit, a typical dark trademark: on the detection of the marking of flowers was based the testing program conducted by Bioagritest. Two thesis were compared: A, standard) treatment with a foliar insecticide, the neonicotinoide acetamiprid, normally used for control of aphids and whiteflies (unlike other neonicotinoides--imidacloprid and thiametoxam--quite selective about B. terrestris) and B, Teppeki) foliar treatment with Teppeki, to the maximum dose indicated on the label. The experimental design included the use of randomized blocks with 4 repetitions (4 plots/thesis with 100 plants each). In every thesis six B. terrestris hives were placed 2 days before treatment: the respective holes remained closed during the treatment and the 12 following hours. In order to verify the pollination, by the detection of the flower marking, 2 flowers per plant were observed, for a total of 200 flowers per plot. The measurements were made on the 3rd (I relief) and 8th day (II relief) after treatment. Statistical analysis was performed by the use of XLSTAT data analysis and statistical software. The analysis of collected data shows that flonicamid has a minor effect of interference with the activity of pollination by B. terrestris, compared to the standard used. 14 days after treatment, 3 hives per thesis were inspected in order to verify the status of the colonies (adults, larvae, eggs, pollen). The colonies appeared generally homogeneous as concerning the number of alive adults--100 for each--all at the end of the development cycle. There was no dead adult. Two colonies, one for thesis, presented evidence of eggs. All colonies had low stocks of pollen. Ultimately, treatment with Teppeki has not given any acute effect on B. terrestris, nor any effect of interference in respect of its pollination activity.

Carfentrazone-ethyl: a new molecule for the control of hazelnut shoots.

Control of the shoots growth is an important practice for the management of hazelnut crop, in order to improve its qualitative and quantitative yield. Bioagritest test facility has set up a trial in order to evaluate the efficacy of the contact herbicide SPOTLIGHT PLUS (carfentrazone 60 g/l) for the control of hazelnut shoots. Experimentation was performed, according to EPPO guidelines and Principles of Good Experimental Practice (GEP), in a family-run hazelnut orchard in Nocera Inferiore (Salerno), southern Italy. Experimental design consisted in random blocks, in 4 repetitions. Two different dosages of SHARK 60EC--0.35 and 0.40 l/hl--were compared with a commercial formulate, Glufosinate-ammonium 2 l/hl (Basta 11.33 SC, BayerCropScience), a non-selective herbicide for use in tree crops, vegetables, non-crop areas and as desiccant of potatoes and other crops. Two applications were realised with an interval of 7 days. 4 scouting's were performed respectively a week, 2 weeks, a month and 2 months after the first application. They consisted in a visual and subjective evaluation on the whole plot of the desiccation of the shoots expressed as a percentage (intensity of necrosis as opposed to any green parts present), according to a scale from 0 to 100. The intensity of desiccation was evaluated together with eventual phytotoxic effects. Statistical analysis was performed by the use of XLSTAT data analysis and statistical software. The chance of an unexpected rain in the hours after the first application produced an interesting result because the two doses of Carfentrazone-ethyl showed a rapid uptake in the tissues of shoots, much higher than that of the standard Glufosinate-ammonium. It is believed that this rapid uptake and, consequently, a fast-desiccation capacity, may represent a value for the use of Carfentrazone-ethyl on hazelnut shoots. Carfentrazone-ethyl provided a good control of hazelnut shoots and, at the highest dose, it induced on all replicates a total dissecation. No effect of phytotoxicity was noticed.

Integrating Plant Essential Oils and Kaolin for the Sustainable Management of Thrips and Tomato Spotted Wilt on Tomato.

Thrips-vectored Tomato spotted wilt virus is one of the most devastating pest complexes affecting tomato in the southern United States and elsewhere. Field trials were conducted over 2 years to determine the effects of volatile plant essential oils and kaolin-based particle films on the incidence of tomato spotted wilt and population dynamics of Frankliniella spp. thrips. The essential oils, geraniol (a monoterpene constituent of various plant essential oils), lemongrass (Cymbopogon flexuosus) oil, and tea tree (Melaleuca alternifolii) oil, were compared with a standard insecticide treatment and an untreated control. All treatments were applied with and without kaolin, in a 5 × 2 factorial design. Although the treatments did not clearly affect the abundance of vector species, there were treatment differences in the incidence of tomato spotted wilt. When combined with kaolin, the three essential oils reduced tomato spotted wilt incidence by 32 to 51% in 2005 and by 6 to 25% in 2006 compared with the control. In both years, tea tree oil plus kaolin reduced tomato spotted wilt as well as the standard insecticide treatments. Overall, kaolin significantly increased yield by over 26% compared to treatments without kaolin. When applied with kaolin, the three essential oils produced yields similar to the insecticide standard. Therefore, naturally occurring products, such as essential oils and kaolin, could be used successfully to reduce insecticide use on tomatoes.

Three years survey of Tomato yellow leaf curl Sardinia virus reservoir weed hosts in southern Italy.

During the period from August 2004 to June 2006 a serious tomato yellow leaf curl epidemic caused by both Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCS) was observed in protected tomato crops in Castrovillari, Calabria Region, in a group of greenhouses where tomato is grown hydroponically. A three years survey for reservoir weed hosts of these viruses was performed during summer period in order to identify where the viruses persist during the host-free period, interesting an area covering a ray of 500 m around the group of greenhouses. About 350 samples were collected from symptomless and symptomatic plants of the following botanic families: Graminaceae, Compositeae, Solanaceae, Portulacaceae, Malvaceae, Chenopodiaceae, Amaranthaceae, Convolvulaceae, Brassicaceae, Labiatae, Plantaginaceae, Asteraceae. Any virus presence was evaluated by DAS ELISA, using a "broad-spectrum" reagent combination detecting different Begomoviruses including TYLCSV and TYLCV. A couple of synthetic oligonucleotides allowing the amplification of the whole coat protein (CP) gene was used for PCR of ELISA positive samples in order to perform the molecular characterisation of the viral isolate responsible of the disease. RFLP analysis performed on the PCR product, 1008 bp long, showed the presence of only TYLCSV in the weeds found infected and belonging to Sonchus asper, Solanum nigrum, Datura stramonium and Cardaria draba species. Similarity analysis performed between the CP of each isolate and the TYLCSV isolate recovered within the greenhouse and responsible of the epidemic in mixed infection with a TYLCV isolate resulted in a value of 100% of identity, thus indicating that there was no variability in TYLCSV population in the surveyed area. S. asper, S. nigrum, D. stramonium and C. draba, as alternative hosts of TYLCSV and nutrient plants of the virus vector, Bemisia tabaci, were found to play an important role in virus ecology and epidemiology in the studied tomato ecosystem. No weed between those investigated has been found to be infected by TYLCV so far. To our knowledge this is the first report of S. asper and C. draba as TYLCSV hosts in natural infection.

In situ characterization of Cymbidium Ringspot Tombusvirus infection-induced posttranscriptional gene silencing in Nicotiana benthamiana.

In plants, posttranscriptional gene silencing (PTGS) is an ancient and effective defense mechanism against viral infection. A number of viruses encode proteins that suppress virus-activated PTGS. The p19 protein of tombusviruses is a potent PTGS suppressor which interferes with the onset of PTGS-generated systemic signaling and is not required for viral replication or for viral movement in Nicotiana benthamiana. This unique feature of p19 suppressor allowed us to analyze the mechanism of PTGS-based host defense and its viral suppression without interfering with other viral functions. In contrast to the necrotic symptoms caused by wild-type tombusvirus, the infection of p19-defective mutant virus results in the development of a typical PTGS-associated recovery phenotype in N. benthamiana. In this report we show the effect of PTGS on the viral infection process for N. benthamiana infected with either wild-type Cymbidium Ringspot Tombusvirus (CymRSV) or a p19-defective mutant (Cym19stop). In situ analyses of different virus-derived products revealed that PTGS is not able to reduce accumulation of virus in primary infected cells regardless of the presence of p19 PTGS suppressor. We also showed that both CymRSV and Cym19stop viruses move systemically in the vasculature, with similar efficiencies. However, in contrast to the uniform accumulation of CymRSV throughout systemically infected leaves, the presence of Cym19stop virus was confined to and around the vascular bundles. These results suggest that the role of p19 is to prevent the onset of mobile signal-induced systemic PTGS ahead of the viral infection front, leading to generalized infection.

Evaluation of glycoalkaloids in tubers of genetically modified virus Y-resistant potato plants (var. Désirée) by non-aqueous capillary electrophoresis coupled with electrospray ionization mass spectrometry (NACE-ESI-MS).

The glycoalkaloid content of transgenic potatoes was evaluated by an optimised method based on non-aqueous capillary electrophoresis coupled on-line with electrospray ionization-mass spectrometry (NACE-ESI-MS). The potato material consisted of tubers from a conventional cv. Désirée and from three lines of modified plants resistant, intermediate and susceptible to infection by potato virus Y (PVY). The main glycoalkaloids were confirmed to be alpha-solanine and alpha-chaconine with parent ion masses m/z 852 and 868, respectively. In addition, an unknown minor peak at m/z 850.6 was found both in conventional (control) and susceptible line potato tubers. Such a compound exhibited an MS(2) spectrum with fragments ions at 704 and 396 m/z derived by loss of two ions, i.e. m/z 146 and 307, most likely corresponding to a rhamnose unit and a [glucose-(rhamnose)(2)] moiety, respectively. Up to 30-80-fold higher concentrations of total glycoalkaloids were found in the peel compared to flesh samples of all tubers examined. TGA content was nearly doubled in peel samples of resistant compared to control lines, and these levels were lower than the limit recommended for food safety, i.e. 20-60 mg of TGA per 100 g fresh weight. Moreover, it was established that tubers produced by virus-resistant clones are substantially equivalent in glycoalkaloid contents to those produced by conventional potato varieties.