The Other Side of Plastics: Bioplastic-Based Nanoparticles for Drug Delivery Systems in the Brain.

Plastics have changed human lives, finding a broad range of applications from packaging to medical devices. However, plastics can degrade into microscopic forms known as micro- and nanoplastics, which have raised concerns about their accumulation in the environment but mainly about the potential risk to human health. Recently, biodegradable plastic materials have been introduced on the market. These polymers are biodegradable but also bioresorbable and, indeed, are fundamental tools for drug formulations, thanks to their transient ability to pass through biological barriers and concentrate in specific tissues. However, this "other side" of bioplastics raises concerns about their toxic potential, in the form of micro- and nanoparticles, due to easier and faster tissue accumulation, with unknown long-term biological effects. This review aims to provide an update on bioplastic-based particles by analyzing the advantages and drawbacks of their potential use as components of innovative formulations for brain diseases. However, a critical analysis of the literature indicates the need for further studies to assess the safety of bioplastic micro- and nanoparticles despite they appear as promising tools for several nanomedicine applications.

Development and Application of Low-Cost and Eco-Sustainable Bio-Stimulant Containing a New Plant Growth-Promoting Strain Kosakonia pseudosacchari TL13.

The use of beneficial microbes as inoculants able to improve fitness, growth and health of plants also in stress conditions is an attractive low-cost and eco-friendly alternative strategy to harmful chemical inputs. Thirteen potential plant growth-promoting bacteria were isolated from the rhizosphere of wheat plants cultivated under drought stress and nitrogen deficiency. Among these, the two isolates TL8 and TL13 showed multiple plant growth promotion activities as production of indole-3-acetic acid (IAA), siderophores, ammonia, and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase production, the ability to solubilize phosphate as well as exerted antimicrobial activity against plant pathogens as Botrytis spp. and Phytophthora spp. The two selected strains were identified as Kosakonia pseudosacchari by sequencing of 16S rRNA gene. They resulted also tolerant to abiotic stress and were able to efficiently colonize plant roots as observed in vitro assay under fluorescence microscope. Based on the best PGP properties, the strain K. pseudosacchari TL13 was selected to develop a new microbial based formulate. A sustainable and environmentally friendly process for inoculant production was developed using agro-industrial by-products for microbial growth. Moreover, the application of K. pseudosacchari TL13- based formulates in pot experiment improved growth performance of maize plants.

Ozone gas as a storage treatment to control Gnomoniopsis castanea, preserving chestnut quality.

BACKGROUND: Chestnuts are gluten-free, low-fat, cholesterol-free products. Postharvest decay reduces chestnut shelf life and can cause severe economic losses. In this study we investigated the effect of ozone (O3 ) gaseous treatment on chestnut rot caused by Gnomoniopsis castanea and the quality parameters of chestnuts. RESULTS: The results showed that ozone treatment (150 ppb during the day, and 300 ppb during the night) reduced the decay of chestnuts and had a fungistatic effect on isolates of G. castanea. The exposure of chestnuts to ozone did not alter weight losses, sugar content and titratable acidity. The concentration of total phenolics decreased during the storage period, both for treated and untreated nuts. However, after 150 days of treatment the polyphenol content of the chestnuts exposed to ozone was significantly higher than in control nuts. CONCLUSIONS: Our results suggested that ozone is an appropriate and economical tool to maximize the quality of chestnut shelf life, enabling it to be stored for long periods. © 2019 Society of Chemical Industry.

Depletion of pentachlorophenol in soil microcosms with Byssochlamys nivea and Scopulariopsis brumptii as detoxification agents.

Pentachlorophenol (PCP) is a toxic compound which is widely used as a wood preservative product and general biocide. It is persistent in the environment and has been classified as a persistent organic pollutant to be reclaimed in many countries. Fungal bioremediation is an emerging approach to rehabilitating areas fouled by recalcitrant xenobiotics. In the present study, we isolated two fungal strains from an artificially PCP-contaminated soil during a long-term bioremediation study and evaluated their potential as bioremediation agents in depletion and detoxification of PCP in soil microcosms. The two fungal strains were identified as: Byssochlamys nivea (Westling, 1909) and Scopulariopsis brumptii (Salvanet-Duval, 1935). PCP removal and toxicity were examined during 28 days of incubation. Bioaugmented microcosms revealed a 60% and 62% PCP removal by B. nivea and S. brumptii, respectively. Co-inoculation of B. nivea and S. brumptii showed a synergetic effect on PCP removal resulting in 95% and 80% PCP decrease when initial concentrations were 12.5 and 25 mg kg-1, respectively. Detoxification in bioaugmented soil and the efficient role of fungi in the rehabilitation of PCP contaminated soil were experimentally proven by toxicity assays. A decrease in inhibition of bioluminescence of Escherichia coli HB101 pUCD607 and an increase of germination index of mustard (Brassica alba) seeds were observed in the decontaminated soils.

Assessing the effectiveness of Byssochlamys nivea and Scopulariopsis brumptii in pentachlorophenol removal and biological control of two Phytophthora species.

Bioremediation and biological-control by fungi have made tremendous strides in numerous biotechnology applications. The aim of this study was to test Byssochlamys nivea and Scopulariopsis brumptii in sensitivity and degradation to pentachlorophenol (PCP) and in biological-control of Phytophthora cinnamomi and Phytophthora cambivora. B. nivea and S. brumptii were tested in PCP sensitivity and degradation in microbiological media while the experiments of biological-control were carried out in microbiological media and soil. The fungal strains showed low PCP sensitivity at 12.5 and 25 mg PCP L(-1) although the hyphal size, fungal mat, patulin, and spore production decreased with increasing PCP concentrations. B. nivea and S. brumptii depleted completely 12.5 and 25 mg PCP L(-1) in liquid culture after 28 d of incubation at 28 °C. Electrolyte leakage assays showed that both fungi have low sensitivity to 25 mg PCP L(-1) and produced no toxic compounds for the plant. B. nivea and S. brumptii were able to inhibit the growth of the two plant pathogens in laboratory studies and reduce the mortality of chestnut plants caused by two Phytophthorae in greenhouse experiments. The two fungal strains did not produce volatile organic compounds able to reduce the growth of two plant pathogens tested.

Chestnut green waste composting for sustainable forest management: Microbiota dynamics and impact on plant disease control.

Making compost from chestnut lignocellulosic waste is a possible sustainable management strategy for forests that employs a high-quality renewable organic resource. Characterization of the microbiota involved in composting is essential to better understand the entire process as well as the properties of the final product. Therefore, this study investigated the microbial communities involved in the composting of chestnut residues obtained from tree cleaning and pruning. The culture-independent approach taken highlighted the fact that the microbiota varied only slightly during the process, with the exception of those of the starting substrate and mature compost. The statistical analysis indicated that most of the bacterial and fungal species in the chestnut compost persisted during composting. The dominant microbial population detected during the process belonged to genera known to degrade recalcitrant lignocellulosic materials. Specifically, we identified fungal genera, such as Penicillium, Fusarium, Cladosporium, Aspergillus and Mucor, and prokaryotic species affiliated with Bacilli, Actinobacteria, Flavobacteria and γ-Proteobacteria. The suppressive properties of compost supplements for the biocontrol of Sclerotinia minor and Rhizoctonia solani were also investigated. Compared to pure substrate, the addition of compost to the peat-based growth substrates resulted in a significant reduction of disease in tomato plants of up to 70 % or 51 % in the presence of Sclerotinia minor or Rhizoctonia solani, respectively. The obtained results were related to the presence of putative bio-control agents and plant growth-promoting rhizobacteria belonging to the genera Azotobacter, Pseudomonas, Stenotrophomonas, Bacillus, Flavobacterium, Streptomyces and Actinomyces in the chestnut compost. The composting of chestnut waste may represent a sustainable agricultural practice for disposing of lignocellulosic waste by transforming it into green waste compost that can be used to improve the fitness of agricultural plants.

Flufuran, an antifungal 3,5-disubstituted furan produced by Aspergillus flavus link.

A 3,5-disubstituted furan, named flufuran, was isolated from a culture filtrate of a strain of Aspergillus flavus obtained from a chestnut compost created in the same orchard. Flufuran was identified by spectroscopic methods, and its structure was confirmed through the preparation of some key derivatives, also used to test the antifungal activity. At a concentration of 0.2 mg/ml, assayed against three Phytophthora species, pathogenic of some forest and agrarian plants, flufuran and especially its acetyl derivative showed significant antifungal activity. Although flufuran appears to be identical to a fungal metabolite isolated previously from some Polyporus spp., its interesting antifungal activity has never been reported before.

Isolation, characterization and structure-elicitor activity relationships of hibernalin and its two oxidized forms from Phytophthora hibernalis Carne 1925.

Three alpha-elicitins, named hibernalin1, hibernalin2 and hibernalin3 (hib1, hib2 and hib3, respectively), were isolated by reverse phase-low-pressure liquid chromatography from culture filtrates of Phytophthora hibernalis Carne 1925, the causal agent of citrus lemon brown rot. Hib1 proved to be identical to syringicin previously isolated from culture filtrates of Phytophthora syringae. Hib2 and hib3 shared the same primary structure with hib1, but contained, at position 50, Met sulphoxide or sulphone, respectively. By SDS-PAGE, the three proteins showed the same electrophoretic mobility, corresponding to about 10 kDa. Exact M(r) values were obtained by MALDI-TOF-MS (10,194.82 for hib1, 10,209.33 for hib2 and 10,223.80 for hib3), while by ESI-MS an M(r) value of 10,194.90 was found for hib1 and no results for hib2 and hib3. The hibernalin forms showed a high propensity to self-association, after exposure to acetonitrile. Hib1 showed to be active in both the hypersensitivity response and electrolytes leakage assays; the sample containing hib1 and hib2 was only weakly active in the first assay and inactive in the second assay, while the sample containing all three hibernalin forms proved to be inactive in both tests. It is proposed that the different activities of the three hibernalin samples could be very likely attributed to both Met50 oxidation and aggregation.

First Report of Phytophthora insolita and P. inflata on Rhododendron in Ohio.

During August 2003, we conducted a statewide survey of rhododendrons to determine if Phytophthora ramorum was present in Ohio ornamental nurseries. In total, 240 samples were randomly collected in 12 nurseries throughout Ohio from rhododendrons showing foliar necrotic lesions and twig dieback symptoms. The samples yielded 51 Phytophthora spp. isolates on PARP-V8 agar. The internal transcribed spacer (ITS) region of all isolates was amplified using the universal primers ITS1 and ITS4 and was sequenced. Consensus sequences from sense and antisense were then blasted against the GenBank database, allowing for the identification to species of ˜80% of all isolates. These identifications, and the ˜20% unknowns, were confirmed using blind morphological tests on the basis of the following parameters: colony morphology; shape and dimensions of sporangia and type of papillae; dimensions of oogonia and oospores; type and position of antheridia; presence or absence of chlamydospores; presence or absence and morphology of hyphal swellings; and growth rate at 35°C according to the Revisited Tabular Key of the species of Phytophthora (1). No P. ramorum was detected among the isolates; however, P. cactorum, P. citricola, P. citrophthora, and P. nicotianae were detected. We also found two occurrences of P. inflata Caros & Tucker and one of P. insolita Ann & Ko. (P. inflata: e-value ≤e-179, identities ≥95%; P. insolita: e-value = 0.0; identities = 95%.) P. inflata was isolated from two tissue types, a dead twig and a necrotic leaf tip. P. insolita was isolated from a necrotic leaf tip. Identity of the two species was confirmed morphologically using the parameters listed above as well as the following measurements (N = 40; all in µm) (1): P. inflata - sporangia: 40 × 24 ([24 to 68] × [18 to 34]); oogonia: 34.6 (28 to 40); oospores: 30.8 (25 to 38); P. insolita - sporangia: 42 × 28 ([34 to 56] × [22 to 38]); oogonia: 32 (26 to 36); oospores: 26 (22 to 30). Koch's postulates were satisfied by inoculating two rhododendron plants (cvs. PJM and Nova Zembla) with the putative pathogens. On each plant, each of three leaves was pierced with a dissecting needle and was inoculated by placing a 0.5-cm-diameter plug of mycelium that was taken from the margin of a colony actively growing on PARP-V8 agar on the wound. The inoculum was retained using clear adhesive tape. A similar procedure was used for twigs. Controls consisted of inoculations with sterile PARP-V8 agar medium. Both cultures of P. inflata and P. insolita produced necrotic lesions in all inoculations on both tissue types within 1 week, and they were reisolated from the margins of lesions on PARP-V8. The lesion margin was at least 2 cm away from the inoculum plug in leaf inoculations and several centimeters in twig inoculations. To our knowledge, this is the first report of P. inflata and P. insolita occurring on rhododendron and the first time P. insolita has been reported outside of Southeast Asia where it has been recovered only from soil. Reference: (1) D. J. Stamps et al. Mycol. Pap. No. 162. CAB Int. Mycol. Inst. Wallingford, UK, 1990.

High-density genetic linkage maps of Phytophthora infestans reveal trisomic progeny and chromosomal rearrangements.

Detailed analysis of the inheritance of molecular markers was performed in the oomycete plant pathogen Phytophthora infestans. Linkage analysis in the sexual progeny of two Dutch field isolates (cross 71) resulted in a high-density map containing 508 markers on 13 major and 10 minor linkage groups. The map showed strong clustering of markers, particularly of markers originating from one parent, and dissimilarity between the parental isolates on linkage group III in the vicinity of the mating-type locus, indicating a chromosomal translocation. A second genetic map, constructed by linkage analysis in sexual progeny of two Mexican isolates (cross 68), contained 363 markers and is thus less dense than the cross 71 map. For some linkage groups the two independent linkage maps could be aligned, but sometimes markers appeared to be in a different order, or not linked at all, indicating chromosomal rearrangements between genotypes. Graphical genotyping showed that some progeny contained three copies of a homologous linkage group. This trisomy was found for several linkage groups in both crosses. Together, these analyses suggest a genome with a high degree of flexibility, which may have implications for evolution of new races and resistance development to crop protection agents.

EST mining and functional expression assays identify extracellular effector proteins from the plant pathogen Phytophthora.

Plant pathogenic microbes have the remarkable ability to manipulate biochemical, physiological, and morphological processes in their host plants. These manipulations are achieved through a diverse array of effector molecules that can either promote infection or trigger defense responses. We describe a general functional genomics approach aimed at identifying extracellular effector proteins from plant pathogenic microorganisms by combining data mining of expressed sequence tags (ESTs) with virus-based high-throughput functional expression assays in plants. PexFinder, an algorithm for automated identification of extracellular proteins from EST data sets, was developed and applied to 2147 ESTs from the oomycete plant pathogen Phytophthora infestans. The program identified 261 ESTs (12.2%) corresponding to a set of 142 nonredundant Pex (Phytophthora extracellular protein) cDNAs. Of these, 78 (55%) Pex cDNAs were novel with no significant matches in public databases. Validation of PexFinder was performed using proteomic analysis of secreted protein of P. infestans. To identify which of the Pex cDNAs encode effector proteins that manipulate plant processes, high-throughput functional expression assays in plants were performed on 63 of the identified cDNAs using an Agrobacterium tumefaciens binary vector carrying the potato virus X (PVX) genome. This led to the discovery of two novel necrosis-inducing cDNAs, crn1 and crn2, encoding extracellular proteins that belong to a large and complex protein family in Phytophthora. Further characterization of the crn genes indicated that they are both expressed in P. infestans during colonization of the host plant tomato and that crn2 induced defense-response genes in tomato. Our results indicate that combining data mining using PexFinder with PVX-based functional assays can facilitate the discovery of novel pathogen effector proteins. In principle, this strategy can be applied to a variety of eukaryotic plant pathogens, including oomycetes, fungi, and nematodes.