The identification of allergen proteins in two different varieties of strawberry by two different approaches: Proteomic and western blotting method

Strawberry (Fragaria ananassa Duch.) is one of most important fruit crops cultivated widely in world. In the present study, we have investigated the proteome variation to different commercially available red strawberry varieties, to gain better insight into the variation in allergen content between the different varieties, as well as their proteome variation. In fact, a combination of proteomic tools have been used to build a complete allergen map of strawberry. The water-soluble fraction of a strawberry extract was precipitated using a phenol-based procedure and separated by 2-DE. Further, all known strawberry allergens were localized on a 2-DE map and they were matched with spots recognized by sera of patients with different allergic patterns. A number of constitutive or differentially accumulated proteins were found. The role of the identified proteins, in particular of the allergen proteins, is discussed in relation to the different variety of strawberry. Keywords: Proteomics, 2-DE, Fragaria ananassa, Mass spectrometry, Allergen protein

Enhancing phytoextraction of HMs at real scale, by combining salicaceae trees with microbial consortia

We applied an integrated phytoremediation system by inoculation of rhizosphere microbes on an area of 2,000 m2 that was contaminated by heavy metals. Hybrid poplar (Populus deltoids × Populus nigra) and willow (Salix purpurea subsp. lambertiana) were planted on four parcels of 500 m2. Of these, two parcels were inoculated with an autochthonous consortium specially selected and the other two were not inoculated. The analyses were carried out on plant and soil samples collected at different times: month 0 (T0), 12 (T1), 24 (T2), and 36 (Tf). Total concentrations of HMs in the roots of Populus and Salix showed significant increase in the plants with rhizosphere microflora consortium. The plants treated with the consortium have an increased enzymatic activity of antioxidants and of stress markers. All enzymes showed an important contribution in enzymatic thiol-dependent ROS scavening mechanisms, through its over-expression at time T1 and T2 and a consequent stabilization at Tf which results in adaptive mechanisms. The plant-fungi-bacterium system was a valid candidate for the implementation of phytoremediation systems in the field, since the plants showed a greater capacity for adaptation and accumulation

Toxic Effects on Thyroid Gland of Male Adult Lizards (Podarcis Siculus) in Contact with PolyChlorinated Biphenyls (PCBs)-Contaminated Soil

Skin exposure is considered a potentially significant but little-studied pathway for PolyChlorinated Biphenyls uptake in terrestrial reptiles. In this study, a native Italian lizard, Podarcis siculus, was exposed to PCBs-contaminated soil for 120 days. Tissues distribution of PCBs, thyroid hormone levels, and thyroid histo-physiopathology were examined. The accumulation of PCBs in skin, plasma, liver, kidney, and brain were highest at 120 days. The alteration of triiodothyronine (T3) and thyroxine (T4) levels after different concentrations and times to exposure of PCBs was accompanied by the changes in the hormones involved in the hypothalamus-pituitary-thyroid (HPT) axis, namely Thyrotropin Releasing Hormone (TRH) and Thyroid Stimulating Hormone (TSH). Moreover, hepatic levels of deiodinase II (5'ORDII) and content of T3 were positively correlated to exposure to PCBs. These results indicated that in lizards, PCBs exposure through the skin has the potential to disrupt the thyroid endocrine system. Overall, the observed results indicate that PCBs could be associated with changes in thyroid homeostasis in these reptiles, through direct interactions with the metabolism of T4 and T3 through the HPT axis or indirect interactions with peripheral deiodination

Immunohistochemical localization of NPY, VIP and 5-HT in the thyroid gland of the lizard, Podarcis sicula.

The thyroid gland of the lizard Podarcis sicula was immunohistochemically studied in adult male specimens using specific antibodies against NPY, VIP and 5-HT and the avidin-biotin peroxidase complex (ABC) procedure to localize the three peptides. Fine beaded VIP-immunoreactive nerve fibers ran between the follicles, and VIP-immunoreactivity was evenly distributed in the apical cytoplasm of follicular cells. NPY-immunoreactive fibers were found around the follicles, and, in the cells, immunoreactivity was localizated only in the cellular apices. Immunoreactivity to 5-HT was observed in the colloid, with a concentration in the follicular lumen exceeding that in the follicular cells. In fact, most follicles showed immunoreactivity in the cytoplasmic bridges formed between the apical portion of the follicular cells and the colloid

The rootstock shape microbial diversity and functionality in the rhizosphere of Vitis vinifera L. cultivar Falanghina

The rhizosphere effect occurring at the root-soil interface has increasingly been shown to play a key role in plant fitness and soil functionality, influencing plants resilience. Here, for the first time, we investigated whether the rootstock genotype on which Vitis vinifera L. cultivar Falanghina is grafted can influence the rhizosphere microbiome. Specifically, we evaluated to which extent the 5BB and 1103P rootstocks are able to shape microbial diversity of rhizosphere environment. Moreover, we explored the potential function of microbial community and its shift under plant genotype influence. We investigated seven vineyards subjected to the same pedo-climatic conditions, similar age, training system and management and collected twelve rhizosphere soil samples for metagenomic analyses and composite soil samples for physical-chemical properties. In this study, we used 16S rRNA gene-based metagenomic analysis to investigate the rhizosphere bacterial diversity and composition. Liner discriminant analysis effect size (LEFSe) was conducted for metagenomic biomarker discovery. The functional composition of sampled communities was determined using PICRUSt, which is based on marker gene sequencing profiles. Soil analyses involved the determination of texture, pH, Cation Exchange Capacity (CSC), Organic Carbon (OC), electrical conductivity (EC), calcium (Ca), magnesium (Mg), potassium (K) content, Phosphorous (P), nitrogen (N). The latter revealed that soil features were quite homogenous. The metagenomic data showed that the bacterial alpha-diversity (Observed OTUs) significantly increased in 1103P rhizosphere microbiota. Irrespective of cultivar, Pseudomonadota was the dominant phylum, followed by Actinomycetota > Bacteroidota > Thermoproteota. However, Actinomycetota was the major marker phyla differentiating the rhizosphere microbial communities associated with the different rootstock types. At the genus level, several taxa belonging to Actinomycetota and Alphaproteobacteria classes were enriched in 1103P genotype rhizosphere. Investigating the potential functional profile, we found that most key enzyme-encoding genes involved in N cycling were significantly more abundant in 5BB rootstock rhizosphere soil. However, we found that 1103P rhizosphere was enriched in genes involved in C cycle and Plant Growth Promotion (PGP) functionality. Our results suggest that the different rootstocks not only recruit specific bacterial communities, but also specific functional traits within the same environment

Soil Metaproteomics for the Study of the Relationships Between Microorganisms and Plants: A Review of Extraction Protocols and Ecological Insights

Soil is a complex matrix where biotic and abiotic components establish a still unclear network involving bacteria, fungi, archaea, protists, protozoa, and roots that are in constant communication with each other. Understanding these interactions has recently focused on metagenomics, metatranscriptomics and less on metaproteomics studies. Metaproteomic allows total extraction of intracellular and extracellular proteins from soil samples, providing a complete picture of the physiological and functional state of the “soil community”. The advancement of high-performance mass spectrometry technologies was more rapid than the development of ad hoc extraction techniques for soil proteins. The protein extraction from environmental samples is biased due to interfering substances and the lower amount of proteins in comparison to cell cultures. Soil sample preparation and extraction methodology are crucial steps to obtain high-quality resolution and yields of proteins. This review focuses on the several soil protein extraction protocols to date to highlight the methodological challenges and critical issues for the application of proteomics to soil samples. This review concludes that improvements in soil protein extraction, together with the employment of ad hoc metagenome database, may enhance the identification of proteins with low abundance or from non-dominant populations and increase our capacity to predict functional changes in soil

Effects of leptin administration on the endocrine pancreas and liver in the lizard Podarcis sicula

In this study, we investigated the presence of leptin receptor in pancreatic islets and the effect of exogenous leptin administration in Podarcis sicula on glucose metabolism. Our data show the presence of leptin receptor immunoreactivity in the endocrine pancreas suggesting that leptin may act at a peripheral level as previously postulated in mammals. The effects of short- and long-term and dose–response treatment with supraphysiological concentrations of leptin on circulating levels of insulin, glucagon and glucose in the blood have been evaluated. Taken together, our results indicate that leptin treatment was followed by an increase in insulin, glucagon and glucose in the blood, depending on the dose of leptin. Moreover, leptin treatment brought about a decrease of glycogen and the appearance of tyrosine-phosphorylated proteins in the liver. This study shows that in the lizard P. sicula leptin is involved in glucose metabolis

Recovery and Valorization of Bioactive and Functional Compounds from the Discarded of <i>Opuntia ficus-indica</i> (L.) Mill. Fruit Peel

This work focused on the recovery and application of bioactive and functional compounds (BFC) from a waste matrix, the Opuntia ficus-indica (L.) Miller fruit peel (OFP), using different extraction methods: maceration and Extractor Naviglio®. The presence of BFC was initially evaluated by following the extraction yields and antioxidant activity using the Folin-Ciocȃlteu reagent and the DPPH method, respectively. The quali-quantitative content of BFC was also verified with UHPLC/UV-ESI-HRMS profile of OFP extracts. The evaluation of the BFC yields is aimed at using extract components, such as betanins, in the technological application as textile dye based on natural pigments: we evaluated the stability of the extracted dyes, tested the dyeing power and assessed the stability of the dye on the textile material. The results showed different compositions depending on the polarities of the solvents. The ethanol-water mixture extracts obtained using Naviglio method revealed an important presence in terms of total phenolic compound yields. Wool, linen and cotton fibers were chosen for the dyeing tests. Rock alum and lemon juice, two etching methods, were used. Results suggest that the OFP may be of great interest as a natural source of BFC for food, nutraceutical and technological applications in the sustainability context

Genetic characterization, micropropagation, and potential use for arsenic phytoremediation of Dittrichia viscosa (L.) Greuter

In the last decade, many scientists have focused their attention on the search for new plant species that can offer improved capacities to reclaim polluted soils and waters via phytoremediation. In this study, seed batches from three natural populations of Dittrichia viscosa, harvested in rural, urban, and industrial areas of central and southern Italy, were used to: (i) evaluate the genetic and morphological diversity of the populations; (ii) develop an efficient protocol for in-vitro propagation from seedling microcuttings; (iii) achieve optimal acclimatization of micropropagated plants to greenhouse conditions; (iv) test the response to arsenic (As) soil contamination of micropropagated plants. The genetic biodiversity study, based on Random Amplification of Polymorphic DNA (RAPD), as well as the morphometric analysis of 20 seedlings from each population revealed some degree of differentiation among populations. Based on these data, the most biodiverse plants from the three populations (10 lines each) were clonally multiplied by micropropagation using microcuttings of in-vitro grown seedlings. Three culture media were tested and Mureshige and Skoog medium was chosen for both seedling growth and micropropagation. The micropropagated plants responded well to greenhouse conditions and over 95% survived the acclimatization phase. Four clones were tested for their capacity to grow on soil spiked with NaAsO2and to absorb and accumulate the metalloid. All clones tolerated up to 1.0 mg As. At the end of the trial (five weeks), As was detectable only in leaves of As-treated plants and concentration varied significantly among clones. The amount of As present in plants (leaves) corresponded to ca. 0.10â\u80\u931.7% of the amount supplied. However, As was no longer detectable in soil suggesting that the metalloid was taken up, translocated and probably phytovolatilized

Plants named “Lotus” in antiquity: Historiography, biogeography, and ethnobotany

In ancient times, several plants were named “lotus.” They assumed very important roles in the religions and art of many cultures, but historiography and descriptions of the various plants called “lotus” have always been poor. The aim of this work is to define what plant species correspond to the ancient name “lotus.” Through analysis of classical texts and other historiographical sources, three types of “lotus” have been identified: “arboreal lotus,” “herbaceous lotus,” and “aquatic lotus.” From the sources examined, several botanical species have been identified for each “lotus” category. In the “arboreal lotus” category there are two species of Ziziphus: Z. lotus and Z. spina-christi. The “herbaceous lotus” include several species in Fabaceae that have been called “lotus,” more specifically in the genera Melilotus, Lotus, Trifolium, and Trigonella. In the last category, “aquatic lotus,” are two species of Nymphaea L. (N. lotus and N. nouchali var. caerulea) and the sacred Indian lotus (Nelumbo nucifera). The attributions of these species have been validated by research on their biogeography and ethnobotanical uses.Peer reviewe