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In last decades, biotechnologies have enabled a deep development of knowledge in all fields of life sciences, allowing the collection of a wide range of data. Furthermore, the relationship between scientific investigations and conservation actions of cultural heritage is increasingly consolidated and now it is one of the peculiar points both for sustainable restoration and use of the cultural asset, also evaluating the cultural assets as source of biological information. In this study, Sicilian wolf (Canis lupus) specimens conserved in Sicilian Natural Museums, become a source of biological information on Sicilian biodiversity, establishing specific conservative protocol developed cooperatively with Sicilian Natural Museums. Using Next-Generation Sequencing technique, ancient DNA (aDNA) molecules extracted from nail were utilized to perform the complete mitochondrial DNA genome sequence. Phylogenetic analyses showed that this genome, which was aligned with a number of historical and extant complete wolf and dog mtDNAs sampled worldwide, was closely related to an Italian wolf mtDNA genome. These mtDNA findings suggest that wolves probably colonized Sicily from southern Italy toward the end of the last Pleistocene glacial maximum, when the Strait of Messina was almost totally dry. Furthermore, a conservative restoration of this peculiar natural specimen (taxidermal preparation) has been performed, basing on: recovery of unglued or unstitched parts; revitalization of the colors of the mucous membranes; restoration of deformed or broken parts of the body, washing of the coat and polishing of the fur; specific treatments against woodworms, moths, and mites; brushing and repositioning of the hair; cleaning, restoration, or creation of adequate supports or display case. The conservative intervention that should be periodically performed on each taxidermized specimen allows the protection against biodeterioration or re-infestation by parasites over time.

Olive leaf spot (OLS), caused by Venturia oleaginea, is one of the most common and serious diseases of olive trees in the Mediterranean region. Understanding the pathogen life cycle is important for the development of effective control strategies. Current knowledge is incomplete owing to a lack of effective detection methods. It is extremely difficult to culture V. oleaginea in vitro, so primers were designed to amplify and sequence the internal transcribed spacer ITS1-5.8S-ITS2 region of the fungus directly from infected olive leaves. Sanger sequencing indicated a unique ITS region present in the European strains screened, confirming the appropriateness of the target region for developing a quantitative PCR (qPCR) assay. Furthermore, high-throughput sequencing of the same region excluded the presence of other Venturia species in the olive phyllosphere. The qPCR assay proved very specific and sensitive, enabling the detection of approximately 26 copies of target DNA. The analysis of symptomless leaves during early stages of the epidemic from the end of winter through spring revealed a similar quantity of pathogen DNA regardless of the leaf growth stage. In contrast, the pathogen titer changed significantly during the season. Data indicated that leaf infections start earlier than expected over the season and very young leaves are as susceptible as adult leaves. These findings have important practical implications and suggest the need for improved scheduling of fungicide treatments. The qPCR assay represents a valuable tool providing quantitative results and enables detection of V. oleaginea in all olive organs, including those in which OLS cannot be studied using previously available methods.

Exposure to atmospheric particulate matter and nitrogen dioxide has been linked to SARS-CoV-2 infection and death. We hypothesized that long-term exposure to farming-related air pollutants might predispose to an increased risk of COVID-19-related death. To test this hypothesis, we performed an ecological study of five Italian Regions (Piedmont, Lombardy, Veneto, Emilia-Romagna and Sicily), linking all-cause mortality by province (administrative entities within regions) to data on atmospheric concentrations of particulate matter (PM2.5 and PM10) and ammonia (NH3), which are mainly produced by agricultural activities. The study outcome was change in all-cause mortality during March–April 2020 compared with March–April 2015–2019 (period). We estimated all-cause mortality rate ratios (MRRs) by multivariate negative binomial regression models adjusting for air temperature, humidity, international import-export, gross domestic product and population density. We documented a 6.9% excess in MRR (proxy for COVID-19 mortality) for each tonne/km2 increase in NH3 emissions, explained by the interaction of the period variable with NH3 exposure, considering all pollutants together. Despite the limitations of the ecological design of the study, following the precautionary principle, we recommend the implementation of public health measures to limit environmental NH3 exposure, particularly while the COVID-19 pandemic continues. Future studies are needed to investigate any causal link between COVID-19 and farming-related pollution.

The impact of primer choice on results of metabarcoding studies was experimentally evaluated by analyzing fungal communities associated with leaves of four plant species. Significant differences in target specificity of primers were highlighted by a percentage of plant reads ranging from almost nothing to 30 to 35% of the total detected sequences. Overall, primer sets targeting the internal transcribed spacer 1 (ITS1) region proved to be more specific than those targeting the ITS2 region. A comparable taxa coverage was revealed for all investigated primer sets. However, each primer set detected only around 50% of the overall detected taxa highlighting that a consistent part of the actual fungal diversity remains undetected in studies conducted using a single couple of primers. The coverage was increased to 70 to 80% by combining results from two different primer sets. Some fungal taxa were preferentially or exclusively detected by certain primer sets and this association between primers and taxa was generally recurrent on several plant hosts. Data highlighted that a perfect set of primers to investigate the whole fungal diversity does not exist and that whatever the choice, only a fraction of the actual microbial diversity will be investigated. However, provided information may be valuable to select the best primers according to the objective of the analysis.

Olive leaf spot (OLS), caused by

Approximately 150,000 potted mandevillas (Apocynaceae) are produced each year in the Etna District of eastern Sicily. Since 2004, leaf chlorosis, wilt, and sudden collapse of the entire plant associated with root and basal stem rot of 6- to 12-month-old potted mandevillas, including Mandevilla × amabilis ‘Alice du Pont’, M. splendens, and M. sanderi ‘Alba’, ‘My Fair Lady’, and ‘Scarlet Pimpernel’, have been observed in six nurseries. Incidence of affected plants varied from 5 to 40%. Four Phytophthora species were consistently isolated from rotted roots and stems on a selective medium (2). Pure cultures of the first species produced colonies with a camellia pattern on potato dextrose agar and grew between 10 and 37°C with an optimum of 27°C. On V8 juice agar they produced ellipsoid to obpyriform (length/breadth [l/b] 1.45:1), nonpapillate sporangia with internal proliferation, coralloid, spherical hyphal swellings and both terminal and intercalary chlamydospores. In dual cultures with A1 and A2 isolates of P. nicotianae, all isolates produced oogonia with amphyginous antheridia only with A2 isolates. Isolates of the second species formed petaloid colonies, had an optimum growth temperature of 25°C, and produced mono- and bipapillate, ovoid to limoniform sporangia (l/b 1.40:1); they did not produce gametangia. Isolates of the third species formed colonies with a slight petaloid pattern and grew between 2 and 30°C with an optimum of 25°C. Sporangia were obpyriform (l/b 1.48:1), nonpapillate, and proliferous. All isolates were A2 mating type. The isolates of the fourth species formed arachnoid colonies, grew between 8 and 38°C with an optimum of 30°C, and produced mono- and bipapillate, ellipsoid, and obpyriform (l/b 1.3:1) sporangia and apical chlamydospores. All isolates were A2 mating type. DNA was extracted from mycelium and amplified by PCR using the ITS 4/ITS 6 primers (1). Blast search of the rDNA-ITS sequence of isolate IMI 397618 (GenBank Accession No. GQ388261) of the first species showed 100% identity with the ITS sequence of an isolate of P. cinnamomi var. parvispora (EU748548). The sequences (GQ463703 and GQ463704) of isolates IMI 397471 and IMI 397472 of the second species showed 99% similarity with the sequences of a P. citrophthora isolate (EU0000631). The sequence of isolate IMI 397473 (GQ463702) of the third species showed 99% similarity with the sequence of a P. cryptogea isolate (AY659443.1), while the sequence of isolate IMI 397474 (GU723474) of the fourth species showed 99% similarity with the sequence of a P. nicotianae isolate (EU331089). The pathogenicity of individual isolates IMI 397618, IMI 397471, IMI 397472, IMI 397473, and IMI 397474 was tested on 3-month-old potted plants (10 plants per isolate) of mandevilla ‘Alice du Pont’ by applying 10 ml of a suspension (2 × 104 zoospores/ml) to the root crown. Plants were maintained at 25°C and 95 to 100% relative humidity. All inoculated plants wilted after 4 weeks, while noninoculated control plants remained healthy. The four Phytophthora spp. were subsequently reisolated only from symptomatic plants. To our knowledge, this is the first report of P. cinnamomi var. parvispora in Italy and on mandevilla worldwide. In recent years, Phytophthora root and stem rot has become the most serious disease of potted mandevillas in Sicily. References: (1) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) H. Masago et al. Phytopathology 67:425, 1977.

The growth of molds represents a major problem during table olive fermentation. Molds are recognized as spoilage agents and may reduce the product safety for their ability to produce mycotoxins. Yeasts, instead, are usually found in table olive processing and are generally considered desirable microorganisms for their technological properties. In the present study, a model system to select anti‐mold yeasts usable as protective adjunct cultures in table olive fermentation was developed. Two hundred and ninety‐nine strains of yeasts, isolated from cheese, olive, vinegar, and wine, were tested in vitro for their growth characteristics, fermentative activity, salt tolerance, and antagonistic activity against 10 mold strains isolated from table olives and olive brines. Experimental steps led to the selection of a strain of Yarrowia lipolytica exhibiting all the previously listed characteristics. PRACTICAL APPLICATIONS: A strain of Yarrowia lipolytica was selected because of its high potentiality as adjunct culture in table olive fermentation to contrast mold growth and increase the product quality and safety. The experimental model system developed in this study may be easily applied to select yeasts usable as adjunct cultures in table olive fermentation in order to contrast mold growth and increase the product safety. These outcomes showed how the use of selected yeasts constitutes a promising way to control mold growth during table olive fermentation.

Orange Peel Waste (OPW) is a widely produced residue whose management is complicated by its physical and chemical properties. Anaerobic digestion (AD), which is commonly used for the treatment and exploitation of many biodegradable wastes, is inefficient on OPW due to the presence of essential oils (mainly d -Limonene) as well as the low pH, which cause the process to be unstable. Here we explore the effect of alkaline pre-treatment of OPW and of the addition of granular activated carbon (GAC) and Zero Valent Iron (ZVI) in improving AD in two semi-continuous reactors at a laboratory scale. The addition and pre-treatment of ZVI/GAC were shown to help process stability up to a loading of 3 kgVS·m−3·d−1 and to increase methane production even at a sub-optimal pH. The investigation of the bacterial community, by high-throughput sequencing, has also increased our insight on their involvement in AD in the presence of ZVI, including its biotic oxidation. In addition, direct interspecies electron transfer was shown to play a role in the reactor supplemented with ZVI.

It is essential to investigate to which extent and how specifically soil&ndash, plant&ndash, microbe interactions can be conditioned by different agricultural practices. Legumes such as Vicia faba is one of the essential functional group in intercropping and crop rotations due to its higher N fixing capacity. Hence, it is important to study the living microbial community of this legume. Further, it is also expected that fluctuations in soil microbial diversity and composition could be complemented by plant phenological stages and different fertilizer amendments. Thus, we investigated bacterial community composition in soil treated with phosphate-based inorganic and organic fertilizers, in the presence of Vicia faba plants at flowering and fruiting time using NGS 16S rRNA gene amplicon sequencing. Further, the evaluation of plant biomass parameters under different fertilizer treatments was also carried out. The presence of the Vicia faba plant increased the abundance of N fixing bacterial such as Bardyrhizobium, Microvirga (Rhizobiales), Arthrobacter, and Psuedoarthrobacter (Actinomycetales) in soil. Fluctuation in composition and diversity of bacterial community was further supplemented by plant phenological stages. These alterations could be due to changes that occurred in the plant nutrient requirement and varied root exudation patterns at a specific phenological stage. Further, fertilizer treatments also have a profound effect on the diversity and structure of the bacterial community. Organic fertilizers, especially vegetable tanned leather waste (VTLW), have a stronger effect on the composition and diversity of bacterial community compared to inorganic fertilizer (PT&mdash, triple superphosphate). Alpha-diversity was significantly decreased by both organic and inorganic amendments, especially a species evenness because each fertilizer tends to stimulate the growth of distinctive microbes that dominated the community of amended soil. Proteobacteria, Actinobacteria, and Cyanobacteria were the most abundant phyla, and Chelatococcus, Cyanobacteria, Sphingomonas, and Microvirga were a most abundant genus that contributed most in co-occurrence pattern, which suggests that these generalists are adapted to a variety of environments. These indicate that plant presence was a key, dominating factor, followed by fertilizers and time, in affecting soil bacterial diversity and composition. Plant recruits system (fertilization and time) -specific taxa due to differences in available nutrients and energy sources among different treatments during different growth stages. Further, fertilizer treatments did not have a stronger effect on plant production as compared to the effect on microbial community, which highlights that organic fertilizers did not tend to increase plant production. Thus, organic and inorganic amendments with matched macronutrients could have a similar impact on crop yields.

Pancratium maritimum L. (Amaryllidaceae) is a geophyte occurring in the Mediterranean region, from the Black Sea to part of the Atlantic coast. This plant is receiving much attention from the international scientific community due to its value as a bioindicator, the potential industrial value of its chemical compounds, and its use as a commercial ornamental plant. Plant morphometry and sequences of three plastid DNA regions (rbcL, matK, trnH-psbA) were used to assess the phenotypic and genetic variability of this taxon and its closest congeneric species (in particular Pancratium linosae, from the volcanic island of Linosa) in the Central Mediterranean (Sicily, Tunisia and surrounding islands). Pancratium maritimum and P. linosae cannot be distinguished based on morphological and genetic data and should belong to the same taxon. Our results also highlight a diversified gene pool in P. maritimum that is worth preserving. The lectotypes of the names Halmira stellaris, Pancratium angustifolium and Pancratium foetidum are here designated.

Chamaerops humilis is decreasing in abundance in Mediterranean Europe, which has induced the European community to call for its protection in Special Areas of Conservation. However, information about its genetic and morphological variability, which is crucial to the development of any conservation strategies, is insufficient. The present study aimed to investigate the genetic and morphological variability of C. humilis in Sicily, which was selected as a model because of the high number of dense populations. The relationships between morphological traits and climatic variables were studied to highlight patterns of adaptation to the environment, along with the genetic similarity among the populations. Ten natural populations were sampled, analyzed using 28 specifically designed SSR primers, and evaluated based on 29 morphological traits. The populations were clustered similarly based on genetic and morphological traits. Heterozygosity was high and inbreeding coefficients were low. These results, along with higher intra- than inter-population differentiation, suggest that C. humilis populations in Sicily differentiated from a common ancestor and that inter-population variation arose from secondary evolution processes induced by ecological adaptation. The correlations between climatic variables and morphological traits suggest that the morphological adaptation to arid environments depends more on summer temperatures than on evapotranspiration or rainfall and that autumn and winter temperatures are determinants of the species establishment at new sites. Considering the response of C. humilis to seasonal temperatures, the present results indicate this species as a candidate for tracking climatic changes in Europe. Further studies are needed to highlight the adaptation of C. humilis to cold environments. Palaeo-climatological and -ecological studies could help clarify its strategies for the colonization of new sites. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176, 66–81.

In order to offer hints for new research, the agronomic techniques used to manageOrobanche crenata in Sicily in the late 19 th and early 20 th Century are examined; the publications in which they were originally described can be obtained only with difficulty. Hand pulling, resistant cultivars and late sowing are the techniques used nowadays. Screening for resistance to broomrape of the traditional cultivars of broad bean, and for the influence of sowing density on broomrape infestations, is still to be done.

The systematic relationships between the five different subspecies actually recognized within Dianthus rupicola (subsp. aeolicus , subsp. bocchorianus , subsp. hermaensis, subsp. lopadusanus and subsp. rupicola ), were assessed by means of morphometric and molecular analysis. Our results highlighted the molecular differences between the five subspecies of D. rupicola and a morphological differentiation of four groups being D. rupicola subsp. aeolicus and D. rupicola subsp. rupicola partly overlapping. At the present state of knowledge, a subspecific status seems the most appropriate for all the recognized subspecies of D. rupicola . The names D. aeolicus , D. bisignanii var. virescens , D. hermaeensis , D. rupicola are here lectotypified respectively on specimens preserved at PAL, NAP, P, and BOLO, while D. bisignanii s.str. is lectotypified on a Tenore’s image and a epitype (at K) is designated. Concerning the name D. rupicola subsp. bocchorianus the holotype is traced.