Onion Fusarium Basal Rot Disease Control by Arbuscular Mycorrhizal Fungi and Trichoderma harzianum.

Soilborne pathogens reduce 60% of the yield of onion crops. A common fungal pathogen causing wilt disease and severe losses is Fusarium basal rot (FBR). In this study, the combination of Arbuscular Mycorrhizal Fungi (AMF) with Trichoderma harzianum was investigated against FBR. Onion samples were collected from the Ankara-Polatli region. Among the isolates, isolate S6 was identified as F. oxysporum f. sp. cepae (FOC) using morphological and molecular methods and pathogenicity tests. Different combinations of AMF (Funneliformis mosseae pure strain and the commercial AMF) and T. harzianum were inoculated on susceptible onion cultivars (Seç, Gence, and Sampiyon). The effects of the treatments on FOC biocontrol were studied under growth chamber conditions. The results showed that Sampiyon was the most resistant, while Gence was the most susceptible to basal rot disease. Different colonization rates (8.91-24%), spore densities (16.4-50.4 spore/10 g soil), and the extent to which a plant needs mycorrhizal conditions to grow to its maximum potential (i.e., mycorrhizal dependencies-18.3-51.9%) were recorded by treatment. Both single and combined applications of AMF and Trichoderma applications suppressed FOC. Suppressive effects were more pronounced when the F. mosseae pure strain was used alone (when F. mosseae was used, disease severity decreased from 90 to 68%, p < 0.05). The F. mosseae pure strain also showed the best plant growth promotion and phosphorus content release. The results indicate an interesting potential use of F. mosseae and the combination of AMF with T. harzianum in the management of FOC in onions.

Ellenberg Indicator Values Disclose Complex Environmental Filtering Processes in Plant Communities along an Elevational Gradient.

Ellenberg indicator values (EIVs) express plant preferences for temperature, light, continentality, soil moisture, pH, and soil nutrients, and have been largely used to deduce environmental characteristics from plant communities. However, EIVs might also be used to investigate the importance of filtering mechanisms in shaping plant communities according to species ecological preferences, a so far overlooked use of EIVs. In this paper, we investigated how community-weighted means (CWM), calculated with EIVs, varied along an elevational gradient in a small mountain in Central Italy. We also tested if species abundances varied according to their ecological preferences. We found that the prevalence of thermophilous species declines with elevation, being progressively replaced by cold-adapted species. Heliophilous species prevail at low and high elevations (characterized by the presence of open habitats), whereas in the middle of the gradient (occupied by the beech forest), sciophilous species predominate. Variations for moisture and soil nutrient preferences followed a similar pattern, probably because of the high moisture and nutrient levels of forest soils with a lot of humus. No distinct pattern was detected for EIVs for pH and continentality since these factors are subject to more local variations. These results highlight the possible role of EIVs to investigate how environmental gradients shape plant communities.

Wild and Micropropagated Artemisia eriantha Infusions: In Vitro Digestion Effects on Phenolic Pattern and Antioxidant Activity.

This study investigated the in vitro simulated gastrointestinal digestion (GID) effects on wild and micropropagated Apennines Genepì infusions. Wild and micropropagated infusions were compared for their antioxidant activity, phenolic contents, and polyphenolic profiles before and after GID. Before digestion, the wild infusions had higher amounts of phenolic compounds and antioxidant activity than the micropropagated ones. Instead, after digestion, the differences in the total phenolic content (TPC) and antioxidant activity between wild and micropropagated infusions were less pronounced. The changes in the TPC and phenolic profiles revealed the presence of several chemical transformations and rearrangements that resulted in compounds with different reactivity and antioxidant potential. Without enzyme actions, the wild infusion digest undergoes higher modifications than those obtained from the micropropagated ones. The current study offers the first concrete proof of the impact of GID on the polyphenolic chemicals present in infusions of wild and micropropagated Apennines Genepì and their antioxidant properties. Our findings are essential for future in-depth analyses of Apennine Genepì infusions and their potential impacts on human health.

Nanoporous silica gel can compete with the flower stigma in germinating and attracting pollen tubes.

To find nanoporous substrates with hydrodynamic properties useful for pollen hydration and germination, we used the glassy Silica gel and Vycor scales and pollen with different morphological and physiological traits, that of Crocus vernus, and that of Narcissus poeticus. For in vitro tests, the scales were spread on microscope slides, hand pollinated, and incubated. Pollen germination was evaluated with the stereomicroscope and the tube growth was explored with scanning electron microscopy (SEM). The in vivo tests were carried out by sprinkling the stigmas of the Crocus plants with Silica gel scales and immediately after having pollinated them by hand, the plants were incubated. Three hours later, the stigmas were removed and treated for observation with SEM. In vitro the pollen of both species germinated on Silica gel with percentages similar to those of the in vivo and in vitro controls, accumulating fibrillary material at the interface. The tubes grew perpendicular to the surface of the scales, trying to penetrate the scales to the point of flattening with the apex. On Crocus stigmas sprinkled with Silica gel scales, pollen developed tubes that grew to the scales rather than penetrating the papillae. The results underline the close interaction of pollen with nanoporous artificial material, so much so that its pollen tubes are attracted to the Silica scales more than to the stigma papillae that arises from a mechanism of natural selection.

Use of Lichens to Evaluate the Impact of Post-Earthquake Reconstruction Activities on Air Quality: A Case Study from the City of L'Aquila.

Lichens are widely used as bioindicators of air quality because of their ability to absorb chemical pollutants. We used the Lichen Diversity Value (LDV) index to assess the effects of the urban reconstruction activities in the city of L'Aquila ten years after the 2009 earthquake on air quality. Sampling was conducted from the city centre (still mostly under reconstruction and closed to traffic) to suburban areas (where reconstruction is minimal). We tested if the LDV index varied with distance from the city centre because of the presence of air pollutants produced by reconstruction works. We also used Energy-Dispersive X-ray Spectroscopy (EDS) to detect the main pollutants accumulated in the sampled lichens. The LDV increased from the city centre towards suburban areas. EDS revealed high concentrations of pollutants related to demolition and reconstruction activities, such as aluminium and silicon (used in the manufacture of concrete), in the more central areas. These results suggest that the LDV index can be a useful tool to monitor air quality, even on a small scale, and in urban environments subject to building demolition and reconstruction. Moreover, EDS could represent a good preliminary analytical technique to identify the air pollutants associated with all of these activities.

Fusarium Oxysporum f. sp. Cannabis Isolated from Cannabis Sativa L.: In Vitro and In Planta Biocontrol by a Plant Growth Promoting-Bacteria Consortium.

Industrial hemp (Cannabis sativa L.) is a multipurpose plant used in several fields. Several phytopathogens attack hemp crops. Fusarium oxysporum is a common fungal pathogen that causes wilt disease in nurseries and in field cultivation and causes high losses. In the present study, a pathogenic strain belonging to F. oxysporum f. sp. cannabis was isolated from a plant showing Fusarium wilt. After isolation, identification was conducted based on morphological and molecular characterizations and pathogenicity tests. Selected plant growth-promoting bacteria with interesting biocontrol properties-Azospirillum brasilense, Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae and Burkholderia ambifaria-were tested against this pathogen. In vitro antagonistic activity was determined by the dual culture method. Effective strains (in vitro inhibition > of 50%) G. diazotrophicus, H. seropedicae and B. ambifaria were combined in a consortium and screened for in planta antagonistic activity in pre-emergence (before germination) and post-emergence (after germination). The consortium counteracted Fusarium infection both in pre-emergence and post-emergence. Our preliminary results show that the selected consortium could be further investigated as an effective biocontrol agent for the management of this pathogen.

Variations in Plant Richness, Biogeographical Composition, and Life Forms along an Elevational Gradient in a Mediterranean Mountain.

Despite the increasing interest in elevational patterns in biodiversity, few studies have investigated variations in life forms and biogeographical composition, especially in the Mediterranean biome. We investigated elevational patterns in species richness, biogeographical composition (chorotypes) and life forms (Raunkiaer classification) along an elevational gradient in a Mediterranean mountain (Central Italy). We found a general hump-shaped pattern of species richness, which can be explained by harsher conditions at the lowest and highest elevations. This pattern is distinctly related to prevalence at mid elevations of species with European and Euro-Asiatic distribution, which are favored by a temperate climate. Phanerophytes and geophytes (which are mainly associated with woods) were concentrated at mid elevations where woodlands prevail. Hemicryptophytes increased with elevation, consistently with their ability to cope with high altitude climatic conditions. Mediterranean species declined with elevation because they are negatively affected by decreasing temperatures. Chamaephytes showed a U-shaped pattern, suggesting they are able to cope with arid and cold conditions at the extremes of the gradient. Endemics increased with elevation because of their association with mountainous areas as key places for endemism evolution. These results illustrate how elevational patterns in species richness, biogeographical composition and life forms are interrelated and demonstrate reciprocal insights for understanding current vegetation settings.

In vitro and in planta antagonistic effects of plant growth-promoting rhizobacteria consortium against soilborne plant pathogens of Solanum tuberosum and Solanum lycopersicum.

Potatoes (Solanum tuberosum L.) and tomatoes (Solanum lycopersicum L.), among the main crops belonging to the Solanaceae family, are attacked by several pathogens. Among them Fusarium oxysporum f. sp. radicis-lycopersici and Rhizoctonia solani are very common and cause significant losses. Four plant growth-promoting rhizobacteria, Azospirillum brasilense, Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae and Burkholderia ambifaria were tested against these pathogens. In vitro antagonistic activities of single strains were assessed through dual culture plates. Strains showing antagonistic activity (G. diazotrophicus, H. seropedicae and B. ambifaria) were combined and, after an in vitro confirmation, the consortium was applied on S. lycopersicum and S. tuberosum in a greenhouse pot experiment. The bioprotection was assessed in pre-emergence (infection before germination) and post-emergence (infection after germination). The consortium was able to successfully counteract the infection of both F. oxysporum and R. solani, allowing a regular development of plants. The biocontrol of the fungal pathogens was highlighted both in pre-emergence and post-emergence conditions. This selected consortium could be a valid alternative to agrochemicals and could be exploited as biocontrol agent to counteract losses due to these pathogenic fungi.

Chemical Characterization of Narcissus poeticus from Sirente -Velino (Apennines - Italy): Galantamine Accumulation and Distribution of Allergenic Compounds in the Flower.

Species of Narcissus (family Amaryllidaceae) are a potential source for large-scale extraction of alkaloids and fragrances. The bulbs typically accumulate a large number of alkaloids, including galantamine, a benzazepine alkaloid proven to be a cholinesterase inhibitor and which is used in the treatment of Alzheimer's disease. The presence of galantamine in N. poeticus L. collected in Abruzzo (Italy) was assessed and several levels of alkaloid were found in all parts of the plant (flower, stem, bulb and root) and not only in the bulb. The amount of galantamine obtained was tested by using two different extraction solvents. Extraction of N. poeticus absolute from the flowers was also performed, as this product is an important floral note in perfumery, and the distribution of allergenic compounds in the coronas and in the tepals was assessed. Moreover, the in vitro propagation of N.:Poeticus was tested as it may be a valuable resource from which to produce biomolecules, as an alternative to chemical synthetic processes.

Volatiles fingerprint of Artemisia umbelliformis subsp. eriantha by headspace-solid phase microextraction GC-MS.

Artemisia umbelliformis subsp. eriantha is a protected species, whose essential oil is used in liqueur industry. Volatile profiles of fresh leaves and flowers from wild plants in comparison with regenerated in vitro plants introduced in experimental fields within an Italian national park were evaluated by headspace-solid phase microextraction gas chromatography mass spectrometry (HS-SPME-GC-MS). The chromatographic profiles appear to be qualitatively similar. The content of thujones, the characteristic metabolites of this species, is comparable with that obtained by analysis of essential oils. Principal component analysis of the HS-SPME-GC-MS data supports the possibility of differentiating scent blends of genetically identical plants, and even flowers and leaves from the same individual. HS-SPME-GC-MS is shown to be a very efficient method to analyse and to describe the pattern of components of A. umbelliformis subsp. eriantha cultivars' scents. It represents a rapid screening method highly recommended for the study of protected species, because it is non-destructive and it only requires small amounts of fresh material.

Volatile fingerprints of artemisinin-rich Artemisia annua cultivars by headspace solid-phase microextraction gas chromatography/ mass spectrometry.

The cultivar Anamed (A3) is a hybrid of Artemisia annua with a high content of the secondary metabolite artemisinin, a well-known antimalarial drug. Here we report for the first time the volatile profile of fresh leaves of this hybrid in comparison with that of Artemisia annua L. wild-type species. Evaluation and comparison of the volatile profiles of A. annua genotypes with different content in artemisinin were carried out by headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography/mass spectrometry (GC/MS) that was performed on fresh leaves of the plants under investigation using a polydimethylsiloxane (PDMS) fiber. The chromatograms obtained from hybrids with a high content of artemisinin (A. annua cv. Anamed A3 and A. annua cv. Artemis F2) reveal the total absence of artemisia ketone, one of the major and characteristic compounds of the wild-type A. annua L., along with a significantly lower variety of volatile compounds. In conclusion, HS-SPME coupled with GC/MS is a very useful, non-destructive and efficient method to describe the volatile pattern of Artemisia annua cultivars. It represents a rapid screening method for the evaluation of volatile biomarkers like artemisia ketone, whose absence is typical of artemisinin-rich A. annua cultivars.

Detection of lead in Zea mays by dual-energy X-ray microtomography at the SYRMEP beamline of the ELETTRA synchrotron and by atomic absorption spectroscopy.

This study is related to the application of the X-ray dual-energy microradiography technique together with the atomic absorption spectroscopy (AAS) for the detection of lead on Zea mays stem, ear, root, and leaf samples. To highlight the places with lead intake, the planar radiographs taken with monochromatic X-ray radiation in absorption regime with photon energy below and above the absorption edge of a given chemical element, respectively, are analyzed and processed. To recognize the biological structures involved in the intake, the dual-energy images with the lead signal have been compared with the optical images of the same Z. mays stem. The ear, stem, root, and leaf samples have also been analyzed with the AAS technique to measure the exact amount of the hyperaccumulated lead. The AAS measurement revealed that the highest intake occurred in the roots while the lowest in the maize ears and in the leaf. It seems there is a particular mechanism that protects the seeds and the leaves in the intake process.

A rapid method for the quantification of artemisinin in Artemisia annua L. plants cultivated for the first time in Burundi.

We describe a simple, rapid combined method for extracting the antimalarial compound artemisinin from the leaves of Artemisia annua L. cultivated for the first time in Burundi, and quantitating the active principle by high-performance liquid chromatography-electrospray mass spectrometry.