Hemp (Cannabis sativa L.) is a multifunctional crop that is capable of prompt environmental adaptation. In this study, a monoecious cultivar (Futura 75) and a dioecious one (Finola) were tested in a mountain area in Valsaviore (Rhaetian Alps, Italy; elevation: 1,100 m a.s.l.) during the growing season 2018. Phytochemical behavior was evaluated by different analytical approaches: HPLC-high-resolution mass spectrometry, SDS-PAGE LC-MS/MS, HS-SPME GC-MS, and GC-FID in order to obtain complete profile of two varieties cultivated in altitude. CSR functional strategy used for ecological evaluation revealed that both genotypes are mainly competitors, although Finola is more stress tolerator (C:S:R = 57:26:17%) than Futura (C:S:R = 69:15:16%). The Finola inflorescences were characterized by higher quantities of β-ocimene and α-terpinolene, while α- and ß-pinene accompanied by extremely high ß-myrcene were found as predominant in Futura. Both varieties were rich in sesquiterpenes (45 recognized) among which trans-caryophyllene and α-humulene were the most abundant. Total tetrahydrocannabinol level was lower than 0.1%, while the most abundant cannabinoid was cannabidiolic acid (CBDA): 2.3% found in Finola vs. 2.7% revealed for Futura. The level of corresponding neutral form, cannabidiol, varied drastically: 0.27% (Finola) vs. 0.056% (Futura). Finola showed the unique cannabinoid profile with unexpectedly high cannabidivarin, 2-fold higher that corresponding acidic analogue, whereas the particularity of Futura 75 was the occurrence of cannabigerolic acid (CBGA) in the quantities that was double than those exposed for Finola. The seeds from both chemovars proved to be rich in polyunsaturated fatty acids, and Finola showed a higher ratio ω6/ω3. No difference was found in the protein content, and the SDS-PAGE profile was similar. The most abundant protein was edestin, followed by heat shock protein 70, ß-conglycinin, and vicilin. In conclusion, comprehensive phytochemical and ecological study of two fiber-type varieties cultivated in Italian Alps displayed specific, legal, and safe cannabinoids profile, followed by particular terpene composition, polyunsaturated fatty acids content, and favorable protein profile. This postulates that geographical provenience of hemp should be considered in selecting a variety that would be suitable for a specific end-use nutraceutical application.
Propolis is used as folk medicine due to its spectrum of alleged biological and pharmaceutical properties and it is a complex matrix not still totally characterized. Two batches of propolis coming from two different environments (plains of Po Valley and the hilly Ligurian–Piedmont Apennines) of Northern Italy were characterized using different analytical methods: Spectrophotometric analysis of phenols, flavones and flavonols, and DPPH radical scavenging activity, HPLC, NMR, HSPME and GC–MS and HPLC–MS Orbitrap. Balsam and moisture content were also considered. No statistical differences were found at the spectrophotometric analysis; balsam content did not vary significantly. The most interesting findings were in the VOCs composition, with the Po Valley samples containing compounds of the resins from leaf buds of Populus nigra L. The hills (Appennines) samples were indeed characterize by the presence of phenolic glycerides already found in mountain environments. HPLC–Q-Exactive-Orbitrap®–MS analysis is crucial in appropriate recognition of evaluate number of metabolites, but also NMR itself could give more detailed information especially when isomeric compounds should be identified. It is necessary a standardized evaluation to protect and valorize this production and more research on propolis characterization using different analytical techniques.
The phytochemical profiling of hemp inflorescences of clonal plants growing in different conditions related to altitude was investigated. Four strains of industrial hemp (Cannabis sativa L., family Cannabaceae) of Kompolti variety were selected and cloned to provide genetically uniform material for analyses of secondary metabolites (terpenes, cannabinoids, and flavonoids) at two different elevations: mountain (Alagna Valsesia 1200 m ASL) and plains (Vercelli Province 130 m ASL). Environmental conditions influenced by elevation have proven to be important factors inducing variations in hemp inflorescences’ secondary metabolite composition. In fact, all plants grown at altitude exhibited a higher total amount of terpenes when compared with plains counterparts, with β-Myrcene, trans-Caryophyllene and α-Humulene as the main contributors. A metabolomic, un-targeted approach performed by HPLC-Q-Exactive-Orbitrap®-MS platform with subsequent data processing performed by Compound Discoverer™ software, was crucial for the appropriate recognition of many metabolites, clearly distinguishing mountain from plains specimens. Cannabidiolic acid CBDA was the most abundant phytocannabinoid, with significantly higher concentrations in the mountain samples. The metabolic pathway of CBGA (considered as the progenitor/precursor of all cannabinoids) was also activated towards the production of CBCA, which occurs in considerably 3 times higher quantities than in the clones grown at high altitude. Isoprenoid flavones (Cannaflavins A, B, and C) were correspondingly upregulated in mountain samples, while apigenin turned out to be more abundant in plains samples. The possibility to use hemp inflorescences in pharmaceutical/nutraceutical applications opens new challenges to understand how hemp crops respond in terms of secondary metabolite production in various environments. In this regard, our results with the applied analytical strategy may constitute an effective way of phytochemical profiling hemp inflorescences.
Once a very important crop, the cultivation of hemp has seen a significant reduction in Europe for a long time, but it is now subject to direct payments and promotion initiatives. Italy used to be an important producer and exporter of hemp textiles until the Second World War but currently information is lacking regarding the hemp production chain and legislation on the issue is often misunderstood by producers. Moreover, there has been an important development of economic activities connected with hemp, such as the so-called “hemp shops” or “grow shops” and the market of a product called “cannabis light”, the dried inflorescences of industrial hemp. The aim of the study was to investigate 30 Italian hemp farms in order to identify the characteristics of the production chain and the uses of the crop. Some considerations on this sector in the Italian economic and legislative contest are made and an anonymous web survey on the commercial activities associated with hemp (grow shops) was conducted. Most farms are multifunctional, 83% have been set up recently (in the last ten years) and directly as hemp producers. They are run by young entrepreneurs (57% holders under 35) and allow the use of marginal abandoned territories (43% of the farms). The 30 farms cultivate 460 ha of hemp and the extension of the field crops is very variable, from small patches in the mountains of 0,001 ha to more than 100 ha farms in the plains (in particular in Campania region). Almost all the farms use the crop to produce more than one end-product (seeds, flour, decorticated seeds, hemp-beer, seeds for animal nutrition and food oil from seeds, seeds and inflorescences harvested for cosmetic use, herbal use and extraction of active ingredients). In some cases, “technical use” linked to selling of the dry top inflorescences of industrial hemp for smoking was declared and it was found that there has been a significant increase in grow shops in Italy, from 4 in 2002 to more than 700 in 2018. As emerges from the analyses of European and Italian legislation, there is a need for clear regulations and a system of control by regulatory organizations considering the actual criticalities. At the same time, the renewed appeal of this crop derives from the multiple possibilities of use of the plant and from growing consumer demand for eco-compatible and sustainable products.
Cannabis sativa L. has been cultivated throughout the world for industrial and medical purposes and is the most controversial plant ever exploited, with considerable discrepancies in the praise and disapproval it receives. Medical Cannabis prescriptions are on the increase in several countries where its therapeutic use is authorised due to its positive role in treating several pathologies even if it represents a multifaceted reality in terms of application. There are at least 550 identified compounds in C. sativa L., including more than 100 phytocannabinoids and 120 terpenes. The chemical complexity of its bioactive constituents highlights the need for standardised and well-defined analytical approaches able to characterise plant chemotype and herbal drug quality as well as to monitor the quality of pharmaceutical cannabis extracts and preparations. This research highlights the potential of using different analytical procedures involving the combination of headspacesolid-phase microextraction (HS-SPME) coupled to GC-MS and accelerated solvent extraction (ASE) coupled to high resolution mass-spectrometry (HPLC-Q Orbitrap®) for the indepth profiling of quality traits in authorised medical varieties of Cannabis sativa L. flos (Bediol®) and corresponding macerated oil preparations. This approach could add new knowledge to the field of "omic" analytical applications which are fundamental nowadays for Cannabis used for therapeutic remedies.
Landraces are an agri-food and historical-cultural heritage but are undergoing losses worldwide. Italy is taking action to counteract this problem by following European guidelines. One of the most important measures is the Agrobiodiversity National Register (ANR), but 12 Italian regions currently appear without any landraces and around 80% of the landraces listed are trees, with less detailed data on herbaceous species. The aim of this study is to investigate the situation for Italian herbaceous landraces preserved on farms (in situ) by merging and analyzing data contained in the main databases on plant agrobiodiversity in Italy. Data were georeferenced, organized by botanic families and Italian regions, and analyzed by GIS and R. A total of 1615 herbaceous landraces were found (versus the 416 recorded in the ANR). Poaceae, Fabaceae, and Solanaceae together comprise 70% of all herbaceous landraces and are mostly preserved/grown in areas between 150 and 800 m a.s.l. Some hilly and sub-mountain areas of the Apennines and the Alps are hotspots of herbaceous landraces due to anthropic and environmental factors. The results of this research will be useful to enrich the ANR and trigger actions of characterization, conservation, and promotion of these plant resources.
Cannabis sativa L. (hemp) is a versatile plant which can adapt to various environmental conditions. Hempseeds provide high quality lipids, mainly represented by polyunsaturated acids, and highly digestible proteins rich of essential aminoacids. Hempseed composition can vary according to plant genotype, but other factors such as agronomic and climatic conditions can affect the presence of nutraceutic compounds. In this research, seeds from two cultivars of C. sativa (Futura 75 and Finola) grown in a mountain environment of the Italian Alps were analyzed. The main purpose of this study was to investigate changes in the protein profile of seeds obtained from such environments, using two methods (sequential and total proteins) for protein extraction and two analytical approaches SDS-PAGE and 2D-gel electrophoresis, followed by protein identification by mass spectrometry. The fatty acids profile and carotenoids content were also analysed. Mountain environments mainly affected fatty acid and protein profiles of Finola seeds. These changes were not predictable by the sole comparison of certified seeds from Futura 75 and Finola cultivars. The fatty acid profile confirmed a high PUFA content in both cultivars from mountain area, while protein analysis revealed a decrease in the protein content of Finola seeds from the experimental fields.
Alpine pastures and meadows are agroecosystems of biological, cultural-historical, and economic importance that are undergoing profound imbalances and which are in a rapid decline due to changes in management and/or abandonment. The European Union is making efforts to protect this heritage and resource. However, the dialog among the different professionals in charge of studying and managing these agroecosystems needs to be as easy and comprehensible as possible for grasslands conservation/restoration actions to be successful. This research introduces VegeT, an easy-to-use tool to facilitate information transfer between botanists and practitioners responsible for providing guidelines for the correct management of mountain grasslands. VegeT is a Microsoft Excel® worksheet that allows the classification of seminatural grasslands and dynamically connected vegetation (shrublands and forests) of the Alps employing two ecological indexes: the index of nutrients (N) and the index of mowing tolerance (MV). VegeT was elaborated upon the floristic-ecological analysis of the vegetation of Taleggio Valley (Italian Alps) performed applying multivariate analysis techniques. From the analyses, it emerged that N and MV are the main variables on which to base a classification system of alpine mountain grasslands and dynamically connected vegetation able to facilitate the interpretation of floristic-vegetation data and to return useful information for management decisions.
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