ABSTRACT
Epigenetic changes, host-gut microbiota interactions, and environmental factors contribute to inflammatory bowel disease (IBD) onset and progression. A healthy lifestyle may help to slow down the chronic or remitting/relapsing intestinal tract inflammation characteristic of IBD. In this scenario, the employment of a nutritional strategy to prevent the onset or supplement disease therapies included functional food consumption. Its formulation consists of the addition of a phytoextract enriched in bioactive molecules. A good candidate as an ingredient is the Cinnamon verum aqueous extract. Indeed, this extract, subjected to a process of gastrointestinal digestion simulation (INFOGEST), exhibits beneficial antioxidant and anti-inflammatory properties in an in vitro model of the inflamed intestinal barrier. Here, we deepen the study of the mechanisms related to the effect of digested cinnamon extract pre-treatment, showing a correlation between transepithelial electrical resistance (TEER) decrement and alterations in claudin-2 expression under Tumor necrosis factor-α/Interleukin-1ß (TNF-α/IL-1) ß cytokine administration. Our results show that pre-treatment with cinnamon extract prevents TEER loss by claudin-2 protein level regulation, influencing both gene transcription and autophagy-mediated degradation. Hence, cinnamon polyphenols and their metabolites probably work as mediators in gene regulation and receptor/pathway activation, leading to an adaptive response against renewed insults.
Subject(s)
Cinnamomum zeylanicum , Inflammatory Bowel Diseases , Humans , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Claudin-2 , Interleukin-1beta/genetics , Plant Bark/metabolism , Inflammatory Bowel Diseases/drug therapy , Inflammatory Bowel Diseases/metabolism , Gene ExpressionABSTRACT
Flowers have many traits to appeal to pollinators, including ultraviolet (UV) absorbing markings, which are well-known for attracting bees at close proximity (e.g., <1 m). While striking UV signals have been thought to attract pollinators also from far away, if these signals impact the plant pollinia removal over distance remains unknown. Here, we report the case of the Australian orchid Diuris brumalis, a nonrewarding species, pollinated by bees via mimicry of the rewarding pea plant Daviesia decurrens. When distant from the pea plant, Diuris was hypothesized to enhance pollinator attraction by exaggeratedly mimicking the floral ultraviolet (UV) reflecting patterns of its model. By experimentally modulating floral UV reflectance with a UV screening solution, we quantified the orchid pollinia removal at a variable distance from the model pea plants. We demonstrate that the deceptive orchid Diuris attracts bee pollinators by emphasizing the visual stimuli, which mimic the floral UV signaling of the rewarding model Daviesia. Moreover, the exaggerated UV reflectance of Diuris flowers impacted pollinators' visitation at an optimal distance from Da. decurrens, and the effect decreased when orchids were too close or too far away from the model. Our findings support the hypothesis that salient UV flower signaling plays a functional role in visual floral mimicry, likely exploiting perceptual gaps in bee neural coding, and mediates the plant pollinia removal at much greater spatial scales than previously expected. The ruse works most effectively at an optimal distance of several meters revealing the importance of salient visual stimuli when mimicry is imperfect.
ABSTRACT
Cinnamon bark is widely used for its organoleptic features in the food context and growing evidence supports its beneficial effect on human health. The market offers an increasingly wide range of food products and supplements enriched with cinnamon extracts which are eliciting beneficial and health-promoting properties. Specifically, the extract of Cinnamomum spp. is rich in antioxidant, anti-inflammatory and anticancer biomolecules. These include widely reported cinnamic acid and some phenolic compounds, such asproanthocyanidins A and B, and kaempferol. These molecules are sensitive to physical-chemical properties (such as pH and temperature) and biological agents that act during gastric digestion, which could impair molecules' bioactivity. Therefore, in this study, the cinnamon's antioxidant and anti-inflammatory bioactivity after simulated digestion was evaluated by analyzing the chemical profile of the pure extract and digested one, as well as the cellular effect in vitro models, such as Caco2 and intestinal barrier. The results showed that the digestive process reduces the total content of polyphenols, especially tannins, while preserving other bioactive compounds such as cinnamic acid. At the functional level, the digested extract maintains an antioxidant and anti-inflammatory effect at the cellular level.
ABSTRACT
Age-related injuries are often connected to alterations in redox homeostasis. The imbalance between free radical oxygen species and endogenous antioxidants defenses could be associated with a growing risk of transient ischemic attack and stroke. In this context, a daily supply of dietary antioxidants could counteract oxidative stress occurring during ischemia/reperfusion injury (I/R), preventing brain damage. Here we investigated the potential antioxidant properties of coffee-derived circulating metabolites and a coffee pulp phytoextract, testing their efficacy as ROS scavengers in an in vitro model of ischemia. Indeed, the coffee fruit is an important source of phenolic compounds, such as chlorogenic acids, present both in the brewed seed and in the discarded pulp. Therefore, rat brain endothelial cells, subjected to oxygen and glucose deprivation (OGD) and recovery (ogR) to mimic reperfusion, were pretreated or not with coffee by-products. The results indicate that, under OGD/ogR, the ROS accumulation was reduced by coffee by-product. Additionally, the coffee extract activated the Nrf2 antioxidant pathway via Erk and Akt kinases phosphorylation, as shown by increased Nrf2 and HO-1 protein levels. The data indicate that the daily intake of coffee by-products as a dietary food supplement represents a potential nutritional strategy to counteract aging.
Subject(s)
Antioxidants/pharmacology , Coffea/chemistry , NF-E2-Related Factor 2/agonists , Phenols/pharmacology , Plant Extracts/pharmacology , Reperfusion Injury/therapy , Animals , Antioxidants/chemistry , Brain Ischemia/metabolism , Brain Ischemia/therapy , Cell Line , NF-E2-Related Factor 2/metabolism , Oxidative Stress/drug effects , Phenols/chemistry , Plant Extracts/chemistry , Rats , Reperfusion Injury/metabolismABSTRACT
This study is aimed at valorizing artichoke (Cynara cardunculus var. scolymus L.) by-products as source of inulin, a fiber showing relevant prebiotic properties, through the realization of a waste value chain. Starting from artichoke by-products, the inulin fraction was assessed both in terms of total amount and degree of polymerization as a function of the harvest season and storage conditions. These parameters have been found significant at influencing inulin yield of extraction. For the first time, artichoke wastes were proposed to be exploited taking into account the optimal conditions to preserve their high-added chemical value. Our data suggest that Italian farms could obtain from their wastes a total amount of 16 t/year of inulin with an average polymerization degree higher than 40 and would allow the development of a circular economy process within the artichoke supply chain, by exploiting its wastes representing 70% of the total artichoke biomass.
Subject(s)
Cynara scolymus , Cynara , Scolymus , Cynara scolymus/chemistry , Inulin/chemistry , Polymerization , PrebioticsABSTRACT
Cinnamon polyphenols are known as health-promoting agents. However, their positive impact depends on the extraction method and their bioaccessibility after digestion. In this work, cinnamon bark polyphenols were extracted in hot water and subjected to an in vitro enzymatic digestion. After a preliminary characterization of total polyphenols and flavonoids (respectively 520.05 ± 17.43 µgGAeq/mg and 294.77 ± 19.83 µgCATeq/mg powder extract), the extract antimicrobial activity was evidenced only against Staphylococcus aureus and Bacillus subtilis displaying a minimum inhibition growth concentration value of 2 and 1.3 mg/mL, respectively, although it was lost after in vitro extract digestion. The prebiotic potential was evaluated on probiotic Lactobacillus and Bifidobacterium strains highlighting a high growth on the in vitro digested cinnamon bark extract (up to 4 × 108 CFU/mL). Thus, the produced SCFAs and other secondary metabolites were extracted from the broth cultures and determined via GC-MSD analyses. The viability of healthy and tumor colorectal cell lines (CCD841 and SW480) was assayed after the exposition at two different concentrations (23 and 46 µgGAeq/mL) of the cinnamon extract, its digested, and the secondary metabolites produced in presence of cinnamon extract or its digested, showing positive protective effects against a tumorigenic condition.
ABSTRACT
In the context of the global need to move towards circular economies, microbial cell factories can be employed thanks to their ability to use side-stream biomasses from the agro-industrial sector to obtain additional products. The valorization of residues allows for better and complete use of natural resources and, at the same time, for the avoidance of waste management to address our needs. In this work, we focused our attention on the microbial valorization of cinnamon waste material after polyphenol extraction (C-PEW) (Cinnamomum verum J.Presl), generally discarded without any additional processing. The sugars embedded in C-PEW were released by enzymatic hydrolysis, more compatible than acid hydrolysis with the subsequent microbial cultivation. We demonstrated that the yeast Rhodosporidium toruloides was able to grow and produce up to 2.00 (±0.23) mg/L of carotenoids in the resulting hydrolysate as a sole carbon and nitrogen source despite the presence of antimicrobial compounds typical of cinnamon. To further extend the potential of our finding, we tested other fungal cell factories for growth on the same media. Overall, these results are opening the possibility to develop separate hydrolysis and fermentation (SHF) bioprocesses based on C-PEW and microbial biotransformation to obtain high-value molecules.
Subject(s)
Carotenoids , Cinnamomum zeylanicum , Fermentation , Rhodotorula , Waste ProductsABSTRACT
It is estimated that about 7000 plant species and a large number of cultivars and varieties have been cultivated for consumption in human history. However, <0.5% of these currently provide the majority of human food energy needs worldwide (e.g., rice, wheat, maize, and potato). Global issues such as climate change, diffusion of pests, and resistance to agrochemical treatments are posing great concern about the sustainable cultivation of these major staples, especially in equatorial and tropical countries, such as Sub Saharan Africa. In addition, most of these regions contain malnutrition and micronutrient deficiencies, and the sum of such problems create serious implications at social, political, and economic levels. A possible solution relies on the exploitation of plant biodiversity and particularly on the so-called NUS (Neglected and Underutilized Species). These plants are traditionally grown in their centres of origin and continue to be maintained by sociocultural preferences, however they remain inadequately documented and neglected by formal research and conservation programs. Although they are important in terms of micronutrients and the ability to grow in harsh conditions, these species are falling into disuse due to agronomic, genetic, economic, and cultural reasons. To promote and spread their cultivation at the global scale, along with knowledge on their suitability for human nutrition, reliable identification systems are necessary to guarantee adequate authenticity along the entire supply chain and distribution network. A precise identification of the different species and their varieties is fundamental both to retrieve information on their origin and authenticate the raw materials (i.e., seeds, leaves and fruit) and related processed products that can be distributed at the local or global scale. DNA-based techniques can help achieve this mission. In particular, the DNA barcoding approach has gained a role of primary importance due to its universality and versatility. Here, we discuss the advantages in using DNA barcoding for the identification of some of the most representative NUS species, as well as their traceability and conservation of cultural practices around them.
Subject(s)
DNA Barcoding, Taxonomic , Nutritive Value , Plants/chemistry , Biodiversity , Conservation of Natural Resources , Food Supply , Fruit/chemistry , Micronutrients/analysis , Nutritional Status , Plant Leaves/chemistry , Seeds/chemistryABSTRACT
Coffee is the second traded food commodity in the world. Beyond roasted seeds, the most part of the original fruit -and in particular pulp- is discarded as waste, with severe environmental and economic consequences in many developing countries. Our research focused on developing an eco-friendly extraction protocol of phytocomplexes from coffee pulp and evaluating their bioactivity and beneficial effects to human health as food supplements. Antioxidant activity assays (Folin-Ciocalteu and DPPH assays) were adopted to select the most effective extraction technique and results show antioxidant activity of coffee pulp extracts. After analysis of cytotoxicity on human epithelial gastric cells, measurements of IL-8 release of treated or pre-treated cells were performed. Results showed that the use of soft technical equipment and sustainable solvents (i.e. maceration process, aqueous extraction) can extract phytocomplexes with antioxidant properties. Moreover, IL-8 measurements showed impairment of this chemokine release at concentrations that may be reached in vivo in the gastrointestinal tract, following consumption of reasonable amount of extract. Pre-treatments analysis demonstrated the ability of coffee pulp extracts to prevent IL-8 release by gastric epithelial cells. Chemical evaluation performed by liquid chromatography mass spectrometry showed that quinic acid derivatives are abundant in coffee pulp extract together with procyanidins derivatives: those compounds might be responsible for the high biological activity. This evidence supports future applications of coffee pulp extracts as food supplement with high added value, starting from a waste that can be valorized through simple yet efficient extraction methods.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Antioxidants/pharmacology , Coffee/chemistry , Dietary Supplements , Food Handling/methods , Gastric Mucosa/drug effects , Inflammation Mediators/metabolism , Interleukin-8/metabolism , Plant Extracts/pharmacology , Seeds/chemistry , Anti-Inflammatory Agents/isolation & purification , Anti-Inflammatory Agents/toxicity , Antioxidants/isolation & purification , Antioxidants/toxicity , Cell Line , Gastric Mucosa/immunology , Gastric Mucosa/metabolism , Humans , Plant Extracts/isolation & purification , Plant Extracts/toxicityABSTRACT
A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.
ABSTRACT
ABSRACT: The most anthropized regions of the world are characterized by an impressive abundance of invasive plants, which alter local biodiversity and ecosystem services. An alternative strategy to manage these species could be based on the exploitation of their fruits in a framework of bioprospecting to obtain high-added value compounds or phytocomplexes that are useful for humans. Here we tested this hypothesis on three invasive plants (Lonicera japonica Thunb., Phytolacca americana L., and Prunus serotina Ehrh.) in the Po plain (northern Italy) which bear fruits that are highly consumed by frugivorous birds and therefore dispersed over large distances. Our biochemical analyses revealed that unripe fruit shows high antioxidant properties due to the presence of several classes of polyphenols, which have a high benchmark value on the market. Fruit collection for phytochemical extraction could really prevent seed dispersal mediated by frugivorous animals and produce economic gains to support local management actions.
Subject(s)
Bioprospecting/methods , Ecosystem , Fruit/physiology , Introduced Species , Plants , Seed Dispersal , Animals , BirdsABSTRACT
Grape berries harbor a wide range of microbes originating from the vineyard environment, many of which are recognized for their role in the must fermentation process shaping wine quality. To better clarify the contribution of the microbiome of grape fruits during wine fermentation, we used high-throughput sequencing to identify bacterial and fungi communities associated with berries and musts of Cannonau. This is the most important cultivar-wine of Sardinia (Italy) where most vineyards are cultivated without phytochemical treatments. Results suggested that microbiomes of berries collected at four different localities share a core composition characterized by Enterobacteriales, Pseudomonadales, Bacillales, and Rhodospirillales. However, any area seems to enrich berries microbiome with peculiar microbial traits. For example, berries belonging to the biodynamic vineyards of Mamoiada were rich in Bacillales typical of manure (i.e. Lysinibacillus, Bacillus, and Sporosarcina), whereas in the Santadi locality, berries showed soil bacteria such as Pasteurellales and Bacteroidales as well as Rhodospirillales and Lactobacillales which are commonly involved in wine fermentation. In the case of fungi, the most abundant taxa were Dothioraceae, Pleosporaceae, and Saccharomycodaceae, and although the proportion of these families varied among localities, they occurred ubiquitously in all vineyards. During vinification processes performed at the same wine cellar under controlled conditions and without using any yeast starter, more than 50% of bacteria groups of berries reached musts, and each locality had its own private bacteria signature, even if Saccharomyces cerevisiae represented the most abundant fungal species. This work suggests that natural berries microbiome could be influenced by pedoclimatic and anthropologic conditions (e.g., farming management), and the fruits' microorganisms persist during the fermentation process. For these reasons, a reliable wine genotyping should include the entire holobiont (plant and all its symbionts), and bioprospecting activities on grape microbiota could lead to improved viticulture yields and wine quality.
Subject(s)
Environment , Microbiota , Vitis/microbiology , Wine , Agriculture , Bacteria/classification , Bacteria/genetics , Biodiversity , Cluster Analysis , DNA Barcoding, Taxonomic , Fungi/classification , Fungi/genetics , Geography , Metagenome , Metagenomics/methodsABSTRACT
Nowadays organizations have to cope with two related challenges: maintaining an engaged and highly performing workforce and, at the same time, protecting and increasing employees' well-being and job satisfaction under conditions of a generalized increase of job demand, in an increasingly growing older population. According to the motivational process of the JD-R model, a work environment with many organizational resources will foster work engagement, which in turn will increase the likelihood of positive personal and organizational outcomes, such as job satisfaction, performance, and intention to stay. However, it is not clear how this motivational process could work in different age cohorts, as older workers may have different priorities to those of younger colleagues. Postulating the existence of a gain-cycle in the relationship between work engagement and outcomes, in this study we tested a longitudinal moderated mediation model in which job satisfaction increases over time through an increment in work engagement. We hypothesized that this process is moderated by job demand and aging. We collected data in public administrations in Northern Italy in order to measure work engagement and job satisfaction. 556 workers aged between 50 and 64 replied to the survey twice (the first time and 8 months later). The findings confirmed a moderated mediation model, in which job satisfaction at time 1 increased work engagement, which in turn fostered job satisfaction 8 months later, confirming the hypothesized gain-cycle. This relationship was shown to be moderated by the joint influence of job demand intensity and age: higher job demands and younger age are related to the maximum level of level gain cycle, while the same high level of job demands, when associated with older age, appears unable to stimulate a similar effect. The results confirm that, on one hand, older workers cannot be seen as a homogeneous group and, on the other hand, the importance of considering the role played by the gain cycle of resources. Our findings show that age matters, and that greater consideration should be devoted to age differences in order to design appropriate human resources practices that foster work engagement and satisfaction.
ABSTRACT
Understanding the genetic background of complex behavioral traits, showing multigenic control and extensive environmental effects, is a challenging task. Among such traits, migration is known to show a large additive genetic component. Yet, the identification of specific genes or gene regions explaining phenotypic variance in migratory behavior has received less attention. Migration ultimately depends on seasonal cycles, and polymorphism at phenological candidate genes may underlie variation in timing of migration or other aspects of migratory behavior. In this study of a Nearctic-Neotropical migratory songbird, the Wilson's warbler (Cardellina pusilla), we investigated the association between polymorphism at two phenological candidate genes, Clock and Adcyap1, and two aspects of the migratory phenotype, timing of spring migration through a stopover site and inferred latitude of the breeding destination. The breeding destination of migrating individuals was identified using feather deuterium ratio (δ (2)H), which reliably reflects breeding latitude throughout the species' western breeding range. Ninety-eight percent of the individuals were homozygous at Clock, and the rare heterozygotes did not deviate from homozygous migration phenology. Adcyap1 was highly polymorphic, and allele size was not significantly associated with migration date. However, Adcyap1 allele size significantly positively predicted the inferred breeding latitude of males but not of females. Moreover, we found a strong positive association between inferred breeding latitude and Adcyap1 allele size in long-distance migrating birds from the northern sector of the breeding range (western Canada), while this was not the case in short-distance migrating birds from the southern sector of the breeding range (coastal California). Our findings support previous evidence for a role of Adcyap1 in shaping the avian migratory phenotype, while highlighting that patterns of phenological candidate gene-phenotype associations may be complex, significantly varying between geographically distinct populations and even between the sexes.
ABSTRACT
In the last decades, food science has greatly developed, turning from the consideration of food as mere source of energy to a growing awareness on its importance for health and particularly in reducing the risk of diseases. Such vision led to an increasing attention towards the origin and quality of raw materials as well as their derived food products. The continuous advance in molecular biology allowed setting up efficient and universal omics tools to unequivocally identify the origin of food items and their traceability. In this review, we considered the application of a genomics approach known as DNA barcoding in characterizing the composition of foodstuffs and its traceability along the food supply chain. Moreover, metabolomics analytical strategies based on Nuclear Magnetic Resonance (NMR) and Mass Spectroscopy (MS) were discussed as they also work well in evaluating food quality. The combination of both approaches allows us to define a sort of molecular labelling of food that is easily understandable by the operators involved in the food sector: producers, distributors, and consumers. Current technologies based on digital information systems such as web platforms and smartphone apps can facilitate the adoption of such molecular labelling.
Subject(s)
DNA Barcoding, Taxonomic , Food Analysis , Metabolomics , Quality Control , Humans , Magnetic Resonance Spectroscopy , Mass SpectrometryABSTRACT
In the present study, we investigated DNA barcoding effectiveness to characterize honeybee pollen pellets, a food supplement largely used for human nutrition due to its therapeutic properties. We collected pollen pellets using modified beehives placed in three zones within an alpine protected area (Grigna Settentrionale Regional Park, Italy). A DNA barcoding reference database, including rbcL and trnH-psbA sequences from 693 plant species (104 sequenced in this study) was assembled. The database was used to identify pollen collected from the hives. Fifty-two plant species were identified at the molecular level. Results suggested rbcL alone could not distinguish among congeneric plants; however, psbA-trnH identified most of the pollen samples at the species level. Substantial variability in pollen composition was observed between the highest elevation locality (Alpe Moconodeno), characterized by arid grasslands and a rocky substrate, and the other two sites (Cornisella and Ortanella) at lower altitudes. Pollen from Ortanella and Cornisella showed the presence of typical deciduous forest species; however in samples collected at Ortanella, pollen of the invasive Lonicera japonica, and the ornamental Pelargonium x hortorum were observed. Our results indicated pollen composition was largely influenced by floristic local biodiversity, plant phenology, and the presence of alien flowering species. Therefore, pollen molecular characterization based on DNA barcoding might serve useful to beekeepers in obtaining honeybee products with specific nutritional or therapeutic characteristics desired by food market demands.
Subject(s)
Bees/physiology , DNA Barcoding, Taxonomic , Plants/classification , Pollen/genetics , Animals , DNA, Plant/genetics , Molecular Sequence Data , Plants/genetics , Pollen/chemistry , Pollination , Sequence Analysis, DNAABSTRACT
We investigated the effects of 1 and 10 mg L(-1) AgNPs on germinating Triticum aestivum L. seedlings. The exposure to 10 mg L(-1) AgNPs adversely affected the seedling growth and induced morphological modifications in root tip cells. TEM analysis suggests that the observed effects were due primarily to the release of Ag ions from AgNPs. To gain an increased understanding of the molecular response to AgNP exposure, we analyzed the genomic and proteomic changes induced by AgNPs in wheat seedlings. At the DNA level, we applied the AFLP technique and we found that both treatments did not induce any significant DNA polymorphisms. 2DE profiling of roots and shoots treated with 10 mg L(-1) of AgNPs revealed an altered expression of several proteins mainly involved in primary metabolism and cell defense.
Subject(s)
Nanoparticles/toxicity , Silver/toxicity , Triticum/drug effects , Amplified Fragment Length Polymorphism Analysis , DNA Damage/drug effects , Electrophoresis, Gel, Two-Dimensional , Germination/drug effects , Mass Spectrometry , Microscopy, Electron, Transmission , Plant Roots/drug effects , Plant Roots/genetics , Plant Roots/physiology , Plant Shoots/drug effects , Plant Shoots/genetics , Plant Shoots/physiology , Proteomics , Seedlings/drug effects , Seedlings/genetics , Seedlings/physiology , Stress, Physiological , Triticum/genetics , Triticum/physiology , Triticum/ultrastructureABSTRACT
BACKGROUND: Stress evaluation is a field of strong interest and challenging due to several methodological aspects in the evaluation process. The aim of this study is to propose a study protocol to test a new method (i.e., the Stress Assessment and Research Toolkit) to assess psychosocial risk factors at work. DESIGN: This method addresses several methodological issues (e.g., subjective vs. objective, qualitative vs quantitative data) by assessing work-related stressors using different kinds of data: i) organisational archival data (organisational indicators sheet); ii) qualitative data (focus group); iii) worker perception (questionnaire); and iv) observational data (observational checklist) using mixed methods research. In addition, it allows positive and negative aspects of work to be considered conjointly, using an approach that considers at the same time job demands and job resources. DISCUSSION: The integration of these sources of data can reduce the theoretical and methodological bias related to stress research in the work setting, allows researchers and professionals to obtain a reliable description of workers' stress, providing a more articulate vision of psychosocial risks, and allows a large amount of data to be collected. Finally, the implementation of the method ensures in the long term a primary prevention for psychosocial risk management in that it aims to reduce or modify the intensity, frequency or duration of organisational demands.
ABSTRACT
BACKGROUND: Identification keys are decision trees which require the observation of one or more morphological characters of an organism at each step of the process. While modern digital keys can overcome several constraints of classical paper-printed keys, their performance is not error-free. Moreover, identification cannot be always achieved when a specimen lacks some morphological features (i.e. because of season, incomplete development or miss-collecting). DNA barcoding was proven to have great potential in plant identification, while it can be ineffective with some closely related taxa, in which the relatively brief evolutionary distance did not produce differences in the core-barcode sequences. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we investigated how the DNA barcoding can support the modern digital approaches to the identification of organisms, using as a case study a local flora, that of Mt. Valerio, a small hill near the centre of Trieste (NE Italy). The core barcode markers (plastidial rbcL and matK), plus the additional trnH-psbA region, were used to identify vascular plants specimens. The usefulness of DNA barcoding data in enhancing the performance of a digital identification key was tested on three independent simulated scenarios. CONCLUSIONS/SIGNIFICANCE: Our results show that the core barcode markers univocally identify most species of our local flora (96%). The trnH-psbA data improve the discriminating power of DNA barcoding among closely related plant taxa. In the multiparametric digital key, DNA barcoding data improves the identification success rate; in our simulation, DNA data overcame the absence of some morphological features, reaching a correct identification for 100% of the species. FRIDA, the software used to generate the digital key, has the potential to combine different data sources: we propose to use this feature to include molecular data as well, creating an integrated identification system for plant biodiversity surveys.
Subject(s)
DNA Barcoding, Taxonomic/methods , DNA, Plant/genetics , Plants/classification , Plants/genetics , Genetic Markers , Italy , Polymerase Chain ReactionABSTRACT
The plant exposures are one of the most frequent poisonings reported to poison control centres. The diagnosis of intoxicated patients is usually based on the morphological analysis of ingested plant portions; this procedure requires experience in systematic botany, because the plant identification is based on few evident traits. The objective of this research is to test DNA barcoding approach as a new universal tool to identify toxic plants univocally and rapidly. Five DNA barcode regions were evaluated: three cpDNA sequences (trnH-psbA, rpoB and matK) and two nuclear regions (At103 and sqd1). The performance of these markers was evaluated in three plant groups: (1) a large collection of angiosperms containing different toxic substances, (2) congeneric species showing different degrees of toxicity and (3) congeneric edible and poisonous plants. Based on assessments of PCR, sequence quality and resolution power in species discrimination, we recommend the combination of plastidial and nuclear markers to identify toxic plants. Concerning plastidial markers, matK and trnH-psbA showed consistent genetic variability. However, in agreement with CBOL Plant Working Group, we selected matK as the best marker, because trnH-psbA showed some problems in sequences sizes and alignments. As a final and relevant observation, we also propose the combination of matK with a nuclear marker such as At103 to distinguish toxic hybrids form parental species. In conclusion, our data support the claim that DNA barcoding is a powerful tool for poisonous plant identifications.