•High allelic diversity, 445 different allelic variants detected•Genetic structure of common bean from Serbia corresponded to gene pool affiliation•Detected additional variation had genetic basis ...within Andean gene pool•Structure according to seed form within Andean and Mesoamerican gene pool•Congruency in clustering according to gene pool applying SSR and phaseolin markers
Genetic diversity and structure of common bean (Phaseolus vulgaris L.) germplasm from Serbia, comprising 118 landraces and 18 cultivars, was assessed with the application of 27 Single Sequence Repeats (SSR) markers. Thirteen accessions from Agricultural Institute of Slovenia were used as references for gene pool determination. Main parameters of genetic diversity were calculated for each SSR loci, i.e. number of different and rare alleles, number of effective alleles, Shannon's information index, observed and expected heterozygosity and polymorphic information content. A total of 445 allelic variants, with 16.5 alleles per locus on average, were detected. Mean gene diversity (He = 0.79) indicated sufficient reservoir of genetic variation preserved in studied bean germplasm. Landraces displayed higher variability compared to cultivars (405 in relation to 233 allelic variants). Genetic structure and relatedness of accessions was assessed by model-based method and hierarchical clustering method in combination with genetic distance calculation. The Bayesian clustering model implemented in STRUCTURE software, on the primary level (K = 2), revealed clear separation of accessions into two groups, corresponding to gene pool affiliation. Mesoamerican gene pool (M) was represented with 23.5% of accessions, while Andean (A) was larger, composed of 68.4% of studied germplasm. Small group (8.1%) showed admixed genetic structure between two gene pools. Additional variation in respect to two recognized gene pools was revealed (K = 3), whose basis was acknowledged to be within Andean gene pool. Further subdivision of accessions (K = 8), mainly according to the seed forms, was observed. Genetic distance analysis associated with Neighbour-joining clustering method revealed grouping pattern of landraces and cultivars corresponding to the gene pool and their seed phenotypes. Classification and structuring of the bean accessions according to and beyond the gene pool of origin should facilitate conservation strategies and breeding of this material. Combining the information of phenotypic variation obtained in previous research and molecular data reveled in this study will assist in selection of parental components for breeding, or in the choice of smaller sample in order to further acknowledge their breeding value. In addition, obtained results of this work should serve as an additional information on common bean germplasm variation in Western Balkans and beyond, in Europe.
The quality of molecular imaging by means of MeV primary ion-induced secondary ion mass spectrometry by coating with gold was evaluated on different reference organic molecules and plant samples. The ...enhancement of the secondary ion yield was evident for the majority of the studied analytes, reaching the highest values at gold thicknesses between 0.5 and 2 nm, and increased the intensity up to 5-fold for reference samples and >2-fold for specific peaks within the plant sample. Improved propagation of the electric field due to the target potential on otherwise electrically insulating plant samples was also evident through improved image resolution and by reducing the background in mass spectra. However, detection of several molecules was significantly decreased at even at 1 nm thick gold layer. The results indicated that an optimized sequence of analysis is required to reliably interpret results.
We report on the analysis of the onset and the intensity of metastable secondary ions, which are desorbed from the target material as a result of bombarding organic samples with MeV primary ions. The ...bimodal time-of-flight mass spectrometer, which can analyze secondary ion in linear and reflectron modes, was used for detection and characterization of such ions. The use of the bimodal mass spectrometer for this specific purpose is demonstrated on amino acid arginine, where three main fragments were detected. We have also analyzed the influence of the primary ion beam on the intensity of metastable ion signal. Results from chlorine ion beams with energies between 3 and 10 MeV have exhibited the importance of electronic sputtering on the product/precursor ion peak intensity ratio, which is significantly decreased when using primary ions with higher energy.
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•Fragmented metastable ions from MeV-SIMS can be observed as long tails of molecular peaks in mass spectra.•Quick characterization of fragments is obtained through linear mode, and more precise identification by reflectron mode.•Fragments with intensities fragment/parent >0.01 can be detected.•Higher energy primary ion beams induce less metastable secondary ions.•Fragmentation pathways are not affected by the primary ion beam energy.
In the present work we have investigated the chemical sensitivity of a mass spectrometry imaging method MeV – SIMS. Primary ion beam within the MeV energy range domain was employed to bombard samples ...of two organic compounds; amino acid arginine, and peptide hormone angiotensin II (human), with average molecular weights of 174.2 u and 1046.2 u respectively. Secondary ion yield was measured as a function of number of molecules per area unit, and the detection limit was determined. For both molecular compounds and two different energies of primary 35Cl ion beam, the secondary ion yield exhibited a significant decrease below the area density of 1015 molecules/cm2, while the density of 1013 molecules/cm2 resulted in molecular peak / background ratio being lesser than 3, which is below the commonly used sensitivity threshold in other techniques. Other experiment related drawbacks to sensitivity were also discussed.
The present work reports on cationization characteristics of organic molecules in alkali metal environment. Samples of arginine and polyethylene glycol polymers with average masses of 600, 1000 and ...1500 were mixed with sodium- and potassium trifluoroacetate of various concentrations, and analyzed through secondary ion mass spectrometry. Bombardment proceeded with ions within both keV and MeV energy domains. Added salts positively affected the secondary ion yield through attachment of Na or K to organic molecule. Ionization differences between swift (MeV) ions and cluster ions, commonly used for secondary ion mass spectrometry analysis, were studied for arginine and PEG 600. While for arginine, secondary ion yield enhancement was only by a factor of 2, when analysing with MeV ions, and no enhancement was observed with keV Bi3 clusters, all polyethylene glycol samples showed increase of the secondary ion yield by factors between 3 and 40, depending on the amount of potassium or sodium that was mixed into the matrix. Additionally, higher amounts of salts also resulted in decreased fragmentation probability for organic molecules, reducing the intensities of specific fragments by more than two orders of magnitude.
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•Added salts significantly increase secondary ion yield for both keV and MeV primary ions.•Ionization increase due to salt addition is greater with keV SIMS.•Added matrix reduces fragmentation – more with MeV-SIMS.•With secondary ion yields of approx. 10−2, submicron molecular imaging is feasible.
Legumes are a rich source of valuable nutrients thus represent important component in human and animal nutrition. The most important and often a limiting factor in legume production is the presence ...of seed pests, such as the bean weevil Acanthoscelides obtectus (Say, 1831). This work tested the suitability of three different legume species (common bean, faba bean and grass pea), the species with a growing interest in the human diet, for the development of the bean weevil, aiming to provide a reliable forecast of its population growth. After four months, been weevils consumed the highest percentage of the common bean kernels (70.79%), followed by the grass pea (53.13%), and faba bean (0.42%). The progeny production and population growth were significantly affected by the tested legume species. After each month, the total number of adults was the highest on the common bean, indicating its best suitability for the weevil’s development. Based on the number of the emerged specimens after each month of the observation, the bean weevil development was unhampered and continuous also on the grass pea. The lowest number of emerged adults, in all observation periods, was in faba bean, indicating its low preference and suitability for the weevil’s development. The population growth of the bean weevil was the highest on the common bean, followed by grass pea, and it fitted best to the quadratic equation model that enabled the prediction of the population growth of the bean weevil for each legume species in the next generations.
This study presents a tentative analysis of the lipid composition of 47 legume samples, encompassing species such as
spp.,
spp.,
spp., and
spp. Lipid extraction and GC/MS (gas chromatography with ...mass spectrometric detection) analysis were conducted, followed by multivariate statistical methods for data interpretation. Hierarchical Cluster Analysis (HCA) revealed two major clusters, distinguishing beans and snap beans (
spp.) from faba beans (
), peas (
), and grass peas (
). Principal Component Analysis (PCA) yielded 2D and 3D score plots, effectively discriminating legume species. Linear Discriminant Analysis (LDA) achieved a 100% accurate classification of the training set and a 90% accuracy of the test set. The lipid-based fingerprinting elucidated compounds crucial for discrimination. Both PCA and LDA biplots highlighted squalene and fatty acid methyl esters (FAMEs) of 9,12,15-octadecatrienoic acid (C18:3) and 5,11,14,17-eicosatetraenoic acid (C20:4) as influential in the clustering of beans and snap beans. Unique compounds, including 13-docosenoic acid (C22:1) and γ-tocopherol, O-methyl-, characterized grass pea samples. Faba bean samples were discriminated by FAMEs of heneicosanoic acid (C21:0) and oxiraneoctanoic acid, 3-octyl- (C18-ox). However, C18-ox was also found in pea samples, but in significantly lower amounts. This research demonstrates the efficacy of lipid analysis coupled with multivariate statistics for accurate differentiation and classification of legumes, according to their botanical origins.
Phenotypic diversity of common bean (
Phaseolus vulgaris
L.) landraces and reference cultivars were evaluated based on variability of twelve traits, chosen according to international descriptor list. ...Landraces were collected form 53 locations in Republic of Serbia. Origin and dissemination of local common bean germplasm was assessed based on variation of phaseolin types. Phaseolin analysis showed that landraces of both Mesoamerican and Andean origin are grown in Serbia, with higher frequency of Andean types. Ratio of three phaseolin types found in studied common bean germplasm suggested its introduction in Serbia from northern borders. Shannon–Weaver index was used as indicator of phenotypic diversity, showing high diversity level among studied germplasm. HOMALS analysis was performed in order to determine discriminative power of traits and structure of studied germplasm. Seed and flower traits, as well as growth habit type had highest, opposed to phaseolin type with lowest discriminative power among genotypes. Genotypes were phenotypically individualized and classified in five homogenous groups according to their profiles, displaying differentiation on the gene pool level, as well.
Epidermis stem cells have a crucial role through the processes of proliferation and differentiation, to replace cells that are constantly lost during tissue turnover or following injury. On the other ...hand, thyroid hormones regulate the proliferation and differentiation of epidermal cells and thus significantly influence the homeostasis of the skin. It is well known that maternal hypothyroidism during pregnancy leads to impaired development of many organ systems in their offspring. However, there is a lack of data about the influence of maternal subclinical hypothyroidism during pregnancy and lactation on the development of the skin and its derivatives in the litter. The aim of this study was to investigate the effects of maternal thyroid dysfunction on the development of the skin and its derivatives in their offspring in the early postnatal period. Antithyroid substance 6-n-propyl-2-thiouracil was added into the drinking water to female Albino Oxfords rats from the beginning of pregnancy and during lactation, with the aim to induce subclinical and overt form of hypothyroidism. Skin samples were taken from male pups within twenty-four hours and seven days after birth. The main findings of this investigation were that both forms of maternal hypothyroidism lead to serious damage of the epidermis in pups in terms of pronounced hyperkeratosis and reduction of the germinal layer along with a reduced number of hair follicles and their delayed morphogenesis. Epidermal impairments were more pronounced in pups with the overt form of hypothyroidism while offspring with the subclinical form had impairments that were less pronounced and delayed in occurrence.
•Detailed phenolic profile of seven legumes' herb and root extracts.•High levels of isoflavones genistein and daidzein in the all examined species.•Good FRAP reducing power and moderate ability to ...scavenge DPPH and NO radicals.•Low anti-inflammatory activity of the examined extracts.
Herb and root parts of legumes are not commonly used in human consumption, since they are traditionally considered waste material. Although inedible, these plant parts have a complex chemical composition, with potential biological activities. They are especially rich in polyphenols, such as flavonoids, isoflavonoids and phenolic acids. In this paper, phenolic profile of the seven selected legumes (soybean (Glycine max L.), common bean (Phaseolus vulgaris L.), pea (Pisum sativum L.), broad bean (Vicia faba L.), chickpea (Cicer arietinum L.), white lupin (Lupinus albus L.) and grass pea (Lathyrus sativus L.)) was examined using LC–MS/MS method. In the examined herb and root extracts 33 phenolic compounds were identified and quantified. High levels of isoflavones genistein and daidzein were found in G. max, P. vulgaris and L. albus root extracts (1.02–1.53, 0.309–0.648 and 2.81mg/g, respectively). C. arietinum, L. albus and P. vulgaris herb extracts were found to be the richest sources of flavonoid compounds (13.1–19.4, 5.98 and 2.36–3.64mg/g, respectively). Since polyphenols have redox properties, antioxidant activity of the abovementioned plants was investigated using DPPH, NO, and FRAP assay. All samples demonstrated the ability to scavenge both DPPH and NO radical, and exhibited a good FRAP reducing power. V. faba exhibited the highest antioxidant activity (DPPH IC50 11.7–20.7μg/mL, NOIC30 0.080–0.34 mg/mL, FRAP 32.6–66.9 AAE/g dw). Anti-inflammatory potential was examined using in vitroCOX-1/12-LOX pathway inhibition assay, resulting in moderate activity of all samples (IC50 2.09–8.00 for all metabolites). The obtained results indicate a strong potential of waste legume material as a promising source for isolation of bioactive antioxidant and anti-inflammatory compounds for their use as therapeutic and dietary supplements in pharmaceutical and food industry.
