Are Portuguese Cowpea Genotypes Adapted to Drought? Phenological Development and Grain Quality Evaluation.

Along with population growth, global climate change represents a critical threat to agricultural production, compromising the goal of achieving food and nutrition security for all. It is urgent to create sustainable and resilient agri-food systems capable of feeding the world without debilitating the planet. The Food and Agriculture Organization of the United Nations (FAO) refers to pulses as a superfood, as one of the most nutritious crops with high health benefits. Considered to be low-cost, many can be produced in arid lands and have an extended shelf-life. Their cultivation helps reduce greenhouse gases and increases carbon sequestration, also improving soil fertility. Cowpea, Vigna unguiculata (L.) Walp. is particularly drought tolerant, with a wide diversity of landraces adapted to different environments. Considering the importance of knowing and valuing the genetic variability of this species in Portugal, this study assessed the impact of drought on four landraces of cowpea (L1 to L4) from different regions of the country and a national commercial variety (CV) as a reference. The development and evaluation of morphological characteristics were monitored in response to terminal drought (imposed during the reproductive phase), and its effects were evaluated on the yield and quality of the produced grain, namely the weight of 100 grains, color, protein content, and soluble sugars. Under drought conditions, the landraces L1 and L2 showed early maturation as a strategy to avoid water deficit. Morphological alteration of the aerial part of the plants was evident in all genotypes, with a rapid reduction in the number of leaves and a reduction in the number of flowers and pods by between 44 and 72%. The parameters of grain quality, the weight of 100 grains, color, protein, and soluble sugars did not vary significantly, except for sugars of the raffinose family that is associated with the adaptive mechanisms of plants to drought. The performance and maintenance of the evaluated characteristics reflect the adaptation acquired in the past by exposure to the Mediterranean climate, highlighting the potential agronomic and genetic value, still little exploited, that could contribute to production stability, preserved nutritional value, and food safety under water stress.

Genetic Diversity and Population Structure of Cowpea (Vigna unguiculata (L.) Walp.) Landraces from Portugal and Mozambique.

Cowpea (Vigna unguiculata (L.) Walp.) is currently a legume crop of minor importance in Europe but a highly relevant staple crop in Africa and the second most cultivated legume in Mozambique. In Portugal and Mozambique, cowpea's phenotypic and genetic variation has been maintained locally by farmers in some areas. We used the molecular markers SSR, SilicoDArT and SNP to analyze the genetic diversity and population structure of 97 cowpea accessions, mainly from Portugal (Southern Europe) and Mozambique (Southern Africa). As far as we know, this is the first time that the genetic variation and the relationship between cowpea landraces collected in Portugal with those originated in Mozambique is reported. Despite the shared historical past, the Portuguese landraces did not share a common genetic background with those from Mozambique, and two different gene pools were revealed. Knowledge of the genetic structure of cowpea landraces offers an opportunity for individual selection within landraces adapted to particular eco-physiological conditions and suggests the existence of a valuable gene pool for exploitation in future Portugal-PALOP (Portuguese-speaking African countries) cowpea breeding programs.

Effect of Addition of Dimethyl Sulfoxide to Simplified Adhesives on Dentin Bond Durability after Three Years of Water Storage.

PURPOSE: To evaluate the effect of inclusion of two dimethyl sulfoxide (DMSO) concentrations in simplified etch-and-rinse adhesives on dentin bonding durability after three years of water storage. MATERIALS AND METHODS: Forty-two caries-free third molars were divided into six experimental groups (n = 7) according to the following factors: 1) adhesive (Adper Single Bond 2 [SB], 3M Oral Care; Prime&Bond 2.1 [PB], Dentsply Sirona); 2) concentration of DMSO (control group: 0.0% DMSO; addition of 0.2% DMSO [0.2] and 2% DMSO [2.0]). After completing restoration, specimens were stored in water (37°C) for 24 h, sectioned into adhesive-dentin sticks (0.8 mm2), tested for microtensile bond strength (µTBS) at 0.5 mm/min, and examined for nanoleakage (NL) using SEM immediately thereafter or after three years of water storage. Data were subjected to a three-way repeated-measures ANOVA and Tukey's test (α = 0.05) for each property evaluated. RESULTS: After three years of water storage, for both adhesives, the incorporation of 2% DMSO maintained the µTBS when compared to immediate µTBS (p > 0.05). In general, SB resulted in a statistically significantly higher mean of µTBS compared to PB, independent of the DMSO concentration after water storage (p < 0.05). Furthermore, the amount of NL was lower and practically limited to the hybrid layer given the concentrations of 0.2% and 2% DMSO for both tested adhesives after three years. CONCLUSION: The incorporation of DMSO in simplified etch-and-rinse adhesives maintains the long-term stability of the dentin bond.

Using microbial seed coating for improving cowpea productivity under a low-input agricultural system.

BACKGROUND: Plant-growth-promoting rhizobacteria (PGPR) and arbuscular mycorrhizal (AM) fungi have the ability to enhance the growth, fitness, and quality of various agricultural crops, including cowpea. However, field trials confirming the benefits of microbes in large-scale applications using economically viable and efficient inoculation methods are still scarce. Microbial seed coating has a great potential for large-scale agriculture through the application of reduced amounts of PGPR and AM fungi inocula. Thus, in this study, the impact of seed coating with PGPR, Pseudomonas libanensis TR1 and AM fungus, Rhizophagus irregularis (single or multiple isolates) on grain yield and nutrient content of cowpea under low-input field conditions was evaluated. RESULTS: Seed coating with P. libanensis + multiple isolates of R. irregularis (coatPMR) resulted in significant increases in shoot dry weight (76%), and in the number of pods and seeds per plant (52% and 56%, respectively) and grain yield (56%), when compared with non-inoculated control plants. However, seed coating with P. libanensis + R. irregularis single-isolate (coatPR) did not influence cowpea grain yield. Grain lipid content was significantly higher (25%) in coatPMR plants in comparison with control. Higher soil organic matter and lower pH were observed in the coatPMR treatment. CONCLUSIONS: Our findings indicate that cowpea field productivity can be improved by seed coating with PGPR and multiple AM fungal isolates under low-input agricultural systems. © 2019 Society of Chemical Industry.

Phytochemical profiling of underexploited Fabaceae species: Insights on the ontogenic and phylogenetic effects over isoflavone levels.

There is an increasing trend towards finding alternative sources of valued phytochemicals due to their diverse potentialities in food industry and pharmaceutical applications. Phenolic compounds, in particular, have been the focus of several profiling reports, but isoflavones characterization has been studied in fewer cases and in a very limited group of plant species. Despite their acknowledged bioactivity, there's actually a strict number of plants validated for their isoflavones contents. In a previous report, we have identified nine Leguminosae species (from genera Biserrula, Lotus, Ornithopus and Scorpiurus) as potential alternative sources of these phenolic compounds. However, the isoflavone profiles are highly modulated by the ontogenic stage. Therefore, the present study was conducted in the same Leguminosae species, but harvested at three sequential vegetative development stages: vegetative elongation, late bud and late flowering, with the main purpose of assessing the evolution of isoflavones content throughout the plant development. In general, the plant species from Biserrula and Lotus genera showed the highest potential as new natural sources of isoflavones, especially owing their high levels of biochanin A. Independently of the plant species, it was possible to identify the phenologic stages where each of the quantified isoflavones is maximized. These findings are useful to predict isoflavone yields according to harvesting time, validating the potential use of the studied plants in innovative food formulations.

European cowpea landraces for a more sustainable agriculture system and novel foods.

BACKGROUND: Genetic diversity is fundamental to breeding programs and consequently has an important role in obtaining new varieties. To properly use the genetic diversity present in germplasm collections, a good knowledge of the agro-morphological traits of each accession is needed. The aim of this study was to explore the production capacity of 24 cowpea landraces from southern Europe, through phenotypic characterization and evaluation in three different locations in Greece and Portugal. RESULTS: Most qualitative parameters tested showed a high stability among the three locations. A wide difference was observed among the three locations with respect to number of days to flowering, ranging from 55 to 99 days. Quantitative traits showed a higher genotype × environment than genetic variance component. In general, an inverse relationship between σ2ge /σ2g ratio (where σ2ge is genotype × genotype interaction and σ2g is genotype impact) and heritability value was observed. Principal component analysis was able to group accessions based on their origin. The first two principal components explained 97.52% of variation, being the number of seeds per plant, plant height and seed protein content, the traits which contributed most to variability. CONCLUSION: The results show that sufficient variation exists in different traits within landraces in the studied cowpea germplasm to pursue a breeding program. However, the quantitative traits showed a higher genotype × environment component. © 2017 Society of Chemical Industry.

Comparison of RAPD, ISSR, and AFLP Molecular Markers to Reveal and Classify Orchardgrass (Dactylis glomerata L.) Germplasm Variations.

Three different DNA-based techniques, Random Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Amplified Fragment Length Polymorphism (AFLP) markers, were used for fingerprinting Dactylis glomerata genotypes and for detecting genetic variation between the three different subspecies. In this study, RAPD assays produced 97 bands, of which 40 were polymorphic (41.2%). The ISSR primers amplified 91 bands, and 54 showed polymorphism (59.3%). Finally, the AFLP showed 100 bands, of which 92 were polymorphic (92%). The fragments were scored as present (1) or absent (0), and those readings were entered in a computer file as a binary matrix (one for each marker). Three cluster analyses were performed to express--in the form of dendrograms--the relationships among the genotypes and the genetic variability detected. All DNA-based techniques used were able to amplify all of the genotypes. There were highly significant correlation coefficients between cophenetic matrices based on the genetic distance for the RAPD, ISSR, AFLP, and combined RAPD-ISSR-AFLP data (0.68, 0.78, 0.70, and 0.70, respectively). Two hypotheses were formulated to explain these results; both of them are in agreement with the results obtained using these three types of molecular markers. We conclude that when we study genotypes close related, the analysis of variability could require more than one DNA-based technique; in fact, the genetic variation present in different sources could interfere or combine with the more or less polymorphic ability, as our results showed for RAPD, ISSR and AFLP markers. Our results indicate that AFLP seemed to be the best-suited molecular assay for fingerprinting and assessing genetic relationship among genotypes of Dactylis glomerata.

Medicago spp. as potential sources of bioactive isoflavones: Characterization according to phylogenetic and phenologic factors.

A high variety of plant species are often proposed as potential natural sources of specific bioactive components, with emphasis in phenolic compounds. However, the ability to produce a determined phytochemical might be variable, even among species with close phylogeny. Furthermore, the metabolic dynamics vary greatly according to phenologic factors. Herein, it was verified whether isoflavone production in Medicago spp. is more associated with phylogenetic or phenologic determinants, to define the optimal productive conditions. Isoflavone profiles were characterized in field-grown Medicago species in three phenologic stages. Isoflavones were extracted by matrix solid-phase dispersion method and analyzed using high-performance liquid chromatography coupled with a diode-array detector. The obtained data were evaluated by a generalized linear model (GLM) and linear discriminant analysis (LDA). Formononetin, genistein and irilone were the most abundant isoflavones, reaching values higher than those present in acknowledged plant sources like soy or red clover. Outputs from GLM and LDA indicate that the phylogenetic factors are the most defining criteria. This study promotes Medicago spp. as potential isoflavone sources, particularly because the effects of these compounds are highly dependent on their type and concentration, with potential application as foodstuff, feedstuff, or in the nutraceutical and pharmaceutical industry.

Advances in isoflavone profile characterisation using matrix solid-phase dispersion coupled to HPLC/DAD in Medicago species.

INTRODUCTION: Analytical methods used in phytochemistry analysis are limited by the sample preparation step, which should ideally be fast, accurate, ecofriendly and achievable using low quantities of the sample. Matrix solid-phase dispersion (MSPD) may be a good alternative for combining extraction and purification procedures, thereby reducing the indicated limitations. OBJECTIVE: Applying an MSPD extraction procedure coupled to high-performance liquid chromatography diode-array detection (HPLC/DAD) as an alternative methodology to evaluate isoflavone profiles. METHODS: Isoflavone profiles were determined for the leaves of nine species of Medicago in the late flower phenological stage (one or more nodes with 50% open flowers, no seed pods). Extraction was performed following MSPD, and isoflavone profiles were characterised using HPLC/DAD. The quantified amounts were compared with previous results in different species commonly recognised as good sources of isoflavones. RESULTS: Formononetin was the major isoflavone in most species, except M. polymorpha and M. truncatula. The isoflavone amounts were significantly different among the assayed species, with M. orbicularis and M. arabica as the major isoflavone sources, while M. rigidula presented the lowest contents. Furthermore, the detected differences allow electing the best species as a primary source of a specific isoflavone. CONCLUSION: The MSPD allowed good extraction efficiency, reproducibility and recovery. Some of the species showed relevant isoflavone contents, even when compared with acknowledged plant sources such as soy or red clover. To the best of our knowledge the results presented are reported for the first time in these species.

Impact of colorectal cancer on patient and family: implications for care.

The quality of life of a colorectal cancer patient and his family is deeply modified when faced with this diagnosis as a result of physical and psychological changes induced by it. The aim of this study was to identify and assess the impact of colorectal cancer patients and their families. The study was a descriptive cross-sectional study and the sample consisted of 153 patients and 96 informal caregivers. In terms of quality of life, those patients who had a stoma reported lower overall quality of life, lower body image, poorer health related quality of life and poor social activity when compared with colorectal cancer patients who had no stoma. The stoma patients also had significantly higher depression and anxiety level were related to the caregivers' burden. The caregivers of stoma patients had more depression and anxiety when compared with the caregivers of non-stoma patients. To sum up, it seems unquestionable that the presence of a stoma significantly alters the patients' quality of life, psychological morbidity, sexuality and body image. The stoma patients' caregivers also suffer from high levels of anxiety and depression.