Nutraceutical Potential of Leafy Vegetables Landraces at Microgreen, Baby, and Adult Stages of Development.

Nutraceutical compounds present in leafy vegetables have gained substantial attention due to the health benefits they offer beyond their nutritional value. The biosynthesis, composition, and concentration of these compounds vary widely among leafy vegetables and carry the influence of genetic, agronomic, and environmental factors. Recently, micro-vegetables are gaining importance among consumers worldwide and are used in gastronomy at different development stages. Another tendency is the utilization of local genetic resources as an integral component of agricultural biodiversity crucial for sustainable production. The present study identifies the nutraceutical potential of 10 leafy vegetables at the microgreen, baby, and adult development stages using local genetic resources from the Spanish Vegetable Genebank (CITA, Aragón). Specifically, two landraces for each of the following crops were used: chard (Beta vulgaris), spinach (Spinacia oleracea), lettuce (Lactuca sativa), borage (Borago officinalis), and chicory (Cichorium intybus). The results reinforce the value of traditional local genetics and demonstrate the potential of these leafy vegetables as a source of functional compounds (fatty acids, vitamin C, carotenoids, polyphenols, antioxidant activity, and tocopherols). The observed variability depending on the crop and the developmental stage recommends the necessity of having a varied diet, since each leafy vegetable product offers a unique nutritional profile.

Bioactive compounds, antioxidant activity, and mineral content of bróquil: A traditional crop of Brassica oleracea var. italica.

Brassicaceae edible plants are rich in bioactive compounds and promote health benefits. However, there is less interest in expanding knowledge about the Brassica cultivars to date. In particular, underutilized species and local cultivars could constitute a source of agrodiversity in adapting to the territory with likely higher contents of nutraceutical compounds. In this context, Bróquil (Brassica oleracea var. italica) is a traditional Brassicaceae crop grown in the Spanish region of Aragón. Currently, it is cultivated mainly in family orchards for autoconsumption and, in minority, in small farms for local markets. This study evaluates a collection of 13 bróquil landraces from the Spanish Vegetable Genebank of the Agrifood Research and Technology Center of Zaragoza (BGHZ-CITA), describing their mineral contents, bioactive compounds, and antioxidant activities, including a broccoli commercial variety "Parthenon" as the control. The study reports data on the health-promoting nutrients and antioxidants of bróquil for the first time. Under our experimental conditions, we found that bróquil has a great variability for these compounds that showed on average similar or higher levels than the broccoli control. The different bróquil landraces also revealed variability in both intraccessions and interaccessions due to the lack of a formal breeding selection. Despite this variability, we highlight accession HB5 that corresponds to Headed Bróquil BGHZ6685. In particular, we can stand out its antioxidant activity of 87.07 ± 0.81%I, total phenolic content of 13.21 ± 0.53 mg GAE g-1 dw, total flavonoid content of 14.50 ± 1.29 mg QE g-1 dw, total glucosinolate content of 43.70 ± 1.09 mg SnE g-1 dw, and vitamin C content of 7.21 ± 0.13 mg AA g-1 dw. Regarding bróquil mineral composition, K was the highest macroelement (22.66-33.62 mg g-1 dw), followed by Ca, P, and S whose values were relatively lower compared to K. Mg and Na showed the lowest values. Among the microelements evaluated (Mn, Zn, and Fe), iron was the most abundant detected, higher in all bróquil accessions than in broccoli, except for one accession. Therefore, the results reported for bróquil landraces show promising nutritional quality. This could lead to an increase in agrobiodiversity and contribute to a more diversified and healthy diet.

Evaluation of Borage (Borago officinalis L.) Genotypes for Nutraceutical Value Based on Leaves Fatty Acids Composition.

Borage (Borago officinalis L.) is a traditional vegetable grown and consumed in some Spanish regions. The objective of this study was to determine the variability and evolution of fatty acid composition in a borage germplasm collection formed by wild types, breeding lines, commercial varieties, and landraces. Fatty acids were analysed in petioles, the commonly edible part of the leaves, and the leaf blades, the by-product of the borage industry, in two growth stages: at the optimal harvest period (120 days after sowing) and at the end of the harvest period (150 days after sowing). The results showed that for each of the eight fatty acids identified, there were significant differences among the twelve borage genotypes depending on the developmental plant stage at sampling date and the part of the leaf analysed, the interaction effect also being statistically significant. The main polyunsaturated fatty acids identified were: linoleic acid (18:2 n6, LA), α-linolenic acid (18:3 n3, ALA), γ-linolenic acid (18:3 n6, GLA), and stearidonic acid (SDA, 18:4, n-3), account for approximately 70% of polyunsaturated fatty acids. Blue-flowered genotypes differ from white-flowered genotypes by their high content of ALA and SDA, which can be exploited in borage breeding programs. Petioles from young plants present higher n6 fatty acids, while older plants produce a great amount of n3 fatty acids. Besides, the higher content of ALA in the leaf blades gives them a good dietary potential. All these fatty acids, with multiple health benefits, support the nutraceutical interest of borage leaves (both petioles and leaf blades) for human consumption, animal feeding, medicine, and pharmacy.

Assessment of Capsaicinoid and Capsinoid Accumulation Patterns during Fruit Development in Three Chili Pepper Genotypes (Capsicum spp.) Carrying Pun1 and pAMT Alleles Related to Pungency.

Quantification, using an accurate analytical approach, of capsinoids and capsaicinoids was performed on three chili pepper (Capsicum spp.) genotypes: "Chiltepín", "Tampiqueño 74", and "Bhut Jolokia" at various stages of fruit development. The accumulation of capsinoids, in all these peppers started between 10 to 20 days post-anthesis (dpa), increased and reached the highest capsinoid amount at 40 dpa, and then decreased until 60 dpa. Conversely, capsaicinoids could already be determined at 10 dpa in "Bhut Jolokia" and their accumulation pattern was different from that of the capsinoids in this genotype. The capsiate/dihydrocapsiate ratio presented a higher variation between genotypes and developmental stages than the capsaicin/dihydrocapsaicin ratio. Capsinoid ratios (4-24%) and Pun1/pAMT genotyping were determined. These results provide information on the progress of the accumulation of capsinoids in the aforementioned pungent and superhot cultivars and could support future breeding studies toward the understanding of the factors affecting their accumulation.

Quantitation of capsiate and dihydrocapsiate and tentative identification of minor capsinoids in pepper fruits (Capsicum spp.) by HPLC-ESI-MS/MS(QTOF).

A highly sensitive, selective method has been developed and validated for determination of capsiate and dihydrocapsiate for the first time using HPLC-ESI/MS(QTOF). For both capsinoids, LODs and LOQs were 0.02 and 0.05 µmol/l, respectively. The intra- and interday repeatability values (RSD %) were 0.26-0.41% for retention time, and 2.25-2.11% for peak area. Recoveries were up to 98% and 97% for capsiate and dihydrocapsiate, respectively. This method was successfully applied to quantify capsiate and dihydrocapsiate in eight pepper fruit accessions. Capsinoids were found in the range of 1.21-544.59 µg/g DW for capsiate and of 0.61-81.95 µg/g DW for dihydrocapsiate. In the 'Tabasco' accession, capsiate and dihydrocapsiate were quantified for the first time, ranging from 3.09 to 58.76 and 1.80 to 6.94 µg/g DW, respectively. Additionally, the ESI-MS/MS(QTOF) analysis has allowed the tentative identification of two other minor capsinoids by exact mass and fragmentation pattern, in the 'Bhut Jolokia' accession.

Spanish traditional tomato. Effects of genotype, location and agronomic conditions on the nutritional quality and evaluation of consumer preferences.

Traditional tomatoes are highly valued for their organoleptic quality and cultural links with a territory. At present, strong competition has put these crops at risk, and it is necessary to differentiate the local cultivars and improve their nutritional value. This work focused on the nutritional study of four selected lines of a local tomato grown in two locations and in two agronomic conditions to nutritionally characterize the tomatoes and to study the effect of location and cultivation on nutritional parameters. Data on nutritional characterization revealed significant effects of location and treatment in most compounds. Tomatoes grown in traditional areas showed a significantly higher concentration of some phenolic acids and beta-carotene. Lycopene contents were not location dependent. The open field test showed significant differences in all the components. Regarding the best nutritional genotypes, all the components were dependent on lines, and significant differences were confirmed between them.

Plant Genebanks: Present Situation and Proposals for Their Improvement. the Case of the Spanish Network.

Genebanks were created by the middle of the twentieth century to preserve cultivated biodiversity when landraces began to be substituted by modern varieties. This move was generally accepted as a necessary step to safeguard the future. After about 75 years of collecting and maintaining genetic resources, the increasing ability of biotechnology to create new variability brings the roles of genebanks in the present and near future into question. As a continuation of several workshops that started in 2014, staff of some representative genebanks have met to discuss how the Spanish Plant Genetic Resources Network can be improved, identifying the following major shortcomings: lack of efficient coordination in the distribution of species among genebanks; too many genebanks; existence of detected and undetected duplicates; insufficient rate of regeneration; insufficient phenotyping, genotyping, and epiphenotyping; unsatisfactory rate of use by end users; and, insufficient funding. As a considerable increase in public funding is unlikely, we propose some strategies to increase the efficiency of the system. The most urgent tasks are to strengthen the rationalization of the network by establishing a clear hierarchy and functions, to improve the information in the base collection by deep characterization including not only phenotypes but also uses and utilities, to progressively replace the active collections with focused core collections constructed to meet users' needs, to optimize regeneration protocols, to limit new collecting expeditions of Spanish crop wild relatives to those growing in threatened habitats, and to develop user-friendly platforms to access germplasm documentation, including a unified system of descriptors and classification categories. Current advances in biotechnology, and especially those in gene editing will have without doubt an impact on the role of genebanks. However, the high number of genes and gene combinations created by evolution they hold cannot be produced by these techniques at present. So, these reservoirs of variability will continue to be indispensable for the near-medium future while the function of all the genes is unveiled. In turn, biotechnologies and gene editing will allow us to take advantage of the information held in genebanks in a more efficient and fast way, contributing to a better rationalization and functioning.

Recovery of a Common Bean Landrace (Phaseolus vulgaris L.) for Commercial Purposes.

The "Caparrona" bean is a landrace that was grown largely in Monzón, and for that reason, it is also known by the name of "Caparrona de Monzón." Historical references mention that in the thirties of the last century, Caparrona beans reached a production higher than 200,000 kg. Nevertheless, the increasing modernization of agriculture at the end of the 20th century enhanced its replacement by newer varieties. As a result, only a few local growers continued producing Caparrona beans mainly for family use. However, in recent years, the high demand for local products, grown with environmentally friendly farming techniques, has reawakened interest in this local bean. In order to recover the Caparrona bean crop, a study was conducted with the aim of assessing this landrace, along with all the processes, from collecting seeds to securing the in situ and ex situ conservation. Six bean samples were initially collected from local farmers and the traditional knowledge was also recorded. After the first seed-borne virus test, two samples were rejected because of the positive results for Bean Common Mosaic Virus (BCMV). The four remaining samples were evaluated in a randomized complete block design with three replications at two locations. All through the growth phase of the plants, samples were taken for a virus test. Two samples tested positive for BCMV and were discarded. Between the two healthy seed samples, regarding morphology, chemical composition, and agronomic data, no significant statistical differences were found. Therefore, both samples were selected for commercial production. The seeds obtained from the assays were transferred to a recently created producers' association, which registered a private label to commercialize the Caparrona beans as a gourmet product. Seeds are also available from the Spanish BGHZ-CITA public genebank.

Ontogenetic Variation of Individual and Total Capsaicinoids in Malagueta Peppers (Capsicum frutescens) during Fruit Maturation.

The ontogenetic variation of total and individual capsaicinoids (nordihydrocapsaicin (n-DHC), capsaicin (C), dihydrocapsaicin (DHC), homocapsaicin (h-C) and homodihydrocapsaicin (h-DHC)) present in Malagueta pepper (Capsicum frutescens) during fruit ripening has been studied. Malagueta peppers were grown in a greenhouse under controlled temperature and humidity conditions. Capsaicinoids were extracted using ultrasound-assisted extraction (UAE) and the extracts were analyzed by ultra-performance liquid chromatography (UHPLC) with fluorescence detection. A significant increase in the total content of capsaicinoids was observed in the early days (between 12 and 33). Between day 33 and 40 there was a slight reduction in the total capsaicinoid content (3.3% decrease). C was the major capsaicinoid, followed by DHC, n-DHC, h-C and h-DHC. By considering the evolution of standardized values of the capsaicinoids it was verified that n-DHC, DHC and h-DHC (dihydrocapsaicin-like capsaicinoids) present a similar behavior pattern, while h-C and C (capsaicin-like capsaicinoids) show different evolution patterns.

Doubled haploid production from Spanish onion (Allium cepa L.) germplasm: embryogenesis induction, plant regeneration and chromosome doubling.

The use of doubled haploids in onion breeding is limited due to the low gynogenesis efficiency of this species. Gynogenesis capacity from Spanish germplasm, including the sweet cultivar Fuentes de Ebro, the highly pungent landrace BGHZ1354 and the two Valenciana type commercial varieties Recas and Rita, was evaluated and optimized in this study. The OH-1 population, characterized by a high gynogenesis induction, was used as control. Growing conditions of the donor plants were tested with a one-step protocol and field plants produced a slightly higher percentage of embryogenesis induction than growth chamber plants. A one-step protocol was compared with a two-step protocol for embryogenesis induction. Spanish germplasm produced a 2-3 times higher percentage of embryogenesis with the two-step protocol, Recas showing the highest percentage (2.09%) and Fuentes de Ebro the lowest (0.53%). These percentages were significantly lower than those from the OH-1 population, with an average of 15% independently of the protocol used. The effect of different containers on plant regeneration was tested using both protocols. The highest percentage of acclimated plants was obtained with the two-step protocol in combination with Eco2box (70%), whereas the lowest percentage was observed with glass tubes in the two protocols (20-23%). Different amiprofos-methyl (APM) treatments were applied to embryos for chromosome doubling. A similar number of doubled haploid plants were recovered with 25 or 50 µM APM in liquid medium. However, the application of 25 µM in solid medium for 24 h produced the highest number of doubled haploid plants. Somatic regeneration from flower buds of haploid and mixoploid plants proved to be a successful approach for chromosome doubling, since diploid plants were obtained from the four regenerated lines. In this study, doubled haploid plants were produced from the four Spanish cultivars, however further improvements are needed to increase their gynogenesis efficiency.

New insights into Capsicum spp relatedness and the diversification process of Capsicum annuum in Spain.

The successful exploitation of germplasm banks, harbouring plant genetic resources indispensable for plant breeding, will depend on our ability to characterize their genetic diversity. The Vegetable Germplasm Bank of Zaragoza (BGHZ) (Spain) holds an important Capsicum annuum collection, where most of the Spanish pepper variability is represented, as well as several accessions of other domesticated and non-domesticated Capsicum spp from all over the five continents. In the present work, a total of 51 C. annuum landraces (mainly from Spain) and 51 accessions from nine Capsicum species maintained at the BGHZ were evaluated using 39 microsatellite (SSR) markers spanning the whole genome. The 39 polymorphic markers allowed the detection of 381 alleles, with an average of 9.8 alleles per locus. A sizeable proportion of alleles (41.2%) were recorded as specific alleles and the majority of these were present at very low frequencies (rare alleles). Multivariate and model-based analyses partitioned the collection in seven clusters comprising the ten different Capsicum spp analysed: C. annuum, C. chinense, C. frutescens, C. pubescens, C. bacatum, C. chacoense and C. eximium. The data clearly showed the close relationships between C. chinense and C. frutescens. C. cardenasii and C. eximium were indistinguishable as a single, morphologically variable species. Moreover, C. chacoense was placed between C. baccatum and C. pubescens complexes. The C. annuum group was structured into three main clusters, mostly according to the pepper fruit shape, size and potential pungency. Results suggest that the diversification of C. annuum in Spain may occur from a rather limited gene pool, still represented by few landraces with ancestral traits. This ancient population would suffer from local selection at the distinct geographical regions of Spain, giving way to pungent and elongated fruited peppers in the South and Center, while sweet blocky and triangular types in Northern Spain.

Potential Sources of Resistance for Melon to Nonpersistently Aphid-borne Viruses.

Two hundred and sixty-eight Cucumis melo and wild relative accessions were evaluated for resistance to Cucumber mosaic virus (CMV), Papaya ringspot virus strain W (PRSV-W), Watermelon mosaic virus (WMV), and Zucchini yellow mosaic virus (ZYMV). Symptom development and systemic infection based on double antibody sandwich enzyme-linked immunosorbent assays were monitored. Sources of resistance were scarce. However, a number of them were found among the C. melo accessions tested. Thus, the accession C-189 behaved similarly to PI 161375 and showed resistance to "common" CMV strains although it was infected by a "song" type CMV. For WMV, the accessions C-768 and C-425, although infected, exhibited very mild symptoms, and recovery from infection occurred in some occasions in the former. A similar trait was also found in C-105, although in this case symptoms rarely appear and part of the plants are not infected. In addition, C-105 was highly resistant to virus transmission by Aphis gossypii, similarly to PI 161375. Accessions C-885 and C-769 exhibited resistance to PRSV-W, WMV, and ZYMV; therefore they are potential sources of multiple resistance. Resistance traits were also found in wild relatives that could be exploited when interspecific barriers with C. melo can be circumvented.