First report of Colletotrichum incanum causing leaf spots on common bean in Europe (Slovenia).

A Phaseolus vulgaris L. leaf showing necrotic spots was collected in an experimental bean field in central Slovenia in August 2021. The field contained diverse common bean lines sourced from genebank collections, with each line represented by 10 plants. While symptomatic leaves were seen across various lines, the reported species derived exclusively from a Huasca Huallaga Colorado plant (single-seed descent, USDA accession PI153714, doi: 10.18730/H7P9N), a Peruvian landrace. After incubating the leaf for 2 d at ambient temperature in a moist chamber, setose acervuli developed producing curved, distally tapering and proximately truncated conidia. Single-spore cultures developed equally-shaped conidia measuring 14.5-21.5 (avg. 18.5) × 3-4 (avg. 3.5) µm (n=60) on corn meal agar when mounted in lactic acid. Obtained morphological characters and sequences of the partial actin (GenBank accession, OR208162), beta-tubulin (OR208164), and histone 3 (OR208165) gene identified the isolate as Colletotrichum incanum H.-C. Yang, J.S. Haudenshield & G.L. Hartman. Sequences were identical to those from CBS 133485 (= NRRL 62592, IL6A), ex-type strain of C. incanum (KC110823, KC110814, and KC110796). Partial sequences of the chitin synthase (CHS) gene (OR208163), not available for the ex-type strain, was identical to sequences of other C. incanum strains reported from China (KP145539, ON189040, and OQ613679-OQ613686) or differed in two nucleotide positions (OL471268 and OL471269). The strain from Slovenia was deposited in the CBS biobanks of the Westerdijk Fungal Biodiversity Institute (Utrecht, The Netherlands) as CBS 150848. Pathogenicity of the strain was tested by spraying ca. 3×105 conidia as a watery spore suspension onto each leaf of 6 greenhouse-grown and 3 wk-old common bean plantlets (cv. KIS Amand). Nonsterile commercial substrate (Potgrond H, AGRO-FertiCrop) was used and natural light conditions at ambient temperatures (18-23°C) applied. Sterile water was sprayed on 6, equally grown negative control plants. Treated plants showed small brownish spots after 3 wks similar to those described by Yang et al. (2014) on soybean. Setose acervuli formed within 5 days after detached leaves were incubated in moist chambers. No acervuli formed on negative control plants. Conidia re-isolated from these acervuli and obtained cultures were morphologically identical to originally obtained conidia and cultures and those used for performing the pathogenicity test. Anthracnose is an important disease of common bean attributed to various races of C. lindemuthianum (Sacc. & Magnus) Briosi & Cavara (Nunes et al. 2021). Reporting an additional agent potentially able to cause diseases in common bean and so far not known to occur in Europe is of high relevance as the various genetic bean lines used in Europe may show alternative susceptibility levels to it. However, symptoms caused by C. incanum seem to be less severe as those caused by C. lindemuthianum and the species belongs to the C. spaethianum species complex, whose members have so far not been considered as pathogens of economic importance (Talhinhas & Baroncelli 2021). Yang et al. (2014) based C. incanum on isolates from soybean petioles (USA) and associated it with common bean by re-identifying strain ATCC 64682 obtained by Tu (1990) in Canada. Database queries revealed that it was encountered also on sugar beet (USA; Hanson et al. 2023) and on various crop hosts in China (e.g., chili; Diao et al. 2017), but not in Europe. The work was funded by the Ministry of Agriculture, Forestry and Food and conducted as part of research programs P4-0072 and P4-0431, financed by the Slovenian Research and Innovation Agency ARIS, and the Horizon 2020 project INCREASE funded by the European Union.

Factors Influencing the Emergence of Heterogeneous Populations of Common Bean (Phaseolus vulgaris L.) and Their Potential for Intercropping.

The common bean is an important legume valued for its protein-rich seeds and its ability to fix nitrogen, making it a key element of crop rotation. In conventional agriculture, the emphasis is on uniformity and genetic purity to optimize crop performance and maximize yields. This is due to both the legal obligations to register varieties and the challenges of implementing breeding programs to create genetically diverse varieties. This paper focuses on the factors that influence the occurrence of heterogeneous common bean populations. The main factors contributing to this diversity have been described, including local adaptations, variable weather conditions, different pollinator species, and intricate interactions between genes controlling seed coat colour. We also discuss the benefits of intercropping common beans for organic farming systems, highlighting the improvement in resistance to diseases, and adverse environmental conditions. This paper contributes to a better understanding of common bean seed heterogeneity and the legal obligation to use heterogeneous populations.

How Nutritious Are French Beans (Phaseolus vulgaris L.) from the Citizen Science Experiment?

French beans are tender, immature, edible pods that are harvested early in the plant's growth cycle and are usually eaten cooked. The growth habits of French beans were studied for the first time in a Citizen Science experiment, and 19 pod samples were collected for further nutritional analysis. Various macronutrients (e.g., protein, ash, fat, carbohydrates, amino acids) and multi-element profiles were determined. A survey of their growing habits revealed that beans are usually planted once or twice a year in May and June at a length of 5-10 m, with a predominance of dwarf beans cultivation over climbing varieties, and pest resistance and stringless pods are the most important characteristics when deciding on a bean. Homogenised freeze-dried pod samples contained 16.1-23.1% protein, 4.5-8.2% ash, 0.1-1.1% fat, and 62.0-70.6% carbohydrates and had a caloric value of 337-363 kcal/100 g. Of the 17 free amino acids identified, 8 were essential (histidine, threonine, methionine, valine, lysine, isoleucine, leucine, phenylalanine) and 9 were non-essential (cysteine, aspartic acid, serine, glutamic acid, glycine, arginine, alanine, proline, tyrosine); meanwhile, of the 12 elements, 5 were macroelements and 7 were microelements. The predominant free amino acids were aspartic acid, glutamic acid, and serine. In the multiple comparisons (Box and Whisker plot), the parameters caloric value and iron showed the strongest response. A very strong positive significant Pearson correlation (≥0.95) was found for five pairs of variables within the free amino acids. Comparison of the nutrient data obtained in the pods showed near-perfect or high complementarity (85.2-103.4%) with the food composition databases for half of the parameters, suggesting that the home-grown French beans from the Citizen Science experiment are a highly nutritious vegetable.

Agro-Morphological and Molecular Characterization Reveal Deep Insights in Promising Genetic Diversity and Marker-Trait Associations in Fagopyrum esculentum and Fagopyrum tataricum.

Characterisation of genetic diversity is critical to adequately exploit the potential of germplasm collections and identify important traits for breeding programs and sustainable crop improvement. Here, we characterised the phenotypic and genetic diversity of a global collection of the two cultivated buckwheat species Fagopyrum esculentum and Fagopyrum tataricum (190 and 51 accessions, respectively) using 37 agro-morphological traits and 24 SSR markers. A wide range of variation was observed in both species for most of the traits analysed. The two species differed significantly in most traits, with traits related to seeds and flowering contributing most to differentiation. The accessions of each species were divided into three major phenoclusters with no clear geographic clustering. At the molecular level, the polymorphic SSR markers were highly informative, with an average polymorphic information content (PIC) of over 0.65 in both species. Genetic diversity, as determined by Nei's expected heterozygosity (He), was high (He = 0.77 and He = 0.66, respectively) and differed significantly between species (p = 0.03) but was homogeneously distributed between regions, confirming the lack of genetic structure as determined by clustering approaches. The weak genetic structure revealed by the phenotypic and SSR data and the low fixation indices in both species suggested frequent seed exchange and extensive cultivation and selection. In addition, 93 and 140 significant (p < 0.05) marker-trait associations (MTAs) were identified in both species using a general linear model and a mixed linear model, most of which explained >20% of the phenotypic variation in associated traits. Core collections of 23 and 13 phenotypically and genetically diverse accessions, respectively, were developed for F. esculentum and F. tataricum. Overall, the data analysed provided deep insights into the agro-morphological and genetic diversity and genetic relationships among F. esculentum and F. tataricum accessions and pointed to future directions for genomics-based breeding programs and germplasm management.

Comparative and population genomics of buckwheat species reveal key determinants of flavor and fertility.

Common buckwheat (Fagopyrum esculentum) is an ancient crop with a world-wide distribution. Due to its excellent nutritional quality and high economic and ecological value, common buckwheat is becoming increasingly important throughout the world. The availability of a high-quality reference genome sequence and population genomic data will accelerate the breeding of common buckwheat, but the high heterozygosity due to the outcrossing nature has greatly hindered the genome assembly. Here we report the assembly of a chromosome-scale high-quality reference genome of F. esculentum var. homotropicum, a homozygous self-pollinating variant of common buckwheat. Comparative genomics revealed that two cultivated buckwheat species, common buckwheat (F. esculentum) and Tartary buckwheat (F. tataricum), underwent metabolomic divergence and ecotype differentiation. The expansion of several gene families in common buckwheat, including FhFAR genes, is associated with its wider distribution than Tartary buckwheat. Copy number variation of genes involved in the metabolism of flavonoids is associated with the difference of rutin content between common and Tartary buckwheat. Furthermore, we present a comprehensive atlas of genomic variation based on whole-genome resequencing of 572 accessions of common buckwheat. Population and evolutionary genomics reveal genetic variation associated with environmental adaptability and floral development between Chinese and non-Chinese cultivated groups. Genome-wide association analyses of multi-year agronomic traits with the content of flavonoids revealed that Fh05G014970 is a potential major regulator of flowering period, a key agronomic trait controlling the yield of outcrossing crops, and that Fh06G015130 is a crucial gene underlying flavor-associated flavonoids. Intriguingly, we found that the gene translocation and sequence variation of FhS-ELF3 contribute to the homomorphic self-compatibility of common buckwheat. Collectively, our results elucidate the genetic basis of speciation, ecological adaptation, fertility, and unique flavor of common buckwheat, and provide new resources for future genomics-assisted breeding of this economically important crop.

Influence of Hulling, Cleaning and Brushing/Polishing of (Pseudo)Cereal Grains on Compositional Characteristics.

(Pseudo)cereal grains have been the basis of human nutrition for thousands of years. The various types of cereals are usually harvested by grain harvesters and must be technologically processed in different ways before consumption. In addition to genotype and growing conditions, the compositional characteristics of the (pseudo)cereal grains are highly dependent on the processes used. In the present study, the effects of hulling, cleaning and brushing/polishing wheat, spelt, oat, barley, common and Tartary buckwheat grains and their fractions on physical parameters (thousand kernel weight, kernel width, fractional yield) and nutritional characteristics (protein, fat, ß-glucan, macro- and microelements) were investigated. Grain samples contained 22.7-148.5 mg/g protein, 4.5-69.6 mg/g fat and 0.5-54.4 mg/g ß-glucan. The content of macro- (K, Mg, P, S, Ca) and microelements (Mn, Fe, Zn, Na, Cu, Cr, Mo) varied considerably among the studied (pseudo)cereals and their grain fractions. Analysis of variance showed that species and fractions significantly influenced most of the analyzed characteristics. However, the composition of the edible fractions was not significantly dependent on the brushing/polishing process.

Compositional Traits of Grains and Groats of Barley, Oat and Spelt Grown at Organic and Conventional Fields.

Barley, oats, or spelt consumed as minimally processed whole grains provide several health benefits, especially when grown under organic field management conditions. Therefore, the effects of organic and conventional farming on the compositional traits (protein, fibre, fat, and ash) of barley, oat, and spelt grains and groats were compared using three winter barley varieties ('Anemone', 'BC Favorit', and 'Sandra'), two spring oat varieties ('Max' and 'Noni'), and three spelt varieties ('Ebners Rotkorn', 'Murska bela', and 'Ostro'). Groats were produced from harvested grains by a combination of threshing, winnowing, and brushing/polishing. Multitrait analysis showed significant differences between species, field management practices, and fractions, with clear compositional differences between organic and conventional spelt. Barley and oat groats had a higher thousand kernel weight (TKW) and ß-glucan, but lower crude fibre, fat, and ash contents than the grains. The composition of the grains of the different species differed significantly for more traits (TKW, fibre, fat, ash, and ß-glucan) than that of the groats (TKW and fat), while field management only affected the fibre content of the groats and the TKW, ash, and ß-glucan contents of the grains. The TKW, protein, and fat contents of the different species differed significantly under both conventional and organic growing conditions, while the TKW and fibre contents of grains and groats differed under both systems. The caloric value of the final products of barley, oats, and spelt groats ranged from 334-358 kcal/100 g. This information will be useful for not only the processing industry, but also for breeders and farmers, and last, but not least, for consumers.

Nutrients, Phytic Acid and Bioactive Compounds in Marketable Pulses.

Pulses are edible seeds of plants belonging to the legume family, which are of great importance for human and animal nutrition. In this study, several nutrients, antinutrients and bioactive compounds were quantified in the seeds of ten pulses, i.e., common and runner beans, field peas, lupins (white, blue and yellow), faba beans, lentils (brown and red) and chickpeas. Homogenised, air-dried seed samples were analysed for various parameters: protein (18.0-43.1%), fat (0.6-18.5%) and phytic acid content (507-2566 mg/100 g dry weight (DW)), phenolic profile (27 phenolic compounds in total) and multi-mineral composition. The analysed phenolic compounds mainly belong to phenolic acids (hydroxybenzoic acids and hydroxycinnamic acids) and/or flavonoids (flavones, flavonols and flavanols). Total phenolic content (TPC) ranged from 719 µg/g DW in chickpeas to 5012 µg/g DW in common beans. A total of ten elements belonging to macro- (Mg, P, S, K and Ca) and micro-minerals (Cr, Mn, Fe, Zn and Mo) were determined. Using cluster analysis, pulses were divided into three groups according to the parameters studied: 1. common and runner bean; 2. field pea, white and blue lupin, faba bean, red and brown lentil and chickpea; and 3. yellow lupin. The most varying phytochemicals in terms of their content in the analysed pulses were phytic acid, quinic acid, catechin and TPC. A perfect positive significant Pearson correlation (1.00) was observed for six pairs of variables within the group of phenolic compounds.

Determination of Some Elements in Legumes Using ICP-MS and EDXRF Methodology Applications.

The current study involves two analytical research techniques, inductively coupled plasma-mass spectrometry (ICP-MS) and energy dispersive X-ray fluorescence (EDXRF) spectroscopy, used to determine the elemental composition of different legumes usually produced and consumed in Slovenia. Results indicate that data obtained using these methods are in agreement with certified reference materials. In total, nineteen elements were determined from twenty legume samples. An intercomparison between four macro- (P, S, K, Ca) and three microelements (Fe, Zn, Mo) measured using ICP-MS and EDXRF methods showed a strong correlation. The EDXRF was found to be a cheaper, simpler and more environmentally friendly method for determination of elements P, S, Cl, K, Ca, Fe, Zn, Mo, Sr, Rb, Ti and Br in legumes, while for the identification and determination of Na, Mg, V, Cr, Mn, Co and Cu content ICP-MS was the method of choice due to its excellent sensitivity and accuracy. Using principal component analysis (PCA), the samples of the studied legumes were classified into four groups according to their elemental composition.

Diversification and genetic structure of the western-to-eastern progression of European Phaseolus vulgaris L. germplasm.

BACKGROUND: Common bean (Phaseolus vulgaris L.) is the most important food legume for direct human consumption around the world, as it represents a valuable source of components with nutritional and health benefits. RESULTS: We conducted a study to define and explain the genetic relatedness and diversification level of common bean (Phaseolus vulgaris L.) germplasm from Portugal to Ukraine, along a western-to-eastern line of southern European countries, including Poland. This was based on the P. vulgaris genetic structure, and was designed to better describe its distribution and domestication pathways in Europe. Using the multi-crop passport descriptors that include geographic origin and different phaseolin types (corresponding to the Mesoamerican and Andean gene pools), 782 accessions were obtained from nine gene banks and 12 geographic origins. We selected 33 genome/ gene-related/ gene-pool-related nuclear simple sequence repeat markers that covered the genetic diversity across the P. vulgaris genome. The overall polymorphic information content was 0.800. Without specifying geographic origin, global structure cluster analysis generated 10 genetic clusters. Among the PvSHP1 markers, the most informative for gene pool assignment of the European P. vulgaris germplasm was PvSHP1-B. Results of AMOVA show that 89% of the molecular variability is shared within the 782 accessions, with 4% molecular variability among the different geographic origins along this western-to-eastern line of southern Europe (including Poland). CONCLUSIONS: This study shows that the diversification line of the European P. vulgaris germplasm followed from the western areas of southern Europe (Portugal, Spain, Italy, Slovenia) to the more eastern areas of southern Europe. This progression defines three geographically separated subgroups, as the northern (Poland, Ukraine, Romania), southern (Albania, Bulgaria), and central (Bosnia and Herzegovina, Serbia, Hungary) areas of eastern Europe.

Morphological Seed Characterization of Common (Phaseolus vulgaris L.) and Runner (Phaseolus coccineus L.) Bean Germplasm: A Slovenian Gene Bank Example.

Genetic resources comprised of 953 accessions of common (Phaseolus vulgaris L.) and 47 accessions of runner (Phaseolus coccineus L.) bean from the national Slovene gene bank were characterized using fourteen morphological seed descriptors. Seeds of each accession were evaluated for six quantitative characteristics: seed length, seed thickness, seed width, seed length/width ratio, seed width/thickness ratio, and 100 or 10 seed weight. Furthermore, seeds were evaluated using eight qualitative characteristics: seed colour; number of seed colours; primary/main seed colour; predominant secondary seed colour; distribution of secondary seed colour; seed veining; seed shape; and seed colour (primary and secondary) and coat pattern. For each, common, and runner bean collection, first four components within principal component analysis explained 75.03% and 80.16% of morphological variability, respectively. Regarding Ward's method and squared Euclidian distance, three clusters with the most distinct characteristics were established for each species. The results of morphological seed characterization indicate the origin (Andean, Mesoamerican, putative hybrids between gene pools) and domestication pathways of common and runner bean. This is the first study describing morphological seed characteristics of the entire common and runner bean germplasm conserved in one of the Central European bean collections. The results obtained in this study are serving as the useful information on genetic diversity of common and runner bean accessions at the Slovene gene bank, which could be used for development of new bean varieties for studied seed characteristics.