Genome Variability in Artificial Allopolyploid Hybrids of Avena sativa L. and Avena macrostachya Balansa ex Coss. et Durieu Based on Marker Sequences of Satellite DNA and the ITS1-5.8S rDNA Region.

Artificial hybrids between cultivated Avena species and wild Avena macrostachya that possess genes for resistance to biotic and abiotic stresses can be important for oat breeding. For the first time, a comprehensive study of genomes of artificial fertile hybrids Avena sativa × Avena macrostachya and their parental species was carried out based on the chromosome FISH mapping of satellite DNA sequences (satDNAs) and also analysis of intragenomic polymorphism in the 18S-ITS1-5.8S rDNA region, using NGS data. Chromosome distribution patterns of marker satDNAs allowed us to identify all chromosomes in the studied karyotypes, determine their subgenomic affiliation, and detect several chromosome rearrangements. Based on the obtained cytogenomic data, we revealed differences between two A. macrostachya subgenomes and demonstrated that only one of them was inherited in the studied octoploid hybrids. Ribotype analyses showed that the second major ribotype of A. macrostachya was species-specific and was not represented in rDNA pools of the octoploids, which could be related to the allopolyploid origin of this species. Our results indicate that the use of marker satDNAs in cytogenomic studies can provide important data on genomic relationships within Avena allopolyploid species and hybrids, and also expand the potential for interspecific crosses for breeding.

A universal karyotypic system for hexaploid and diploid Avena species brings oat cytogenetics into the genomics era.

BACKGROUND: The identification of chromosomes among Avena species have been studied by C-banding and in situ hybridization. However, the complicated results from several cytogenetic nomenclatures for identifying oat chromosomes are often contradictory. A universal karyotyping nomenclature system for precise chromosome identification and comparative evolutionary studies would be essential for genus Avena based on the recently released genome sequences of hexaploid and diploid Avena species. RESULTS: Tandem repetitive sequences were predicted and physically located on chromosomal regions of the released Avena sativa OT3098 genome assembly v1. Eight new oligonucleotide (oligo) probes for sequential fluorescence in situ hybridization (FISH) were designed and then applied for chromosome karyotyping on mitotic metaphase spreads of A. brevis, A. nuda, A. wiestii, A. ventricosa, A. fatua, and A. sativa species. We established a high-resolution standard karyotype of A. sativa based on the distinct FISH signals of multiple oligo probes. FISH painting with bulked oligos, based on wheat-barley collinear regions, was used to validate the linkage group assignment for individual A. sativa chromosomes. We integrated our new Oligo-FISH based karyotype system with earlier karyotype nomenclatures through sequential C-banding and FISH methods, then subsequently determined the precise breakage points of some chromosome translocations in A. sativa. CONCLUSIONS: This new universal chromosome identification system will be a powerful tool for describing the genetic diversity, chromosomal rearrangements and evolutionary relationships among Avena species by comparative cytogenetic and genomic approaches.

Genomic relationships among sixteen species of Avena based on (ACT)6 trinucleotide repeat FISH.

Knowledge of the locations of repeat elements could be very important in the assembly of genome sequences and their assignment to physical chromosomes. Genomic and species relationships among 16 species were investigated using fluorescence in situ hybridization (FISH) with the Am1 and (ACT)6 probes. The Am1 oligonucleotide probe was particularly enriched in the C genomes, whereas the (ACT)6 trinucleotide repeat probe showed a diverse distribution of hybridization patterns in the A, AB, C, AC, and ACD genomes but might not be present in the B and D genomes. The hybridization pattern of Avena sativa was very similar to that of A. insularis, indicating that this species most likely originated from A. insularis as a tetraploid ancestor. Although the two FISH probes failed to identify relationships of more species, this proof-of-concept approach opens the way to the use of FISH probes in assigning other signature elements from genomic sequence to physical chromosomes.

Genome size variation in the genus Avena.

Genome size is an indicator of evolutionary distance and a metric for genome characterization. Here, we report accurate estimates of genome size in 99 accessions from 26 species of Avena. We demonstrate that the average genome size of C genome diploid species (2C = 10.26 pg) is 15% larger than that of A genome species (2C = 8.95 pg), and that this difference likely accounts for a progression of size among tetraploid species, where AB < AC < CC (average 2C = 16.76, 18.60, and 21.78 pg, respectively). All accessions from three hexaploid species with the ACD genome configuration had similar genome sizes (average 2C = 25.74 pg). Genome size was mostly consistent within species and in general agreement with current information about evolutionary distance among species. Results also suggest that most of the polyploid species in Avena have experienced genome downsizing in relation to their diploid progenitors. Genome size measurements could provide additional quality control for species identification in germplasm collections, especially in cases where diploid and polyploid species have similar morphology.

Use of Repetitive Sequences for Molecular and Cytogenetic Characterization of Avena Species from Portugal.

Genomic diversity of Portuguese accessions of Avena species--diploid A. strigosa and hexaploids A. sativa and A. sterilis--was evaluated through molecular and cytological analysis of 45S rDNA, and other repetitive sequences previously studied in cereal species--rye subtelomeric sequence (pSc200) and cereal centromeric sequence (CCS1). Additionally, retrotransposons and microsatellites targeting methodologies--IRAP (inter-retrotransposon amplified polymorphism) and REMAP (retrotransposon-microsatellite amplified polymorphism)--were performed. A very high homology was detected for ribosomal internal transcribed sequences (ITS1 and ITS2) between the species analyzed, although nucleolar organizing regions (NOR) fluorescent in situ hybridization (FISH) analysis revealed distinct number of Nor loci between diploid and hexaploid species. Moreover, morphological diversity, evidenced by FISH signals with different sizes, was observed between distinct accessions within each species. pSc200 sequences were for the first time isolated from Avena species but proven to be highly similar in all genotypes analyzed. The use of primers designed for CCS1 unraveled a sequence homologous to the Ty3/gypsy retrotransposon Cereba, that was mapped to centromeric regions of diploid and hexaploid species, being however restricted to the more related A and D haplomes. Retrotransposon-based methodologies disclosed species- and accessions-specific bands essential for the accurate discrimination of all genotypes studied. Centromeric, IRAP and REMAP profiles therefore allowed accurate assessment of inter and intraspecific variability, demonstrating the potential of these molecular markers on future oat breeding programs.

Diversity of oat varieties in eliciting the early inflammatory events in celiac disease.

PURPOSE: Celiac disease (CD) is an autoimmune enteropathy, triggered by dietary gluten. The only treatment is a strict gluten-free diet. Oats are included in the list of gluten-free ingredients by European Regulation, but the safety of oats in CD is still a matter of debate. The present study examined the capability of different oat cultivars of activating the gliadin-induced transglutaminase-2 (TG2)-dependent events in some in vitro models of CD. In addition, we compared this capability with the electrophoresis pattern of peptic-tryptic digests of the proteins of the oat cultivars. METHODS: K562(S) cells agglutination, transepithelial electrical resistance of T84-cell monolayers, intracellular levels of TG2 and phosphorylated form of protein 42-44 in T84 cells were the early gliadin-dependent events studied. RESULTS: The results showed that the Nave oat cultivar elicited these events, whereas Irina and Potenza varieties did not. The ability of a cultivar to activate the above-described events was associated with the electrophoretic pattern of oat proteins and their reactivity to anti-gliadin antibodies. CONCLUSION: We found significant differences among oat cultivars in eliciting the TG2-mediated events of CD inflammation. Therefore, the safety of an oat cultivar in CD might be screened in vitro by means of biochemical and biological assays, before starting a clinical trial to definitely assess its safety.

Study on the sensitivity of three oat varieties to the saddle gall midge, haplodiplosis margina ta (von Roser) (Diptera: cecidomyiidae).

The saddle gall midge, Haplodiplosis marginota (von Roser, 1840) is a univoltine pest of cereals which occurs in Europe. The larvae feed on stems and attractive saddle-shaped depressions, driving to important yield losses when the galls are numerous. After 40 years without any reporting, large populations of H. marginata and important damage have been observed since 2010 in wheat crops in Belgium, especially in the Flemish Polders where clay soils and intensive farming of cereals favour heavy infestations. According to some research conducted in the 1960s during the last outbreak, oat (Avena sativa L.) is known to be one of the less favourable hosts to the saddle gall midge. Our study was performed in order to assess the host sensitivity of three oat varieties currently grown in Belgium: EVITA, EFFEKTIv and FREDDY. Therefore, oat varieties were sown on infested soil in two separate enclosures in a glasshouse. In the first enclosure, only the three oat varieties were grown; in the second one, these three oat varieties were grown together with two varieties of spring wheat: GRANNY and KWS CHAMSIN. TWO parameters were measured: the percentage of leaves with laid eggs, and the number of galls per stem. The percentage of leaves with eggs showed that the infestation was significantly lower on oats when they were in presence of wheat. The egg infestation was also significantly higher on wheat than on oat, which means oat is a much less favourable host plant than spring wheat for egg laying. Oat varieties were significantly different from each other regarding the number of galls per stem, but with very little damage compared to wheat. The FREDDY variety even seemed to be completely resistant to saddle gall midge, as no galls were observed although there was a similar percentage of leaves with eggs for the three oat varieties. Cropping oat could thus contribute to reduce infestations of H. morginato.

Use of tyramide-fluorescence in situ hybridization and chromosome microdissection for ascertaining homology relationships and chromosome linkage group associations in oats.

The physical mapping of single locus sequences by tyramide-fluorescence in situ hybridization (Tyr-FISH) and the analysis of sequences obtained from microdissected chromosomes were assayed as potential tools for (1) determining homology and homoeology among chromosome regions of Avena species, and (2) establishing associations between linkage groups and specific chromosomes. Low copy number probes, derived from resistance gene analogues (RGAs) and 2.8-4.5 kb long, successfully produced hybridization signals on specific chromosomes. Four sets of homoeologous chromosome regions were identified in the hexaploids using 3 probes that produced 4 single locus markers in A. strigosa and 2 in A. eriantha. Laser capture microdissection of metaphase I cells of A. sativa monosomic lines allowed the isolation of critical univalents. Sequences derived from 2 RGAs were successfully amplified in DNA extracted from univalents. In one instance, it was possible to map a nucleotide polymorphism specific for 1 chromosome. An association was established between this chromosome and its linkage groups in 2 hexaploid genetic maps. The results indicate that Tyr-FISH is useful in the characterization of homoeologous chromosome segments in hexaploids, whereas chromosome microdissection, as employed in this work, needs to be improved before it can routinely be used with meiotic chromosomes.

Diversity in oat potential immunogenicity: basis for the selection of oat varieties with no toxicity in coeliac disease.

BACKGROUND AND AIMS: Coeliac disease (CD) is triggered by an abnormal reaction to gluten. Peptides resulting from partially digested gluten of wheat, barley or rye cause inflammation of the small intestinal mucosa. Previous contradictory studies suggest that oats may trigger the abnormal immunological response in patients with CD. Monoclonal antibodies (moAbs) against the main immunotoxic 33-mer peptide (A1 and G12) react strongly against wheat, barley and rye but have less reactivity against oats. The stated aim of this study is to test whether this observed reactivity could be related to the potential toxicity of oats for patients with CD. METHODS: In the present study, different oat varieties, controlled for their purity and by their distinct protein pattern, were used to examine differences in moAb G12 recognition by ELISA and western blot. Immunogenicity of oat varieties was determined by 33-mer concentration, T cell proliferation and interferon γ production. RESULTS: Three groups of oat cultivars reacting differently against moAb G12 could be distinguished: a group with considerable affinity, a group showing slight reactivity and a third with no detectable reactivity. The immunogenicity of the three types of oats as well as that of a positive and negative control was determined with isolated peripheral blood mononuclear T cells from patients with CD by measurement of cell proliferation and interferon γ release. A direct correlation of the reactivity with G12 and the immunogenicity of the different prolamins was observed. CONCLUSIONS: The results showed that the reactivity of the moAb G12 is proportional to the potential immunotoxicity of the cereal cultivar. These differences may explain the different clinical responses observed in patients suffering from CD and open up a means to identify immunologically safe oat cultivars, which could be used to enrich a gluten-free diet.

The world's first glyphosate-resistant case of Avena fatua L. and Avena sterilis ssp. ludoviciana (Durieu) Gillet & Magne and alternative herbicide options for their control.

Avena fatua and A. ludoviciana (commonly known as wild oats) are the most problematic winter grass species in fallows and winter crops in the northeast region of Australia. A series of experiments were conducted to evaluate the performance of glyphosate and alternative post-emergence herbicides on A. fatua and A. ludoviciana. This study reports the world's first glyphosate-resistant (GR) biotypes of A. fatua and A. ludoviciana. The glyphosate dose required to kill 50% of the plants (LD50) and to reduce 50% of the biomass (GR50) for the GR biotype of A. fatua was 556 g a.e./ha and 351 g a.e./ha, respectively. These values for A. ludoviciana were 848 g a.e./ha and 289 g a.e./ha. Regardless of the growth stage (3-4 or 6-7 leaf stages), clethodim (120 g a.i./ha), haloxyfop (78 g a.i./ha), pinoxaden (20 g a.i./ha), and propaquizafop (30 g a.i./ha) were the best alternative herbicide options for the control of A. fatua and A. ludoviciana. The efficacy of butroxydim (45 g a.i./ha), clodinafop (120 g a.i./ha), imazamox + imazapyr (36 g a.i./ha), and paraquat (600 g a.i./ha) reduced at the advanced growth stage. Glufosinate (750 g a.i./ha), flamprop (225 g a.i./ha), and pyroxsulam + halauxifen (20 g a.i./ha) did not provide effective control of Avena species. This study identified alternative herbicide options to manage GR biotypes of A. fatua and A. ludoviciana.

The evolution pattern of rDNA ITS in Avena and phylogenetic relationship of the Avena species (Poaceae: Aveneae).

Ribosomal ITS sequences are commonly used for phylogenetic reconstruction because they are included in rDNA repeats, and these repeats often undergo rapid concerted evolution within and between arrays. Therefore, the rDNA ITS copies appear to be virtually identical and can sometimes be treated as a single gene. In this paper we examined ITS polymorphism within and among 13 diploid (A and C genomes), seven tetraploid (AB, AC and CC genomes) and four hexaploid (ACD genome) to infer the extent and direction of concerted evolution, and to reveal the phylogenetic and genome relationship among species of Avena. A total of 170 clones of the ITS1-5.8S-ITS2 fragment were sequenced to carry out haplotype and phylogenetic analysis. In addition, 111 Avena ITS sequences retrieved from GenBank were combined with 170 clones to construct a phylogeny and a network. We demonstrate the major divergence between the A and C genomes whereas the distinction among the A and B/D genomes was generally not possible. High affinity among the A(d) genome species A. damascena and the ACD genome species A. fatua was found, whereas the rest of the ACD genome hexaploids and the AACC tetraploids were highly affiliated with the A(l) genome diploid A. longiglumis. One of the AACC species A. murphyi showed the closest relationship with most of the hexaploid species. Both C(v) and C(p) genome species have been proposed as paternal donors of the C-genome carrying polyploids. Incomplete concerted evolution is responsible for the observed differences among different clones of a single Avena individual. The elimination of C-genome rRNA sequences and the resulting evolutionary inference of hexaploid species are discussed.

Volatile Profiles from Traditional Chinese Oat Meal Varied Significantly from Oat Porridge and Differed with Cultivars and Locations.

Volatile profiles of oat-based foods are mainly analyzed on the oat flakes and porridge as snack or breakfast, whereas the volatile characteristics of the traditional Chinese oat meal (TCOM), a popular main food in some regions of northern China, with special strong aroma, are not known. Here, we compared the volatile profiles from headspace solid phase microextraction gas chromatography-mass spectrometry analysis of oat porridge (OP) and TCOM, which were of different processing and cooking methods, from those of different cultivars, and analyzed the effect of cultivation locations on oat volatile features. Apart from the 35 volatiles shared by OP and TCOM, there were 23 and 24 volatiles specific to OP and TCOM, respectively, with the later showing more toasting and frying-related volatiles due to the dry frying process of the grains before milling. Principle component analysis of the volatiles of OP and TCOM from 16 cultivars showed that they were clustered into two groups, and four cultivars were clustered together, independent of processing and cooking methods. The oat volatile profiles of cultivars grown in three regions of north China were dependent on the cultivation locations rather than cultivars, regardless of OP or TCOM, with those from Datong of Shanxi Province and Zhangjiakou of Hebei Province clustered together. The location effect could be due to significant less precipitation in the two regions above than the other region Ulanqab of Inner Mongolia. PRACTICAL APPLICATION: The volatile compounds in oat are closely related to cultivation regions, which could be applied as a key factor by oat producers for marketing. The four cultivars showed similar and stable volatile profiles, which could be used as reference cultivars for breeding of high-quality oat with better flavor.

The Pros and Cons of Using Oat in a Gluten-Free Diet for Celiac Patients.

A therapeutic gluten-free diet often has nutritional limitations. Nutritional qualities such as high protein content, the presence of biologically active and beneficial substances (fiber, beta-glucans, polyunsaturated fatty acids, essential amino acids, antioxidants, vitamins, and minerals), and tolerance by the majority of celiac patients make oat popular for use in gluten-free diet. The health risk of long-time consumption of oat by celiac patients is a matter of debate. The introduction of oat into the diet is only recommended for celiac patients in remission. Furthermore, not every variety of oat is also appropriate for a gluten-free diet. The risk of sensitization and an adverse immunologically mediated reaction is a real threat in some celiac patients. Several unsolved issues still exist which include the following: (1) determination of the susceptibility markers for the subgroup of celiac patients who are at risk because they do not tolerate dietary oat, (2) identification of suitable varieties of oat and estimating the safe dose of oat for the diet, and (3) optimization of methods for detecting the gliadin contamination in raw oat used in a gluten-free diet.

Naked Oat (Avena nuda L.) Oligopeptides:Immunomodulatory Effects on Innate and AdaptiveImmunity in Mice via Cytokine Secretion, AntibodyProduction, and Th Cells Stimulation.

The study aimed to investigate the immunomodulatory activity of oligopeptides derivedfrom oat (Avena Nuda L.) (OOPs). Healthy female BALB/c mice were randomly assigned to fivegroups, given deionized water (control) and 0.25, 0.5, 1.0, and 2.0 g/kg body weight (BW) of OOPsdaily by intragastric administration. Seven assays were performed to determine theimmunomodulatory effects of OOPs on immune organ ratios, cellular and humoral immuneresponses, macrophage phagocytosis, and natural killer (NK) cell activity. Spleen T lymphocytesubpopulations (by flow cytometry), serum cytokine and immunoglobulin levels (by multiplexsandwich immunoassays) were determined to evaluate how OOPs affected the immune system.Our results showed that OOPs could significantly improve innate and adaptive immune responsesin mice through the enhancement of cell-mediated and humoral immunity, macrophagephagocytosis capacity, and NK cell activity. We concluded that the immunomodulatory effectsmight be attributed to increased T and Th cell percentages, serum interferon (IFN)-γ, interleukin(IL)-1 α, IL-2, IL-6, IL-10, IL-12, tumor necrosis factor (TNF)- α, and granulocyte-macrophagecolony-stimulating factor (GM-CSF) secretions as well as immunoglobulin (Ig) A, IgG, and IgMproductions. These results indicate that dietary OOPs could be considered as promisingimmunomodulators with dosages ranging from 0.25 to 2.0 g/kg BW.

Identification and Quantification of Avenanthramides and Free and Bound Phenolic Acids in Eight Cultivars of Husked Oat ( Avena sativa L) from Finland.

Finland is the second largest oat producer in Europe. Despite the existing knowledge of phenolics in oat, there is little information on the phenolic composition of oats from Finland. The aim of the study was to investigate the concentrations of free and bound phenolic acids, as well as avenanthramides in eight Finnish cultivars of husked oat ( Avena sativa L.). Seven phenolic acids and one phenolic aldehyde were identified, including, in decreasing order of abundance: p-coumaric, ferulic, cinnamic, syringic, vanillic, 2,4-dihydroxybenzoic, and o-coumaric acids and syringaldehyde. Phenolic acids were mostly found as bound compounds. Significant varietal differences ( p < 0.05) were observed in the cumulative content of phenolic acids, with the lowest level found in cv. 'Viviana' (1202 ± 52.9 mg kg-1) and the highest in cv. 'Akseli' (1687 ± 80.2 mg kg-1). Avenanthramides (AVNs) 2a, 2p, and 2f were the most abundant. Total AVNs levels ranged from 26.7 ± 1.44 to 185 ± 12.5 mg kg-1 in cv. 'Avetron' and 'Viviana', respectively.

Herbicidal effects on nontarget vegetation: investigating the limitations of current pesticide registration guidelines.

The impact of herbicide exposure on nontarget vegetation within agroecosystems has sparked extensive research that revealed that current pesticide registration guidelines may be inadequate at predicting the effects of herbicides on wild plants and habitats. This study extends the current interest by presenting three experiments highlighting some of the limitations to current phytotoxicity testing guidelines. Several crops and wild plant species were grown under greenhouse conditions following standard protocol for phytotoxicity testing. Plants were sprayed with five different herbicides at the four- to six-leaf stage, and biomass was recorded at 28 d after spray. Results showed that current regulatory protocol will likely underestimate herbicide phytotoxicity if testing does not include data for the complete tank-mix formulation. The present study also showed that the range in herbicide sensitivity among cultivars of the same crop can be quite extensive and that, depending on the cultivar included in a risk assessment, conclusions regarding the phytotoxicity of any given herbicide may differ. Although no significant differences in sensitivity were found between crops and related wild species, results revealed that current guidelines are too rigid in terms of species selection. Considering the variability among crop cultivars, coupled with the ecological importance and the ease of germination of many noncrop plant species, pesticide regulatory guidelines would be improved if wild species were included in testing. Findings of the present study indicate that current pesticide regulatory guidelines require modifications to ensure a more accurate assessment of herbicide effects on nontarget plant species.

Assessment of Avenins from Different Oat Varieties Using R5-Based Sandwich ELISA.

Gluten derived from wheat and related triticeae cereals possesses distinct amino acid sequences that provoke the immunopathogenic features of celiac disease (CD) in genetically susceptible individuals. However, the role of oat-derived gluten, or avenins, in CD pathogenesis remains a disputed matter, as evidenced by a lack in harmonized legislation regarding gluten classification in relation to gluten-free labeling. In this study, we have analyzed a panel of pure oat cultivars using a sandwich ELISA based on the R5 monoclonal antibody (mAb), which binds to canonical epitopes occurring within celiagenic peptides present in triticeae-derived gluten but reportedly not present in avenins. We have identified three varieties of oats that reproducibly bind R5 antibodies and levels indicating the presence of gluten at more than the 20 ppm gluten regulatory threshold. Nested assessment using Western blot analysis and alternative gluten detection systems corroborated these results. Collectively, these data suggest that select oat varieties may prove problematic to patients with CD and to food companies and regulatory agencies and will extend our basic understanding of current gluten detection systems.

Wide Hybridization Between Oat and Pearl Millet.

Wide hybridization is a one of the important techniques in plant breeding. Oat (Avena sativa L.) and pearl millet (Pennisetum glaucum L.) belong to different subfamilies of Poaceae. In generally, such distant relative species show uniparental chromosome elimination after successful fertilization. However, all seven pearl millet chromosomes are retained beside the genome of oat during embryogenesis. Hybrid seedlings develop, but show necrosis after light irradiation. Here, a detailed protocol for wide hybridization between oat and pearl millet is described.

Multi-substrate chromosome preparations for high throughput comparative FISH.

BACKGROUND: A modification of a standard method of fluorescence in situ hybridisation (FISH) is described, by which a combination of several substrates and probes on single microscope slides enables more accurate comparisons of the distribution and abundance of chromosomal sequences and improves the relatively low throughput of standard FISH methods. RESULTS: The utility and application of multi-colour, multi-substrate FISH is illustrated by the simultaneous physical mapping of retrotransposon sequences to three species of Avena, and single locus BAC (bacterial artificial chromosome) clones and rDNA probes to three species of Brachypodium, demonstrating how this would enable better understanding of complex phylogenetic relationships among some of the species belonging to these two genera. CONCLUSION: The results show that use of multi-substrate chromosome preparations significantly increases the utility of FISH in comparative analyses of the distribution and abundance of chromosomal sequences in closely related plant species.

Promoting the Use of Common Oat Genetic Resources through Diversity Analysis and Core Collection Construction.

The assessment of diversity and population structure and construction of a core collection is beneficial for the efficient use and management of germplasm. A unique collection of common oat landraces, cultivated in the temperate climate of central Europe until the end of the twentieth century, is preserved in the Polish gene bank. It consists of 91 accessions that have never been used in breeding programs. In order to optimise the use of this genetic resource, we aimed to: (1) determine genetic and agro-morphological diversity, (2) identify internal genetic variation of the tested accessions, (3) form a core collection and (4) recognise the accessions useful for breeding programs or re-release for cultivation. The collection was screened using ISSR markers (1520 loci) and eight agro-morphological traits. Uniquely, we performed molecular studies based on 24 individuals of every accession instead of bulk samples. Therefore, assessment of the degree of diversity within each population and the identification of overlapping gene pools were possible. The observed internal diversity (Nei unbiased coefficient) was in the range of 0.17-0.31. Based on combined genetic and agro-morphological data, we established the core collection composed of 21 landraces. Due to valuable compositions of important traits, some accessions were also identified as useful for breeding programs. The population structure and principal coordinate analysis revealed two major clusters. Based on the previous results, the accessions classified within the smaller one were identified as obsolete varieties instead of landraces. Our results show that the oat landraces are, in general, resistant to local races of diseases, well adapted to local conditions and, in some cases, yielding at the level of modern varieties. Therefore, in situ conservation of the landraces in the near future may be satisfactory for both farmers and researchers in terms of the genetic resources preservation.