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Tiller number is a key component of plant architecture with direct effect on grain yield of wheat. Interspecific hybridization using wild relative species are promising approach for increasing wheat genetic diversity, including yield potential. Agropyron glael, a perennial hybrid of Thinopyrum intermedium and Th. ponticum has five subgenomes (J, Jst, Jvs, Jr, and St), making A. glael an important gene source for transferring useful agronomical traits into wheat. This work reports on development of a wheat-A. glael translocation line WT153397. Sequential in situ hybridizations with J-, St-, and D-genomic DNA probes and pSc119.2, Afa family, pTa71, and (GAA)7 DNA repeats, as well as an analysis using molecular markers specific for the wheat 6D chromosome, revealed the presence of a 6DS.6Jvs centric fusion in the translocation line. Field trials in low-input and high-input breeding nurseries over three growing seasons demonstrated high compensating ability of the Agropyron chromosome arm for the missing 6DL as spike morphology and fertility of WT153397 did not differ significantly from those of wheat parents. Moreover, the introgressed 6Jvs chromosome arm significantly increased the number of productive tillers, which manifested in the significantly higher grain yield potential relative to the parental wheat cultivars. The translocated chromosome could be flow-sorted in high purity providing an opportunity to employ chromosome genomics to identify Agropyron gene variant(s) responsible for the tillering capacity. The translocation line WT153397 represents an important genetic stock for functional genetic studies of tiller formation and useful breeding material to increase wheat yield potential.

Wheat-rye T1BL.1RS translocation is widespread worldwide as the genes on 1RS arm have positive effect on stress resistance, grain yield and adaptation ability of wheat. Nowadays, the T1BL.1RS wheat cultivars have become susceptible to rust diseases because of the monophyletic (‘Petkus’) origin of 1RS. Here we report and discuss the production and detailed investigation of a new T1BL.1RS translocation line carrying 1RS with widened genetic base originating from Secale cereanum. Line ‘179’ exhibited improved spike morphology traits, resistance against stripe rust and leaf rust, as well as higher tillering capacity, fertility and dietary fiber (arabynoxylan) content than the parental wheat genotype. Comparative analyses based on molecular cytogenetic methods and molecular (SSR and DArTseq) makers indicate that the 1RS arm of line ‘179’ is a recombinant of S. cereale and S. strictum homologues, and approximately 16% of its loci were different from that of ‘Petkus’ origin. 162 (69.5%) 1RS-specific markers were associated with genes, including 10 markers with putative disease resistance functions and LRR domains found on the subtelomeric or pericentromeric regions of 1RS. Line ‘179’ will facilitate the map-based cloning of the resistance genes, and it can contribute to healthy eating and a more cost-efficient wheat production.

The chromosomal constitution of 9 dwarf (D) and 8 semidwarf (SD) lines derived by crossing hexaploid Triticale line NA-75 (AABBRR, 2n = 6x = 42) with Triticumaestivum (AABBDD, 2n = 6x = 42) cv. Chinese Spring was investigated using molecular cytogenetic techniques: fluorescence in situ hybridization and genomic in situ hybridization. A wheat-rye translocation (T4DS.7RL), 8 substitution lines, and a ditelosomic addition line (7RSdt) were identified. In the substitution lines, 1, 2, or 4 pairs of wheat chromosomes, belonging to the A, B, or D genome, were replaced by rye chromosomes. Substitutions between chromosomes belonging to different wheat genomes [5B(5A), 1D(1B)] also occurred. The lines were genetically stable, each carrying 42 chromosomes, except the wheat-rye ditelosomic addition line, which carried 21 pairs of wheat chromosomes and 1 pair of rye telocentric chromosomes (7RS). The chromosome pairing behavior of the lines was studied during metaphase I of meiosis. The chromosome pairing level and the number of ring bivalents were different for each line. Besides rod bivalents, univalent and multivalent associations (tri- and quadrivalents) were also detected. The main goal of the experiment was to develop genetically stable wheat/Triticale recombinant lines carrying chromosomes/chromatin fragments originating from the R genome of Triticale line NA-75. Introgression of rye genes into hexaploid wheat can broaden its genetic diversity, and the newly developed lines can be used in wheat breeding programs.

Summary Broadening the genetic base of crops is crucial for developing varieties to respond to global agricultural challenges such as climate change. Here, we analysed a diverse panel of 371 domesticated lines of the model crop barley to explore the genetics of crop adaptation. We first collected exome sequence data and phenotypes of key life history traits from contrasting multi‐environment common garden trials. Then we applied refined statistical methods, including some based on exomic haplotype states, for genotype‐by‐environment (G×E) modelling. Sub‐populations defined from exomic profiles were coincident with barley's biology, geography and history, and explained a high proportion of trial phenotypic variance. Clear G×E interactions indicated adaptation profiles that varied for landraces and cultivars. Exploration of circadian clock‐related genes, associated with the environmentally adaptive days to heading trait (crucial for the crop's spread from the Fertile Crescent), illustrated complexities in G×E effect directions, and the importance of latitudinally based genic context in the expression of large‐effect alleles. Our analysis supports a gene‐level scientific understanding of crop adaption and leads to practical opportunities for crop improvement, allowing the prioritisation of genomic regions and particular sets of lines for breeding efforts seeking to cope with climate change and other stresses., Significance Statement We analysed exome sequence data and life history traits measured in multi‐environment trials of a diverse panel of 371 domesticated barley lines to explore the genetics of crop adaptation. G×E interactions indicated adaptation profiles that varied for sub‐populations defined from exomic data. Exploration of circadian clock‐related genes associated with days to heading revealed complexities in G×E effect directions, and the importance of latitudinally based genic context in the expression of large effect alleles. Analysis provides practical information for future crop breeding.

Photosynthetic responses of Aegilops comosa genotypes were compared to those of wheat Mv9kr1 and Chinese spring in order to verify whether Ae. comosa TA2760 and MvGB1039 genotypes are potentially suitable gene sources for improving the drought and salt tolerance of bread wheat. Although there are some differences between the non-stressed plants and the measure of the decrease of the net-photosynthesis (PN), it was strongly inhibited by water deficit. Salt stress had similar effect on PN but at the highest (300 mM) NaCl concentration PN of the genotypes showed some activity. Severe drought induced a strong decrease in the effective quantum yield of PS II (ɸPSII) in the genotypes, while it was moderate in the case of salt treatment. Moreover, ɸPSII was unaffected by the increase of NaCl concentration in wheat lines. Parallel with the decrease in ɸPSII, the photoprotective mechanisms were enhanced in the wheat and wild wheat genotypes during water deficit. These results suggest that the Ae. comosa genotypes seem to respond to these stress factors with similar photosynthetic activity to the wheat lines. Thus, based on the above-mentioned facts, the examined Ae. comosa lines are not particularly good candidates for improving drought tolerance of wheat.

The chromosomal constitution of 9 dwarf (D) and 8 semidwarf (SD) lines derived by crossing hexaploid Triticale line NA-75 (AABBRR, 2n = 6x = 42) with Triticumaestivum (AABBDD, 2n = 6x = 42) cv. Chinese Spring was investigated using molecular cytogenetic techniques: fluorescence in situ hybridization and genomic in situ hybridization. A wheat-rye translocation (T4DS.7RL), 8 substitution lines, and a ditelosomic addition line (7RSdt) were identified. In the substitution lines, 1, 2, or 4 pairs of wheat chromosomes, belonging to the A, B, or D genome, were replaced by rye chromosomes. Substitutions between chromosomes belonging to different wheat genomes [5B(5A), 1D(1B)] also occurred. The lines were genetically stable, each carrying 42 chromosomes, except the wheat-rye ditelosomic addition line, which carried 21 pairs of wheat chromosomes and 1 pair of rye telocentric chromosomes (7RS). The chromosome pairing behavior of the lines was studied during metaphase I of meiosis. The chromosome pairing level and the number of ring bivalents were different for each line. Besides rod bivalents, univalent and multivalent associations (tri- and quadrivalents) were also detected. The main goal of the experiment was to develop genetically stable wheat/Triticale recombinant lines carrying chromosomes/chromatin fragments originating from the R genome of Triticale line NA-75. Introgression of rye genes into hexaploid wheat can broaden its genetic diversity, and the newly developed lines can be used in wheat breeding programs.

The photosynthetic responses to salt stress were examined in a wheat (Triticum aestivum L. cv. Asakaze)–barley (Hordeum vulgare L. cv. Manas) 7H addition line having elevated salt tolerance and compared to the parental wheat genotype. For this purpose, increasing NaCl concentrations up to 300 mM were applied and followed by a 7-day recovery period. Up to moderate salt stress (200 mM NaCl), forcible stomatal closure, parallel with a reduction in the net assimilation rate (P N), was only observed in wheat, but not in the 7H addition line or barley. Since the photosynthetic electron transport processes of wheat were not affected by NaCl, the impairment in P N could largely be accounted for the salt-induced decline in stomatal conductance (g s), accompanied by depressed intercellular CO2 concentration and carboxylation efficiency. Both, P N and nonstomatal limitation factors (Lns) were practically unaffected by moderate salt stress in barley and in the 7H addition line due to the sustained g s, which might be an efficient strategy to maintain the efficient photosynthetic activity and biomass production. At 300 mM NaCl, both P N and g s decreased significantly in all the genotypes, but the changes in P N and Lns in the 7H addition line were more favourable similar to those in wheat. The downregulation of photosynthetic electron transport processes around PSII, accompanied by increases in the quantum yield of regulated energy dissipation and of the donor side limitation of PSI without damage to PSII, was observed in the addition line and barley during severe stress. Incomplete recovery of P N was observed in the 7H addition line as a result of declined PSII activity probably caused by enhanced cyclic electron flow around PSI. These results suggest that the better photosynthetic tolerance to moderate salt stress of barley can be manifested in the 7H addition line which may be a suitable candidate for improving salt tolerance of wheat.

There is a wealth of resistance genes in the Mexican wild relative of cultivated Solanum, but very few of these species are sexually compatible with cultivated Solanum tuberosum. The most devastating disease of potato is late blight caused by the oomycete Phytophthora infestans (Pi). The wild hexaploid species S. demissum, which it is able to cross with potato, was used to transfer eleven race-specific genes by introgressive hybridization that were subsequently widely used in potato breeding. However, there are now more virulent races of Pi that can overcome all of these genes. The most sustainable strategy for protecting potatoes from late blight is to pyramid or stack broad-spectrum resistance genes into the cultivars. Recently four broad-spectrum genes (Rpi) conferring resistance to Pi were identified and cloned from the sexually incompatible species S. bulbocastanum: Rpi-blb1 (RB), Rpi-blb2, Rpi-blb3, and Rpi-bt1. For this research, a resistant S. bulbocastanum accession was selected carrying the genes Rpi-blb1 and Rpi-blb3 together with race-specific R3a and R3b genes. This accession was previously used to produce a large number of somatic hybrids (SHs) with five commercial potato cultivars using protoplast electrofusion. In this study, three SHs with cv. ‘Delikat’ were selected and backcross generations (i.e., BC1 and BC2) were obtained using cvs. ‘Baltica’, ‘Quarta’, ‘Romanze’, and ‘Sarpo Mira’. Their assessment using gene-specific markers demonstrates that these genes are present in the SHs and their BC progenies. We identified plants carrying all four genes that were resistant to foliage blight in greenhouse and field trials. Functionality of the genes was shown by using agro-infiltration with the effectors of corresponding Avr genes. For a number of hybrids and BC clones yield and tuber number were not significantly different from that of the parent cultivar ‘Delikat’ in field trials. The evaluation of agronomic traits of selected BC2 clones and of their processing qualities revealed valuable material for breeding late blight durable resistant potato. We show that the combination of somatic hybridization with the additional use of gene specific markers and corresponding Avr effectors is an efficient approach for the successful identification and introgression of late blight resistance genes into the potato gene pool.

Wild relatives of wheat, such as Aegilops spp. are potential sources of genes conferring tolerance to drought stress. As drought stress affects seed composition, the main goal of the present study was to determine the effects of drought stress on the content and composition of the grain storage protein (gliadin (Gli), glutenin (Glu), unextractable polymeric proteins (UPP%) and dietary fiber (arabinoxylan, β-glucan) components of hexaploid bread wheat (T. aestivum) lines containing added chromosomes from Ae. biuncialis or Ae. geniculata. Both Aegilops parents have higher contents of protein and β-glucan and higher proportions of water-soluble arabinoxylans (determined as pentosans) than wheat when grown under both well-watered and drought stress conditions. In general, drought stress resulted in increased contents of protein and total pentosans in the addition lines, while the β-glucan content decreased in many of the addition lines. The differences found between the wheat/Aegilops addition lines and wheat parents under well-watered conditions were also manifested under drought stress conditions: Namely, elevated β-glucan content was found in addition lines containing chromosomes 5Ug, 7Ug and 7Mb, while chromosomes 1Ub and 1Mg affected the proportion of polymeric proteins (determined as Glu/Gli and UPP%, respectively) under both well-watered and drought stress conditions. Furthermore, the addition of chromosome 6Mg decreased the WE-pentosan content under both conditions. The grain composition of the Aegilops accessions was more stable under drought stress than that of wheat, and wheat lines with the added Aegilops chromosomes 2Mg and 5Mg also had more stable grain protein and pentosan contents. The negative effects of drought stress on both the physical and compositional properties of wheat were also reduced by the addition of these. These results suggest that the stability of the grain composition could be improved under drought stress conditions by the intraspecific hybridization of wheat with its wild relatives.

PubMedID: 31043760 For more than 10,000 years, the selection of plant and animal traits that are better tailored for human use has shaped the development of civilizations. During this period, bread wheat (Triticum aestivum) emerged as one of the world’s most important crops. We use exome sequencing of a worldwide panel of almost 500 genotypes selected from across the geographical range of the wheat species complex to explore how 10,000 years of hybridization, selection, adaptation and plant breeding has shaped the genetic makeup of modern bread wheats. We observe considerable genetic variation at the genic, chromosomal and subgenomic levels, and use this information to decipher the likely origins of modern day wheats, the consequences of range expansion and the allelic variants selected since its domestication. Our data support a reconciled model of wheat evolution and provide novel avenues for future breeding improvement. © 2019, The Author(s), under exclusive licence to Springer Nature America, Inc. 339728 Kyoto University California Department of Fish and Game: SFB924 Wageningen UR University of Dundee 2819103915 Région Auvergne-Rhône-Alpes European Research Council 23000816 SRESRI 2015 Seventh Framework Programme: FP7-613556, FP7/ 2007–2013 23000892 SYMBIOSE 2016 Agence Nationale de la Recherche: 14-CE02-0002 Çukurova Üniversitesi: FUA-2016–6033 The authors wish to thank the INRA Biological Resources Center on small grain cereals (https://www6.ara.inra.fr/umr1095_eng/Teams/Research/Biological-Resources-Centre) for providing seeds and passport data, and for establishing a wheat biorepository. The authors thank the Federal ex situ Genbank Gatersleben, Germany (IPK), the N. I. Vavilov All-Russian Research Institute of Plant Industry, Russia (VIR), Centre for Genetic Resources, WUR, Netherlands (CGN), Kyoto University, National Bioresource Project, Japan (NBRP), the Australian Winter Cereal Collection Tamworth, Australia (AWCC), the National Plant Germplasm System, USA (USDA-ARS), the International Center for Agriculture Research in the Dry Areas (ICARDA), the Max Planck Institute for Plant Breeding Research Cologne, Germany (MPIPZ), Germplasm Resource Unit at the John Innes Centre UK (JIC) and the Wheat and Barley Legacy for Breeding Improvement (WHEALBI) consortium for providing plant material and passport data. The research leading to these results has received funding from the European Community’s Seventh Framework Programme (FP7/ 2007–2013) under grant agreement FP7-613556, Whealbi project (http://www.whealbi.eu/project/). R.W. and J.R. also acknowledge support from the Scottish Government Research Program and R.W. from the University of Dundee. H.O. acknowledges support from Çukurova University (FUA-2016–6033). K.F.X.M. acknowledges support from the German Federal Ministry of Food and Agriculture (2819103915) and the DFG (SFB924). T.L. acknowledges supports from the Agence Nationale pour la Recherche (BirdIslandGenomic project 14-CE02-0002), European Research Council (TREEPEACE project, grant agreement 339728) and the bioinformatics platform from Toulouse Midi-Pyrénées (Bioinfo Genotoul) for providing computing and storage resources. J.S. acknowledges support from the Région Auvergne-Rhône-Alpes and FEDER Fonds Européens de Développement Régional (23000816 SRESRI 2015), the CPER contrat de plan État-région (23000892 SYMBIOSE 2016) and AgreenSkills fellowship (applicant ID 4146).

The cytomolecular discrimination of the Am- and A-genome chromosomes facilitates the selection of wheat-Triticum monococcum introgression lines. Fluorescence in situ hybridisation (FISH) with the commonly used DNA probes Afa family, 18S rDNA and pSc119.2 showed that the more complex hybridisation pattern obtained in T. monococcum relative to bread wheat made it possible to differentiate the Am and A chromosomes within homoeologous groups 1, 4 and 5. In order to provide additional chromosomal landmarks to discriminate the Am and A chromosomes, the microsatellite repeats (GAA)n, (CAG)n, (CAC)n, (AAC)n, (AGG)n and (ACT)n were tested as FISH probes. These showed that T. monococcum chromosomes have fewer, generally weaker, simple sequence repeat (SSR) signals than the A-genome chromosomes of hexaploid wheat. A differential hybridisation pattern was observed on 6Am and 6A chromosomes with all the SSR probes tested except for the (ACT)n probe. The 2Am and 2A chromosomes were differentiated by the signals given by the (GAA)n, (CAG)n and (AAC)n repeats, while only (GAA)n discriminated the chromosomes 3Am and 3A. Chromosomes 7Am and 7A could be differentiated by the lack of (GAA)n and (AGG)n signals on 7A. As potential landmarks for identifying the Am chromosomes, SSR repeats will facilitate the introgression of T. monococcum chromatin into wheat.

The processing quality of 37 wheat varieties grown in Hungary and Austria (2011–2013) were assessed under organic and conventional low input management. The varieties studied were developed using three breeding strategies (conventional, organic and their combination: BFOA). The aim was to evaluate the effect of the field management and to assess the performance of varieties developed using different breeding methods, based on their quality traits under different managements. Furthermore, properties were identified that could characterize wheat quality and be used effectively for selection under both types of growing conditions. Strong year and genotype effects were found for all the quality traits (protein, starch, gluten, GI, Zeleny, Farinograph water absorption, development time, stability and quality number, falling number, flour yield, hardness index) of the studied varieties, while the effect of the management was significant for the physical properties (test weight, thousand-kernel weight, hardness) and gluten quality characters (gluten spread, GI, dough stability) of the grain. The standard deviation of the gluten quality traits characterized the differences between the breeding strategies. It proved possible to pre select organic varieties for quality traits with high broad-sense heritability under conventional growing conditions, but direct selection in organic fields is suggested for gluten quality characters.

A Thinopyrum intermedium × Thinopyrum ponticum synthetic hybrid wheatgrass is an excellent source of leaf and stem rust resistance produced by N.V.Tsitsin. Wheat line Mv9kr1 was crossed with this hybrid (Agropyron glael) in Hungary in order to transfer its advantageous agronomic traits into wheat. As the wheat parent was susceptible to leaf rust, the transfer of resistance was easily recognizable in the progenies. Three different partial amphiploid lines with leaf rust resistance were selected from the wheat/Thinopyrum hybrid derivatives by multicolour genomic in situ hybridization. Chromosome counting on the partial amphiploids revealed 58 chromosomes (18 wheatgrass) in line 194, 56 (14 wheatgrass) in line 195 and 54 (12 wheatgrass) in line 196. The wheat chromosomes present in these lines were identified and the wheatgrass chromosomes were characterized by fluorescence in situ hybridization using the repetitive DNA probes Afa-family, pSc119.2 and pTa71. The 3D wheat chromosome was missing from the lines. Molecular marker analysis showed the presence of the Lr24 leaf rust resistance gene in lines 195 and 196. The morphological traits were evaluated in the field during two consecutive seasons in two different locations.

Interspecific hybridization between bread wheat (Triticum aestivum, 2n = 42) and related species allows the transfer of agronomic and quality traits, whereby subsequent generations comprise an improved genetic background and can be directly applied in wheat breeding programmes. While wild relatives are frequently used as sources of agronomically favourable traits, cultivated species can also improve wheat quality and stress resistance. A salt-tolerant ‘Asakaze’/‘Manas’ 7H disomic addition line (2n = 44) with elevated β-glucan content, but with low fertility and an unstable genetic background was developed in an earlier wheat-barley prebreeding programme. The aim of the present study was to take this hybridization programme further and transfer the favourable barley traits into a more stable genetic background. Taking advantage of the breakage-fusion mechanism of univalent chromosomes, the ‘Rannaya’ winter wheat 7B monosomic line was used as female partner to the 7H addition line male, leading to the development of a compensating wheat/barley Robertsonian translocation line (7BS.7HL centric fusion, 2n = 42) exhibiting higher salt tolerance and elevated grain β-glucan content. Throughout the crossing programme, comprising the F1-F4 generations, genomic in situ hybridization, fluorescence in situ hybridization and chromosome-specific molecular markers were used to trace and identify the wheat and barley chromatin. Investigations on salt tolerance during germination and on the (1,3;1,4)-β-D-glucan (mixed-linkage glucan [MLG]) content of the seeds confirmed the salt tolerance and elevated grain MLG content of the translocation line, which can be directly applied in current wheat breeding programmes.

Barley chromosome 5H, carrying important QTLs for plant adaptation and tolerance to abiotic stresses, is extremely instable in the wheat genetic background and is eliminated in the early generations of wheat-barley crosses. A spontaneous wheat-barley 5HS-7DS.7DL translocation was previously obtained among the progenies of the Mv9kr1 x Igri hybrid. The present work reports on the transfer of the 5HS-7DS.7DL translocation into a modern wheat cultivar, Mv Bodri, in order to use it in the wheat breeding program. The comparison of the hybridization bands of DNA repeats HvT01, pTa71, (GAA)n and the barley centromere-specific (AGGGAG)n in Igri barley and the 5HS-7DS.7DL translocation, together with the visualization of the barley chromatin made it possible to determine the size of the introgressed barley segment, which was approximately 74% of the whole 5HS. Of the 29 newly developed PCR markers, whose source ESTs were selected from the Genome Zipper of barley chromosome 5H, 23 were mapped in the introgressed 1-0.26 FL 5HS bin, three were located in the missing C-0.26 FL region, while three markers were specific for 5HL. The translocation breakpoint was flanked by markers Hv7502 and Hv3949. A comparison of the parental wheat cultivars and the wheat-barley introgression lines indicated that the presence of the translocation improved tillering ability in the Mv9kr1 and Mv Bodri genetic background. The similar or better yield components under high- or low-input cultivation environments, respectively, indicated that the 5HS-7DS.7DL translocation had little or no negative effect on yield components, making it a promising genotype to improve wheat genetic diversity. These results promise to accelerate functional genomic studies on barley chromosome 5H and to support pre-breeding and breeding research on wheat.

The Hungarian rye (Secale cereale L., 2n = 2x = 14, RR) cultivar Lovaszpatonai possesses numerous agronomically useful traits that can be exploited in wheat breeding. This study reports on the production, molecular cytogenetic identification, and evaluation of resistance to wheat stripe (or yellow) rust (Puccinia striiformis f. sp. tritici) under field conditions of a newly developed wheat-rye (Mv9kr1-‘Lovaszpatonai’) disomic 4R addition line. Analysis of grain quality properties showed that the transfer of ‘Lovaszpatonai’ chromosome 4R into wheat significantly increased the total arabinoxylan and protein content. This genetic material proved to be resistant to stripe rust in 2014 and 2015 when epidemics broke out in Hungary. It is assumed that this addition line carries an effective, new resistance gene different from that on rye chromosome arm 1RS of the 1RS.1BL translocation. The Mv9kr1-‘Lovaszpatonai’ 4R addition line is a promising pre-breeding material being the first step towards creating translocation lines in order to transfer stripe rust resistance as well as high protein and dietary fibre content into cultivated wheat without deleterious effects on yield and grain quality.

Salinity-induced osmotic, ionic and oxidative stress responses were investigated on Asakaze/Manas wheat/barley addition lines 7H, 7HL and 7HS, together with their barley (salt-tolerant) and wheat (relatively salt-sensitive) parents. Growth, photosynthetic activity, chlorophyll degradation, proline, glycine betaine accumulation, sugar metabolism, Na+ and K+ uptake and transport processes and the role of polyamines and antioxidants were studied in young plants grown in hydroponic culture with or without salt treatment. Changes in plant growth and photosynthetic activity of plants demonstrated that the salt tolerance of the addition lines 7H and 7HL was similar to that of barley parent cv. Manas, while the sensitivity of the addition line 7HS was similar to that of the wheat parent cv. Asakaze. The Na accumulation in the roots and shoots did not differ between the addition lines and wheat parent. The activation of various genes related to Na uptake and transport was not correlated with the salt tolerance of the genotypes. These results indicated that the direct regulation of Na transport processes is not the main reason for the salt tolerance of these genotypes. Salt treatment induced a complex metabolic rearrangement in both the roots and shoots of all the genotypes. Elevated proline accumulation in the roots and enhanced sugar metabolism in the shoots were found to be important for salt tolerance in the 7H and 7HL addition lines and in barley cv. Manas. In wheat cv. Asakaze and the 7HS addition line the polyamine metabolism was activated. It seems that osmotic adjustment is a more important process in the improvement of salt tolerance in 7H addition lines than the direct regulation of Na transport processes or antioxidant defence.

Cereal grain fiber is an important health-promoting component in the human diet. One option to improve dietary fiber content and composition in wheat is to introduce genes from its wild relatives Aegilops biuncialis and Aegilops geniculata. This study showed that the addition of chromosomes 2Ug, 4Ug, 5Ug, 7Ug, 2Mg, 5Mg, and 7Mg of Ae. geniculata and 3Ub, 2Mb, 3Mb, and 7Mb of Ae. biuncialis into bread wheat increased the seed protein content. Chromosomes 1Ug and 1Mg increased the proportion of polymeric glutenin proteins, while the addition of chromosomes 1Ub and 6Ub led to its decrease. Both Aegilops species had higher proportions of β-glucan compared to arabinoxylan (AX) than wheat lines, and elevated β-glucan content was also observed in wheat chromosome addition lines 5U, 7U, and 7M. The AX content in wheat was increased by the addition of chromosomes 5Ug, 7Ug, and 1Ub while water-soluble AX was increased by the addition of chromosomes 5U, 5M, and 7M, and to a lesser extent by chromosomes 3, 4, 6Ug, and 2Mb. Chromosomes 5Ug and 7Mb also affected the structure of wheat AX, as shown by the pattern of oligosaccharides released by digestion with endoxylanase. These results will help to map genomic regions responsible for edible fiber content in Aegilops and will contribute to the efficient transfer of wild alleles in introgression breeding programs to obtain wheat varieties with improved health benefits. Key Message: Addition of Aegilops U- and M-genome chromosomes 5 and 7 improves seed protein and fiber content and composition in wheat.

Cereal grain fiber is an important health-promoting component in the human diet. One option to improve dietary fiber content and composition in wheat is to introduce genes from its wild relatives Aegilops biuncialis and Aegilops geniculata. This study showed that the addition of chromosomes 2Ug, 4Ug, 5Ug, 7Ug, 2Mg, 5Mg, and 7Mg of Ae. geniculata and 3Ub, 2Mb, 3Mb, and 7Mb of Ae. biuncialis into bread wheat increased the seed protein content. Chromosomes 1Ug and 1Mg increased the proportion of polymeric glutenin proteins, while the addition of chromosomes 1Ub and 6Ub led to its decrease. Both Aegilops species had higher proportions of β-glucan compared to arabinoxylan (AX) than wheat lines, and elevated β-glucan content was also observed in wheat chromosome addition lines 5U, 7U, and 7M. The AX content in wheat was increased by the addition of chromosomes 5Ug, 7Ug, and 1Ub while water-soluble AX was increased by the addition of chromosomes 5U, 5M, and 7M, and to a lesser extent by chromosomes 3, 4, 6Ug, and 2Mb. Chromosomes 5Ug and 7Mb also affected the structure of wheat AX, as shown by the pattern of oligosaccharides released by digestion with endoxylanase. These results will help to map genomic regions responsible for edible fiber content in Aegilops and will contribute to the efficient transfer of wild alleles in introgression breeding programs to obtain wheat varieties with improved health benefits. Key Message: Addition of Aegilops U- and M-genome chromosomes 5 and 7 improves seed protein and fiber content and composition in wheat.

The 3HS.3BL spontaneous Robertsonian translocation obtained from the progenies of wheat–barley (Chinese Spring × Betzes) hybrids backcrossed with wheat line Mv9kr1 was transferred into the modern Martonvásár wheat cultivar Mv Bodri. The translocation was identified with molecular cytogenetic methods. The inheritance of the translocation was traced using genomic in situ hybridization. Fluorescence in situ hybridization using barley subtelomeric (HvT01) and centromere-specific [(AGGGAG)4] repetitive DNA probes confirmed that the complete barley chromosome arm was involved in the Robertsonian translocation. The wheat-specific repetitive DNA probes identified the presence of the whole wheat genome, except the short arm of the 3B chromosome. Genotypes homozygous for the centric fusion were selected, after which morphological analysis was performed on the plants and the yield components were measured in the field during two consecutive vegetative seasons. The introgression of the 3HS.3BL translocation into the modern wheat cultivar Mv Bodri significantly reduced the plant height due to the incorporation of the dwarfing allele RhtD1b. The presence of the 3HS.3BL translocation in the Mv9kr1 and Mv Bodri wheat background improved tillering and seeds per plant productivity in field experiments carried out in Martonvásár and Keszthely, Hungary.

The salt stress responses of wheat–barley addition lines (2H, 3H, 3HS, 4H, 6H, 7H and 7HL) were compared to those of the parental genotypes wheat cv. Asakaze and barley cv. Manas and two other wheat genotypes [Chinese Spring (CS) and Mv9kr1] during germination and in young plants grown in hydroponic culture with or without salt treatment. Among the wheat genotypes frequently used for interspecific hybridization, Asakaze possesses relatively high salt tolerance, as indicated by the less pronounced reduction in germination % and in root and shoot growth and the retention of high leaf water content and photosynthetic activity, as compared to CS and Mv9kr1. The barley cv. Manas showed better salt tolerance than wheat cv. Asakaze, although Manas accumulated more Na in the root, but its transport to the shoots is restricted. Among the addition lines tested, the disomic addition line 7H and ditelosomic line 7HL exhibited higher salt tolerance both during germination and in the early developmental stages than the wheat parent, which may be related to the elevated osmotic adjustment capacity of these addition lines, similar to that found for barley cv. Manas. The paper also discusses the effects of other chromosomes on the salt stress response.

Wheat–barley addition (2H, 3H, 4H, 6HS, 7H) and translocation (7D-5HS) lines developed from a winter wheat ‘Martonvasari 9 kr1’ × winter barley ‘Igri’ cross were examined to determine how the added barley chromosomes (or segments) influence morphological and agronomic traits. The aim of our study was to investigate whether the advantageous traits of barley can be transferred to wheat by hybridization of the two species. The field experiment was carried out at UP Georgikon Faculty, Keszthely, during 2010–2011. Plants carrying the barley chromosomes had altered ontogenesis: days to 50% flowering, date of maturity, plant height, length and morphology of ear, flag leaf area, yield and components of grain yield. Regarding anthesis and maturity, the 7H addition line was the earliest, whereas the 4H addition was the latest to mature. Plant height and the length of spikes were decreased by the addition of 3H, 4H and 7H. Grain yield was decreased in addition lines 7H and 6HS and increased in addition 3H. Thousand grain weight was decreased in addition lines 2H, 4H and 6HS.

The established karyotype was generated by genomic in situ hybridization (GISH) using total barley genomic DNA as labelled probe on mitotic metaphase bread wheat chromosomes. GISH produced specific banding signals on 16 of the 21 chromosome pairs. The following chromosomes showed distinctive banding patterns: 2A, 3A, 4A, 5A, 6A, 7A, 1D, 2D, 7D and all of the B chromosomes. The remaining chromosomes showed either faint bands or no hybridization signals at all. The in situ hybridization patterns corresponded to the GAA-satellite sequence, which is similar to the N-banding pattern in wheat. In situ hybridization by labelling total barley genomic DNA made it possible to identify most of the bread wheat chromosomes. The present paper describes a GISH-banding method for hexaploid wheat chromosomes. It is a valuable alternative method for fast chromosome selection without using FISH repetitive DNA clones.

A recently developed synthetic amphiploid, Triticum timococcum Kost., nom. nud. (2n = 6x = 42, AtAtGGAmAm) is described in the present study. This hexaploid taxon was developed by colchicine treatment in Martonvasar from the hybrid of a selected accession of Triticum timopheevii Zhuk. (2n = 4x = 28, AtAtGG) and a prebred semi-dwarf line of Triticum monococcum L. (2n = 2x = 14, AmAm). A detailed cytomolecular examination was carried out using the sequential multicolour fluorescence and genomic in situ hybridization techniques (FISH and mcGISH). It was proved that T. timococcum has 42 chromosomes originating from its parents. The chromosomes of the A genomes of T. monococcum and T. timopheevii could be distinguished in the amphiploid using FISH. The successful discrimination of the chromosomes was supported by the karyotypes of the three genomes and the successful optimization of the mcGISH technique for the A and G chromosomes achieved in the present study. A phenotypic evaluation was also carried out under natural and artificial growing conditions in 2012 and 2013. Based on the results, T. timococcum has intermediate characteristics in terms of spike (spikelet) shape and plant height, while it is similar to the female parent, T. timopheevii regarding pubescence. Like its parents, T. timococcum showed outstanding resistance to the main fungal diseases of wheat. T. timococcum headed later and developed longer and looser spikes, fewer tillers and only a third as many seeds than its parents. The third generation of T. timococcum was able to develop an acceptable number of seeds, even taking into account the reduced germination ability in the field.

Osmotic stress responses of water content, photosynthetic parameters and biomass production were investigated in wheat-Aegilops biuncialis amphiploids and in wheat genotypes to clarify whether they can use to improve the drought tolerance of bread wheat. A decrease in the osmotic pressure of the medium resulted in considerable water loss, stomatal closure and a decreased CO2 assimilation rate for the wheat genotypes, while the changes in these parameters were moderate for the amphiploids. Maximal assimilation rate was maintained at high level even under severe osmotic stress in the amphiploids, while it decreased substantially in the wheat genotypes. Nevertheless, the effective quantum yield of PS II was higher and the quantum yield of non-photochemical quenching of PS II and PS I was lower for the amphiploids than for the wheat cultivars. Parallel with this, higher cyclic electron flow was detected in wheat than in the amphiploids. The elevated photosynthetic activity of amphiploids under osmotic stress conditions was manifested in higher biomass production by roots and shoots as compared to wheat genotypes. These results indicate that the drought-tolerant traits of Ae. biuncialis can be manifested in the wheat genetic background and these amphiploids are suitable genetic materials for improving drought tolerance of wheat.

Chromosomes 5A (u) , 5S and 5D can be isolated from wild progenitors, providing a chromosome-based approach to develop tools for breeding and to study the genome evolution of wheat. The three subgenomes of hexaploid bread wheat originated from Triticum urartu (A(u)A(u)), from a species similar to Aegilops speltoides (SS) (progenitor of the B genome), and from Ae. tauschii (DD). Earlier studies indicated the potential of chromosome genomics to assist gene transfer from wild relatives of wheat and discover novel genes for wheat improvement. This study evaluates the potential of flow cytometric chromosome sorting in the diploid progenitors of bread wheat. Flow karyotypes obtained by analysing DAPI-stained chromosomes were characterized and the contents of the chromosome peaks were determined. FISH analysis with repetitive DNA probes proved that chromosomes 5A(u), 5S and 5D could be sorted with purities of 78-90 %, while the remaining chromosomes could be sorted in groups of three. Twenty-five conserved orthologous set (COS) markers covering wheat homoeologous chromosome groups 1-7 were used for PCR with DNA amplified from flow-sorted chromosomes and genomic DNA. These assays validated the cytomolecular results as follows: peak I on flow karyotypes contained chromosome groups 1, 4 and 6, peak II represented homoeologous group 5, while peak III consisted of groups 2, 3 and 7. The isolation of individual chromosomes of wild progenitors provides an attractive opportunity to investigate the structure and evolution of the polyploid genome and to deliver tools for wheat improvement.

3MbTriticum aestivum L. (Mv9kr1) – Aegilops biuncialis Vis. (MvGB642) addition lines were crossed with the Chinese Spring ph1b mutant genotype (CSph1b) to produce 3Mb–wheat chromosome rearrangements. In the F3generation, 3Mb(4B) substitution lines and 3Mb.4BS centric fusions were identified with in situ hybridization using repetitive and genomic DNA probes, and with SSR markers. Grain micronutrient analysis showed that the investigated Ae. biuncialis accession MvGB382 and the parental line MvGB642 are suitable gene sources for improving the grain micronutrient content of wheat, as they have higher K, Zn, Fe, and Mn contents. The results suggested that the Ae. biuncialis chromosome 3Mbcarries genes determining the grain micronutrient content, as the 3Mb.4BS centric fusion had significantly higher Zn and Mn contents compared with the recipient wheat cultivar. As yield-related traits, such as the number of tillers, the length of main spike, and spikelets per main spike, were similar in the 3Mb.4BS centric fusion and the parental wheat genotype, it can be concluded that this line could be used in pre-breeding programs aimed at enriching elite wheat cultivars with essential micronutrients.

This work is a detailed study of the infection of fungal biotrophic pathogens causing powdery mildew diseases on introgression lines originating from the intergeneric hybridisation between wheat and barley (Triticum aestivum L. × Hordeum vulgare L.). Powdery mildew fungi are among the most widespread biotrophic pathogens of plants also and infect dicot and monocot species. Most powdery mildew species are strictly host specific. They colonize only a narrow range of species or one particular host species. The intergeneric hybridisation between wheat and barley could result in expansions of host ranges of the barley powdery mildew. Our experiments covered natural infections in the field and artificial infections under greenhouse conditions. Formae speciales of powdery mildew were identified on the basis of the sequencing results of ribosomal internal transcribed spacer sequences (rDNA-ITS). We identified Blumeria graminis f.sp. tritici isolate 14 (HM484334) on the wheat parent and all wheat-barley introgression lines and B. g. f. sp. hordei isolate MUMH1723 (AB 273556) on the barley parent, respectively. The wheat-barley introgression lines were inoculated with barley powdery mildew under greenhouse conditions. According to our results the added barley chromosomes (or segments) do not cause host range expansion of barley powdery mildew.

This paper reports detailed FISH-based karyotypes for three diploid wheatgrass species Agropyron cristatum (L.) Beauv., Thinopyrum bessarabicum (Savul.&Rayss) A. Love, Pseudoroegneria spicata (Pursh) A. Love, the supposed ancestors of hexaploid Thinopyrum intermedium (Host) Barkworth & D.R.Dewey, compiled using DNA repeats and comparative genome analysis based on COS markers. Fluorescence in situ hybridization (FISH) with repetitive DNA probes proved suitable for the identification of individual chromosomes in the diploid JJ, StSt and PP genomes. Of the seven microsatellite markers tested only the (GAA)n trinucleotide sequence was appropriate for use as a single chromosome marker for the P. spicata AS chromosome. Based on COS marker analysis, the phylogenetic relationship between diploid wheatgrasses and the hexaploid bread wheat genomes was established. These findings confirmed that the J and E genomes are in neighbouring clusters.

Several useful alien gene transfers have been reported from related species into wheat (Triticum aestivum), but very few publications have dealt with the development of wheat/barley (Hordeum vulgare) introgression lines. An overview is given here of wheat × barley hybridization over the last forty years, including the development of wheat × barley hybrids, and of addition and translocation lines with various barley cultivars. A short summary is also given of the wheat × barley hybrids produced with other Hordeum species. The meiotic pairing behaviour of wheat × barley hybrids is presented, with special regard to the detection of wheat–barley homoeologous pairing using the molecular cytogenetic technique GISH. The effect of in vitro multiplication on the genome composition of intergeneric hybrids is discussed, and the production and characterization of the latest wheat/barley translocation lines are presented. An overview of the agronomical traits (β-glucan content, earliness, salt tolerance, sprouting resistance, etc.) of the newly developed introgression lines is given. The exploitation and possible use of wheat/barley introgression lines for the most up-to-date molecular genetic studies (transcriptome analysis, sequencing of flow-sorted chromosomes) are also discussed.

The heading characters and morphological traits of two partial sets of wheat–barley disomic addition lines, namely Mv9kr1/Igri and Asakaze/Manas, were evaluated under controlled environmental conditions in a phytotron under long-day, short-day and non-vernalised conditions and in field-sown experiments. The winter barley chromosome additions significantly influenced the flowering time of wheat both in the controlled environment test and under field-sown conditions. Of all the barley addition lines, the effect of the 4H and 7H additions was the most characteristic. The 7H addition lines were the earliest in both cultivar combinations in each treatment. In the Mv9kr1/Igri combination the 4H addition was the latest under all the environmental conditions. In the Asakaze/Manas combination 4H addition was the latest under short-day and long-day illumination in the phytotron but the 6H addition was the latest without vernalisation and in the field in 2012. There was 12 and 11 days difference between the flowering times of the 7H and 4H Mv9kr1/Igri and Asakaze/Manas addition lines in the field in 2012, which increased to 52 and 44 days under short-day illumination in the phytotron. In the winter wheat background, the addition of 2H carrying the photoperiod sensitivity gene Ppd-H1 decreased the flowering time under the short photoperiod regime, but had a very strong delaying effect under field-grown conditions. Considering the yield components under field conditions, 4H was the most fertile of the addition lines, while 7H showed the highest tillering capacity, and Igri 3H had good tillering capacity and the highest number of seeds per plant.

as ´ ar, Hungary Abstract The dietary fibre (1,3;1,4)-β-D-glucan (β-glucan), is a major quality parameter of cereals. Grain β-glucan content is the most important factor from the aspect of human health maintenance. The grain of barley (Hordeum vulgare )i s one of the most important β-glucan sources having a β-glucan content 10 times higher than that of wheat (Triticum aestivum). Winter wheat/winter barley 'Mv9kr1'/'Igri' 1HS ditelosomic and 'Mv9kr1'/'Igri' 7H disomic addition lines carrying the HvCslF9 and HvCslF6 barley genes, respectively, were used to investigate the additive effect of barley cellulose synthase-like genes on the wheat β-glucan content. A significantly higher β-glucan level was detected in the leaves and grains of the wheat/barley 1HS and 7H addition lines compared to the control wheat line. The expression of the HvCslF9 and HvCslF6 genes in the genetic background of wheat was also determined by quantitative RT- PCR. The expression pattern of the HvCslF9 gene transcript showed a gradual increase throughout grain development, while the HvCslF6 gene was normally transcribed at relatively high levels. In leaves, the transcript of the HvCslF9 gene could not be detected at the end of tillering, while the HvCslF6 gene was still strongly expressed at this time in the 7H addition line. This study mapped the HvCslF9 gene to the short arm of the 1H chromosome. The HvGlb1 barley gene, encoding (1,3;1,4)-β-D-glucan endohydrolase isoenzyme EI, is possibly involved in the regulation of the β-glucan level during grain development. Previously this was also mapped to the barley 1H chromosome, and this study suggested that it was located on the 1HL chromosome arm. It was also concluded that the independent expression of the HvCslF9 gene in the wheat background resulted in a slight increase in the β-glucan content. The results provide new insights into the expression and regulation of the HvCslF genes in the genetic background of wheat and indicate that cisgenesis can be used to increase the leaf and grain β-glucan content in wheat.

A spontaneous wheat-barley translocation line was previously detected in the progenies of the Mv9kr1 × 'Igri' wheat-barley hybrid and the translocation was identified as 5HS-7DS.7DL. Multicolor genomic in situ hybridization (mcGISH) with D and H genomic DNA probes and three-color fluorescence in situ hybridization (FISH) with repetitive DNA probes (Afa-family, pSc119.2, and pTa71) were performed to characterize the rearranged chromosome. The effect of 5HS and the deleted 7DS fragment on the morphological traits (plant height, fertility, yield, and spike characteristics) of wheat was assessed. Despite the non-compensating nature of the translocation, the plants showed good viability. The aim of the study was to physically localize SSR markers to the telomeric and subtelomeric regions of the 7DS chromosome arm. Of the 45 microsatellite markers analyzed, ten (Xbarc0184, Xwmc0506, Xgdm0130, Xgwm0735, Xgwm1258, Xgwm1123, Xgwm1250, Xgwm1055, Xgwm1220, and Xgwm0635) failed to amplify any 7DS-specific fragments, signaling the elimination of a short chromosome segment in the telomeric region. The breakpoint of the 5HS-7DS.7DL translocation appeared to be more distal than that of reported deletion lines, which provides a new physical landmark for future deletion mapping studies.

In recent years 56 accessions of Triticum timopheevii Zhuk. (2n=4x=28, AtAtGG) were characterized for the main phenotypic and resistance characters. Among these accessions 38 originated from the base species together with subspecies and varietas forms thereof, and 18 belong to the subspecies armeniacum group. After the evaluation of field assessment data gathered over 12 years, the most promising 11 accessions were selected for a crossability trial with cultivated einkorn. As a result of this trial, the accession with the highest seed set (Acc. No.: MVGB845) was chosen for the development of a new synthetic amphiploid using the same semi-dwarf line of diploid cultivated einkorn (Triticum monococcum L. ssp. monococcum 1T-1, 2n=2x=14, AmAm) as in the crossability trial. This einkorn line was bred in Martonvásár, and has both outstanding resistance and other promising phenotypic and agronomic characters.After crossing the accession MVGB845 with 1T-1, the triploid hybrids were treated with colchicine to obtain fertile progenies with a doubled genome. The newly developed synthetic hexaploid wheat breeding stock (named Triticum timococcum Kost., 2n=6x=42, AtAtGGAmAm) could ease the introgression of valuable resistance genes into bread wheat at the hexaploid level (bridge-crossing).The aim of the present research was to redevelop Triticum timococcum based on a detailed characterization of gene bank accessions, and to introduce this new material into wheat breeding.

The 4H(4D) wheat/barley substitution line was crossed with the ‘Chinese Spring’ ph1b mutant genotype in order to induce wheat-barley homoeologous recombinations. F3 and F4 seeds of the 4H(4D) × ‘Chinese Spring’ ph1b mutant cross were analysed using genomic in situ hybridization, and a Robertsonian translocation was detected in monosomic form. Disomic centric fusions were selected among the self-fertilized progenies. The presence of the long arm of 4H was confirmed with SSR markers. The long arm of the 5D wheat chromosome in the Robertsonian translocation was identified using fluorescent in situ hybridization with the help of three DNA probes: pSc119.2, Afa family and pTa71. The wheat/barley centric fusion was identified as a 4HL.5DL translocation. This line exhibited supernumerary spikelet character, but the number of seeds/plant did not increase. The 4HL.5DL centric fusion line is suitable genetic material to study the expression of genes located on 4HL in a wheat genetic background.

Salix viminalis L. (2n = 38) is a diploid dicot species belonging to the Salix genus of the Salicaceae family. This short-rotation woody crop is one of the most important renewable bioenergy resources worldwide. In breeding for high biomass productivity, limited knowledge is available on the molecular cytogenetics of willow, which could be combined with genetic linkage mapping. The present paper describes the adaptation of a fluorescence in situ hybridisation (FISH) protocol as a new approach to analyse the genomic constitution of Salix viminalis using the heterologous DNA clones pSc119.2, pTa71, pTa794, pAs1, Afa-family, pAl1, HT100.3, ZCF1 and the GAA microsatellite marker. Three of the nine probes showed unambiguous signals on the metaphase chromosomes. FISH analysis with the pTa71 probe detected one major 18S-5.8S-26S rDNA locus on the short arm of one chromosome pair; however, the pTa794 rDNA site was not visible. One chromosome pair showed a distinct signal around the centromeric region after FISH with the telomere-specific DNA clone HT100.3. Two chromosome pairs were found to have pAs1 FISH signals, which represent a D-genome-specific insert from Aegilops tauschii. Based on the FISH study, a set of chromosomes with characteristic patterns is presented, which could be used to establish the karyotype of willow species.

Chromosome pairing in the meiotic metaphase I of wheat-rye hybrids has been characterized by sequential genomic and fluorescent in situ hybridization allowing not only the discrimination of wheat and rye chromosomes, but also the identification of the individual wheat and rye chromosome arms involved in the chromosome associations. The majority of associations (93.8%) were observed between the wheat chromosomes. The largest number of wheat-wheat chromosome associations (53%) was detected between the A and D genomes, while the frequency of B-D and A-B associations was significantly lower (32 and 8%, respectively). Among the A-D chromosome associations, pairing between the 3AL and 3DL arms was observed with the highest frequency, while the most frequent of all the chromosome associations (0.113/cell) was found to be the 3DS-3BS. Differences in the pairing frequency of the individual chromosome arms of wheat-rye hybrids have been discussed in relation to the homoeologous relationships between the constituent genomes of hexaploid wheat.

The aim of the study was to select wheat-Aegilops biuncialis addition lines carrying Aegilops biuncialis chromosomes differing from those which were introgressed into the wheat-Ae. biuncialis addition lines produced earlier in Martonvasar, Hungary. In the course of the experiments new wheat-Ae. biuncialis addition lines carrying chromosomes 2Ub, 6Mb, 6Ub; 5Ub, 3Ub, 7Ub; 5Mb, 6Mb and 7Mb were selected. The 2Ub disomic addition line is relatively stable, as 91% of the progenies contain this chromosome pair. The 6Mb disomic addition line proved to be dwarf and sterile, but it still exists as a monosomic addition line. Progenies analysed from the 6Ub monosomic addition line did not carry the 6Ub chromosome. One plant containing the 5Ub, 3Ub and 7Ub chromosomes and one plant carrying 5Mb, 6Mb and 7Mb chromosomes showed very low fertility. Each of the plants produced a single seed, but seeds of the parent plants are still available. Line No. 49/00 carried a submetacentric Ae. biuncialis chromosome pair and the chromosome number 44 has been constant for several generations. After FISH no hybridisation site was observed on the Ae. biuncialis chromosome pair using the pSc119.2 and Afa family repetitive DNA probes, so it was not possible to identify the Ae. biuncialis chromosome pair. However, the use of wheat SSR markers and the (GAA)n microsatellite DNA probe allowed it to be characterised more accurately. These new lines facilitate gene transfer from Ae. biuncialis into cultivated wheat and the selection of U and M genome-specific wheat SSR markers.