PI-15 - Rubén Gómez RUFO - Genetic structure of Mediterranean wheat landraces assessed with SNP markers

Genetic structure of Mediterranean wheat landraces assessed with SNP markers

Rubén Gómez RUFO¹, Fanny ÁLVARO¹, Conxita ROYO¹ and José Miguel SORIANO¹

¹ Av. Alcalde Rovira Roure, 191, 25198 Lleida

Corresponding author: rubenrufogomez@gmail.com

Wheat is one of the most important grain crops in the world, being the hexaploid wheat (Triticum aestivum L.) the 95% of world wheat production. Landraces represent a valuable source of genetic diversity and are well adapted to local environmental conditions. Landraces may provide new alleles for the improvement of commercially valuable traits, being very useful especially in breeding for suboptimal environments as the Mediterranean Basin. Knowledge of genetic diversity is essential for understanding the relationships between cultivars and for the use of landraces in breeding programmes.

In this study, the genetic structure of a collection of 170 wheat landraces from 24 Mediterranean countries was ascertained using the wheat 15K SNP array. A total of 10,458 markers resulted polymorphic among the 170 genotypes. A model-based cluster method (Bayesian algorithm) using STRUCTURE software allocated 83.5% of the accessions in three subpopulations related to their geographic origin: North Africa and West of Mediterranean, East of Mediterranean and South Europe and North of Mediterranean.

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**PI-16 - Andrii FATIUKHA** - Genomic architecture of morphological and phenological traits associated with domestication syndrome in durum × wild emmer wheat RIL population

# Genomic architecture of morphological and phenological traits associated with domestication syndrome in durum × wild emmer wheat RIL population

Andrii FATIUKHA<fnref n="1" />, Tzion FAHIMA<fnref n="1" />, Valentina KLYMIUK<fnref n="1" />, Tamar KRUGMAN<fnref n="1" /> and Abraham B KOROL<fnref n="1" />

Corresponding author: avfatyha@gmail.com

Domestication and subsequent evolution under domestication of wheat caused substantial genetic changes, which affected plant morphology, physiology and phenology. Morphological characters, such as compactness of spikes, the number of side shoots, can be mentioned as domestication related traits in cereals. We suggest to consider the angle of side shoots (Ash) as a novel trait associated with the domestication syndrome. The objective of this study is to provide a better understanding of the antagonism between natural and man-made selection of the traits under domestication in order to identify the significant changes in phenology and morphology of wheat during domestication. We used a recombinant inbred line (RIL) population derived from a cross between *Triticum durum* (cv. Langdon) and *Triticum dicoccoides* (acc. G18-16) for mapping of quantitative trait loci (QTL) of five morphological and three phenological traits. A total of 36 QTL effects were identified that were co-located in 21 loci. Eight of these loci showed pleiotropic effects on the studied traits (including phenology). A major QTL effect of Ash, co-located with strong phenological effect, was identified on chromosome 2BL. We found that phenological loci affected the duration of flowering and development of wheat in different manners. The duration of the reproductive stage in cereals affects the development of apical meristem and many other morphological traits, such as the number of spikelets per spike and the number of side shoots. These results shed more light on shaping of wheat plant architecture and development during its evolution under domestication.

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<footnote n="1">¹ Institute of Evolution and the Department of Evolutionary and Environmental Biology, Faculty of Science and Science Education, University of Haifa</footnote>
**PI-17 - Anna HAWLICZEK-STRULAK** - Investigation of genetic diversity patterns and search for selective sweeps in a worldwide collection of rye

# Investigation of genetic diversity patterns and search for selective sweeps in a worldwide collection of rye

Anna HAWLICZEK-STRULAK<fnref n="1" />, Katarzyna TOFIL<fnref n="1" />, Ewa BORZECKA<fnref n="1" />, Piotr GAWRONSKI<fnref n="1" />
and Hanna BOLIBOK-BRAGOSZEWSKA<fnref n="1" />

Corresponding author: ahawliczek@gmail.com

Rye (Secale cereale L.) is an important cereal in Poland and other countries of Central and Eastern Europe. There are over 80 rye gene banks worldwide, with over 21,000 accessions. A better knowledge about the genetic variation contained in rye ex situ collections would facilitate both the conservation and management of genetic resources and their exploitation in breeding programs.

We compiled a rye germplasm collection consisting of over 600 accessions, representing major gene banks, different improvement status and various geographic origins. So far, 269 accessions were genotyped using Genotyping by Sequencing (DArTSeq). The plant material included 44 wild accessions, 105 landraces, 83 historical varieties, 14 modern varieties and 25 accessions of unknown improvement status. Based on 7,664 SilicoDArT and 2,559 SNP markers (missing data < 0.05, minor allele freq. >0.05) we conducted hierarchical clustering, principal coordinates analysis, analysis of molecular variance, and assignment tests. Distribution of polymorphism along rye chromosomes was also analyzed.

Obtained results allow us to conclude about the influence of improvement status, accessions source and geographical origin on the genetic diversity structure. Our aim is also an identification and characterization of genome regions targeted by selection during the domestication and breeding of rye.

This research was founded by the Polish National Science Centre grant No. DEC-2014/14/E/NZ9/00285

**Keywords:** rye, genetic diversity, genotyping by sequencing, DArTSeq, population structure, genetic resources

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<footnote n="1">¹ Department of Plant Genetics, Breeding and Biotechnology, Warsaw University of Life Sciences, Warsaw, Poland</footnote>
**PI-18 - Arantxa MONTEAGUDO** - Detecting useful genetic diversity from Spanish landraces to improve an elite cultivar

# Detecting useful genetic diversity from Spanish landraces to improve an elite cultivar

Arantxa MONTEAGUDO<fnref n="1" />, Ana M CASAS<fnref n="1" />, M Pilar GRACIA<fnref n="1" /> and Ernesto IGARTUA<fnref n="1" />

Corresponding author: amonteagudo@eead.csic.es

Spanish Barley Core Collection (SBCC) has shown genetic variability with potential interest for breeding, particularly aiming at adaptability to Mediterranean environments. Cierzo, a successful elite barley cultivar selected under Spanish Mediterranean conditions, was chosen as recurrent parent of a backcross scheme. The donor parent, SBCC073, was selected among the SBCC lines due to its good yield performance under drought conditions. Under an augmented design with incomplete blocks, yield and drought-related traits and flowering time were evaluated in two field trials in Zaragoza (Spain). Raw data were spatially corrected using a moving average correction approach. Genotyping of 264 lines was carried out using the Barley Illumina iSelect 50k SNPchip. A genetic map was generated with 12,667 SNPs covering the seven linkage groups. Using the results of two years of field trials (2014-2015 and 2015-2016), we identified seven regions with stable QTLs. Among them, SBCC073 contributed favorable alleles for vegetative growth, measures in different moments of development, with a QTL in the region of 5H 138.5 cM, related also to flowering. The candidate of this region could be the response-regulator PRR95, component of the circadian clock. Another important region was found on 6H, at 59.1 cM, with QTLs for plant height, thousand-kernel weight and yield, being the SBCC073 allele responsible of increasing plant height and thousand-kernel weight, and reducing yield. No candidate gene has been found for this region yet. The QTLs found would be useful to develop improved breeding strategies based on exploiting the SBCC genetic diversity.

**Keywords:** Landraces, Yield, drought, flowering, QTLs

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<footnote n="1">¹ Aula Dei Experimental Station (EEAD-CSIC), Avda. Montañana 1005, E-50059 Zaragoza, Spain</footnote>
**PI-19 - Hanna Bolibok-Bragoszewska** - Highly parallel amplicon sequencing for characterization of genetic diversity in rye germplasm

Highly parallel amplicon sequencing for characterization of genetic diversity in rye
germplasm

Anna HAWLICZEK-STRULAK<fnref n="1" />, Bradley TILL<fnref n="2" />, Joanna JANKOWICZ-CIESLAK<fnref n="2" />, Ewa BORZECKA<fnref n="1" />,
Katarzyna TOFIL<fnref n="1" />, Adam KRAL<fnref n="1" /> and Hanna BOLIBOK-BRAGOSZEWSKA<fnref n="1" />

Corresponding author: [hanna_bolibok_bragoszewska@sggw.pl](mailto:hanna_bolibok_bragoszewska@sggw.pl)

Improved plant cultivars will be required to feed the world's growing population, while preserving natural resources, and to face climatic changes. Novel, favorable alleles can be found within the crop - mostly in landraces and also in the wild relatives. The aim of the project was to develop a low-cost amplicon sequencing approach employing high sample pooling suitable for assessment of genetic diversity in heterozygous and heterogenous rye accessions.
We performed a pilot screen involving 95 diverse accessions of different origin and improvement status (landraces, historical and contemporary varieties, wild species). Each accession was represented by a pooled DNA sample representing 96 individuals, thus 9,120 plants were analyzed in total. Six genes: aluminium activated citrate transporter (AACT1), taumatin-like protein (TLP), fructose-biphosphate aldolase (FBA), prolamine binding factor (PBF), secaloindoline-b (SEC-B) and grain softness protein (GSP1), ranging in length from 456 to 4,638 bp, were amplified from the 95 pooled samples. Amplicon sequencing was done using an Illumina MiSeq with 2x300PE reads. Due to small scale of the experiment a high depth of coverage was obtained (above 1000x for the lowest represented amplicon). So far, 869 polymorphic sites with 990 variant alleles were identified using the GATK SNP-discovery tool. Allele frequencies varied from 0.001 to 1.0 and 25% of all values were in the range 0.1 – 0.2. Among detected variants, 28% (243) appeared in only one accession from the set, and the number of polymorphic sites per accession ranged from 111 to 337. In a validation experiment, based on a small number of individual plants (10-20) from selected accessions and information on restriction enzyme site gain/loss provided by PARSESNP, we were able to confirm SNPs occurring with frequencies above 0.1. Currently, evaluation of nucleotide variation is being carried out using SNVer, FreeBayes and CAMBa. Nucleotide changes are further evaluated for their effect on gene function.
We conclude that the proposed experimental design allows for reliable, high-throughput SNP discovery and accurate variant frequency estimation in large rye germplasm sets.

This research was founded by the Polish National Science Centre grant No. DEC-2014/14/E/NZ9/00285

**Keywords:** rye, NGS, genetic diversity, Ecotilling, amplicon sequencing

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<footnote n="1">¹ Department of Plant Genetics, Breeding and Biotechnology, Warsaw University of Life Sciences, Warsaw, Poland</footnote>

<footnote n="2">² International Atomic Energy Agency, Vienna International Centre, Vienna, Austria</footnote>
**PI-20 - Ian DAWSON** - Exploring local adaptation in barley landraces through latitudinal differences in days to heading

Exploring local adaptation in barley landraces through latitudinal differences in
days to heading

Ian DAWSON<fnref n="1" />, Daniela BUSTOS-KORTS<fnref n="2" />, D CAMMARANO<fnref n="1" />, Fred A VAN EEUWIJK<fnref n="2" />, Alessandro TONDELLI<fnref n="3" />,
Laura ROSSINI<fnref n="4" />, S KYRIAKIDIS<fnref n="1" />, Luigi CATTIVELLI<fnref n="3" />, R WAUGH<fnref n="1" /> and JR RUSSELL<fnref n="1" />

Corresponding author: joanne.russell@hutton.ac.uk

Landraces are defined as being adapted to their local production systems, but actual tests of their adaptation are limited,
despite the importance of the issue for designing crop-breeding strategies that respond appropriately to anthropogenic
climate change. Here, we explore the issue of local adaptation through assessing the heading dates of spring-type
barley landraces and formally bred cultivars sampled from across a wide latitudinal range in spring-planted field
experiments including those of the current WHEALBI initiative. Our tests provide some evidence for local adaptation
in barley landraces that supports conventional – but frequently unproven – wisdom, but reveal a number of caveats
and limitations in the applied approach. We discuss more optimal approaches to explore adaptation more widely.

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<footnote n="1">¹ James Hutton Institute, Invergowrie, Dundee, Scotland</footnote>

<footnote n="2">² Biometris, Wageningen University & Research Centre, Wageningen, The Netherlands</footnote>

<footnote n="3">³ Consiglio per la ricerca in agricoltura e l'analisi dell'economia agraria (CREA), Research Centre for Genomics and Bioinformatics, Fiorenzuola, Italy</footnote>

<footnote n="4">⁴ Parco Tecnologico Padano, Lodi, Italy</footnote>
**PI-21 - Jane ROCHE** - Coexpression network and phenotypic analysis identify metabolic pathways associated with the effect of warming on grain yield components in wheat

Coexpression network and phenotypic analysis identify metabolic pathways
associated with the effect of warming on grain yield components in wheat

Jane ROCHE<fnref n="1" />, Christine GIROUSSE<fnref n="1" />, Claire GUÉRIN<fnref n="1" />, Jacques LE GOUIS<fnref n="1" />, Sandrine BALZERGUE<fnref n="1" />, Saïd MOUZEYAR<fnref n="1" /> and Mohamed Fouad BOUZIDI<fnref n="1" />

Corresponding author: Jane.Roche@uca.fr

Wheat grains are an important source of human food but current production amounts cannot meet world needs. Several environmental constraints such as heat could potentially negatively affect wheat production. Therefore, it is important to understand the response of grains during the development to an increase of ambient temperature in a global warming context.

Plants from two wheat genotypes have been submitted to two growth temperature regimes. One set has been grown at optimal mean daily temperature of 19°C while the second set of plants has been submitted to warming at 27°C from 2 days after anthesis (daa) to 13 daa. Gene expression was evaluated using micro-array during nine different seed developmental stages and agronomic traits were measured at harvest.

While warming did not affect mean grain number per spike, it significantly reduced other yield-related indicators such as grain width, length, volume and maximal number in the endosperm. Whole genome expression analysis identified 6,258 and 5,220 genes whose expression was affected by temperature in the two genotypes respectively. The greater difference between low and high temperature was displayed at a key stage 120°C days after anthesis (°Cd) which suggest that significant transcriptional changes occur during the transition that encompasses the 120°Cd stage in response to warming.

Co-expression analysis using WGCNA uncovered modules associated with agronomic traits. In particular, modules enriched in genes related to nutrient reservoir and endopeptidase inhibitor activities were found positively associated with cell number in the endosperm. A hypothetical model pertaining to the effects of warming on gene expression and growth in wheat grain is proposed.

Network analyses suggest a negative effect of the expression of genes related to seed storage proteins and starch biosynthesis upon moderately high temperature on the grain size in wheat.

**Keywords:** wheat, elevation of temperature, grain size, grain development, transcriptomic, WGCNA

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<footnote n="1">¹ INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France</footnote>
**PI-22 - José Miguel SORIANO** - Eastern and Western durum wheat Mediterranean landraces differ in marker alleles responsible for yield components and phenology as revealed by association analysis

# Eastern and Western durum wheat Mediterranean landraces differ in marker alleles responsible for yield components and phenology as revealed by association analysis

José Miguel SORIANO<fnref n="1" />, Dolors VILLEGAS<fnref n="1" />, Luis F GARCIA DEL MORAL<fnref n="2" />, Maria Jose ARANZANA<fnref n="3" />, Martina ROSELLÓ<fnref n="1" />, Marcos MALOSETTI<fnref n="4" />, Mark Earl SORRELLS<fnref n="5" /> and Conxita ROYO<fnref n="1" />

Corresponding author: josemiguel.soriano@irta.cat

A panel of 172 durum wheat landraces from 21 Mediterranean countries were phenotyped during 3 years in northern and southern locations from Spain for 14 traits including phenology, biomass, yield and yield components.

The genetic structure of the collection was ascertained with 44 simple sequence repeat markers allocating 90% of the accessions in four subpopulations related to their geographic origin: eastern Mediterranean, eastern Balkans and Turkey, western Balkans and Egypt, and western Mediterranean.

Association mapping was used to identify genome regions affecting yield formation, crop phenology and crop biomass. The panel was genotyped with 1,149 DArT markers. A total of 245 significant marker trait associations (MTAs) were detected. Eighty-six MTAs corresponded with yield and yield component traits, 70 to phenology and 89 to biomass production. Twelve genomic regions harbouring stable MTAs were identified, while five and two regions showed specific MTAs for northern and southern environments, respectively. Sixty per cent of MTAs were located on the B genome and 29% on the A genome.

QTL meta-analysis was performed to compare MTAs with previously identified QTLs. A total of 477 unique QTLs were projected onto a durum wheat consensus map and were condensed to 71 meta-QTLs and left 13 QTLs as singletons. Sixty-one percent of QTLs explained less than 10% of the phenotypic variance confirming the high genetic complexity of the traits analysed.

Using a subset of the landrace panel 23 marker alleles were identified with a different frequency in landraces from east and west of the Mediterranean Basin, which affected important agronomic traits. Eastern landraces had higher frequencies than the western ones of alleles increasing the number of spikes, grains per m2, and grain filling duration. Eastern landraces had higher frequencies of marker alleles associated with reduced cycle length, and lighter grains than the western ones. Breeders may use the molecular markers identified in the current study for improving yield under specific Mediterranean environments.

**Related publication:**

Soriano JM, Villegas D, Aranzana MJ, Garcia del Moral LF, Royo C (2016) Genetic structure of modern durum wheat cultivars and Mediterranean landraces matches with their agronomic performance. PLoS One. 11 (8): e0160983.

Soriano JM, Malosetti M, Roselló M, Sorrells ME, Royo C (2017) Dissecting the old Mediterranean durum wheat genetic architecture for phenology, biomass and yield formation by association mapping and QTL meta-analysis. PLoS ONE 12 (5): e0178290

**Keywords:** Durum wheat, genetic structure, association mapping, meta-QTL, Mediterranean Basin

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<footnote n="1">¹ Sustainable Field Crops Programme, IRTA (Institute for Food and Agricultural Research and Technology), Lleida, Spain</footnote>

<footnote n="2">² Department of Plant Physiology, Institute of Biotechnology, University of Granada, Granada, Spain</footnote>

<footnote n="3">³ Plant and Animal Genomics Programme, IRTA, Centre de Recerca en Agrigenómica, Bellaterra, Barcelona, Spain</footnote>

<footnote n="4">⁴ Biometrics, Wageningen University and Research Centre, Wageningen, The Netherlands</footnote>

<footnote n="5">⁵ Department of Plant Breeding and Genetics, Cornell University, Ithaca, NY, USA</footnote>
**PI-23 - Monika RAKOCZY-TROJANOWSKA** - New aspects of the genetic background of benzoxazinoid biosynthesis in rye (*Secale cereale* L.)

# New aspects of the genetic background of benzoxazinoid biosynthesis in rye (*Secale cereale* L.)

Monika RAKOCZY-TROJANOWSKA<fnref n="1" />, Magdalena ŚWIĘCICKA<fnref n="1" />, Jacek RYMUSZKA<fnref n="1" />, Anna STOCHMAL<fnref n="2" />
and Mariusz KOWALCZYK<fnref n="2" />

Corresponding author: monika_rakoczy_trojanowska@sggw.pl

Benzoxazinoids (BX) are the major protective and allelopathic secondary metabolites that are synthesized in many species of the *Poaceae* and, sporadically, in single species of the dicots. Despite years of research many problems connected with BX remain unsolved. Especially ambiguous is the role of two genes controlling indole synthesis: Bx1 and Igl, both taking part in transformation of indole-3-glycerolphosphate (IGP) to indole. According to the general opinion: (1) Bx1 codes for BX1 enzyme catalyzing the conversion of IGP to indole, which is further used in BX biosynthesis. Igl (its rye orthologue), that has been recently isolated by our team, codes for indole-3-glycerol phosphate lyase (IGL) which, "produces" free indole as a response to the stress agent; (2) the Bx1 gene is under developmental control, and is mainly responsible for BX production, whereas the Igl gene is inducible by stress signals, such as wounding, herbivory, or jasmonates. However, the results of our previous study contradict this concept and show, that the expression of ScBx1 gene increases in the native conditions until the third day after germination and then decreases rapidly, while the BX are still synthesized at a constant level. The situation changed completely after the BSMV infection – the expression of ScBx1 increased nearly 40 fold (!). Quite poorly understood aspect of the genetic background of BX biosynthesis is the regulation of Bx gene expression. The majority of studies were focused on the influence of BX on pests, pathogenic fungi and weeds. The knowledge about the opposite relationship: how different factors affect Bx gene expression is still fragmentary and limited. So far, it has not been investigated whether the induction of BX synthesis in the above ground parts of plants is accompanied by an increase in the production of these compounds in the roots (and vice versa) and what is the mechanism of signalling. Another question concerns the sites of BX synthesis - whether it takes place only in the above ground parts of plants or also in the roots.
With regard to rye, we have been able to clarify some of these doubts. Three rye inbred lines: D33, L318 and D39 characterized, respectively, by high, middle and low content of BX in the above ground parts and roots (measured in the early spring) were used.

The main results of the reported study are presented below.

1. *ScIgL* gene has been isolated from line L318 BAC library and characterized. *ScIgl* is predicted to be composed of seven exons of total length equal 989 bp. Its complete length (including introns) is 1,651 bp. The results of bioinformatic analysis (presence and frequency of specific motifs potentially involved in different abiotic and biotic stress response) suggest that *ScIgL* gene plays role in different defense processes.

2. It was proved that the expression of *ScBx1* and *ScIgL* genes take place both in the above ground parts of rye plants and in roots. It denies the current opinion according to which a step of BX synthesis controlled by *Bx1* is located in chloroplasts.

3. The examination of expression profiles of genes *ScBx1* – *ScBx5* and *ScIgL* in plants affected by allochemicals secreted by berseem clover (Trifolium alexandrinum L.) showed that: (i) compared to control (cultivation without berseem clover), their expression usually decreased after 2 weeks, after the next 2 weeks – it increased and in the sixth week, the significantly heightened expression of the majority of genes (except for *ScBx1*) was observed only in line D39; (ii) the expression level of *ScBx1* decreased while in case of *ScIgL* - it increased in the above parts of plants. (iii) line D39 was characterized by the highest enhance of an expression due to allochemicals of the majority of genes up to the fourth week of co-cultivation with berseem clover (iv) among six investigated genes, *ScIgL* and *ScBx5* were most affected by allochemicals in the case of the above ground parts of plants, and *ScBx3*, *ScBx4* and *ScBx5* in the case of roots, especially after 4 weeks of co-cultivation with berseem clover, (v) an increase in the level of DIBOA

<footnote n="1">¹ Warsaw University of Life Sciences, Nowoursynowska 166, 02-787 Warszawa, Poland</footnote>

<footnote n="2">² Institute of Soil Science and Plant Cultivation, Czartoryskich 8, 24-100 Pulawy</footnote>
synthesis in the above ground parts of plants was observed after 2 weeks of co-culture with berseem clover in case of line D33, in fourth week - for line L318 and in sixth week - for line D39 which not fully correlate with gene expression profiles. Full interpretation of these results will be possible after the completion of biochemical analyses of BX content in roots which are currently in progress.

The obtained results not only broadened the current knowledge about the genetic basis of BX biosynthesis, but they also changed some of the existing dogmas, at least those concerning the places of BX biosynthesis and the regulation of ScBx and ScIgL gene expression by some stresses. Perhaps our findings will change the opinion that only Bx1 takes part in the biosynthesis of BX and is developmentally regulated whereas *Igl* controls the synthesis of free indole emitted by plants as a response to the stress.

The research has been financed by the National Science Centre (Poland) – No UMO-2015/19/B/NZ9/00921

**Keywords:** rye, *Secale cereale* L., *ScBx* gene, *ScIgL* gene, gene expression, allelolopathy, biotic stress, berseem clover, *Trifolium alexandrinum* L.

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**PI-24 - Rasha TARAWNEH - Mapping QTL for drought tolerance in spring wheat genebank accessions using genome wide association approach**

**Mapping QTL for drought tolerance in spring wheat genebank accessions using
genome wide association approach**

Rasha TARAWNEH<fnref n="1" />, Manuela NAGEL<fnref n="1" />, Fruzsina SZIRA<fnref n="2" />, István MONOSTORI<fnref n="2" />, Marion RÖDER<fnref n="1" />
and Andreas BÖRNER<fnref n="1" />

Corresponding author: tarawneh@ipk-gatersleben.de

Drought is one of the most important abiotic stresses affects yield production in cereal crops world-wide. As a result of climate changes, drought severity is expected to increase in the future. In order to understand the genetic variation and to detect quantitative trait loci for drought tolerance, a set of 111 spring wheat genebank accessions from 27 countries and genotyped with a 15k chip was grown in the field in two consecutive years. Drought stress was applied by chemical desiccation using potassium iodide (KI, 0.5% w/v) applied 14 days after anthesis to simulate post anthesis drought stress. In addition, a rain-shelter experiment was initiated at the Center for Agricultural Research in Martonvásár, Hungary during one season. The accessions were evaluated for a number of morphological and agronomical traits such as plant height, spike length, grain number and thousand-grain weight. Genome-wide association study (GWAS) was performed to reveal significant marker-trait associations (MTAs) under drought and control conditions. Analysis showed that drought stress had strong impact on yield parameters. Significant differences between genotypes and GWAS analysis revealed major MTAs under control, chemical desiccation and rain-shelter conditions.

**Keywords:** Drought, Chemical desiccation, Genome wide association

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<footnote n="1">¹ Leibniz Institute of Plant Genetics and Crop Plant Research (IPK Gatersleben), Stadt Seeland, Germany</footnote>

<footnote n="2">² Centre for Agricultural Research, Hungarian Academy of Sciences, Martonvásár, Hungary</footnote>
SESSION II. GENOMICS AND EPIGENOMICS IN THE POST SEQUENCE ERA
PII-25 - Lukas KRONENBERG - Wheat stem elongation dynamics assessed by GWAS: short term growth rate depends on temperature

Wheat stem elongation dynamics assessed by GWAS: short term growth rate
depends on temperature

Lukas KRONENBERG<fnref n="1" />, Steven YATES<fnref n="1" />, Achim WALTER<fnref n="1" /> and Andreas HUND<fnref n="1" />

Corresponding author: lukas.kronenberg@usys.ethz.ch

Stem elongation is a critical phase for yield formation in wheat. During this period, active spike growth takes place as the pike is pushed up the stem. Spike dry weight at anthesis is closely related to grain number per spike, a critical determinant of potential yield in wheat. A prolongation of stem elongation has therefore been proposed to increase dry matter accumulation by the spike, increasing potential yield.

By using terrestrial 3D-Laserscanning mounted on a novel high throughput phenotyping platform, stem elongation dynamics were monitored on a set of > 300 commercial European wheat varieties (GABI wheat panel) in high temporal resolution. Results showed that there is large genotypic variation in duration as well as growing degree-days until start and end of stem elongation and that those traits are partly interdependent. Genome wide association studies revealed stable and highly significant quantitative trait loci (QTL) across three years for start and end of stem elongation. Canopy height measurement in high temporal resolution allowed to investigate short term growth response to temperature during stem elongation. Preliminary results indicate a stable QTL for this trait as well as a connection to final height. Start of stem elongation is a key stage in wheat development and the large genetic variation suggests opportunities for breeding. However, traditional rating of this trait is low in throughput and labor intensive. New phenotyping techniques allow to examine the genetic basis of these difficult-to-phenotype trait and offer opportunities for breeding via marker assisted selection.

**Related publication:**

Kronenberg, L., Yu, K., Walter, A. et al. Euphytica (2017) 213: 157. https://doi.org/10.1007/s10681-017-1940-2

**Keywords:** Wheat, Stem elongation, GWAS, Growth, Phenotyping

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<footnote n="1">1 ETH Zürich</footnote>
PII-26 - Carlos P CANTALAPIEDRA - Unmasking new intra-species diversity through k-mer count analysis

Unmasking new intra-species diversity through k-mer count analysis

Carlos P CANTALAPIEDRA<fnref n="1" />, Ernesto IGARTUA<fnref n="1" />, Ana M CASAS<fnref n="1" /> and Bruno Contreras MOREIRA<fnref n="1" />,<fnref n="2" />

Corresponding author: cpcantalapiedra@eead.csic.es

High-throughput sequencing is often used to examine intra-species diversity. Most studies are focused on calling and genotyping SNPs. Other kinds of genomic variation, such as copy-number variation (CNV), are more rarely exploited despite literature reports linking them to phenotypic differences.

For some loci, it is difficult to identify reliable SNPs. For instance, reads from closely related sequences (e.g. paralog genes) will often map stacked to the same location if some of those loci are absent from the reference sequence. Such piled up mappings produce abundant fake heterozygous SNPs, and thus have been called apparent heterozygous mappings (AHMs).

To avoid wrong conclusions from false positive calls, SNPs from AHMs are often discarded, either in early (e.g. samples expected to be homozygous), or in downstream steps of the analysis (e.g. when incoherent haplotype blocks are identified). This would lead to information loss at certain loci.

AHMs can be seen as a kind of CNV which is specific to non-identical copies. Unmasking such variation could help to i) assess the completeness of a genome or pan-genome reference, ii) confirm results from other CNV genotyping methods, when the copies originate in non-identical loci, iii) provide hints about the history and behavior of duplicating DNA loci, and iv) reveal novel intra-species genetic diversity.

Here we present a software pipeline, kmeleon, available at https://github.com/eead-csic-compbio/kmeleon, designed to identify regions harboring AHMs. kmeleon is based on mappings, and thus it can be used for both homozygous and heterozygous samples. First, the different k-mers (sequences of length k) mapping to a single locus are identified and counted. Then, loci are classified based on the presence or absence of AHMs. From those intervals, it is straightforward to perform comparisons between genotypes, or to translate existing annotation to the regions with AHMs.

We used exome capture data to detect AHMs in a set of barley accessions. We included the cultivar Morex, the genotype of the genome reference, as a control sample. As expected, it had the lowest number of AHMs, although some were still detectable. For all accessions, AHMs were found both in inter- and intragenic loci. Enrichment analysis showed that NBS-LRR proteins were overrepresented at AHMs, whereas PPRs proteins were depleted. Also, we will show that AHMs can be used to infer phylogenetic trees which are congruent to those produced with SNP-based approaches, supporting the information value, of this hidden variability, to describe genetic relationships.

**Keywords:** cnv genome diversity bioinformatics genotyping

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<footnote n="1">¹ Aula Dei Experimental Station - CSIC, Avda Montañana 1005, 50059 Zaragoza, Spain</footnote>

<footnote n="2">² Fundación ARAID, calle María de Luna 11, 50018 Zaragoza, Spain</footnote>
PII-27 - Charles CHEN - Genetic improvement facing climate challenging: integrating classical and novel approaches

Genetic improvement facing climate challenging: integrating classical and novel approaches

Charles CHEN<fnref n="1" />

Corresponding author: charles.chen@okstate.edu

Growing on over six million acres in Oklahoma, production of winter wheat that plays a vital role in both grain production and cattle industry in the US, is subject to the worsening drought condition in the southern Great Plains. A double-haploid (DH) Buster population of 282 DH lines derived from the intercross of Duster and Billings was evaluated under condition with various degrees of water stress. To provide genomic predictors, the release of Buster_hmp_v1.0 contains 213, 940 SNPs from both genotyping-by-sequencing (GBS) and exome capture techniques, after removing non-informative and erroneous SNPs. Genome-wide association mapping (GWAS) identified SNP variants significantly associated both yield and quality phenotypes; however, genetic response to environmental perturbations (G×E) clearly demonstrated impact on individual year's yield potential, with a total of 85 variants significantly showing differential response based on a single locus analysis. A closer look of transcript assemblies revealed five genes in drought tolerant genotype and 82 genes in drought susceptible genotype that, mostly related to photosynthetic activity, exhibit high levels of transcriptional variation in response to drought stress.

Further, significant overinflation in accuracy was found when examining genomic selection (GS) performance across field condition. With a more realistic cross-season validation, our results reckoned a nearly 50% reduction in GS accuracy for grain yield can be expected due to environmental variability; this overinflation was most discernible for wheat protein content amongst the 13 traits examined; showing an overall 45% prediction accuracy, adjusted sedimentation was however the most predictable end-user quality phenotype, despite the difference in water pressure.

Finally, the input genotypic information used in GS algorithms can also be a determining factor for GS predictability. Our results indicate that including SNPs derived from functional genomic components, such as the exome capture technique that uses expressed mRNA as templates for probe construction, could lead to more predictable outcomes in well-watered condition; GBS, however, shows slight advantage when predicting grain yield under severe drought.

**Keywords:** genomic resource, GxE, genomic selection

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<footnote n="1">¹ Department of Biochemistry and Molecular Biology, Oklahoma State University</footnote>
SESSION III. YIELD POTENTIAL AND RESOURCE EFFICIENCY
PIII-28 - Elizabeth CHAPMAN - Combining staygreen and altered phenology to improve wheat productivity

Combining staygreen and altered phenology to improve wheat productivity

Elizabeth CHAPMAN<fnref n="1" />, Jacob LAGE<fnref n="2" /> and Simon GRIFFITHS<fnref n="1" />

Corresponding author: elizabeth.chapman@jic.ac.uk

Staygreen plants demonstrate delayed, or altered, senescence profiles, but need to remain photosynthetically active for longer to be of interest. In wheat and other grain crops, functional staygreens have demonstrated yield benefits, especially under stress, including: - heat, drought, and low nitrogen environments. Often physiological adaptations are associated with the staygreen trait, including altered stomatal conductance, leaf, and root architecture. However, the underlying genetics of senescence, and staygreen, are largely unknown, especially in wheat, aside from NAM genes and certain NAC transcription factors.

Originally three staygreen EMS mutant *Triticum aestivum*, cv Paragon, lines were identified during a nitrogen use efficiency trial, which concluded the staygreen trait to be environmentally stable over a 3-year period. These three lines differ relative to their parental cv Paragon in both their senescence profiles; subtle to extreme staygreens; and yield; nitrogen insensitive to high yielding.

Two single seed descent (SSD) populations were developed per staygreen, one of which was a segregating population designed for mapping the mutation(s) (Staygreen x Paragon). The second, a population segregating for phenology and senescence, through crossing with a triple photoperiod (Ppd) insensitive Paragon near isogenic line (NIL); carrying Ppd-A1a, Ppd-B1a, Ppd-D1a. Within these populations, there exists variation in heading date of up to 3 weeks under UK field conditions, within an isogenic background.

Early heading and grain maturity traits are typically adapted to avoid heat/drought stress. Unfortunately, this can negatively affect pollen viability and impair grain filling, whilst it is hypothesised staygreens are associated with stress adaptation under such environments. Thus, can combining early flowering and staygreen result in synergism to increase wheat productivity? Alternatively, does this unite conflicting life history strategies, potentially negatively affecting yield or quality?

A selection of 'early' and 'late' heading segregants, demonstrating staygreen and normal senescence profiles were phenotyped for phenology, yield and yield components during 2015-16 and 2016-17 in UK and France. For 2016 flowering time, rather than senescence, was responsible for significant differences in yield between groups, attributable to low light, but high rainfall, levels. In contrast, 2017 witnessed prolonged periods of drought, prior to late season rainfall, thoroughly testing stress tolerance of these staygreen, especially regarding the early flowering ± staygreen material grown in France.

What effect does staygreen have on grain filling dynamics? Initial grain filling experiments demonstrated a significant extension in grain fill duration for the staygreen parents relative to Paragon, which mirrored their senescence phenotypes. However, in both 2016 and 2017 certain segregants demonstrated phenotypes indicative of sink limitation, and therefore potential indicating benefits of staygreen may be going unrealised.

Exome capture has been undertaken on the three staygreen parents, and Paragon, to identify EMS mutations for the development of molecular markers to begin to map associated region(s) through QTL analysis. Meanwhile 2017-18 field trials focus on testing the staygreen character under different stress condition such as low nitrogen regimes, climates, and soil types. These trials will be conducted by KWS in the UK, France, and Germany.

**Keywords:** Staygreen, wheat, senescence, NUE, stress-adaption, photoperiod-insensitive

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<footnote n="1">¹ John Innes Centre, Norwich Research Park, Colney Lane, Norwich, NR4 7UH, UK</footnote>

<footnote n="2">² KWS-UK, 56 Church Street, Thriplow, Royston, Herts, SG8 7RE, UK</footnote>
PIII-29 - Nadège BAILLOT - Different growth dynamics within the wheat spike

Different growth dynamics within the wheat spike

Nadège BAILLOT<fnref n="1" />, Christine GIROUSSE<fnref n="2" />, Vincent ALLARD<fnref n="2" />, A LACOINTE<fnref n="2" />, Jacques LE GOUIS<fnref n="2" /> and A PIQUET<fnref n="1" />

Corresponding author: nadege.baillot@gmail.com

European wheat production is stagnating while its demand is in constant evolution. Indeed, maintain or increase its yield in less favorable conditions is a major objective for the future. Grain yield is built during wheat development mainly through two components: the number of grains per square meter until anthesis and the thousand-grain weight during grain filling. Usually only the average grain weight is considered but grain weight may vary largely depending on the position along the spike and within a spikelet. As study was then carried out to explain why the individual weight grain is not homogenous along the spike.

Two different varieties were grown in a greenhouse at daily temperature of 22.6°C and a photoperiod of 16 hr, with water and nutrient solution supplied twice a day. From anthesis to harvest at least 13 samplings of eight spikes of main tillers were conducted for each variety. The ear was divided into three parts (basal, central and apical) in which the two proximal grains of two spikelets were dissected. The individual grains were then dried and weighed. A three-parameter logistic model was fitted to describe grains growth depending on their position on the spike.

At maturity, no significantly difference of dry matter was shown between the two proximal grains for both varieties. They were pooled in the following analyses. Grains in the central part of the spike were the heavier followed by the basal ones, the lighter ones being in the apical part. The logistic model fit well to the data. Differences in final grain weight are mainly explained by a difference of ovary's dry matter at anthesis and grain growth while the duration of grain filling is the same for each part.

This work brings new information to explain individual grain weight within the spike through different growth dynamics.

**Keywords:** wheat, spike, grains, dry matter, grain filling, distribution within spike, yield

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<footnote n="1">¹ Vetagrosup, campus agronomique, 89 avenue de l'Europe, 63370 Lempdes and INRA, UMR GDEC, 5 chemin de Beaulieu, 63039 Clermont-Ferrand Cedex 2</footnote>

<footnote n="2">² INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France</footnote>
PIII-30 - Dragana TRKULJA - Seeding density effects on grain yield in bread wheat

Seeding density effects on grain yield in bread wheat

Dragana TRKULJA<fnref n="1" />, Milan MIROSAVLJEVIĆ<fnref n="1" />, Sanja MIKIĆ<fnref n="1" />, Srbislav DENČIĆ<fnref n="1" />, Ankica KONDIĆ-ŠPIKA<fnref n="1" />
and Vojislava MOMČILOVIĆ<fnref n="1" />

Corresponding author: dragana.trkulja@ifvcns.ns.ac.rs

Seeding rate is considered as one of the cultivation practices that most affect crop grain yield and its components. It is based on planting a predetermined number of live seeds per square metre to achieve a specific plant density optimal for overall health and vigor of the plants. In order to study influence of seeding density on tillering potential and yield performances of bread wheat, a set comprised of 80 elite wheat varieties and 16 breeding lines contrasting for tillering was sown at locality Rimski Sancevi, Novi Sad, Serbia in 2016-17 growing season. A trial was set at randomised block design with three replications for each seeding density (300 and 550 viable seeds per square metre) and size of each plot was five square metres. During the season and after harvest the following traits were recorded: number of tillers, stem height, spike length, grain yield and thousand-kernel weight (TKW). The genetic diversity and population structure of the 96 accessions were assessed by 60 microsatellite markers distributed across all three wheat genomes. The presence of marker-trait associations (MTA) was analysed using mixed linear model. A total of 496 alleles were detected, with average PIC value of 0.613. Within analysed wheat accessions, three subpopulations were determined and the grouping was in accordance with their pedigree data. Analysis of variance showed statistically significant differences for grain yield between two seeding densities, as well as among detected subpopulations. Positive correlations were observed between grain yield and TKW, while tillering was negatively correlated with TKW. The 21 and 22 MTAs (p< 0.01) were obtained for trials with 300 and 550 seeds per square metre, respectively. However, only 12 MTAs were common for both. The presence of unique MTAs could indicate to specific genetic mechanisms involved in plant adaptation to different seeding densities in wheat, which should be further studied.

**Keywords:** sowing rate, *Triticum aestivum* L., tillering

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<footnote n="1">¹ Institute of Field and Vegetable Crops, Maksima Gorkog 30, Novi Sad, Serbia</footnote>
PIII-31 - Milan MIROSAVLJEVIĆ - Stay green traits contributed to recent grain yield improvement in six-rowed winter barley in Serbia

# Stay green traits contributed to recent grain yield improvement in six-rowed winter barley in Serbia

Milan MIROSAVLJEVIĆ¹, Vojislava MOMČILOVIĆ¹, Sanja MIKIĆ¹, Dragana TRKULJA¹, Srbislav DENČIĆ¹, Bojan JOCKOVIĆ¹ and Vladimir AĆIN¹

¹ Institute of Field and Vegetable Crops, Maksima Gorkog 30, 21 000 Novi Sad, Serbia

Corresponding author: milan.mirosavljevic@nsseme.com

Stay green phenotype in small grain cereals is characterized by longer maintenance of green leaf area after anthesis, allowing extended photosynthesis and accumulation of assimilates. Moreover, stay-green traits are related to improved yield stability and adaptation, especially to water limited environments. Climate changes, reflected in the mean temperature increase, precipitation decrease and frequent drought stress occurrence at the end of grain filling period, have had major influence on cropping system in Serbia during past decades. Therefore, the objective of this paper was to examine changes in stay green traits related to grain yield improvement in winter barley cultivars that have been grown in Serbia during last four decades. Grain yield and stay green traits (maximum NDVI, thermal time to the midpoint of senescence and thermal time to senescence nearing conclusion of stay green) were determined in a historical series of 8 Serbian and 8 international cultivars from surrounding countries. The experiment was conducted during two growing seasons with two levels of nitrogen application (0 and 100 kg N ha⁻¹). Results from this study showed that in Serbian and international set of cultivars grain yield was positively related to maximum NDVI and the thermal time to midpoint of senescence. Thermal time to senescence nearing conclusion of stay green was positively related with grain yield only in the international set of cultivars. Consistent increase per year of release in maximum NDVI and the time to midpoint of senescence were observed in both sets of cultivars under two levels of nitrogen fertilization, while correlation coefficients between the time to senescence nearing conclusion of stay green and year of release were not significant in either set. Therefore, our study suggests that further winter barley breeding progress should be more focused on increase of maximum NDVI and prolongation of period to midpoint of senescence.

**Keywords:** Hordeum vulgare L., NDVI, thermal time

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PIII-32 - Bojan JOCKOVIĆ - Breeding progress in yield and grain properties of winter wheat

Breeding progress in yield and grain properties of winter wheat

Bojan JOCKOVIĆ<fnref n="1" />, Velimir MLADENOV<fnref n="2" />, Radivoje JEVTIĆ<fnref n="1" />, Milan MIROSAVLJEVIĆ<fnref n="1" />, Vladimir AĆIN<fnref n="1" />,
Sonja ILIN<fnref n="1" /> and Novica MLADENOV<fnref n="1" />

Corresponding author: bojan.jockovic@nsseme.com

Constant grain yield increase of winter wheat represent one of the major breeding objectives worldwide, due to increase in human population and rapid decrease in the cropping area. Moreover, knowledge of the changes in grain traits associated with genetic gains in grain yield is important for future improvement of wheat breeding programs.
Therefore, the objective of this study was to analyze changes in grain yield and related grain traits in winter wheat cultivars grown in Serbia during the past 50 years. For this study, grain yield, 1,000-grain weight and test weight, were analyzed in historical set of 15 winter wheat cultivars. Trial was conducted during two growing seasons at the Experimental Field of the Institute of Field and Vegetable Crops, southeastern Europe.

Results from this study show that there was positive relation between grain yield and year of cultivars release. The average rate of increase of yield potential per year of release was 93 kg ha-1 year-1. In general, 1,000-grain weight varied between 35.8 and 47.0 g, while average values of test weight ranged from 75.1 to 84.5 kg hl-1. However, there was no significant changes in 1,000-grain and test weight in the last 50 years of wheat breeding at the Institute of Field and Vegetable Crops.

In conclusion, there was a significant yield progress during the examined period, however yield increase was not followed by changes in grain properties (1,000-grain and test weight).

**Keywords:** breeding, yield, grain, wheat

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<footnote n="1">¹ Institute of Field and Vegetable Crops, Novi Sad, Serbia</footnote>

<footnote n="2">² University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia</footnote>
SESSION IV. IMPROVING TOLERANCE TO BIOTIC AND ABIOTIC STRESS
PIV-33 - Valentina SPANIC - Quality preservation in FHB attack due to antioxidant system

Quality preservation in FHB attack due to antioxidant system

Valentina SPANIC<fnref n="1" />, Marija Viljevac VULETIC<fnref n="1" />, Daniela HORVAT<fnref n="1" />, Bojan SARKANJ<fnref n="2" />, Ivan ABICIC<fnref n="1" />
and Tihana MARCEK<fnref n="2" />

Corresponding author: valentina.spanic@poljinos.hr

Biotic stress leads to the formation of ROS which can harm the plants. Due to that plants have evolved an antioxidant defense system that includes non-enzymatic compounds, like ascorbate, glutathione, tocopherol, carotenoids, flavonoids and enzymes such as superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX) and polyphenol oxidase (PPO). Six winter wheat varieties, representing commercial Croatian wheat classes with different intrinsic qualities, were subjected to Fusarium Head Blight (FHB) stress. Sampling was done at the 14th day of the FHB stress when the first symptoms occurred. Antioxidant properties of both treatments (control and FHB) were investigated to document the effect of FHB resistance to protein components (albumins, gliadins, ω-, α-, γ- gliadins, glutenins, HMW-GS and LMW-GS) which were detected by RP-HPLC method. The plants of all six varieties under test showed less or no symptoms of FHB till the 14th day of the FHB stress, when two wheat varieties progressively enlarged symptoms up to 90% till 26th dai (days after inoculation) and due to that they were characterized as FHB-susceptible. Very low FHB severity was obtained by 'Kraljica', 'Olimpija' and 'Vulkan' (20% symptoms at 26th dai), while 'Sana' showed moderate susceptibility (35%).

The decline in CAT activity was in correlation with the increase of H2O2 in more resistant varieties ('Olimpija' and 'Vulkan'). The activity of APX increased with the appearance of symptoms in 'Kraljica', 'Golubica', 'Bc Anica', while the activity of APX declined in 'Olimpija' in FHB treatment in comparison to control. However, there was significant increase in FHB treatment in comparison to control, in respect to phenols, POD and H2O2 for all varieties, except for POD in 'Sana' and H2O2 in 'Golubica'. Varieties 'Golubica' and 'Vulkan' exhibited higher PPO activity in FHB treatment in comparison to control despite the presence of enlarged phenol content in FHB treatment in all varieties. The accumulation of MDA content was higher in two susceptible varieties than in tolerant varieties, with one exception.

After harvest, gliadin level in FHB treatment in comparison to control increased in four varieties ('Kraljica', 'Sana', 'Golubica', 'Anica'). There was a significant effect of FHB on percentage of glutenins showing a respective reduction in comparison to control of 5%, 20% and 16% of 'Kraljica', 'Golubica' and 'Bc Anica', respectively, while HMW subunits decreased in those three varieties (9.93, 14.61 and 20.33%, respectively) including 'Sana' (8.79%). LMW subunits were not largely affected as HMW, but 'Olimpija', 'Golubica' and 'Bc Anica' showed statistically significant decrease.

It is interesting that in both treatments the most susceptible variety had the lowest amount of phenol content and PPO activity among other varieties. Due to the role of polyphenols as secondary plant metabolites that play a role in the protection of plants against pathogen, they were higher in genotypes with better-preserved protein quality. The protein components of 'Vulkan' and 'Olimpija' were untouched which could be due to high PPO activity which could participate in detoxification of phenolic acids induced by pathogen attack.

**Keywords:** FHB, wheat, protein, antioxidant activity

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<footnote n="1">¹ Agricultural Institute Osijek, Juzno Predgradje 17, HR-31000 Osijek, Croatia</footnote>

<footnote n="2">² Faculty of Food Technology Osijek, Franje Kuhaca 20, HR-31000 Osijek, Croatia</footnote>
PIV-34 - Hanane AADELA - Agrobacterium-mediated Transformation of Mature Embryo and immature Tissues of Bread Wheat (*Triticum aestivum* L.) Genotypes

Agrobacterium-mediated Transformation of Mature Embryo and immature Tissues
of Bread Wheat (*Triticum aestivum* L.) Genotypes

Hanane AADELA<fnref n="1" />,<fnref n="2" />, Rabha ABDELWAHD<fnref n="1" />, Sripada. Mahabala UDUPA<fnref n="2" />, Amine EL MOUHTADI<fnref n="1" />,
Khadija AHANSAL<fnref n="1" />, Fatima GABOUN<fnref n="1" />, Allal DOUIRA<fnref n="1" /> and Driss IRAQI<fnref n="1" />

Corresponding author: iraqid@yahoo.fr

Although significant progress has been made on Agrobacterium-mediated wheat transformation, the current methodologies using immature embryos as recipient tissues, is labor intensive, time consuming and expensive. The use mature embryo (ME) as explants is increasingly recognized as a best method for developing regenerable cell lines in wheat transformation. Therefore, we have developed an Agrobacterium-based transformation protocol using mature embryos, adjusting other factors that can influence the yield of transformed plants. The experiment was focused on acetosyringone concentrations, genotypes and different kinds of mature embryos used as explants source for transformation studies. The *A. tumefaciens* strain EHA101 and pTF101.1 plasmid carrying HVA1 gene and bar-selectable marker gene were used. The intact mature embryos and half mature embryos produced a highest number of putative transgenic plantlets on selection medium. However, no plantlets were obtained from fragmented mature embryos. ‘Amal’ and ‘Rajae’ regenerated highest number of putative transgenic plants and 200 M acetosyringone was found to be better concentration for transformation. A total of 47 transgenic plants were selected. 11 plantlets showed resistance to leaf painting. The molecular analysis revealed 1% and 0.66% of T0 regenerated plantlets were successfully transformed and carried the HVA1 gene for the genotypes ‘Amal’ and ‘Rajae’, respectively. Additional analysis showed that the transgene was stably inherited from the T1 generation.

**Related publication:**

Agrobacterium-mediated Transformation of Mature Embryo Tissues of Bread Wheat (*Triticum aestivum* L.) Genotypes. Published in Cereal Research Communications CRC-D-17-00013R4-Agrobacterium-mediated transformation of immature embryo of bread wheat (*Triticum aestivum* L.) genotypes. Published in biologia

**Keywords:** bread wheat; mature embryo; explants; *Agrobacterium tumefaciens*; plasmid pTF101.1; HVA1; acetosyringone; genetic transformation

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<footnote n="1">¹ Biotechnology Unit, Institut National de la Recherche Agronomique (INRA), Avenue de la Victoire, B.P. 415, Rabat, Morocco</footnote>

<footnote n="2">² ICARDA-INRA Cooperative Research Project, International Center for Agricultural Research in the Dry Areas (ICARDA), B.P. 6299, Rabat, Morocco</footnote>

<footnote n="3">³ Laboratory of Botany, Biotechnology and Plant Protection, Ibn Tofail University, Faculty of Sciences, BP: 133, 14000 Kenitra, Morocco</footnote>
PIV-35 - Ildikó KARSAI - How may warm spells during winter influence the plant development of cereals?

How may warm spells during winter influence the plant development of cereals?

Ildikó KARSAI<fnref n="1" />, Tibor KISS<fnref n="1" />, Marianna MAYER<fnref n="1" />, Judit BÁNYAI<fnref n="1" />, Krisztina BALLA<fnref n="1" /> and Ottó VEISZ<fnref n="1" />

Corresponding author: karsai.ildiko@agrar.mta.hu

The climatic conditions during winter influence multiple processes in cereals. In addition to determining the winter survival and the vegetative-generative phase transitions of the apex, these conditions significantly affect the later plant development and thus yield components. Due to the climate change, the occurrence of unusual weather conditions and anomalies increases which can include shorter or longer warm spells during winter. We wanted to study the effect of a warm temperature period on seedlings with various degrees of vernalization requirement saturation. For this purpose 19 wheat cultivars of various origins were treated with a two-week period of warm temperature (25 °C) right after the end of 30-, 45- and 60-day low temperature vernalization in the juvenile stage.

Based on the results, the two weeks of 25 °C at the beginning of the plant development significantly modified both plant phenology and morphology, the direction of this effect, however was strongly dependent on the saturation level of the vernalization requirement and on the genotypes. After a 30-day vernalization, the plant development was delayed significantly by the warm temperature averaged over the 19 wheat cultivars, which was more pronounced at the later stages. As the saturation of the vernalization requirement increased, there was a dramatic change in the effect of warm temperature. After a 45-day vernalization the difference between the control and the 25 °C-treated plants were minimal, especially at the later stages, while after a 60-day vernalization treatment the 25 °C temperature fastened the plant development significantly. The short period of warm temperature in juvenile stage also influenced significantly the plant height, the final leaf number and the number of reproductive tillers.

In conclusion, under inductive photoperiods the effects of a short, warm ambient temperature spell on the juvenile plants with winter growth habit depends on to what degree their vernalization requirements are saturated. If it is not saturated then the plant development becomes delayed. If it is saturated then the warm temperature spell hastens the plant development, the extent of which is strongly genotype dependent.

This research was funded by the following grants: EU-FP7 ADAPTAWHEAT, and GINOP-2.3.2-15-2016-00029

**Keywords:** wheat, vernalization, ambient temperature, plant development

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<footnote n="1">¹ Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences
Martonvásár, Brunszvik street 2, Hungary</footnote>
**PIV-36 - Judit BÁNYAI** - Investigations on the effects of drought stress on the morphology, physiology and yield components of durum wheat in Hungary

# Investigations on the effects of drought stress on the morphology, physiology and yield components of durum wheat in Hungary

Judit BÁNYAI<fnref n="1" />, Tibor KISS<fnref n="1" />, Marianna MAYER<fnref n="1" />, Viola TÓTH<fnref n="1" />, András CSEH<fnref n="1" />, Krisztina BALLA<fnref n="1" />, Klára MÉSZÁROS<fnref n="1" />, Ottó VEISZ<fnref n="1" /> and Ildikó KARSAI<fnref n="1" />

Corresponding author: banyai.judit@agrar.mta.hu

The aim of the work was to identify morphological, physiological, biochemical and yield biological traits exhibiting a close relationship with higher yields even when water supplies are less than satisfactory. For this purpose a total of 188 spring durum wheat genotypes with diverse genetic backgrounds and eight near-isogenic lines of spring durum wheat were sown in field experiments under irrigated and rainfed conditions or under a rain shelter in Martonvásár between 2011 and 2014 in order to investigate which plant traits contribute to an increase in drought tolerance. In the course of the field experiment, the individual genotypes were sown on the same experimental field each year so as to reduce the effect of the environment (soil).

The results proved that the year and the treatment had the greatest effect on the traits tested, even in the case of parameters that have a high heritability (h2) index. The only exception was the thousand-kernel weight, which was shown by the three-year data to have great stability even when exposed to drought stress. It was demonstrated that only plants with a larger number of fertile tillers and with flag-leaves having a longer period of photosynthetic activity were able to produce higher yields even with poor water supplies. The significant positive correlation found between the SPAD and NDVI values recorded in the early stage of waxy ripeness (Z83) and the yield showed that some genotypes have delayed senescence in this phenophase under drought conditions, which is manifested in the form of higher yields. The analyses led to the identification of a cultivar ('Cham-1') that was able to maintain stable photosynthetic activity during drought stress in all the years, thus giving better performance.

Association mapping was used to identify loci responsible for drought tolerance. With the help of the SNP map characterising the 188 cultivars, the population structure of the association panel was analysed, and QTL regions linked with yield, heading date and morphological or physiological traits were identified for different water supply levels. Numerous marker-group regions were found to be significant in specific environments, as a function of year or treatment. In the course of the work, two chromosome regions were identified where QTLs for chlorophyll content and heading date were linked (1B, 5A), and in the case of the 5A chromosome the yield QTL was also linked with the identified region.

The results suggest that the phenotyping of morphological, physiological, biochemical and yield biological traits could be an efficient way of identifying genotypes with better resistance to water deficit. The most important criteria for success are a clearly defined trait that can be objectively measured, a satisfactory statistical model and successful validation.

Funding from the EU FP7-244374 DROPS and GINOP-2.3.2-15-2016-00029 projects

**Keywords:** durum, drought stress, phenology, physiology

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<footnote n="1">¹ Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences
Martonvásár, Brunszvik street 2, Hungary</footnote>
PIV-37 - Krisztina BALLA - Investigation of heat stress tolerance on yield properties in cereals

Investigation of heat stress tolerance on yield properties in cereals

Krisztina BALLA<fnref n="1" />, Ildikó KARSAI<fnref n="1" />, Tibor KISS<fnref n="1" />, Szilvia BENCZE<fnref n="1" />, Judit BÁNYAI<fnref n="1" /> and Ottó VEISZ<fnref n="1" />

Corresponding author: balla.krisztina@agrar.mta.hu

Heat stress is becoming increasingly frequent and often affect wheat plants at any time during vegetative growth and especially around heading in Hungary. Our studies focused on that the effect of heat stress strongly depends on the timing and duration of the heat period during the growing season and the damage caused by heat is greatly influenced by the developmental stage of the plants. Therefore, heat stress experiments were undertaken to determine the effects of heat stress on the physiological processes, as well as yield components in the stage of booting (Zadoks-49) and emergence of inflorescence completed (Zadoks-59). 100 winter wheat varieties of different origins were involved in phytotron where the treatments lasted for 5 (H5), 10 (H10) and 15 (H15) days under control conditions. The plants were exposed to heat stress of 360C/200C for 8 hours. The specific aim of this research project was to identify various heat tolerance sources and to determine the genetic background of the responses given to heat stress.

It was found that the shortest duration of heat stress (H5) had the lowest impact on the examined parameters, while the longest treatment (H15) resulted in the most negative impact on the morphological and yield parameters in both examined phenophases. On the average of the varieties, the plant height (PH) and the last internode length (LIL) were reduced only due to the heat stress treatment applied in Zadoks-49. The heat stress resulted in drastic significant reduction in the grain number (GN), grain yield (GY), harvest index (HI), biomass (B) and the grain number of one spikelet. But the thousand kernel weight (TKW) was increased due to the drastic reduction in the number of grains under heat stress for 5 and 10 days. In Zadoks-59, H5, H10 and H15 treatments caused a significant reduction in the number of productive tillers, LIL, HI, B, GN and GY. The high temperature treatments, however, had no significant effect on the PH, the length of the ear, the grain number of one spikelet and the grain number of the main spike too. TKW was declined only by the H15 treatment significantly. The photosynthetic activity was dropped in response to heat stress in both examined phenophases. The stomatal conductance was declined due to heat stress resulted in lower CO2 binding capacity. The net assimilation (Pn) was reduced by more than 50% due to heat stress treatments (H5) in both stages. The plants tried to survive the heat stress by increasing the transpiration level. The chlorophyll content was changed to the most negative extent when the wheat varieties were exposed to heat stress for 15 days.

The best Pn value was measured in the variety Aura (at Zadoks-49) and in Altay-2000 (Zadoks-59) under the applied longest heat stress. The best GY value was received in the variety Valoris (at Zadoks-49) and Spada (at Zadoks-59) when exposed to H15 treatment.

As a consequence of the changing climate and the higher occurrence of extreme temperatures it is important to breed new wheat varieties with high yield quantity and good quality even under extreme weather conditions. The impact of this analyse was to identify different heat tolerance sources and to produce genetic populations, in which the high heat tolerance properties are incorporated.

This project was supported by a grant from the National Research, Development and Innovation Office (NKFIH-119801) and the János Bolyai Research Scholarship of the HAS.

**Keywords:** heat stress, yield properties, photosynthetic activity, wheat

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<footnote n="1">¹ Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences
Martonvásár, Brunszvik street 2, Hungary</footnote>
PIV-38 - Marianna MAYER - Comparative analysis of stem rust resistance in winter wheat genotypes at Hungarian and East African locations

Comparative analysis of stem rust resistance in winter wheat genotypes at Hungarian and East African locations

Marianna MAYER¹, Ildikó KARSAI¹, László LÁNG¹, Ottó VEISZ¹ and Gyula VIDA¹

¹ Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences
Martonvásár, Brunszvik street 2, Hungary

Corresponding author: mayer.marianna@agrar.mta.hu

Stem rust, caused by the fungus *Puccinia graminis* f. sp. *tritici* Eriks. & E. Henn., is an impending disease for wheat production worldwide hence breeding for stem rust resistance is a very important challenge of wheat breeders all around the world. Although the disease has been under effective control in Hungary through the use of host resistance, there are needs for new sources of resistance because of the threatening new pathogen races in Africa and Middle East. The main objective of this work was to compare the stem rust resistance of winter wheat genotypes under different pathogen pressure in Hungary and Kenya and to identify those genotypes which has acceptable resistance under both conditions.

The stem rust resistance of the 161 collected genotypes were scored in adult plant stage by the modified Cobb scale, evaluating the severity and the host response. From the phenotypical data average coefficient of infection (ACI) values were calculated. The field experiments were set up at two different locations, in the stem rust screening nursery of the Kenya Agriculture and Livestock Research Organization, Njoro in 2017 and in the artificially inoculated stem rust nursery of Agricultural Institute, Centre for Agricultural Research, Martonvásár in 2016 and 2017.

We investigated the frequency of the *Sr1A.1R*, *Sr2*, *Sr15*, *Sr31*, *Sr38* stem rust resistance genes and the *Lr34* leaf rust resistant gene in the collected material, using the gene specific SSR Xscm9 (for *Sr1A.1R* and *Sr31*), csSr2 CAPS (for *Sr2*), STS638 (for *Sr15*), VENTRIUP-LN2 (for *Sr38*) and STS csLV34 (for *Lr34*) molecular markers. Statistical analysis of the data was performed to determine the connection between the investigated *Sr* genes and the phenotypic responses of the genotypes.

Among the 161 genotypes 90 (55.9%) carried the *Sr38*, 36 (22.4%) the *Sr31*, 34 (21.1%) the *Lr34*, 33 (20.5%) the *Sr15*, and 4 (2.5%) carried the *Sr2*, respectively. The *Sr1A.1R* were detected in two (1.2%) genotypes. About one third of the genotypes had very good resistance against both the Hungarian and African pathotypes of the stem rust pathogen. From the results it can be concluded that the resistance of the genotypes against the Hungarian races of *Puccinia graminis* f.sp. *tritici* is primarily due to the presence of the *Sr38* or *Sr31* resistance genes meanwhile in the case of the Ug99 lineage the connection between the investigated *Sr* genes and the phenotypic response is not significantly provable.

We would like to thank the excellent help of Dr. Ruth Wanyera and Dr. Sridhar Bhavani for managing the stem rust resistance field tests in Kenya Agriculture and Livestock Research Organization. This research was funded by the following grant: GINOP-2.3.2-15-2016-00029

**Keywords:** stem rust, resistance breeding, molecular markers, wheat (*T. aestivum* L.)

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**PIV-39 - Valérie CADOT - *Tilletia caries*, causal agent of Common Bunt: study of the most prevalent virulences in order to develop a resistance test for the registration to the National List for common wheat in Organic Farming**

**Tilletia caries, causal agent of Common Bunt: study of the most prevalent virulences
in order to develop a resistance test for the registration to the National List for
common wheat in Organic Farming**

Valérie CADOT<fnref n="1" />, G ORGEUR<fnref n="1" />, T BALDWIN<fnref n="1" />, L LE DARE<fnref n="1" />, C BRACHET<fnref n="1" />, C GALON<fnref n="1" />, J GOMBERT<fnref n="2" />,
L FONTAINE<fnref n="3" />, J BRUYERE<fnref n="4" />, S OSTE<fnref n="4" />, P DU CHEYRON<fnref n="5" />, J CHAMPION<fnref n="6" /> and V GRIMAULT<fnref n="1" />

Corresponding author: valerie.cadot@geves.fr

Common bunt, mainly caused by *Tilletia caries* and *T. foetida*, with a high capacity of dissemination has been a re-
emerging seed-borne cereal disease since 2007. In infected ears, bunt balls cause important yield reductions of up to
80% and a deterioration in seed health quality. Most commonly cultivated wheat cultivars are susceptible, due a lack
of selection tools for resistance but also because chemical seed treatments can be used to control this problem in
conventional farming. In organic farming, only a few seed treatments are available which are not always effective,
thus genetic resistance is an important trait to consider for registration to the French List.

The first aim of program ABBLE was to study the variability in populations of common bunt and to identify the main
virulences present in France. Samples were collected in 2015 from the main production areas and the identification of
*Tilletia* species was performed by GEVES. All 20 collected strains were identified as *T. caries*. For each strain, the
characterization of virulence was done on a range of 15 Host Differentials from Metzger. After 7 months of growth,
symptom expression (bunt ears) was quantified on each host differential showing that the most frequent virulences
were Bt-2, Bt-7 and Bt-15.

The second aim was to develop a miniaturized resistance test to common bunt, by comparison with resistance test at
adult stage in field. A test was carried out in controlled conditions with detection of *Tilletia caries* by qPCR at an early
stage (2 leaves) and compared with symptom expression for similar varieties at milky stage. Currently, the method
allows the differentiation of susceptible and resistant varieties. The qPCR protocol was tested on the range of 15 host
differentials infected with a strain representing the most prevalent virulences in France. The same strain has also been
inoculated to 10 cultivars mainly used in organic farming. The qPCR method will be validated after comparison of
resistance classification between the number of positive plantlets detected by qPCR and the percentage of infected
ears observed in field for these same varieties.

Evaluation of cultivar resistance mainly used in organic farming will improve understanding of the current resistance against the main virulences present in France. The development of an early, rapid and reliable resistant test will provide a new tool for screening resistant varieties for breeders and for registration to the National List.

**Keywords: *Tilletia caries*, virulence, early resistance test, qPCR, Organic farming, National List**

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<footnote n="1">¹ GEVES, France</footnote>

<footnote n="2">² FNAMS, France</footnote>

<footnote n="3">³ ITAB</footnote>

<footnote n="4">⁴ FREDON Nord Pas-de-Calais</footnote>

<footnote n="5">⁵ ARVALIS INSTITUT DU VEGETAL</footnote>

<footnote n="6">⁶ CHAMBRE D'AGRICULTURE de la DROME</footnote>
PIV-40 - Harpinder RANDHAWA - Flowering pathway genes regulating *Fusarium* head blight resistance in wheat

Flowering pathway genes regulating *Fusarium* head blight resistance in wheat

Harpinder RANDHAWA<fnref n="1" />, Raman DHARIWAL<fnref n="1" />, Maria Antonia HENRIQUEZ<fnref n="2" />, Colin HIEBERT<fnref n="2" />
and Curt MCCARTNEY<fnref n="2" />

Corresponding author: Harpinder.Randhawa@agr.gc.ca

*Fusarium* head blight (FHB) caused by *Fusarium graminearum* and other spp. of genus *Fusarium*, is a most devastating disease of wheat affecting most growing areas in Canada. Infection with Fusarium leads to shriveled seed or tombstones, which are white or pink in appearance and known as *Fusarium* Damaged Kernels (FDK). The high proportion of FDK in a commercial wheat crop results in severe yield and quality losses. The fungus also produces a toxin known as deoxynivalenol (DON) in the infected grain which is a major food safety issue, especially in the export market. The use of resistance cultivars offers the most effective approach to manage FHB epidemics. Therefore, a number of studies have been conducted to identify FHB resistance QTL in wheat around the globe, however, molecular mechanisms underlying FHB resistance have not been fully known yet. In this study, a doubled haploid mapping population segregating for flowering pathway genes and FHB resistance have been screened for a number of flowering related traits and FHB followed by SNP genotyping and QTL analysis. The results of present study will be discussed.

**Keywords:** *Fusarium* head blight, wheat

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<footnote n="1">¹ Lethbridge Research and Development Centre, Agriculture and Agri-Food Canada, 5403 1st Av S. Lethbridge, AB, Canada973 INSERM, University Paris Diderot, 35 rue Hélène Brion, Paris 7, 75205, Paris Cedex 13, France</footnote>

<footnote n="2">² Morden Research and Development Centre, Agriculture and Agri-Food Canada</footnote>
**PIV-41 - Aurore BERAL** - Is wheat individual grain characteristics variance a source of genetic variability for water stress tolerance?

# Is wheat individual grain characteristics variance a source of genetic variability for water stress tolerance?

Aurore BERAL<fnref n="1" />, Jacques LE GOUIS<fnref n="1" />, Christine GIROUSSE<fnref n="1" /> and Vincent ALLARD<fnref n="1" />

Corresponding author: aurore.beral@inra.fr

Agronomic performance of wheat genotypes is usually evaluated by studying the mean value of pertinent agronomic variables. In particular, grain yield is analyzed through its different average numeric components (number of plants per m-2, number of spikes per plant, number of spikelets per spike, number of grains per spike, number of grains per m-2, individual grain weight). Nevertheless, the variance of these components has not been so far fully explored. Moreover, number of grains per m-2 is often considered as the most important component impacting grain yield. Yet it has been observed that the bigger the number of grains is, the more the number of small grains is, especially those in apical spikelets or in apical position within a spikelet. The variance associated to the grain size and individual grain composition (nitrogen concentration) is linked to both tillering and spike structure. Its impact on agronomic performance, especially in stressful environments (abiotic stress) is not much known.

The aim of this study is to analyze the genetic and ecophysiological determinism of wheat individual grain characteristics variability and evaluate its impacts on agronomic performance in order to quantify the actual genetic variance potential. To this aim, several issues have to be investigated: (i) understand precisely the structure of the variance associated to individual grain characteristics and (ii) understand causal processes of this variance, especially spatial structure of the spike and spikelets.

The present work is based on a field experiment where 228 genotypes were grown under optimal conditions or under a severe water stress. All yield components were recorded. Out of this panel, four genotypes were selected for in depth analysis of grain traits variance structure. For these genotypes, 10 spikes were randomly chosen in each plot and were individually dissected in order to record the exact position of each grain within the spike structure. Grains originating from 2 spikes among these 10 spikes, nitrogen concentrations per grain were also measured.

Our results show that (i) individual grain weight is strongly affected both by its position within the spike as well as by mean spike characteristics and (ii) variance components are under both genetic and environmental control. These results suggest that studying variable components of grain traits can help defining typologies of wheat genotypes differing by their strategies to set grain number (and thus potential yield) at anthesis. Further studies will be needed in order to test whether these strategies have differential effects on wheat tolerance to abiotic stresses during grain filling.

**Keywords:** Variance, grain weight, water stress, wheat

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<footnote n="1">¹ INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France</footnote>
PIV-42 - Barbara STEINER - Improving *Fusarium* head blight resistance in durum wheat

Improving *Fusarium* head blight resistance in durum wheat

Barbara STEINER<fnref n="1" />, Sebastian MICHEL<fnref n="1" />, Noemie PRAT<fnref n="1" />, Aileen HENZE<fnref n="1" />, Maria BÜERSTMAYR<fnref n="1" />, Marco MACCAFERRI<fnref n="2" />, Roberto TUBEROSA<fnref n="2" />, Marc LEMMENS<fnref n="1" /> and Hermann BÜERSTMAYR<fnref n="1" />

Corresponding author: [barbara.steiner@boku.ac.at](mailto:barbara.steiner@boku.ac.at)

*Triticum durum* is particularly susceptible to infection with pathogenic fungi of the Fusarium genera, which cause Fusarium head blight (FHB), one of the most destructive diseases of wheat worldwide. FHB leads to significant yield losses but the major issue associated with FHB is the contamination of the crop with mycotoxins. Notably, durum wheat is almost exclusively used for human consumption; consequently, the risk of toxin-contaminated grain entering the food chain is especially precarious.

The development of resistant varieties is the most effective approach for managing FHB, but resistance breeding in durum wheat has been hampered by the limited genetic variation in its gene pool and the difficulties in efficiently combining the numerous often small effect resistance conferring alleles in the same breeding line.

To increase genetic diversity for FHB resistance in durum wheat we introgressed resistance alleles from wild and cultivated relatives (*T. aestivum*, *T. dicoccoides*, *T. dicoccum*) and developed a large population of several bi- and multi-parental crosses. The population was phenotyped over several years resulting in a broad variation for FHB resistance including many moderately resistant experimental lines. Markers indicative for the major FHB resistance QTL Fhb1, identified in bread wheat, were clearly associated with FHB severity, confirming its successful implementation into durum wheat.

In addition, we investigated the genetic architecture of FHB resistance in the elite durum wheat gene pool and compared the accuracy and response to selection when using phenotypic and genomic selection strategies. A collection of 228 genotyped durum wheat cultivars was phenotyped for FHB resistance in three years and used for genome-wide association mapping. FHB resistance was strongly influenced by the semi-dwarfing gene RhtB1 as well as one QTL on chromosome 3B, which explained 10% of the phenotypic variance. Interestingly, this QTL was positioned in the same chromosomal interval as reported for Fhb1, but further genomic analysis suggests different genetic control.

Integrating the 3B QTL as fixed effect into genomic prediction models increased the prediction accuracy by 10%; however this extension had merely a minor effect on the actual response to selection. Using a selection index approach, we found that genomic selection had a 50% higher response to selection for FHB resistance than phenotypic selection in two-stage selection, without altering the population mean for plant height i.e. selecting undesirable tall plants.

Furthermore, phenotyping intensity could be reduced by testing only 50% of the most promising lines. Genomic selection among all lines in the population in this scenario gave the same response to selection as testing all lines in the disease nursery with subsequent phenotypic selection.

**Keywords:** *Durum wheat*, *Fusarium* head blight

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<footnote n="1">¹ University of Natural Resources and Life Sciences, Vienna (BOKU), Department of Agrobiotechnology (IFA-Tulln), Konrad-Lorenz-Str. 20, 3430 Tulln, Austria</footnote>

<footnote n="2">² Department of Agricultural Sciences, University of Bologna, Bologna, Italy</footnote>
**PIV-43 - Pawel CZEMBOR - Genome wide association study of seedling leaf rust resistance in European winter wheat cultivars**

# Genome wide association study of seedling leaf rust resistance in European winter wheat cultivars

Pawel CZEMBOR<fnref n="1" />, Magdalena RADECKA-JANUSIK<fnref n="1" /> and Dariusz MAŃKOWSKI<fnref n="1" />

Corresponding author: p.czembor@ihar.edu.pl

Leaf rust caused by the biotrophic fungus *Puccinia triticina* Eriks. (Pt) is one of the most important fungal disease affecting wheat worldwide. Genetic resistance is the most economical, effective and ecologically sustainable method of controlling the disease. Almost 80 leaf rust resistance genes have been characterized. Efficient utilization of genetic resistance relies on an appropriate knowledge of the leaf rust resistance genes and their effectiveness in different environments. We applied a genome wide association study (GWAS) approach to identify resistance genes in 143 winter wheat cultivars recently registered in Poland, Germany, France, Switzerland, Denmark and United Kingdom. Identified significant marker-trait associations (MTAs) were verified on a panel of 36 Thatcher near-isogenic lines containing different Lr genes (Tc-LrNILs). Wheat cultivars and Tc-LrNILs were genotyped on DArTseq platform and phenotyped with 18 single Pt isolates at seedling stage. After filtering, out of 40,035 SNP only 2,983 were used in GWAS using restricted maximum likelihood (REML) approach and information on structure of the analyzed wheat set was included. We identified four resistance genes *Lr3bg* (6B), *Lr9* (6B), *Lr24* (3D) and *Lr26* (translocation T1BL.1RS) and several leaf rust resistance loci not present in Tc-LrNILs set on chromosomes 1B, 2A, 3B, 6B, 7A and 7D. Only one-third of the cultivars contained at least one of the identified resistance genes. For some resistant cultivars we were not able to identify Lr genes that might be attributed to the low resolution of Pt isolates used in the study.

This research was supported financially by the Polish Ministry of Agriculture and Rural Development, program Fundamental Research for Biological Progress in Crop Production (years 2014-2020)

**Keywords:** Brown rust, DArTseq markers, gene postulation, resistance, wheat cultivars

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<footnote n="1">¹ Plant Breeding and Acclimatization Institute - National Research Institute Radzików, 05-870 Błonie, Poland</footnote>
# Resistance of European winter wheat cultivars to *Zymoseptoria tritici* isolate IPO92006

Piotr SŁOWACKI<fnref n="1" />, Pawel CZEMBOR<fnref n="1" /> and Magdalena RADECKA-JANUSIK<fnref n="1" />

Corresponding author: p.czembor@ihar.edu.pl

*Septoria tritici* blotch (STB) of wheat (*Triticum aestivum*), caused by the fungal pathogen *Mycosphaerella graminicola* (anamorph: *Zymoseptoria tritici*, syn. *Septoria tritici*), is present in most wheat-growing areas worldwide. Host resistance is the most economical and safest method of controlling the disease and information on resistance loci is crucial for effective breeding for resistance programs. In the study we used a set of 83 wheat cultivars registered in the Descriptive List of Agricultural Plant Varieties (COBORU 2012), 110 cultivars from other European countries and 25 cultivars/lines with identified STB resistance loci. The wheat genotypes were tested on adult plant stage under polytunnel conditions with watering system. Fully expanded flag leaves were sprayed with spore suspension of IPO92006 *Z. tritici* isolate. After incubation period, the percentage leaf area covered by necrosis (NEC) and covered by pycnidia (PYC) were measured on flag leaf of each wheat cultivar/line that were used in agglomerative hierarchical clustering (AHC) analysis with UPGA algorithm (unweighted pair-group average). Three groups of wheat cultivars/lines were identified and the largest group comprised 170 resistant genotypes with NEC 12.4%–82.1% and PYC 1.1%–53.5%. Within this group, set of 43 highly resistant wheat cultivars were identified (NEC 1.1%–41.8%, PYC 1.1%–31.9%): TE9111, Capone, Lear, Intro, Bombus, Fermi, Heros, KWS Erasmus, Oxal, Tabasco, Elixer, Dacanto, RGT Kilimanjaro, Florett, Sukces, Reciproc, Frument, Desamo, Edgar, Colonia, RGT Djoko, Addict, Forum, Famulus, Pionier, Amifor, Mandub, Memory, Terroir, Julius, Joker, Solitar, Kranich, Grapeli, KWS Dacanto, Eron, Marcopolo and Celebration. In addition in the same subgroup two genotypes with identified resistance loci were classified: Florett (QTL-3B, QTL-6D, Stb6 and Stb15) and TE9111 (Stb11, Stb6 and Stb7). This may suggest that resistance to STB in European cultivars is contributed mainly by quantitative loci and those with main effects. Presented work (phenotyping data) is a part of larger project aiming at identification of resistance genes to *Septoria tritici* blotch in winter wheat and will be used in near future in association mapping approach.

This research was supported financially by the Polish Ministry of Agriculture and Rural Development, program Fundamental Research for Biological Progress in Crop Production (years 2014-2020).

**Keywords:** Resistance, *Septoria tritici* blotch, wheat

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<footnote n="1">¹ Plant Breeding and Acclimatization Institute - National Research Institute Radzików, 05-870 Błonie, Poland</footnote>
**PIV-45 - Éva SZAKÁCS** - Development and characterization of a new recombinant 1RS.1BL translocation line carrying leaf and stripe rust resistance from perennial rye (*Secale cereanum*)

# Development and characterization of a new recombinant 1RS.1BL translocation line carrying leaf and stripe rust resistance from perennial rye (*Secale cereanum*)

Éva SZAKÁCS<fnref n="1" />, István MOLNÁR<fnref n="1" />, Kitti SZÖKE-PÁZSI<fnref n="1" />, Annamária SCHNEIDER<fnref n="2" />, Gyula VIDA<fnref n="1" />,
Marianna RAKSZEGI<fnref n="1" /> and Márta MOLNÁR-LÁNG<fnref n="1" />

Corresponding author: szakacs.eva@agrar.mta.hu

The most intensively used source of rye chromatin in bread wheat has been the 1RS chromosome arm in the form of 1RS.1BL translocation because it provided a source of resistance genes against leaf rust (Lr26), stem rust (Sr31), stripe or yellow rust (Yr9) and powdery mildew (Pm8). Nowadays, the proportion of wheat varieties carrying the 1RS.1BL translocation is decreasing due to the fact that resistance genes Lr26, Pm8 and Yr9 are no longer effective against new biotypes of diseases. Virulence to the Sr31 resistance gene was first reported from Uganda in 2010. Appearance of the Sr31-virulent pathotype in countries where wheat production is based on cultivars carrying the 1RS.1BL chromosome translocation can cause serious problems for agriculture. The genetic vulnerability of 1RS.1BL cultivars is the consequence of the genetic uniformity of the 'Petkus'-derived 1RS arm. Therefore, it is important to involve new rye breeding materials in resistance breeding programmes in order to improve the genetic base of cultivated wheat. In order to widen the genetic diversity of this rye chromosome arm, the winter wheat line Martonvásári 9 kr1 (Mv9kr1) was crossed with a Hungarian S. cereanum cultivar (Kriszta). The F1 hybrids were propagated in tissue culture, the regenerated plants were backcrossed with the parental wheat genotype Mv9kr1 and selfed. Progenies were propagated in the Martonvásár nursery. Selection of disease resistant lines carrying recombinant 1RS.1BL started in 2012. A wheat line, designated as line 179, was grown together with the highly susceptible parental wheat line Mv9kr1 genotypes in a pesticide-free nursery and in the Breeders nursery in three consecutive seasons (2013-2014, 2014-2015 and 2015-2016). Mv9kr1 suffered severe attack by stripe rust, while the leaves of the line '179' did not show any symptoms. Disease resistance tests under greenhouse conditions by artificial inoculations with Puccinia triticina isolates proved that this line was also resistant to leaf rust. Furthermore, this line has elevated arabinoxylan content. Molecular (SSR) marker analyses showed that the 1RS arm present in the line '179' was different from that of Petkus rye. In order to detect altered DNA sequences in the recombinant 1RS chromosome arm, a detailed analysis based on DArTseq and SNP analysis is in progress.

This work was supported by the National Research, Development and Innovation Office - NKFIH, K 119387

**Keywords:** 1RS.1BL translocation, disease resistance, leaf rust, stripe rust, perennial rye (*Secale cereanum*)

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<footnote n="1">¹ Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences
Brunszvik út 2, 2462 Martonvásár, Hungary</footnote>

<footnote n="2">² National Food Chain Safety Office, Directorate of Plant Production and Horticulture</footnote>
**PIV-46 - Lidia KOWALSKA** - Improvement of *Parastagonospora nodorum* blotch resistance in winter triticale by using the in vitro somaclonal and androgenic approaches.

# Improvement of *Parastagonospora nodorum* blotch resistance in winter triticale by using the in vitro somaclonal and androgenic approaches.

Lidia KOWALSKA¹ and Edward ARSENIUK¹

¹ Department of Plant Pathology, Plant Breeding and Acclimatization Institute-National Research Institute

Corresponding author: kowalska@ihar.edu.pl

Leaf and glume blotch caused by *Parastagonospora nodorum* is a widespread triticale disease. It can cause severe losses in grain yield wherever triticale is grown. Resistance breeding is the most acceptable way to control the disease. Plant breeders and pathologists have worked intensively to incorporate resistance to the pathogen in new cultivars. Conventional methods of breeding for resistance can be supported by using the biotechnological ones, i.e. somatic embryogenesis and androgenesis. Therefore, an effort was undertaken to study and to compare variation in *P. nodorum* resistance among winter triticale somaclones, dihaploids and conventional varieties.

For the purpose we have evaluated under controlled environment conditions eight triticale commercial cultivars, twenty one double haploids and seventeen somaclonal lines produced from cultivars varying in resistance to *P. nodorum*. The first seedling leaves were inoculated with a mixture of *P. nodorum* isolates. Spore concentrations were adjusted to 5 x 10⁶ of viable spores per one millilitre. After 10 days of incubation, the disease severity on seedling leaves was rated on a scale, where 100% – susceptible, 0 – resistant. The differences between seedling leaves for all components were statistically significant. Most of the somaclonal and dihaploid lines produced from commercial triticale cultivars showed significantly improved resistance to the pathogen in question. However, resistant parental cultivars produced primarily more resistant somaclones and dihaploids. Higher resistance to *P. nodorum* was observed more often on leaves of somaclonal lines then on dihaploid ones. On average disease severity reached 10% on leaves of somaclones and 12% on leaves of dihaploids. Some of genotypes were showing low leaf infection, e.g. dihaploid DH63 received from F1 plants of Borwo x Tomko. Similarly, lower infection level was observed on a somaclone SE64 produced from F1 plant of Borwo x Cyrkon. In conclusion, we have demonstrated that somaclonal variation might be used as an additional source of triticale resistance to the pathogen and it could be recommended to use in commercial breeding programs. The reported results broaden our knowledge about triticale resistance to *P. nodorum* and bring closer the use of biotechnological methods in breeding for disease resistance of the small grain species.

**Keywords:** *Parastagonospora nodorum*, resistance, somaclonal, androgenic variation, triticale, breeding

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PIV-47 - Sławomir BARTOSIAK - Occurrence of *Parastagonospora nodorum* blotch, *Parastagonospora avenae* blotch and *Zymosetoria tritici* blotch on wheat and triticale in Poland

# Occurrence of *Parastagonospora nodorum* blotch, *Parastagonospora avenae* blotch and *Zymosetoria tritici* blotch on wheat and triticale in Poland

Sławomir F BARTOSIAK<fnref n="1" />, E BEDNARCZYK<fnref n="1" /> and E ARSENIUK<fnref n="1" />

Corresponding author: s.bartosiak@ihar.edu.pl

Septoria diseases belong to the most devastating diseases in central Europe. Common casual agents causing septoria leaf spot in Poland are *Phaeosphaeria nodorum* (E. Müller) (anamorph: *Parastagonospora nodorum* (Berk.)), *Phaeosphaeria avenaria* (G. F. Weber) (anamorph: *Parastagonospora avenae* (A. B. Frank), *Mycosphaerella graminicola* (Fuckel) (anamorph: *Zymoseptoria tritici* (Desm.). Yield losses due to *P. nodorum* wheat infection can reach up to 31%, while *Z. tritici* can cause even 50% yield reduction. *P. avenae* is host-specific to oats, however recently the widening of *P. avenae* host range was observed.

Breeding for resistance is one of key elements of integrated plant protection. It offers the possibility to improve the resistance level to septoria disease complex through phenotyping and selection of resistant cereal types. The aim of this paper is to assess wheat and triticale varieties according to their resistance to septoria leaf spot caused by *P. nodorum*, *P. avenae*, *Z. tritici* and glume blotch caused by *P. nodorum*. Field experiments were set up at height locations in Poland to determine the natural infection of small grain cereal species by the above mentioned pathogens. The so called “septoria disease nursery” in each location consisted of 10 - winter wheat cvs., 10 - winter triticale cvs., 6 - spring wheat cvs. and 6 - spring triticale cvs. After natural infection, disease symptoms development rating was made. The 9-digit scale was used in assessment of disease severity on leaves and glumes. Affected leaves and glumes were collected and mature pycnidia were analysed under binocular 10-45X and inverted microscope at 200X magnification. The length and width of pycnidiospores were measured and plated on solid media to identify pathogen species by using a microscope camera. Dimensions (length by width) of conidia ranged on average for: *P. avenae* f. sp. *tritici* (25-45 x 3-4 µm), *P. nodorum* (15-32 x 2-4 µm) and *Z. tritici* (35-98 x 1-3 µm). The incidence frequency and disease severity on leaves were determined for each species of pathogens. Our investigation has shown that *Z. tritici* was the predominant pathogen of wheat, while *P. nodorum* was the most often encountered on triticale. Our examination revealed that presently *P. avenae* is quite common on triticale and in certain geographical regions it appeared also on wheat. Sources of resistance to examined pathogens among wheat and triticale were identified.

**Keywords:** *Parastagonospora*, *Zymoseptoria*, resistance, wheat, triticale, breeding

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<footnote n="1">¹ Department of Plant Pathology, Plant Breeding and Acclimatization Institute-National Research Institute, Radzikow, 05-870 Blonie, Poland</footnote>
**PIV-48 - Faiza BOUSSORA** - Delaying in the spike development and reduction in spikelet primordia numbers decrease the yield in salt-stressed Tunisian barley

# Delaying in the spike development and reduction in spikelet primordia numbers
decrease the yield in salt-stressed Tunisian barley

Faiza BOUSSORA<fnref n="1" />,<fnref n="2" />, Helmy M YOUSSEF<fnref n="1" />,<fnref n="3" />, Ali FERCHICHI<fnref n="4" />, Thorsten SCHNURBUSCH<fnref n="1" /> and Andreas BÖRNER<fnref n="1" />

Corresponding author: boussora@ipk-gatersleben.de

The big challenges which we face in the twenty-first century are producing sufficient food to meet demands of the increasing global population. Abiotic stresses caused by climate change, such as salinity, drought, low or high temperatures, flooding and frost are predicted to become more severe and widespread. The losses of crop productions due to salinity and drought stresses have increased progressively over the last decades. Common cereal members such as wheat (*Triticum* spp.), maize (*Zea mays* L.), rice (*Oryza sativa* L.) and barley (*Hordeum vulgare* L.) are the main important members of most worldwide nations in food and feed system. Barley is one of the oldest cereal crops known to be cultivated since about 10,000 years. It has a natural tolerance to salinity, drought, and fungal diseases, thus making it a model plant in stress biology research. We aim in this study to understand the effect of salinity stress on the spike developmental stages and the final grain yield of five Tunisian barley cultivars. Experiments were performed at IPK-Gatersleben, Germany. Seeds were sown under controlled condition in green house (LD condition, 16/8 h day/night and ~20/~16°C day/night). A saline treatment with 10gNaCl/l was imposed starting 7 days after sowing and was compared to a nonsaline control treatment. Results obtained show that, in addition to its impact on plant growth, salt has a severe and marked effect on spike development and final yield. It caused delaying in inflorescence developmental stages and decreasing in floral units numbers. Anthesis and flowering also occurred later in treated plants compared with control plants. The measurement of ten main-spike components indicated that salt negatively affected number of spikelets primordia/spike, number of grains/spike, spike length and thousand-grain weight, as well as number of tellers, number of leaves and plant height. Cultivars Ardhaoui and Rihane showed to have faster spike developmental stages and more yield than the other three under salt stress conditions.

**Keywords:** Tunisian barley, salinity, spike development, yield, growth traits

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<footnote n="1">¹ Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany</footnote>

<footnote n="2">² Faculty of Sciences of Bizerte (FSB), Bizerte, Tunisia</footnote>

<footnote n="3">³ Faculty of Agriculture Cairo University Giza Egypt</footnote>

<footnote n="4">⁴ Institut National Agronomique de Tunis, Tunisia</footnote>
**PIV-49 - Klára MÉSZÁROS** - Examination of the resistance of barley genotypes to *Pyrenophora teres f. teres* in the seedling and adult stage

# Examination of the resistance of barley genotypes to *Pyrenophora teres f. teres* in the seedling and adult stage

Klára MÉSZÁROS<fnref n="1" />, Mónika CSEPLÓ<fnref n="1" />, Viola KUNOS<fnref n="1" />, Judit BÁNYAI<fnref n="1" />, Gyula VIDA<fnref n="1" /> and József BAKONYI<fnref n="1" />

Corresponding author: meszaros.klara@agrar.mta.hu

One of the aims of the breeding programs is to elevate abiotic and biotic stress resistance, because this is an essential point of adaptation and sustainable agriculture. Therefore, the development and cultivation of disease resistant/tolerant cultivars is of great importance and the deployment of resistant cultivars is the most economic and eco-friendly method to control plant diseases.

*Pyrenophora teres* Drechsler (PTT) [anamorph *Drechslera teres* (Sacc.) Shoem.] has become an economically important and major fungal pathogen of barley worldwide. The susceptibility of current cultivars and trends in environmental conditions could contribute to the pathogen's increased importance. In Hungary, *P. teres* had become a very harmful causal agent of spring barley by the 1980's, and now is considered to be a major pathogen in both spring and winter barleys. Due to the regional and global diversity in fungal virulence it is necessary to know and breed against the local fungal pathotypes. The efficiency of net blotch resistance genes depends on the developmental stage of barley and the fungal pathotypes present in the pathogen's local population. Screening a wide genetic basis of barley varieties provides a good opportunity to find a resistance or tolerant genotypes which can be the parents of the new cultivars.

Therefore, our aims is to characterise the resistance of the Martonvásár barley collection against the pathogen in seedling and adult plants.

260 barley genotypes were tested under greenhouse and field conditions. The young barley genotypes were inoculated under greenhouse conditions with mycelium suspension of *Pyrenophora teres f. teres* isolate (H-502/1 derived from Hungary).

The barley genotypes were sown in small plots in autumn 2016 in two rows (1.5 m long with 15 cm in-row gap) and two replicates, arranged in randomised blocks. Spreading net blotch-infected straw from previous season. The area under the disease progress curve (AUDPC) was calculated from the lesion types (Tekauz scale) and infected area at various times.

The average seedling infection (AUDPC) was 13.26, ranged from 0,02 of Flagship to 91 of Hundred. In the case of adult resistance the level of infected leaves were also scored. The average AUDPC was 4495 ranged from 1642 of Patriciana to 7443 of Adagio. There were not significant difference among the infection of sensitive control and the 39% of tested varieties. The 27% of the cultivars proved to be less sensitive then the control and further 27% of studied genotypes were moderately tolerant (p=0.01). The correlation between the seedling and adult resistance was 0.32, which indicate the quite independent background of the two types of resistance. The majority of tested varieties were more sensitive in adult stage. The reason of that must be that the seedling test was carry out with one isolate while the adult resistance of genotypes were tested with PTT population. The 7.3% of the varieties were resistance in both growing stages.

This information assists the breeder to choose resistant parents in the crossing programs, but the clarification of the background of net blotch resistance of barley needs further detailed study.

This work was supported by the NRDI Fund; project number: 119276 and GINOP-2.3.2-15-2016-00029

**Related publication:**

Ficsor A, Bakonyi J, Tóth B, Tomcsányi A, Palágyi A, Csősz M, Károlyi-Cséplő M, Mészaros K, Vida G (2010) First report of spot form of net blotch of barley caused by *Pyrenophora teres f. maculata* in Hungary. Plant Dis. 94 (8), p. 1062.

<footnote n="1">¹ Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, H-
2462 Martonvásár, Brunszvik Street 2, Hungary</footnote>
Ficsor A, Bakonyi J, Csősz M, Tomcsányi A, Varga J, Tóth B (2014a) Occurrence of barley pathogenic *Pyrenophora* species and their mating types in Hungary. Cereal Res. Commun. 42 (4), 612–619.

Ficsor A, Tóth B, Varga J, Csősz M, Tomcsányi A, Mészáros K, Kótai É, Bakonyi J (2014b) Variability of *Pyrenophora teres f. teres* in Hungary as revealed by mating type and RAPD analyses. J. Plant Pathol. 96 (3), 515–523.

**Keywords:** barley, *Pyrenophora teres f. teres*, seedling and adult resistance, biotic stress

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# PIV-50 - Jean-Charles DESWARTE - Using grain carbon isotope composition (d13C) at harvest to assess growing conditions and cultivar drought tolerance

## Using grain carbon isotope composition (d13C) at harvest to assess growing conditions and cultivar drought tolerance

Jean-Charles DESWARTE<fnref n="1" />, Katia BEAUCHÊNE<fnref n="1" />, Valérie BONTEMS<fnref n="1" /> and Elise SAILLIOT<fnref n="1" />

Corresponding author: jc.deswarte@arvalis.fr

Although drought is a frequent limiting factor of cereal production across the world, breeding for drought tolerance remains challenging (Blum, 2005). Possible causes have been frequently debated: diversity of drought scenarios, difficulties to run field trials under suitable stressing conditions, large genotype x environment interactions, numerous traits to study and phenotype for (Chenu et al., 2011).

The use of carbon isotope composition (δ13C) or discrimination (Δ13C) in plant tissues as an estimate of water use efficiency has been demonstrated for many years (Farquhar and Richards, 1984); however, its use as a common tool in cereal breeding program seems limited, despite some successful demonstrations (Rebetzke et al., 2002).

The interest of using grain δ13C in West-European bread wheat breeding programs has been explored during an FSOV research project focused on identifying traits leading to drought tolerance and associated evaluation tools (Deswarte et al.). A set of grain samples have been analyzed from various trials, in order to provide basic and practical information to breeders about the way to implement and analyse such data.

The experimental precision of grain δ13C within trial was assessed by analyzing separately samples from 3-4 blocks. Average values were thus completed with error-types. Across a 15-trials network and two check cultivars (Apache and Bermude), average error-type was 0.37‰ (with a range between 0.12 to 0.99‰), with higher plot-to-plot instability under water-stress conditions.

Genetic diversity (70 cvs) for δ13C in grain was assessed through 25 trials, with a single sample per trial and per cultivar. Difference between the highest and the lowest lines was >2‰, and between the top and bottom 20th percentiles was 0.41‰. Within a set of French or European commercial cultivars, genetic diversity for δ13C largely exceeded experimental precision; it is thus feasible to screen extreme parts of populations and identify lines to keep or discard. It is noticeable that under west-European conditions, high-yielding varieties tend to show low water-use efficiencies.

Tissue δ13C can also be used as an indicator of actual water stress experienced by a crop. Average trial grain yield was regressed against estimated crop water deficit (based on soil description and weather data) and averaged δ13C (measured on several known cultivars) across 35 trials with contrasting water supply and yield levels. Correlation between mean δ13C and grain yield was higher (r²=0.58) than the one between estimated crop water deficit and yield (r²=0.38). In particular, the use of δ13C provided a significant modification of interpretation when soil water supply was difficult to assess (chalky soil, ground water table), or climatic conditions were stressful (high temperature and evaporation demand).

δ13C can thus be used either to select high-yielding lines or to qualify growing conditions without extensive soil description and computation of agro-environmental indices. Using grains instead of leaf tissue allows mechanization compared to leaf tissue, and final output remains precise and informative.

This project was funded by FSOV (2012 grant). Partners involved were: Arvalis-Institut du vegetal (JC Deswarte, K Beauchêne), NIAB (E Ober), Saaten Union (T Moittie), Secobra Recherches (C Bedard), R2N (L Duchalais), Limagrain Europe (J Derory), Bayer Crop Science (C Zimmerli), Florimond-Desprez (V Laurent), Unisigma (C Debiton), IPS2 (B Gakière) and INRA (F Baret)

**Related publication:**

Blum, A. (2005). Drought resistance, water-use efficiency, and yield potential- are they compatible, dissonant, or mutually exclusive? Aust. J. Agric. Res. 56, 1159–1168.

<footnote n="1">¹ Arvalis-Institut du végétal</footnote>
Chenu, K., Cooper, M., Hammer, G.L., Mathews, K.L., Dreccer, M.F., and Chapman, S.C. (2011). Environment characterization as an aid to wheat improvement: interpreting genotype x environment interactions by modelling water-deficit patterns in North-Eastern Australia. J. Exp. Bot. 62, 1743-1755.

Deswarte, J., Beauchêne, K., Ober, E.S., Moittie, T., Bedard, C., Duchalais, L., Derory, J., Zimmerli, C., Laurent, V., Debiton, C., et al. Identification des traits de tolérance à la sécheresse et élaboration des outils d'aide à leur évaluation. In Actes de La Rencontre Scientifique Du 23 Mars 2017, (Paris: FSOV), p.

Farquhar, G.D., and Richards, R.A. (1984). Isotopic composition of plant carbon correlates with water-use efficiency of wheat genotypes. Aust. J. Plant Physiol. 11, 539-552.

Rebetzke, G.J., Condon, A.G., Richards, R.A., and Farquhar, G.D. (2002). Selection for Reduced Carbon Isotope Discrimination Increases Aerial Biomass and Grain Yield of Rainfed Bread Wheat. Crop Sci. 42, 739-745.

**Keywords:** drought, carbon isotope composition, growing conditions, wheat, grain tissue

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PIV-51 - Roberto TUBEROSA - Dissecting the root angle QTLome to enhance durum wheat sustainability

# Dissecting the root angle QTLome to enhance durum wheat sustainability

Roberto TUBEROSA<fnref n="1" />, Marco MACCAFERRI<fnref n="1" /> and Danara ORMANBEKOVA<fnref n="1" />

Corresponding author: roberto.tuberosa@unibo.it

The optimisation of root system architecture (RSA) is an important objective for the sustainability of durum wheat grown under different water regimes. In the present study, linkage and association mapping (AM) for RSA evaluated at the seedling stage evidenced 20 clusters of quantitative trait loci (QTLs) for root length and number as well as 30 QTLs for root growth angle (RGA). The most divergent RGA phenotypes observed by seminal root screening were validated by root phenotyping of field-grown adult plants. QTL analysis of RSA and grain yield data indicates RGA as a valuable target to enhance grain yield and yield stability across different soil moisture regimes (Maccaferri et al. 2016, JXB, 67, 1161-1178, 2016). Based on their relative additive effects, allelic distribution in the AM panel and co-location with QTLs for yield, eight RGA QTLs have been prioritised in terms of breeding interest and value. These QTLs were investigated for gene content based on the chromosomal pseudomolecules of Chinese Spring *T. aestivum* and the TriAnnot v4.3 gene prediction and annotation pipeline and the Zavitan *T. dicoccoides* genome assembly (Avni et al. 2017). The chromosome regions contained 25 to 242 predicted genes (123 on average). In six RGA QTLs, from one to four gene annotations were involved in auxin pathways. The comparison between the *T. aestivum* and *T. dicoccoides* gene content indicates the high quality of the *T. dicoccoides* assembly and its usefulness to identify candidates to explore the polymorphism and the structural variation present in the A and B wheat genomes.

**Keywords:** Durum wheat, Root angle, Root system architecture, QTL, Molecular mapping, Candidate gene

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<footnote n="1">¹ Department of Agricultural Sciences (DipSA), University of Bologna, Viale Fanin 44, 40127 Bologna, Italy</footnote>
PIV-52 - Roberto TUBEROSA - Fine-mapping of QSBM.UBO-2BS=SBM2, a major QTL for resistance to soil-borne
cereal mosaic virus (SBCMV)

Fine-mapping of QSBM.UBO-2BS=SBM2, a major QTL for resistance to soil-borne
cereal mosaic virus (SBCMV)

Roberto TUBEROSA<fnref n="1" />, Martina BRUSCHI<fnref n="1" />, Marco MACCAFERRI<fnref n="1" />, Giuseppe SCIARA<fnref n="1" />, Danara ORMANBEKOVA<fnref n="1" />,
Simona CORNETI<fnref n="1" />, Claudio RATTI<fnref n="1" /> and Agata GADALETA<fnref n="2" />

Corresponding author: roberto.tuberosa@unibo.it

QSbm.ubo-2BS=Sbm2, a major QTL controlling the response to Soil-Borne Cereal Mosaic Virus (SBCMV) in durum wheat, was characterized in two recombinant inbred line populations, namely Meridiano (resistant, R) x Claudio (moderately susceptible, MS) and Simeto (susceptible, S) x Levante (R). By means of meta-QTL analysis QSbm.ubo-2BS was mapped as a unique QTL within a 2 cM-wide interval (LOD-2) in the distal region of chromosome arm 2BS (Maccaferri et al., 2012). The addition of the Illumina 90K SNPs array to the durum linkage maps allowed to identify 36 transcripts-associated SNPs tightly associated with the Mendelized QTL (Maccaferri et al., 2014). Nine SNPs from the Illumina 90K wheat array were converted to KASP markers, which provided fluorescent high-throughput assays spanning the QTL region. Marker-assisted selection (MAS) was performed on ~2,000 RILs from the Svevo (R) x Ciccio (S) population (Gadaleta et al., 2009) with two KASP markers flanking the QTL interval, KUBO 9 and KUBO 13. MAS identified 303 lines recombinant between the two markers. These lines were characterized for SBCMV response in the 2016 field nursery under severe and uniform SBCMV infection. The lines were scored for symptom severity (SS) on a 0 to 4 scale, where 0 = very resistant and 4 = very susceptible (Vallega and Rubies Autonell, 1985). They were also genotyped with seven KASP markers distributed along the QTL interval (KUBO 1, KUBO 3, KUBO 27, KUBO 29, KUBO 38, KUBO 40 and KUBO 41) and with the DArT marker wPt-2106, the marker most associated with the phenotype in the Meridiano x Claudio RIL population (Maccaferri et al., 2012). The bimodal distribution of SS reflects the segregation of the resistance trait in the QTL interval: frequency peaks were observed around values of 1.7 and 3.8. The fine-mapping allowed to narrow the most probable support interval to 0,2 cM between KUBO 27 and KUBO 1 based on the Svevo x Ciccio RILs population. Work is in progress to analyze other ~1,000 RILs of the same population in order to confirm the results obtained with the first part of the study and to improve the ongoing fine-mapping of the gene responsible for the resistance.

**Keywords:** durum wheat, SBCMV, QTL, fine-mapping, KASP marker

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<footnote n="1">¹ Department of Agricultural Sciences (DipSA), University of Bologna, Viale Fanin 44, 40127 Bologna, Italy</footnote>

<footnote n="2">² Department of Agricultural and Environmental Sciences (DiSAAT), University of Bari</footnote>
PIV-53 - Tomasz GÓRAL - *Fusarium* head blight and *Fusarium* toxins accumulation in grain of winter wheat breeding lines inoculated with *Fusarium culmorum*

*Fusarium head blight and Fusarium toxins accumulation in grain of winter wheat
breeding lines inoculated with Fusarium culmorum*

Tomasz GÓRAL<fnref n="1" />, Halina WIŚNIEWSKA<fnref n="2" />, Piotr OCHODZKI<fnref n="1" />, Dorota WALENTYN-GÓRAL<fnref n="1" />, Maciej MAJKA<fnref n="1" />
and Iga GRZESZCZAK<fnref n="1" />

Corresponding author: t.goral@ihar.edu.pl

*Fusarium* head blight (FHB) is a disease of cereals caused by fungi of genus *Fusarium*. These fungi produce toxic metabolites – mycotoxins, with phyto-and zootoxic potential. There are several types of resistance to FHB: type I – to infection, type II – to the spread of *Fusarium* in the ear, type III – to the kernel damage by *Fusarium*. Other types are the tolerance to FHB or toxins (DON) (type IV) and type V – resistance to the accumulation of toxins (trichothecenes) in the grain through their chemical modification or blocking of synthesis.

In order to obtain the forms of winter wheat that combine different types of resistance 77 lines were evaluated in field experiments in two locations. Among them there were five resistant checks: '20828[Fhb1-]', 'A40-19-1-2', 'Arina', 'Fregata', 'UNG 136.6.1.1[Fhb1+]'; five lines with Fhb1 gene from crosses of winter wheat cultivars with 'Sumai 3'; six susceptible checks; three lines with high accumulation of trichothecenes; three high yielding cultivars 'Artist', 'Patras' and 'RGT Kilimanjaro'. Wheat heads were inoculated with the spore suspension of *F. culmorum* isolates producing DON, NIV and ZEN. *Fusarium* head blight index (FHBi) was evaluated. The proportion of *Fusarium* damaged kernels (FDK) was determined visually by dividing the sample on healthy looking kernels and with symptoms of *Fusarium* damage. Using the technique of gas chromatography and immunoenzymatic tests the contents of the DON and acetyl derivatives, NIV and ZEN in the grain were analysed.

Average FHBi was 14.6%; at the range of 3.6 – 453.2%. The least infected were lines 'S43[Fhb1+]', 'Fregata' cv., '20828 [Fhb1-]', 'S32[Fhb1+]', 'S38[Fhb1-]', 'A40-19-1-2', 'UNG 136.6.1.1', 'S10[Fhb1+]', STH 032, STH 2041, KOH 275 and the most infected were six susceptible checks. The average value of the FDK amounted to 14.4%; at the range of 1.5 – 41.5%. The least damaged kernels had lines 'S 43[Fhb1+]', 'S38[Fhb1-]', 'S10[Fhb1+]', 'POB 679/03', 'S 32[Fhb1+]', 'POB 170/04'. The most damaged were kernels of susceptible checks and three cultivars 'Artist', 'Patras' and 'RGT Kilimanjaro'. There was a significant linear relationship between the FHB index and the proportion of Fusarium kernel damage – r = 0.771, p < 0.001. Chemical analyses showed the presence of Fusarium toxins DON, 3AcDON, NIV and ZEN in the grain samples. Average concentrations of DON, 3AcDON, NIV and ZEN were 6.777 mg/kg (0.750-19.750 mg/kg), 0.303 mg/kg (0.050-0.950 mg/kg), 5.146 mg/kg (0,750-22.600 mg/kg), 1115 mg/kg (147-6875 mg/kg), respectively. Lines differed significantly in terms of content of toxins in the grain. The lowest concentration of toxins was detected in grain of resistant checks 'S32[Fhb1+]', 'S38[Fhb1-]', 'S10[Fhb1+]', 'S30[Fhb1+]', 'S43[Fhb1+]', '20828[Fhb1-]' and breeding lines 'POB 0616' 'POB 0111', 'POB 0211', 'POB 6793/03', 'NAD 13015', 'POB 170/04', 'POB 457/07', 'NAD 13024'. The highest concentration of toxins was detected in grain of five susceptible checks, cultivar 'Artist' and breeding lines 'DM 2566/11', 'KBP 08 13', 'LAD 463/05', 'C 3779/10'.

It was possible to identify lines combining resistance to FHB (type I + II), to kernel damage (type III) and the low accumulation of *Fusarium* toxins (type V).

**Keywords:** *Fusarium*, wheat, toxins, trichothecenes, zearalenone

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<footnote n="1">¹ Plant Breeding and Acclimatization Institute - National Research Institute, Radzików, 05-870 Błonie, Poland</footnote>

<footnote n="2">² Institute of Plant Genetics of the Polish Academy of Sciences, Strzeszyńska 34 str., 60-479 Poznań, Poland</footnote>
PIV-54 - Marine OLLIER - A fast and reliable method for assessing *Fusarium* damaged kernels and the mycotoxin content by digital picture analysis in wheat and triticale

A fast and reliable method for assessing *Fusarium* damaged kernels and the
mycotoxin content by digital picture analysis in wheat and triticale

Marine OLLIER<fnref n="1" />, Vincent TALLE<fnref n="1" />, Anne-Laure BRISSET<fnref n="1" />, Zoe LE BIHAN<fnref n="1" />, Simon DUERR<fnref n="1" />, Ellen GOUDEMAND<fnref n="2" />,
Olivier ROBERT<fnref n="2" />, Jean-Louis HILBERT<fnref n="3" /> and Hermann BÜERSTMAYR<fnref n="1" />

Corresponding author: marine.ollier@boku.ac.at

*Fusarium* Head Blight (FHB) is a cereal disease that induces significant yield losses and contaminates the kernels with mycotoxins. It is widely admitted that the use of FHB-resistant plants is the best method for controlling the disease. The ultimate goal of resistance breeding is to identify cultivars with low risk of mycotoxin contamination. However, testing large numbers of lines for toxin content is expensive and remains prohibitive. As infected kernels are smaller, shriveled and white to pale pink in color, another method widely used is to visually estimate the proportion of *Fusarium Damaged Kernels (FDK)* as an approximation for *Fusarium* severity. This method is an easy and efficient way to select for low toxin, but is time consuming and labor intensive. Digital picture analysis could be a very good option to simplify FDK evaluation. Our new method is based on the hypothesis that it is possible to select for low mycotoxin accumulating lines by checking the level of whitening of the kernel sample. Pictures are taken with a digital camera under controlled light conditions. We used RGB criteria to segregate the picture's pixels in three categories: background pixels, healthy-grain pixels and diseased-grain pixels. The Whitened Kernel Surface (WKS) is then calculated as the proportion of diseased-grain pixels among all grain pixels. We tested this method on two bread wheat panels gathering 228 grains samples in total, and one triticale panel of 37 samples. We showed a high Pearson correlation between FDK and WKS (r=0.79 and r=0.77 for bread wheat and r=0.81 for triticale), and between mycotoxin content and WKS (r=0.86 and r=0.73 for bread wheat and r=0.61 for triticale). This new notation criterion based on pictures analysis is a promising tool for breeders and researchers. It is as efficient as the traditional visual notation and happens to be faster, easier and more stable. A very economical way to evaluate mycotoxin content and to enable the large scale scoring and ranking needed to select resistant cereal varieties for the future.

<figure><img src="image_46.png" /><figcaption>**Fig. 4.** Infected sample of bread wheat. **A**, Original picture; **B**, Treated picture, with background pixel artificially turned in black, and diseased grain pixels turned in pink.</figcaption></figure>

**Keywords:** Digital picture analysis, *Fusarium* Head Blight, FHB, Wheat, Triticale, FDK, DON, Mycotoxin, WKS

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<footnote n="1">¹ BOKU-University of Natural Resources and Life Sciences Vienna, Department IFA-Tulln, Institute for Biotechnology in Plant Production, Konrad Lorenz Str. 20, 3430 Tulln, Austria</footnote>

<footnote n="2">² EA 7394, USC INRA 1411, Institut Charles Viollette (ICV), Agro-food and biotechnology research institute, Université de Lille, INRA, ISA, Univ. Artois, Univ. Littoral Côte d'Opale, Cité Scientifique, 59655 Villeneuve d'Ascq, France</footnote>

<footnote n="3">³ Florimond-Desprez, 3 rue Florimond-Desprez, BP 41, 59242 Cappelle-en-Pévèle, France</footnote>
**PIV-55 - Jose MORENO-AMORES - Assessing the incorporation of covariates in genomic predictions for *Fusarium* head blight in durum wheat**

# Assessing the incorporation of covariates in genomic predictions for *Fusarium* head blight in durum wheat

Jose MORENO-AMORES¹, Sebastian MICHEL¹ and Hermann BÜERSTMAYR¹

¹ Department for Agrobiotechnology IFA-Tulln, Institute for Biotechnology in Plant Production, University of Natural Resources and Life Sciences Vienna BOKU

Corresponding author: jose.moreno-amores@boku.ac.at

Durum wheat (*Triticum turgidum* var. durum) is an important cereal in Europe especially susceptible to *Fusarium* head blight (FHB) which produces yield and quality reduction due to the grain contamination with mycotoxins. In this study, we used a cross-validation schema to assess the accuracies of the genomic predictions for our trait of interest (FHB mean severity) and its correlation with two covariates: heading date (HD) and plant height (PH). Predictions relied on genomic best linear unbiased prediction GBLUP models using a panel of 184 durum wheat genotypes across five environments alongside with their genome-wide set of 30,611 SNP and DArT markers.

Significant negative correlations were found between the estimations of FHB mean severity and the two covariates. This motivated us to evaluate: (i) two methods to correct the FHB estimated values before running the prediction models; (ii) the covariate(s) incorporation as fixed factor(s) on GBLUP models and (iii) the use of restriction indexes in a multi-trait model context.

We corroborated the increase in FHB mean prediction accuracy (0.54) when genotype-environment interaction was included compared to the one obtained within each trial (ranging from 0.38 to 0.52). Also, accuracy differences were mainly found within environments between the adjustment attempts for covariate HD as follows: correcting FHB values before predictions proved to be the best option for at least not producing any significant decrease in accuracy terms. On the other hand, among adjustments methods for PH, there were no accuracy differences. All of the attempts succeed reducing to non-significant levels the correlation between the predicted FHB and both covariates. Our results describe the impact of incorporating covariates in genomic prediction models and possible approaches to addressee it.

**Keywords:** Genomic prediction, *Fusarium* head blight, durum wheat, covariates, plant height, heading date

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**PIV-56 - Mahbubjon RAHMATOV** - Development and characterization of new stem rust and stripe rust resistance genes in Swedish wheat-rye introgression lines

# Development and characterization of new stem rust and stripe rust resistance genes in Swedish wheat-rye introgression lines

Mahbubjon RAHMATOV<fnref n="1" />, Matthew ROUSE<fnref n="2" />, Mehran PATPOUR<fnref n="3" />, Mogens HOVMØLLER<fnref n="3" />, Brian STEFFENSON<fnref n="4" />
and Eva JOHANSSON<fnref n="1" />

Corresponding author: mahbubjon@gmail.com

The emergence of new stem rust (Puccinia graminis f. sp. tritici) and stripe rust (Puccinia striiformis f. sp. tritici (Pst)) races poses a major threat to Swedish wheat production. To mitigate the threats of these virulent pathogens, the objectives of this study were to characterize new sources of resistance genes in wheat-Secale cereale derivatives. A new 2DS·2RL wheat-rye translocation was obtained through the breakage-fusion mechanism and found to carry a novel stem rust resistance gene designated *Sr59* from Secale cereale. *Sr59* confers a high level of resistance to many widely virulent races including TTKSK, TTTSK, TTKTT, TKTTF, TKTTF+Sr25, TTRTF, TPMKC, RKQQC, RCRSC and two isolates of race TTTTF from USA and Russia. Research is currently being conducted to transfer *Sr59* into adapted elite lines with high yield potential, agronomic adaptation, and end-use quality characteristics. The characterization of *Sr59* and its introgression through 2RL into wheat provides wheat breeders with an additional genetic resource for breeding for stem rust resistance. Moreover, six 4R (4D), 5R (5D) and 7R (7D) wheat-rye multiple disomic substitution lines were found highly resistant to a suite of broadly virulent Pst races (21 in total). Development of wheat-rye translocations are currently being investigated through induced-homoeologous recombination that may enhance the utility of rye resistance genes, thus contributing to the diversification of both stem rust and stripe rust resistance. Rye has been an important donor of rust resistance genes, in addition to contributing to yield potential, adaptability, and tolerance to various biotic and abiotic stresses in wheat. Rye remains a largely untapped reservoir of genes for wheat improvement and should be investigated more thoroughly.

**Related publication:**

Rahmatov M, Rouse MN, Steffenson BJ, Andersson SC, Wanyera R, Pretorius ZA, Houben A, Kumarse N, Bhavani S, Johansson E (2016) Sources of stem rust resistance in wheat-alien introgression lines. Plant Disease 100:1101-1109

Rahmatov M, Rouse MN, Nirmala J, Danilova T, Friebe B, Steffenson BJ (2016) A new 2DS·2RL Robertsonian translocation transfers stem rust resistance gene *Sr59* into wheat. Theor Appl Genet 129

Rahmatov M, Hovmøller MS, Nazari K, Andersson SC, Steffenson BJ, Johansson E (2017) Seedling and adult plant stripe rust resistance in diverse wheat-alien introgression lines. Crop Science 57:2032–2042

**Keywords:** wheat, rye, resistance gene, stem rust, stripe rust, Robertsonian translocation

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<footnote n="1">¹ Department of Plant Breeding, Swedish University of Agricultural Sciences, PO Box 101, 23053 Alnarp, Sweden</footnote>

<footnote n="2">² United States Department of Agriculture, Agricultural Research Service, Cereal Disease Laboratory, St. Paul, MN 55108, USA</footnote>

<footnote n="3">³ Department of Agroecology, Aarhus University, Flakkebjerg, DK4200 Slagelse, Denmark</footnote>

<footnote n="4">⁴ Department of Plant Pathology, University of Minnesota, St. Paul, MN 55108, USA</footnote>
PIV-57 - Andrii FATIUKHA - Detection of functional and non-functional alleles of the stripe rust resistance gene *Yr15* using diagnostic markers

# Detection of functional and non-functional alleles of the stripe rust resistance gene *Yr15* using diagnostic markers

Andrii FATIUKHA<fnref n="1" />, Valentina KLYMIUK<fnref n="1" />, Lin HUANG<fnref n="1" />, Elitsur YANIV<fnref n="2" />, Dina RAATS<fnref n="1" />, Jorge DUBCOVSKY<fnref n="3" />,
Alan H SCHULMAN<fnref n="2" />, Abraham B KOROL<fnref n="1" /> and Tzion FAHIMA<fnref n="1" />

Corresponding author: avfatyha@gmail.com

Stripe rust, caused by the fungus *Puccinia striiformis f.s. tritici* (Pst), is a destructive disease of wheat globally. Depletion of effective resistance to Pst in cultivated wheat has led to search for new resistance genes in the wild relatives of wheat. One of the most promising genes conferring broad-spectrum resistance to stripe rust is *Yr15*, derived from wild emmer wheat (*Triticum dicoccoides*) accession G25. *Yr15*, mapped on chromosome arm 1BS, has recently been cloned by our consortium and designated as Wheat Tandem Kinase 1 (WTK1). We found wtk1 susceptible alleles in most 274 tested durum, bread, and wild emmer wheat lines. Out of 69 tested durum and bread wheat cultivars and lines, only 33 *Yr15* introgression lines contained the functional allele (Wtk1) from G25 and were resistant to Pst. The remaining 36 susceptible lines carried non-functional alleles (wtk1), which included insertions of large transposable elements that resulted in changes in reading frame. Development of reliable molecular markers can facilitate the introgression of *Yr15* into new varieties via marker-assisted selection. Diagnostic markers designed based on the polymorphism between the WTK1 alleles are preferred in order to avoid negative linkage drag. Therefore, we have designed high-throughput co-dominant KASP markers that can differentiate between the functional (Wtk1) and all known non-functional (wtk1) alleles, and can be used in breeding programs for development of modern cultivars with high resistance to stripe rust.

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<footnote n="1">¹ Institute of Evolution and the Department of Evolutionary and Environmental Biology, Faculty of Science and Science Education, University of Haifa</footnote>

<footnote n="2">2 LUKE & University of Helsinki</footnote>

<footnote n="3">³ University of California</footnote>
**PIV-58 - Christophe GIGOT - Using crop-pathogen modeling to identify plant traits to control Zymoseptoria tritici**
epidemics on wheat

# Using crop-pathogen modeling to identify plant traits to control Zymoseptoria tritici epidemics on wheat

Christophe Gigot<fnref n="1" />,<fnref n="2" />, David CLAESSEN<fnref n="2" />, Guillaume GARIN<fnref n="1" />,<fnref n="3" />, Christian FOURNIER<fnref n="4" />, Christophe PRADAL<fnref n="5" />,<fnref n="6" />
and Corinne ROBERT<fnref n="1" />,<fnref n="2" />

Corresponding author: christophe.gigot@inra.fr

Diversification in pathogen control methods to reduce the severity of economically important foliar diseases such as *Zymoseptoria tritici* on wheat is needed. One way is to identify plant physiological and architectural traits that influence disease development and that can be selected in the process of crop breeding. Such traits may be used for improving tolerance or disease escape. Traits favoring disease escape, the focus of our work, may significantly decrease crop epidemics (Robert et al., 2018). However, understanding the role of such traits in crop-pathogen interactions is a daunting task because the interactions are multiple and dynamic in time. To characterize and quantify crop-pathogen interactions, an innovative trait-based and resource-based modeling framework was developed (Precigout et al., 2017). In this framework, the pathosystem is assumed to respond dynamically to both architecture and physiological status of the host canopy. A canopy consists of plenty of small patches, i.e. small functional and infectable units of leaf tissue. Production of new patches, for canopy growth and renewal of photosynthetically active plant tissues, is a function of the available resources produced by the other patches. Pathogen spores can contaminate nearby healthy patches. The definition of patch proximity depends on dispersal abilities of the pathogen and canopy architecture. We used and adapted this modeling framework to quantify the effects of several plant traits on *Zymoseptoria tritici* epidemics for varied climate scenarios. The complex infection cycle of *Z. tritici* characterized by a long symptomless incubation period was implemented in the model. We studied plant architectural traits such as leaf size or stem height, and plant physiological traits such as leaf lifespan or leaf metabolite contents. In our simulations, these traits impacted the epidemics dynamics though their effects on pathogen dispersal and on the amount of resources available for the pathogen. Sensitivity analyses showed how disease severity depended on plant traits and pathogen virulence. The importance of several plant and pathogen traits could be linked to the pathogen's ability to manage the race for the colonization of the canopy in the face of canopy growth. Playing on host traits also made it possible to simulate different wheat varieties - with contrasted heights, pathogen resistance or precocity - to characterize the behavior of the pathosystem of interest for different host ideotypes. We argue that this kind of trait-based modeling approach is a valuable tool to identify plant traits promoting more resilient agroecosystems in particular for crop breeding in a context of innovative and sustainable crop protection.

**Related publication:**

Précigout P-A, Claessen D, Robert C. 2017. Crop fertilization impacts epidemics and optimal latent period of biotrophic fungal pathogens. Phytopathology 107, 1256-1267.

Robert C, Garin G, Abichou M, Houlès V, Pradal C, Fournier C. 2018. Plant architecture and foliar senescence impact the race between wheat growth and *Zymoseptoria tritici* epidemics. Annals of Botany (in press).

**Keywords:** *Septoria tritici* blotch, *Zymoseptoria tritici*, wheat, fertilization, epidemiology, tolerance, trait-based modeling, virulence, consumer-resource dynamics, disease-escaping traits

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<footnote n="1">¹ INRA, UMR 1402 ÉcoSys, 78850 Thiverval-Grignon, France</footnote>

<footnote n="2">² CNRS-ENS-INSERM, UMR 8197, Institut de Biologie de l'École Normale Supérieure, École Normale Supérieure, 46 rue d'Ulm, 75005 Paris, France</footnote>

<footnote n="3">³ ITK, avenue de l'Europe, 34830 Clapiers, France</footnote>

<footnote n="4">⁴ INRA, UMR 759 LEPSE, 34060 Montpellier, France</footnote>

<footnote n="5">⁵ CIRAD, UMR AGAP - INRIA, Virtual Plants, 34398 Montpellier, France</footnote>

<footnote n="6">⁶ Institut de Biologie Computationnelle, 34095 Montpellier, France</footnote>
PIV-59 - Claire GUÉRIN - Genome-wide identification, characterization and expression profiling of NAC genes in *Triticum aestivum*

# Genome-wide identification, characterization and expression profiling of NAC genes in *Triticum aestivum*

Claire GUÉRIN¹, Jane ROCHE¹, Vincent ALLARD¹, Mohamed Fouad BOUZIDI¹ and Saïd MOUZEYAR¹

¹ INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France

Corresponding author: guerin-cla@orange.fr

The bread wheat, *Triticum aestivum*, is an important economic crop worldwide and its yield is continuously affected by the global warming. Insights into wheat adaptability mechanisms of its seed development under different abiotic stresses represent a new challenge to improve wheat yield.

Transcription factors are major actors of molecular and cellular mechanisms involved in the regulation of development under physiological conditions and in response to many biotic and abiotic stresses.

The NACs are a large family of transcription factors, present in the whole plant reign. Some members of this family have been shown to be involved in the development such as flowering, lateral roots division, cell division or seed development; and in response to biotic (i.e. stripe rust) and abiotic (i.e. salinity or drought) stresses in different species.

However, the characterization of this family remains largely unknown in *Triticum aestivum*.

Here, we compared the latest sequences available in the database of the International Wheat Genome Sequencing Consortium (https://www.wheatgenome.org/) with the data gathered by Borrill et al. (2017) in order to improve the quality of the NAC family existent set.

Furthermore, a phylogenic structuration of the enriched family was done; particularly by completing the homeologous groups. Its chromosomal repartition was also studied and showed that: i) NACs are equitably distributed within the three subgenomes (A, B and D) of *Triticum aestivum* and ii) NACs have a different distribution within the seven chromosomes, with a higher abundance on chromosomes 2 and 7.

We hypothesized that some duplication events may explain the physical and phylogenical clustering of several NAC genes within the family.

Secondly, we focused on the expression profile of some key members of the NAC family in bread wheat.

We selected 24 NAC genes that were found to be differentially expressed in different tissues (roots, leaves, seeds) or in response to different abiotic stresses in leaves (heat shock and PEG induced drought at a young stage), based on the data available on the www.wheat-expression.com platform (Borrill et al., 2016). Using real-time quantitative PCR, the behavior of these 24 NAC genes was studied on the genome referenced genotype, Chinese Spring (IWGSC) in the same heat shock conditions. The expression of those 24 NAC genes was also quantified in leaves and seeds at 220 and 450°Cd post-anthesis, under severe drought conditions applied during the seed development of two contrasted genotypes in field (Pheno3C platform, INRA Clermont-Ferrand).

This work may lead to increase knowledge about the NAC family in *Triticum aestivum*. We identified genes which are preferentially expressed in leaves and seeds among which some respond to heat and drought stresses.

These data might further give useful information for breeders to develop adapted varieties to global warming.

**Keywords:** bread wheat, transcription factor, NAC family, phylogenic structuration, gene expression

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**PIV-60 - Ghasemali NAZEMI - The association mapping of short-term heat shock and chronic heat stress tolerance in durum wheat**

**The association mapping of short-term heat shock and chronic heat stress tolerance
in durum wheat**

Ghasemali NAZEMI<fnref n="1" />, Marco MACCAFERRI<fnref n="2" />, Silvio SALVI<fnref n="2" /> and Roberto TUBEROSA<fnref n="2" />

Corresponding author: nazemigh@gmail.com

Heat stress due to increased temperature is an agricultural problem in many areas of the world. Short-term heat stress (acute heat stress) provokes heat shock response, in contrast, long-term heat exposure (chronic heat stress) induces larger scale adaptations. The present work aims to carry out a comprehensive phenotypic analysis of a durum wheat collection for the response to high-temperature followed by an association mapping study to detect Quantitative Trait Loci (QTLs) associated with heat tolerance. The response to chronic heat stress and short-term heat shock has been studied in controlled environments where specific heat treatments may be applied. The panel was profiled with simple sequence repeat, Diversity Arrays Technology and sequence-tagged site markers (957 markers in total). The genetic relationships among the accessions have been investigated using both a genetic-similarity and a model-based Bayesian clustering method. Significant differences among genotypes were observed for all measured heat response traits and thirty four single marker/QTL regions were located in all chromosomes. It is interesting that a higher number of QTLs were detected for constitutive heat response trait (heat shock applied to detached leaves) as compared to the two traits involving acquired responses, measured on living plants pre-adapted to high temperatures. Molecular markers linked to the heat tolerance trait represented a more reliable tool for selecting heat tolerance genotypes in durum wheat breeding programs.

**Related publication:**

Maccaferri M, El-Feki W, Nazemi G, Salvi S, Canè M.A, Colalongo M.C, Stefanelli S, Tuberosa R 2016 Prioritizing quantitative trait loci for root system architecture in tetraploid wheat. Journal of Experimental Botany. 67(4):1161-78.

Nazemi G, Valli F, Maccaferri M, Salvi S, Tuberosa R 2015 Genetic variation for aerenchyma and other root anatomical traits in durum wheat. (2015), Genetic Resource Crop Evolution Journal. DOI 10.1007/s10722-015-0279-6.

**Keywords:** Durum wheat, Heat stress response, association mapping

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<footnote n="1">¹ Young Researchers and Elite Club, Bandar Abbas Branch, Islamic Azad University, Bandar Abbas, Iran</footnote>

<footnote n="2">² Department of Agricultural Sciences (DipSA), University of Bologna, Bologna, Italy</footnote>
PIV-60bis - Johanna LETHIN - Molecular breeding of salt tolerant wheat

Molecular breeding of salt tolerant wheat

<u>Johanna Lethin</u><fnref n="1" />, Sharhiar Shakil<fnref n="2" />, Olof Olsson<fnref n="2" /> and Henrik Aronsson<fnref n="1" />

Corresponding author: johanna.lethin@bioenv.gu.se

Over the last few decades, on average ca 2,000 ha of irrigated agricultural land has been lost every day due to high salinity levels. The size of high salinity areas has been growing from 45 million ha in the early 1990s to its current volume of 62 million ha [1]. Globally, 3.1% of the total land mass is affected by salinity, with over half of the world's countries afflicted [2]. Presently, about one-fifth of the world's irrigated lands are too salty for agriculture. Clearly, this negative trend has to be broken.

Our aim is to develop wheat (Triticum aestivum L.) varieties with increased salinity tolerance. Starting from Gom-
25 [3], a commonly grown in Bangladesh variety already fairly well adapted to salt and heat stress, we introduced ca
1 million mutations per genome by EMS (Ethyl Methane Sulphonate), and raised a population of ca 1800 lines.
Mutated wheat lines with an increased salinity tolerance are now selected from this population. The aim is to reuse
some parts of the 1 million ha area of salt contaminated land in Bangladesh.

**Related publication:**

[1] Qadir *et al* (2009) Salt-induced land and water degradation in the Aral Sea basin: A challenge to sustainable agriculture in Central Asia. Natural Resources Forum. Wiley Online Library, pp 134-149

[2] Corbishley and Pearce (2007) Growing trees on salt-affected land. Australian Centre for International Agricultural Research

[3] Hossain and Da Silva (2013) Wheat production in Bangladesh: its future in the light of global warming. AoB plants 5: pl042

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<footnote n="1">¹ Department of Biological and Environmental Science, Carl Skottsbergs gata 22B, University of Gothenburg, SE-40530 Gothenburg, Sweden</footnote>

<footnote n="2">² Department of Pure and Applied Biochemistry, Lund University, Box 124, SE-22100 Lund, Sweden</footnote>
SESSION V. HIGH QUALITY AND SAFE CEREALS FOR FOOD, FEED AND INDUSTRY
**PV-61 - Catherine RAVEL** - Rapid identification of high-molecular-weight glutenin subunits by SNP markers to breed for high quality

# Rapid identification of high-molecular-weight glutenin subunits by SNP markers to breed for high quality

Catherine RAVEL<fnref n="1" />, Annie FAYE<fnref n="1" />, Marion RANOUX<fnref n="1" />, Céline DUPUIS<fnref n="1" />, Mireille DARDEVET<fnref n="1" />, Florence EXBRAYAT<fnref n="1" />, Charles PONCET<fnref n="1" />, Pierre SOURDILLE<fnref n="1" /> and Gérard BRANLARD<fnref n="1" />

Corresponding author: catherine.grand-ravel@inra.fr

In bread wheat (*Triticum aestivum*), the high-molecular-weight glutenin subunits (HMW-GS) are the most important determinant of technological quality. Known for their high level of diversity, they are encoded by tightly linked genes (*Glu-1-1* & *Glu-1-2*). Their identification is based on elephoresis (SDS-PAGE). However, molecular markers would facilitate the early selection of high-quality HMW-GS alleles in breeding programmes. In this work, we identified sequence polymorphisms and developed 19 SNP markers thanks to an alignment comprising the nucleotidic sequences of the main HMW-GS alleles publicly available. Genotyping and electrophoresis result obtained in a core collection (n=364 lines), which represents the worldwide diversity, were compared. SNP markers developed at *Glu-A1* and *Glu-D1* and SDS-PAGE gave concordant results. Discrepancies were observed mainly at *Glu-B1* locus. However, genetic association showed that markers at *Glu-B1* genes could be more efficient than SDS-PAGE allele to improve rheological properties of the dough. To conclude, this study provided a set of SNP markers, which could constitute a useful tool-box to breed for the end-used value.

**Keywords:** Bread wheat (*Triticum aestivum* L.). End used value. High-molecular-weight glutenin subunit. Single Nucleotide Polymorphism

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<footnote n="1">¹ INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France</footnote>
**PV-62 - Emmanuelle BANCEL - Proteomic changes occuring in einkorn (Triticum monoccocum) grain during filling stages of development**

**Proteomic changes occuring in einkorn (Triticum monoccocum) grain during filling stages of development**

Emmanuelle BANCEL<fnref n="1" />, Titouan BONNOT<fnref n="1" />, Marlène DAVANTURE<fnref n="2" />, Marie PAILLOUX<fnref n="3" />, Michel ZIVY<fnref n="2" />, Pierre MARTRE<fnref n="4" /> and Catherine RAVEL<fnref n="1" />

Corresponding author: emmanuelle.bancel@inra.fr

Albumins and globulins of wheat endosperm represent about 20% of total grain proteins. Nutritionally, the albumins and globulins (non-glutens) have a very good amino acid balance. Many of these proteins are enzymes involved in metabolic activity. Most of these physiologically active proteins can also influence the processing and rheological properties of wheat flour.

Incomplete knowledge of these enzymes and related proteins present in the endosperm of wheat limits our understanding of the biochemical events taking place in the grain during the first stages of its development. So it is necessary to study and evaluate qualitative and quantitative changes occurring during this process. To reach this objective, our study is based on *Triticum monococcum* (einkorn), a diploid wheat species, used as a model species of bread wheat (*T. aestivum*).

We used a proteomic approach to identify the key actors during the filling stages of development in einkorn grain. Five stages were studied (200 to 600°Cd). Large-scale analysis using the gel free proteomic approach allowed us to identify a total of 2,475 proteins. Among these proteins, 512 (20.7%) were accurately quantified, 488 of them presented changes related to stage development, highlighting this dynamic process (Bonnot et al, 2017).

Hierarchical cluster analysis revealed four major expression profiles throughout grain development. The classification of proteins identified by MS/MS in 17 different biochemical functions and nine molecular functions provided a picture of the temporal cellular processes involved. Several proteins differentially expressed and associated with grain development and/or quality were also shown.

A co-accumulation network of all differentially expressed proteins mainly centred on enzymes involved in carbon and protein metabolism was finally established.

**Related publication:**

Bonnot T, Bancel E, Alvarez D, Davanture M, Boudet J, Pailloux M, Zivy M, Ravel C, Martre P (2017) Grain subproteome responses to nitrogen and sulfur supply in diploid wheat *Triticum monococcum* ssp. *monococcum*. Plant J. doi: 10.1111/tpj.13615

**Keywords:** wheat, grain, albumins globulins, proteome, co-accumulation network

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<footnote n="1">¹ INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France</footnote>

<footnote n="2">² UMR GQE, INRA, Université Paris-Sud, CNRS, AgroParisTech, Université Paris-Saclay, 91 190 Gif-sur-Yvette, France</footnote>

<footnote n="3">³ LIMOS, CNRS, Université Clermont Auvergne, 63 173 Aubière, France</footnote>

<footnote n="4">⁴ UMR LEPSE, INRA, Montpellier SupAgro, 34 060 Montpellier, France</footnote>
PV-63 - Julie BOUDET - SPA Heterodimerizing Protein, a bZIP transcription factor, represses glutenin synthesis in *Triticum aestivum*

SPA Heterodimerizing Protein, a bZIP transcription factor, represses glutenin synthesis in *Triticum aestivum*

Julie BOUDET<fnref n="1" />*, Marièle MERLINO<fnref n="1" />*, Anne PLESSIS<fnref n="1" />,<fnref n="5" />, Jean-Charles GAUDIN<fnref n="2" />,<fnref n="4" />, Mireille DARDEVET<fnref n="1" />,
Sibille PERROCHON<fnref n="1" />, David ALVAREZ<fnref n="1" />, Thierry RISACHER<fnref n="3" />,<fnref n="6" />, Pierre MARTRE<fnref n="1" />,<fnref n="7" /> and Catherine RAVEL<fnref n="1" />

Corresponding author: julie.boudet@inra.fr

The quality of the wheat grain is determined by the quantity and the composition of grain storage proteins (GSPs). These GSPs consist of the glutenin of low- (LMW-GS) and high- (HMW-GS) molecular-weight and the gliadin. The synthesis of these proteins is essentially regulated at the transcriptional level and by the availability of nitrogen and sulfur. This network of regulation has been extensively studied in barley. BLZ1 and BLZ2, belonging to the basic leucine Zipper (bZIP) family activate the synthesis of hordeins. To date, in wheat, only the orthologue of BLZ2, SPA or Storage Protein Activator, was identified to play a major role in the regulation of GSP synthesis. Here, the involvement of the orthologous of BLZ1 named SPA Heterodimerizing Protein (SHP) was analyzed on the transcriptional regulation of the genes coding for GSPs. By gel mobility shift assays, we have demonstrated that SHP binds with cis-motifs, known to bind to the bZIP family TFs, on the HMW-GS and LMW-GS promoters (*GluB1-1* and *GluD3*, respectively). Moreover, we showed by transient expression assays in wheat endosperms that SHP acts as a repressor on the activity of these gene promoters. These results were supported by transgenic lines overexpressing SHP gene grown with low and high nitrogen supply. The phenotype of SHP overexpressing lines showed a lower proportion of both LMW-GS and HMW-GS, while the proportion of gliadin was unchanged, whatever the availability in nitrogen. Thus, the gliadin/glutenin ratio was increased, which suggest that gliadin genes may be regulated differently than glutenin genes.

**Keywords:** bZIP transcription factor, gluten, SPA Heterodimerizing Protein, storage proteins, wheat (*Triticum aestivum* L.).

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<footnote n="1">¹ INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France</footnote>

<footnote n="2">2 UR BIA, INRA, 44 316 Nantes, France</footnote>

<footnote n="3">³ Biogemma, Centre de Recherche de Chappes, 63 720 Chappes, France</footnote>

<footnote n="4">⁴ UR AGPF, INRA, 45 075 Orléans, France</footnote>

<footnote n="5">⁵ School of Biological Sciences, Plymouth University, Drake Circus, Plymouth PL4 8AA, UK</footnote>

<footnote n="6">⁶ Limagrain Europe, 63 720 Chappes, France</footnote>

<footnote n="7">⁷ UMR LEPSE, INRA, Montpellier SupAgro, 34 060 Montpellier, France</footnote>

<footnote>* These authors contributed equally to this manuscript</footnote>
**PV-64 - Pierre BERTHOMIEU** - Towards the positional cloning of genes controlling zinc and iron content in durum wheat grain

# Towards the positional cloning of genes controlling zinc and iron content in durum wheat grain

Pierre BERTHOMIEU<fnref n="1" />, Abdoulaye BEYE<fnref n="1" />, Virginie THIRY<fnref n="1" />, Pierre ROUMET<fnref n="2" /> and Jacques DAVID<fnref n="2" />

Corresponding author: pierre.berthomieu@supagro.fr

Wheat grain contains low levels of iron (Fe) and zinc (Zn). In addition, most of the phosphorus in the grain is stored as phytate, which is an antinutrient that chelates Fe and Zn, thereby rendering them unavailable in the human diet. Thus, improving both accumulation and bioavailability of Fe and Zn in wheat grains is desirable. A couple of international breeding programs work in that direction. However, there is up to now very little information on the mechanisms and genes controlling the amount and bioavailability of Fe and Zn in the grain.

Our project aims at identifying genetic loci and associated genes controlling Fe, Zn and phytate accumulation in durum wheat grain. We developed a genome wide association study on an original population (1) obtained by intercrossing a large set of durum, emmer and wild wheat accessions. Preliminary results show that two regions (6 Mb- and 4 Mb-long, respectively) were identified as harbouring loci controlling Fe and Zn content in grain. Interestingly, the haplotype associated to high Fe content seems to come from the wild compartment.

Thanks to the recent availability of the sequence of the durum wheat genome (2), positional cloning is under way.

**Related publication:**

David et al. (2014) Genotyping by sequencing transcriptomes in an evolutionary pre-breeding durum wheat population. Molecular breeding, 34:1531-48

Avni et al. (2017) Wild emmer genome architecture and diversity elucidate wheat evolution and domestication. Science. 357:93-7

**Keywords:** Wheat grain, biofortification, zinc, iron, GWAS, EPO population

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<footnote n="1">¹ UMR BPMP, Montpellier SupAgro, INRA, CNRS, University of Montpellier, France</footnote>

<footnote n="2">² UMR AGAP, INRA, Montpellier SupAgro, CIRAD, University of Montpellier, France</footnote>
SESSION VI. FUTURE CHALLENGES AND INNOVATIONS
PVI-65 - Péter MÍKÓ - Selection of winter cereals for organic agriculture

Selection of winter cereals for organic agriculture

Péter MÍKÓ<fnref n="1" />, Mária MEGYERI<fnref n="1" />, Márta MOLNÁR-LÁNG<fnref n="1" />, Marianna RAKSZEGI<fnref n="1" /> and Gyula VIDA<fnref n="1" />

Corresponding author: miko.peter@agrar.mta.hu

Productivity in agriculture has more than tripled in developed countries since the 1950s. Beyond the success of plant breeding, the increased use of inorganic fertilizers, application of pesticides, and spread of irrigation also contributed to this success. However, impressive yield increases started to decline in the 1980s because of the lack of sustainability. One of the most beneficial ways to increase sustainability is organic agriculture. In such systems the prerequisite of successful farming is the availability of crop genotypes that perform well. However, selection of winter cereals for sub-optimal growing conditions is still neglected, and the organic seed market also lacks of information on credibly tested varieties suitable for organic agriculture.

Quality and agronomic performance of a diverse set of 37 winter bread wheat varieties and 14 winter durum varieties were examined in organic and conventional management systems of Austria, France (only durum) and Hungary through 3 years to identify traits sensitive to management systems.

Based on the results of the winter wheat trial, seven traits showed significant management × genotype interaction. Heading date, sensitivity to leaf rust and powdery mildew had high heritability making it reasonable to select them for organic agriculture also in conventional field in early generations. However, it is suggested to select for the other four traits (grain yield, test weight, leaf-inclination and vigorous plant growth) later under organic growing conditions.

In the case of winter durum, heading time, wet gluten content, semolina yield and grain protein content are traits that showed genotype-dependent significant differences between the two management systems examined. Therefore, breeding for these traits could result in specifically adapted genotypes for organic agriculture in the different countries.

Based on strong or moderate significant correlations between the winter durum traits, gluten index and plant height could also be specifically selected in an indirect way. The need for environmentally specific selection for grain yield in later generations was also demonstrated.

Our findings provide evidence for the influence of the selection environment, whether it is the management system or the growing region. Based on these findings, at least a partly separate winter cereal selection program is recommended for organic and low input agriculture in each country, where targeted selection could be carried out more effectively.

Knowledge gained from these experiments will be used to design efficient cultivar testing strategies for organic farming in a European network. This will support farmers in their choice of cultivar with stable yield and high quality and thus support the emerging movement towards sustainable farming systems.

This research received funding from the European Community's Seventh Framework Programme (FP7/2007-2013) under Grant Agreement No. 245058-SOLIBAM (supplementary Hungarian project EU_BONUS_12-1-2012-0032) and from the European Union's Horizon 2020 research and innovation programme under grant agreement No 727230 (LIVESEED). The information contained in this communication only reflects the author's view. The Research Executive Agency is not responsible for any use that may be made of the information provided.

**Related publication:**

Mikó, P., Vida, G., Rakszegi, M., Lafferty, J., Lorentz, B., Longin, C.F.H., Megyeri, M. (2017): Selection of winter durum genotypes grown under conventional and organic conditions in different European regions. Euphytica, 213: 169; DOI 10.1007/s10681-017-1953-x

Mikó, P., Löschenberger, F., Hiltbrunner, J., Aebi, R., Megyeri, M., Kovács G., Molnár-Láng, M., Vida, G., Rakszegi, M. (2014): Comparison of bread wheat varieties with different breeding origin under organic and low input management. Euphytica, 199(1-2): 69-80; DOI: 10.1007/s10681-014-1171-8

**Keywords:** wheat, durum wheat, organic breeding, sustainability, variety performance

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<footnote n="1">¹ Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences
Martonvásár, PO Box 19. H-2462</footnote>
**PVI-66 - Rita ARMONIENE** - Identification of novel alleles of starch biosynthesis genes responsible for amylose variation in winter wheat TILLING population

# Identification of novel alleles of starch biosynthesis genes responsible for amylose variation in winter wheat TILLING population

Rita ARMONIENE<fnref n="1" /> and Gintaras BRAZAUSKAS<fnref n="1" />

Corresponding author: rita.armoniene@lammc.lt

Starch is a main component of wheat grain and consists of two glucan polymers amylose and amylopectin with the ratio ranging between 25 and 75%, respectively. The amylose/amylopectin ratio has a major influence over the physiochemical properties of starch and determines its optimal application in the industry. Recently, amylose-free (Waxy) and high-amylose wheats, consisting up to 100% apylopectin and 70% amylose, respectively, were produced through the development of new biotechnology techniques. The starches of these wheats provide the unique starch functional properties that are desirable for food and non-food industries.

Targeting Induced Local Lesions in Genomes (TILLING) population of the two winter wheat cultivars ('Kena DS' and 'Gaja DS') were developed using ethylmethane sulfonate (EMS) in order to induce mutations in genes of interest. Mutation density of one mutation per 37.84 Kb of the DNA was observed in this TILLING population while screening freezing tolerance associated genes. In this study, pilot experiment to evaluate amylose content variation in our material was performed. Twenty M3 generation genotypes of TILLING-population and its wild type cultivar 'Kena DS' and Waxy wheat breeding line as control were chosen to identify variation for amylose content using traditional Iodine-Potassium Iodide (I2-KI) solution. Low blue colour intensity indicating low amylose content was observed in three M3 genotypes. This indicates our TILLING population will serve as a valuable source of novel alleles of starch biosynthesis genes in order to develop winter wheat cultivars with various amylose content. Further work will be: (1) to identify amylose content variation in 756 genotypes of M3 TILLING population using traditional I2-KI dyeing method; (2) to estimate precise amylose content in selected genotypes by spectrometry; (3) to make sequence analysis of key starch biosynthesis genes of selected genotypes in order to identify novel alleles.

**Keywords:** Starch, TILLING, *Triticum aestivum* L. Waxy wheat

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<footnote n="1">¹ Institute of Agriculture, Lithuanian Research Centre for Agriculture and Forestry, Instituto a. 1, LT58344 Akademija, Kėdainiai distr., Lithuania</footnote>
PVI-67 - Sarah BEN SADOUN - Multi-trait selection in wheat: contribution of genomic selection tools

Multi-trait selection in wheat: contribution of genomic selection tools

Sarah BEN SADOUN<fnref n="1" />, Sophie BOUCHET<fnref n="1" />, Jérôme AUZANNEAU<fnref n="2" />, François-Xavier OURY<fnref n="1" />, Bernard ROLLAND<fnref n="3" />,
Emmanuel HEUMEZ<fnref n="4" /> and Gilles CHARMET<fnref n="1" />

Corresponding author: sarah.ben-sadoun@inra.fr

Genomic selection has shown its effectiveness in improving complex traits for livestock and for some crop species. It has been shown to improve genetic gain per unit of time and cost through prediction of performance before or without phenotypic characterization of new candidates. Most of genomic selection studies in plant science have focused on single-trait analyses.

The objective of my PhD program is to explore multi-trait genomic selection methods that were specially designed to exploit the information brought by correlated traits. It has been shown that prediction accuracy of traits with low heritability can be improved by a multi-trait model using correlated traits with higher heritability. In addition, prediction accuracy of difficult or expensive to measure traits can be improved by a multi-trait model using less expensive or easier to measure correlated traits with higher number of phenotypes.

A reference population of 760 lines from the INRA-AO breeding program that was evaluated for many traits between 2000 and 2016 will be analyzed as a proof of concept. It was genotyped for 180,000 SNPs developed within the Breedwheat project. This PhD program integrates with the objectives of metaprogram Selgen INRA (http://www selgen.inra.fr/).

**Keywords:** multi-trait model, genomic prediction, wheat

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<footnote n="1">¹ INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France</footnote>

<footnote n="2">2 Agri-Obtentions, Ferme de Gauvilliers 78660 Orsonville - France</footnote>

<footnote n="3">³ INRA-APBV, Domaine de la Motte, BP 35327, 35653 Le Rheu Cedex, France</footnote>

<footnote n="4">⁴ INRA UE Lille, 2 chaussée Brunehaut, Estrées-Mons, BP 50136, 80203 Péronne Cedex, France</footnote>
proWeizen – the German wheat research and breeding alliance

Tanja GERJETS¹

¹ German Federation for Plant Innovation (GFPi), proWeizen Alliance, Kaufmannstr. 71, 53115 Bonn, Germany

Corresponding author: tanja.gerjets@bdp-online.de

Wheat is one of the most important crops and Germany is an important wheat producer. In Germany, 16 breeding companies are running independent wheat breeding programmes.

The German Wheat Research and Breeding Alliance was founded in 2012 by the German wheat breeders to combine the scientific excellence in wheat research and breeding expertise in Germany. As a public-private partnership, the proWeizen alliance acts to foster wheat breeding and research on a national and international level as well as a platform for communication and coordination. The proWeizen platform (www.proweizen.de) is equally open to scientists and companies working in wheat breeding and research.

Currently, 14 research projects, funded by the German Federal Ministry of Food and Agriculture (BMEL) as well as the German Federal Ministry of Education and Research (BMBF), are run within the proWeizen alliance and focus on breeding for yield increase and stability, better adaptation to environmental stresses and utilization of heterosis. In these projects, German universities and research institutes are working in close collaboration with wheat breeders who are vital partners and plan to implement project results in their future breeding programmes.

In addition to support in project management and coordination, proWeizen liases with wheat researchers and breeders and participates in wheat research and breeding on national and international levels, respectively. proWeizen also helps with mobilizing funding opportunities and to gain public interest and awareness of wheat research and breeding.

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# BreedWheat: Breeding for sustainable wheat varieties, an integrated project from genomics to selection

The BreedWheat Consortium<fnref n="1" />, Emmanuelle LAGENDIJK<fnref n="2" /> and Jacques LE GOUIS<fnref n="3" />

Corresponding author: breedwheat@inra.fr

The challenge for wheat breeding is to deliver safe, high-quality, and health-promoting food and feed in a sustainable manner across environments affected by global change. In that context, the BreedWheat project gathers 28 public and private partners in research and breeding to ensure that the knowledge, resources, and methods are translated rapidly into products and varieties. BreedWheat will not only participate in new technological developments of markers and phenotyping methods, but will also integrate high throughput genotyping, phenotyping, physiological studies and modeling to decipher the genetic and ecophysiological bases of major traits for Northern Europe. This long-term project (9 years) includes the sequencing of chromosome 1B, detection of new structural genetic polymorphisms, large scale SNP production, genetic and physical mapping of those SNP, and the generation of more than 2 billion genotyping data points. A large experimental network was developed to phenotype association panels for nitrogen use efficiency, grain proteins composition, drought tolerance, Septoria leaf blotch and Fusarium head blight resistance. About 5,000 wheat lines from the INRA genetic resources centre were characterized and used to construct a new association panel. Moreover nine advanced backcross populations were created to introgress stress tolerance diversity in French elite material. Innovative statistical tools, including genomic selection, are developed within a real size selection program, in order to address both technical and socio-economic aspects of the implementation of novel breeding strategies in seed companies. Finally, a robust bioinformatics platform enabling efficient data storage and breeder friendly access is established.

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<footnote n="1">1 https://breedwheat.fr/</footnote>

<footnote n="2">2 INRA-Transfert, Paris, France</footnote>

<footnote n="3">3 INRA, UCA, UMR 1095 Génétique, Diversité et Ecophysiologie des Céréales (GDEC), 5 chemin de Beaulieu, 63 039 Clermont-Ferrand cedex2, France</footnote>
Keynote lecture list

<table><tr><td>Raphael DUMAIN - Global context for wheat, Innovation & Bayer Activities</td><td>xvii</td></tr><tr><td>Andreas GRANER - Ex situ collections: getting dressed up for another green revolution</td><td>2</td></tr><tr><td>Scott Allenn JACKSON - Sources of variation in crop legumes - genetic, epigenetic and structural</td><td>10</td></tr><tr><td>John FOULKES - Exploiting genetic diversity for enhancing yield potential and resource efficiency in wheat</td><td>18</td></tr><tr><td>Heribert HIRT - Next green revolution: Plant endophytes to enhance stress tolerance and yield of crops</td><td>25</td></tr><tr><td>Tatsuya IKEDA - Importance of international collaboration on wheat quality and safety</td><td>34</td></tr><tr><td>Peter ROGOWSKY - CRISPR-Cas9 technology in plant science</td><td>39</td></tr></table>

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EUCARPIA oral presentation list

<table>
  <tbody>
    <tr>
      <td>Davide GUERRA</td>
      <td>Studying the genetic basis of barley adaptation to contrasting environments in the WHEALBI collection</td>
      <td>3</td>
    </tr>
    <tr>
      <td>Ernesto IGARTUA</td>
      <td>Association with high-resolution climate data reveals selection footprints in the genomes of barley landraces across the Iberian Peninsula</td>
      <td>4</td>
    </tr>
    <tr>
      <td>Marco MACCAFERRI</td>
      <td>SNP-based characterization of a wide collection of tetraploid wheat reveals extensive domestication sweeps and haplotypes associated to traits of breeding interest</td>
      <td>5</td>
    </tr>
    <tr>
      <td>Helmy M YOUSSEF</td>
      <td>Natural Diversity of Inflorescence Architecture Traces Cryptic Domestication Genes in Barley (Hordeum vulgare L.)</td>
      <td>7</td>
    </tr>
    <tr>
      <td>Constance LAVERGNE</td>
      <td>Introduction of Aegilops sharonensis cytoplasm into common wheat and production of addition / wheat - Ae. sharonensis lines translocation lines</td>
      <td>8</td>
    </tr>
    <tr>
      <td>Luigi CATTIVELLI</td>
      <td>Durum wheat genome reveals the signature of 10,000 years of selection</td>
      <td>11</td>
    </tr>
    <tr>
      <td>Caroline JUERY</td>
      <td>Linking genomic and epigenomic features in hexaploid wheat genome</td>
      <td>13</td>
    </tr>
    <tr>
      <td>Cécile MONAT</td>
      <td>Hi-C and chromosome-scale assembly to detect large chromosomal rearrangements in wheat genomes</td>
      <td>14</td>
    </tr>
    <tr>
      <td>Maria BÜERSTMAYR</td>
      <td>High-resolution mapping of the peri-centromeric region on wheat chromosome arm 5AS harboring the Fusarium head blight resistance QTL Qfhs.ifa-5A</td>
      <td>15</td>
    </tr>
    <tr>
      <td>Romain DE OLIVEIRA</td>
      <td>Deciphering structural variations in the wheat genome using resequencing data</td>
      <td>16</td>
    </tr>
    <tr>
      <td>Bruno CONTRERAS-MOREIRA</td>
      <td>Agronomic advantage of presence of PpdH2 in winter barleys proven through a natural selection approach using GBS in pools of field-grown populations</td>
      <td>19</td>
    </tr>
    <tr>
      <td>Gaëtan TOUZY</td>
      <td>Improving water use efficiency in bread wheat by multi-trait multi-environment genome-wide association studies</td>
      <td>20</td>
    </tr>
    <tr>
      <td>Eric OBER</td>
      <td>Implementing large-scale field phenotyping in genomic selection to accelerate wheat breeding</td>
      <td>21</td>
    </tr>
    <tr>
      <td>Kerstin NEUMANN</td>
      <td>Non-invasive phenotyping reveals stress-adaptive and constitutive biomass QTL in cereals</td>
      <td>22</td>
    </tr>
    <tr>
      <td>Ulrike LOHWASSER</td>
      <td>Searching for frost tolerance in wheat (Triticum aestivum L.) - a genome wide association study</td>
      <td>23</td>
    </tr>
    <tr>
      <td>Ewen MULLINS</td>
      <td>Intensive field phenotyping to support breeding initiatives against septoria tritici blotch (STB) disease of wheat</td>
      <td>26</td>
    </tr>
    <tr>
      <td>Yvan MOËNNE-LOCCOZ</td>
      <td>Interactions of plant-beneficial rhizosphere bacteria according to cereal genotype</td>
      <td>27</td>
    </tr>
    <tr>
      <td>Laeticia WILLOCQUET</td>
      <td>Developing phenotyping methods for quantitative host plant resistance using simulation modelling and ROC curves</td>
      <td>28</td>
    </tr>
    <tr>
      <td>Hermann BÜERSTMAYR</td>
      <td>Genomics assisted improvement of Fusarium head blight resistance in bread wheat, durum wheat and triticale</td>
      <td>29</td>
    </tr>
    <tr>
      <td>Javier SANCHEZ-MARTIN</td>
      <td>GWAS reveals new race-specific resistance genes to powdery mildew in wheat</td>
      <td>30</td>
    </tr>
    <tr>
      <td>Ali NIAZ</td>
      <td>Novel rye-wheat translocations in Thinopyrum intermedium wheat introgression lines enhancing Wheat streak mosaic virus resistance</td>
      <td>31</td>
    </tr>
    <tr>
      <td>Pierre-Antoine PRÉCIGOUT</td>
      <td>Questioning the sustainability of quantitative physiological resistance: epidemiological and evolutionary responses of foliar fungal pathogens to changes in wheat plant traits</td>
      <td>32</td>
    </tr>
    <tr>
      <td>Sebastian MICHEL</td>
      <td>Multiple to conquer: Simultaneous selection for grain yield and baking quality in genomics-assisted wheat breeding</td>
      <td>35</td>
    </tr>
    <tr>
      <td>Suong Cu</td>
      <td>Genome-wide association studies of zinc, iron, copper, manganese and phosphorus in wheat (Triticum aestivum L.) grain and rachis at two developmental stages</td>
      <td>36</td>
    </tr>
    <tr>
      <td>Simon GRIFFITHS</td>
      <td>Biochemical and genetic analysis of arabinoxylan fibre in wheat grain</td>
      <td>37</td>
    </tr>
    <tr>
      <td>Fred VAN EEUWIJK</td>
      <td>Statistical methodology for modelling genotype by environment interactions in QTL mapping and genomic prediction, with applications to barley and wheat</td>
      <td>40</td>
    </tr>
  </tbody>
</table>
<table><tr><td>Andreas HUND</td><td>- Zooming into GxE by means of modern field phenotyping techniques</td><td>41</td></tr><tr><td>C Friedrich H LONGIN</td><td>- Optimum breeding schemes for line breeding in wheat</td><td>42</td></tr><tr><td>Daniela BUSTOS-KORTS</td><td>- A synthesis of crop growth and statistical genetic modelling to evaluate phenotyping strategies in wheat</td><td>43</td></tr><tr><td>Renaud RINCENT</td><td>- Genomic prediction of genotype by environment interactions by coupling genetic and physiological modelling</td><td>44</td></tr><tr><td>Philipp BOEVEN</td><td>- Rare and wanted! How to breed for favorable male floral traits required for hybrid wheat?</td><td>45</td></tr><tr><td>Funmi LADEJOBI</td><td>- Differentially penalized regression improves genomic prediction of wheat flowering time</td><td>46</td></tr></table>

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Posters list

<table>
  <tbody>
    <tr>
      <td>PI-01 - Hermann BÜERSTMAYR - Comparative genetic analysis of common bunt and dwarf bunt resistance in winter wheat</td>
      <td>49</td>
    </tr>
    <tr>
      <td>PI-02 - Nicholas FRADGLEY - The NIAB Diverse MAGIC wheat population: a community resource for QTL mapping</td>
      <td>50</td>
    </tr>
    <tr>
      <td>PI-03 - Mariateresa LAZZARO - From heritage to modern germplasm: evaluation of competitive ability, molecular characterization and genome-wide association in common wheat</td>
      <td>51</td>
    </tr>
    <tr>
      <td>PI-04 - Natalia TIKHENKO - Reconstruction of the triticale genome in the progeny of incompatible crosses</td>
      <td>52</td>
    </tr>
    <tr>
      <td>PI-05 - Mathilde LAINÉ - BreedWheat GWAS data in GnpIS information system</td>
      <td>53</td>
    </tr>
    <tr>
      <td>PI-06 - Laura PASCUAL - Genomic variability of Spanish Durum wheat landraces</td>
      <td>54</td>
    </tr>
    <tr>
      <td>PI-07 - Davide GUERRA - Allele mining for frost tolerance in barley unveiled untapped alleles and copy number variants of CBF and Vrn-H1 genes</td>
      <td>55</td>
    </tr>
    <tr>
      <td>PI-08 - Jonas ANDEREGG - Hyperspectral assessment of senescence dynamics and its relationship to grain yield and grain protein content in a high-yielding environment</td>
      <td>56</td>
    </tr>
    <tr>
      <td>PI-09 - Salar SHAAF - Integrated genetic approaches to modify canopy architecture for improved barley biomass and yield</td>
      <td>57</td>
    </tr>
    <tr>
      <td>PI-10 - Gianluca BRETANI - Genome scan to detect loci involved in culm morphological traits in a diverse barley collection</td>
      <td>58</td>
    </tr>
    <tr>
      <td>PI-11 - Edina TÜRKÖSI - 7BS.7HL wheat_b barley robertsonian translocation line with increased salt tolerance and β -glucan content</td>
      <td>59</td>
    </tr>
    <tr>
      <td>PI-12 - Martina ROSELLÓ - Dissecting the QTLome for protein content in durum wheat Mediterranean landraces</td>
      <td>61</td>
    </tr>
    <tr>
      <td>PI-13 - Roberto TUBEROSA - Development and characterization of a whole-genome radiation hybrid panel from reference tetraploid wheat cultivar Svevo</td>
      <td>62</td>
    </tr>
    <tr>
      <td>PI-14 - Radim SVAČINA - Ph2 gene mapping through development and phenotyping of deletion lines in bread wheat</td>
      <td>63</td>
    </tr>
    <tr>
      <td>PI-15 - Rubén Gómez RUFO - Genetic structure of Mediterranean wheat landraces assessed with SNP markers</td>
      <td>64</td>
    </tr>
    <tr>
      <td>PI-16 - Andrii FATIUKHA - Genomic architecture of morphological and phenological traits associated with domestication syndrome in durum × wild emmer wheat RIL population</td>
      <td>65</td>
    </tr>
    <tr>
      <td>PI-17 - Anna HAWLICZEK-STRULAK - Investigation of genetic diversity patterns and search for selective sweeps in a worldwide collection of rye</td>
      <td>66</td>
    </tr>
    <tr>
      <td>PI-18 - Arantxa MONTEAGUDO - Detecting useful genetic diversity from Spanish landraces to improve an elite cultivar</td>
      <td>67</td>
    </tr>
    <tr>
      <td>PI-19 - Hanna Bolibok-Bragoszewska - Highly parallel amplicon sequencing for characterization of genetic diversity in rye germplasm</td>
      <td>68</td>
    </tr>
    <tr>
      <td>PI-20 - Ian DAWSON - Exploring local adaptation in barley landraces through latitudinal differences in days to heading</td>
      <td>69</td>
    </tr>
    <tr>
      <td>PI-21 - Jane ROCHE - Coexpression network and phenotypic analysis identify metabolic pathways associated with the effect of warming on grain yield components in wheat</td>
      <td>70</td>
    </tr>
    <tr>
      <td>PI-22 - José Miguel SORIANO - Eastern and Western durum wheat Mediterranean landraces differ in marker alleles responsible for yield components and phenology as revealed by association analysis</td>
      <td>71</td>
    </tr>
    <tr>
      <td>PI-23 - Monika RAKOCZY-TROJANOWSKA - New aspects of the genetic background of benzoxazinoid biosynthesis in rye (Secale cereale L.)</td>
      <td>72</td>
    </tr>
    <tr>
      <td>PI-24 - Rasha TARAWNEH - Mapping QTL for drought tolerance in spring wheat genebank accessions using genome wide association approach</td>
      <td>74</td>
    </tr>
    <tr>
      <td>PII-25 - Lukas KRONENBERG - Wheat stem elongation dynamics assessed by GWAS: short term growth rate depends on temperature</td>
      <td>76</td>
    </tr>
    <tr>
      <td>PII-26 - Carlos P CANTALAPIEDRA - Unmasking new intra-species diversity through k-mer count analysis</td>
      <td>77</td>
    </tr>
    <tr>
      <td>PII-27 - Charles CHEN - Genetic improvement facing climate challenging: integrating classical and novel approaches</td>
      <td>78</td>
    </tr>
    <tr>
      <td>PIII-28 - Elizabeth CHAPMAN - Combining staygreen and altered phenology to improve wheat productivity</td>
      <td>80</td>
    </tr>
    <tr>
      <td>PIII-29 - Nadège BAILLOT - Different growth dynamics within the wheat spike</td>
      <td>81</td>
    </tr>
  </tbody>
</table>
<table>
  <tbody>
    <tr>
      <td>PIII-30 - Dragana TRKULJA - Seeding density effects on grain yield in bread wheat</td>
      <td>82</td>
    </tr>
    <tr>
      <td>PIII-31 - Milan MIROSAVLJEVIĆ - Stay green traits contributed to recent grain yield improvement in six-rowed winter barley in Serbia</td>
      <td>83</td>
    </tr>
    <tr>
      <td>PIII-32 - Bojan JOCKOVIĆ - Breeding progress in yield and grain properties of winter wheat</td>
      <td>84</td>
    </tr>
    <tr>
      <td>PIV-33 - Valentina SPANIC - Quality preservation in FHB attack due to antioxidant system</td>
      <td>86</td>
    </tr>
    <tr>
      <td>PIV-34 - Hanane AADELA - Agrobacterium-mediated Transformation of Mature Embryo and immature Tissues of Bread Wheat (Triticum aestivum L.) Genotypes</td>
      <td>87</td>
    </tr>
    <tr>
      <td>PIV-35 - Ildikó KARSAI - How may warm spells during winter influence the plant development of cereals?</td>
      <td>88</td>
    </tr>
    <tr>
      <td>PIV-36 - Judit BÁNYAI - Investigations on the effects of drought stress on the morphology, physiology and yield components of durum wheat in Hungary</td>
      <td>89</td>
    </tr>
    <tr>
      <td>PIV-37 - Krisztina BALLA - Investigation of heat stress tolerance on yield properties in cereals</td>
      <td>90</td>
    </tr>
    <tr>
      <td>PIV-38 - Marianna MAYER - Comparative analysis of stem rust resistance in winter wheat genotypes at Hungarian and East African locations</td>
      <td>91</td>
    </tr>
    <tr>
      <td>PIV-39 - Valérie CADOT - Tilletia caries, causal agent of Common Bunt: study of the most prevalent virulences in order to develop a resistance test for the registration to the National List for common wheat in Organic Farming</td>
      <td>92</td>
    </tr>
    <tr>
      <td>PIV-40 - Harpinder RANDHAWA - Flowering pathway genes regulating Fusarium head blight resistance in wheat</td>
      <td>93</td>
    </tr>
    <tr>
      <td>PIV-41 - Aurore BERAL - Is wheat individual grain characteristics variance a source of genetic variability for water stress tolerance?</td>
      <td>94</td>
    </tr>
    <tr>
      <td>PIV-42 - Barbara STEINER - Improving Fusarium head blight resistance in durum wheat</td>
      <td>95</td>
    </tr>
    <tr>
      <td>PIV-43 - Pawel CZEMBOR - Genome wide association study of seedling leaf rust resistance in European winter wheat cultivars</td>
      <td>96</td>
    </tr>
    <tr>
      <td>PIV-44 - Piotr SŁOWACKI - Resistance of European winter wheat cultivars to Zymoseptoria tritici isolate IPO92006</td>
      <td>97</td>
    </tr>
    <tr>
      <td>PIV-45 - Éva SZAKÁCS - Development and characterization of a new recombinant 1RS.1BL translocation line carrying leaf and stripe rust resistance from perennial rye (Secale cereanum)</td>
      <td>98</td>
    </tr>
    <tr>
      <td>PIV-46 - Lidia KOWALSKA - Improvement of Parastagonospora nodorum blotch resistance in winter triticale by using the in vitro somaclonal and androgenic approaches</td>
      <td>99</td>
    </tr>
    <tr>
      <td>PIV-47 - Sławomir BARTOSIAK - Occurrence of Parastagonospora nodorum blotch, Parastagonospora avenae blotch and Zymosetoria tritici blotch on wheat and triticale in Poland</td>
      <td>100</td>
    </tr>
    <tr>
      <td>PIV-48 - Faiza BOUSSORA - Delaying in the spike development and reduction in spikelet primordia numbers decrease the yield in salt-stressed Tunisian barley</td>
      <td>101</td>
    </tr>
    <tr>
      <td>PIV-49 - Klára MÉSZÁROS - Examination of the resistance of barley genotypes to Pyrenophora teres f. teres in the seedling and adult stage</td>
      <td>102</td>
    </tr>
    <tr>
      <td>PIV-50 - Jean-Charles DESWARTE - Using grain carbon isotope composition (d13C) at harvest to assess growing conditions and cultivar drought tolerance</td>
      <td>104</td>
    </tr>
    <tr>
      <td>PIV-51 - Roberto TUBEROSA - Dissecting the root angle QTLome to enhance durum wheat sustainability</td>
      <td>106</td>
    </tr>
    <tr>
      <td>PIV-52 - Roberto TUBEROSA - Fine-mapping of QSBM.UBO-2BS=SBM2, a major QTL for resistance to soil-borne cereal mosaic virus (SBCMV)</td>
      <td>107</td>
    </tr>
    <tr>
      <td>PIV-53 - Tomasz GÓRAL - Fusarium head blight and Fusarium toxins accumulation in grain of winter wheat breeding lines inoculated with Fusarium culmorum</td>
      <td>108</td>
    </tr>
    <tr>
      <td>PIV-54 - Marine OLLIER - A fast and reliable method for assessing Fusarium damaged kernels and the mycotoxin content by digital picture analysis in wheat and triticale</td>
      <td>109</td>
    </tr>
    <tr>
      <td>PIV-55 - Jose MORENO-AMORES - Assessing the incorporation of covariates in genomic predictions for Fusarium head blight in durum wheat</td>
      <td>110</td>
    </tr>
    <tr>
      <td>PIV-56 - Mahbubjon RAHMATOV - Development and characterization of new stem rust and stripe rust resistance genes in Swedish wheat-rye introgression lines</td>
      <td>111</td>
    </tr>
    <tr>
      <td>PIV-57 - Andrii FATIUKHA - Detection of functional and non-functional alleles of the stripe rust resistance gene Yr15 using diagnostic markers</td>
      <td>112</td>
    </tr>
    <tr>
      <td>PIV-58 - Christophe GIGOT - Using crop-pathogen modeling to identify plant traits to control Zymoseptoria tritici epidemics on wheat</td>
      <td>113</td>
    </tr>
    <tr>
      <td>PIV-59 - Claire GUÉRIN - Genome-wide identification, characterization and expression profiling of NAC genes in Triticum aestivum</td>
      <td>114</td>
    </tr>
    <tr>
      <td>PIV-60 - Ghasemali NAZEMI - The association mapping of short-term heat shock and chronic heat stress tolerance in durum wheat</td>
      <td>115</td>
    </tr>
  </tbody>
</table>
<table><tr><td>PIV-60bis - Johanna LETHIN - Molecular breeding of salt tolerant wheat</td><td>116</td></tr><tr><td>PV-61 - Catherine RAVEL - Rapid identification of high-molecular-weight glutenin subunits by SNP markers to breed for high quality</td><td>118</td></tr><tr><td>PV-62 - Emmanuelle BANCEL - Proteomic changes occuring in einkorn (Triticum monococcum) grain during filling stages of development</td><td>119</td></tr><tr><td>PV-63 - Julie BOUDET - SPA Heterodimerizing Protein, a bZIP transcription factor, represses glutenin synthesis in Triticum aestivum</td><td>120</td></tr><tr><td>PV-64 - Pierre BERTHOMIEU - Towards the positional cloning of genes controlling zinc and iron content in durum wheat grain</td><td>121</td></tr><tr><td>PVI-65 - Péter MIKÓ - Selection of winter cereals for organic agriculture</td><td>123</td></tr><tr><td>PVI-66 - Rita ARMONIENE - Identification of novel alleles of starch biosynthesis genes responsible for amylose variation in winter wheat TILLING population</td><td>124</td></tr><tr><td>PVI-67 - Sarah BEN SADOUN - Multi-trait selection in wheat: contribution of genomic selection tools</td><td>125</td></tr><tr><td>PVI-68 - Tanja GERJETS - proWeizen - the German wheat research and breeding alliance</td><td>126</td></tr><tr><td>Emmanuelle LAGENDIJK - BreedWheat: Breeding for sustainable wheat varieties, an integrated project from genomics to selection</td><td>127</td></tr></table>

Table of content
Author's index

A

<table><tr><td>Aadela Hanane</td><td>87</td></tr><tr><td>Aasen Helge</td><td>56</td></tr><tr><td>Abdelwahd Rabha</td><td>87</td></tr><tr><td>Abicic Ivan</td><td>86</td></tr><tr><td>Aćin Vladimir</td><td>83, 84</td></tr><tr><td>Adam-Blondon Anne-Françoise</td><td>53</td></tr><tr><td>Ahansal Khadija</td><td>87</td></tr><tr><td>Alaux Michael</td><td>53</td></tr><tr><td>Allard Vincent</td><td>81, 94, 114</td></tr><tr><td>Alvarez David</td><td>120</td></tr><tr><td>Álvaro Fanny</td><td>61, 64</td></tr><tr><td>Amado Carolina Rivera</td><td>18</td></tr><tr><td>Ametz Christian</td><td>35</td></tr><tr><td>Amores Jose Esteban Moreno</td><td>29</td></tr><tr><td>Anderegg Jonas</td><td>41, 56</td></tr><tr><td>Arana-Ceballos Fernando</td><td>23</td></tr><tr><td>Aranzana Maria Jose</td><td>71</td></tr><tr><td>Armoniene Rita</td><td>124</td></tr><tr><td>Aronsson Henrik</td><td>116</td></tr><tr><td>Arseniuk Edward</td><td>99, 100</td></tr><tr><td>Ashling Stephen</td><td>8</td></tr><tr><td>Auzanneau Jérôme</td><td>125</td></tr><tr><td>Avni Raz</td><td>11</td></tr><tr><td>Ayoub Mohammad A</td><td>7</td></tr></table>

B

<table><tr><td>Badeck F</td><td>55</td></tr><tr><td>Bagnaresi Paolo</td><td>11</td></tr><tr><td>Baillot Nadège</td><td>81</td></tr><tr><td>Bakonyi József</td><td>102</td></tr><tr><td>Baldwin T</td><td>92</td></tr><tr><td>Balfourier François</td><td>16, 53</td></tr><tr><td>Balla Krisztina</td><td>88, 89, 90</td></tr><tr><td>Balzergue Sandrine</td><td>70</td></tr><tr><td>Bancel Emmanuelle</td><td>119</td></tr><tr><td>Bányai Judit</td><td>88, 102</td></tr><tr><td>Barabaschi Delfina</td><td>15</td></tr><tr><td>Bartoš Jan</td><td>63</td></tr><tr><td>Bartosiak Sławomir F</td><td>100</td></tr><tr><td>Baruch Kobi</td><td>14</td></tr><tr><td>Beauchêne Katia</td><td>20, 104</td></tr><tr><td>Bednarczyk E</td><td>100</td></tr><tr><td>Bedo Zoltan</td><td>37</td></tr><tr><td>Beguería Santiago</td><td>4</td></tr></table>

<table><tr><td>Benavente Elena</td><td>54</td></tr><tr><td>Bencze Szilvia</td><td>90</td></tr><tr><td>Bennett Malcolm</td><td>18</td></tr><tr><td>Bentley Alison</td><td>21, 46, 50</td></tr><tr><td>Beral Aurore</td><td>94</td></tr><tr><td>Berthomieu Pierre</td><td>121</td></tr><tr><td>Beye Abdoulaye</td><td>121</td></tr><tr><td>Biyiklioglu Sezgi</td><td>11</td></tr><tr><td>Boer Martin</td><td>40, 43</td></tr><tr><td>Boeven Philipp</td><td>45</td></tr><tr><td>Bogard Matthieu</td><td>46</td></tr><tr><td>Bolibok-Bragoszewska Hanna</td><td>66, 68</td></tr><tr><td>Bonnot Titouan</td><td>119</td></tr><tr><td>Bontems Valérie</td><td>104</td></tr><tr><td>Booth A</td><td>3</td></tr><tr><td>Börner Andreas</td><td>23, 52, 74, 101</td></tr><tr><td>Borzecka Ewa</td><td>66, 68</td></tr><tr><td>Bouchet Sophie</td><td>125</td></tr><tr><td>Boudet Julie</td><td>120</td></tr><tr><td>Bouffaud Marie-Lara</td><td>27</td></tr><tr><td>Boussora Faiza</td><td>101</td></tr><tr><td>Bouzidi Mohamed Fouad</td><td>70, 114</td></tr><tr><td>Brachet C</td><td>92</td></tr><tr><td>Branlard Gérard</td><td>118</td></tr><tr><td>Brazauskas Gintaras</td><td>124</td></tr><tr><td>Bretani G</td><td>55</td></tr><tr><td>Brisset Anne-Laure</td><td>109</td></tr><tr><td>Brown James KM</td><td>26</td></tr><tr><td>Brugger Klaus</td><td>15</td></tr><tr><td>Bruschi Martina</td><td>62, 107</td></tr><tr><td>Bruyere J</td><td>92</td></tr><tr><td>Budak Hikmet</td><td>11</td></tr><tr><td>Büerstmayr Hermann</td><td>15, 29, 35, 49, 95, 109, 110</td></tr><tr><td>Büerstmayr Maria</td><td>15, 29, 95</td></tr><tr><td>Bull H</td><td>58</td></tr><tr><td>Buntjer Jaap</td><td>21</td></tr><tr><td>Bustos-Korts Daniela</td><td>3, 40, 43, 69</td></tr><tr><td>Byrne Stephen</td><td>26</td></tr></table>

C

<table><tr><td>Cadot Valérie</td><td>92</td></tr><tr><td>Çakır E</td><td>3</td></tr><tr><td>Cammarano D</td><td>69</td></tr><tr><td>Cantalapiedra Carlos P</td><td>4, 19, 77</td></tr><tr><td>Caroline Juery</td><td>13</td></tr><tr><td>Casas Ana M</td><td>4, 19, 58, 67, 77</td></tr><tr><td>Cattivelli Luigi</td><td>3, 5, 11, 15, 51, 55, 58, 69</td></tr></table>