Littérature scientifique sur le sujet « Genetic characterization wheat mycorrhizae QTL »

Fusarium head blight (FHB) is one of the most important fungal wheat diseases worldwide. Understanding the genetics of FHB resistance is the key to facilitating the introgression of different FHB resistance genes into adapted wheat. The objectives of the present study were to detect and map quantitative trait loci (QTL) associated with FHB resistance genes and characterize the genetic components of the QTL in a doubled-haploid (DH) spring wheat population using both single-locus and two-locus analysis. A mapping population, consisting of 174 DH lines from the cross between DH181 (resistant) and AC Foremost (susceptible), was evaluated for type I resistance to initial infection during a 2-year period in spray-inoculated field trials, for Type II resistance to fungal spread within the spike in 3 greenhouse experiments using single-floret inoculation, and for resistance to kernel infection in a 2001 field trial. One-locus QTL analysis revealed 7 QTL for type I resistance on chromosome arms 2DS, 3AS, 3BS, 3BC (centromeric), 4DL, 5AS, and 6BS, 4 QTL for type II resistance on chromosomes 2DS, 3BS, 6BS, and 7BL, and 6 QTL for resistance to kernel infection on chromosomes 1DL, 2DS, 3BS, 3BC, 4DL, and 6BS. Two-locus QTL analysis detected 8 QTL with main effects and 4 additive by additive epistatic interactions for FHB resistance and identified novel FHB resistance genes for the first time on chromosomes 1DL, 4AL, and 4DL. Neither significant QTL by environment interactions nor epistatic QTL by environment interactions were found for either type I or type II resistance. The additive effects of QTL explained most of the phenotypic variance for FHB resistance. Marker-assisted selection for the favored alleles at multiple genomic regions appears to be a promising tool to accelerate the introgression and pyramiding of different FHB resistance genes into adapted wheat genetic backgrounds.Key words: Triticum aestivum, Fusarium graminearum, microsatellite, additive effect, additive by additive epistatic effect.

Durum wheat is one of the most important cultivated cereal crops, providing nutrients to humans and domestic animals. Durum breeding programs prioritize the improvement of its main agronomic traits; however, the majority of these traits involve complex characteristics with a quantitative inheritance (quantitative trait loci, QTL). This can be solved with the use of genetic maps, new molecular markers, phenotyping data of segregating populations, and increased accessibility to sequences from next-generation sequencing (NGS) technologies. This allows for high-density genetic maps to be developed for localizing candidate loci within a few Kb in a complex genome, such as durum wheat. Here, we review the identified QTL, fine mapping, and cloning of QTL or candidate genes involved in the main traits regarding the quality and biotic and abiotic stresses of durum wheat. The current knowledge on the used molecular markers, sequence data, and how they changed the development of genetic maps and the characterization of QTL is summarized. A deeper understanding of the trait architecture useful in accelerating durum wheat breeding programs is envisioned.

Leaf and stripe rust are major threats to wheat production worldwide. The effective, multiple rust resistances present in the Brazilian cultivar Toropi makes it an excellent choice for a genetic study of rust resistance. Testing of DNA from different seed lots of Toropi with 2,194 polymorphic 90K iSelect single nucleotide polymorphism markers identified significant genetic divergence, with as much as 35% dissimilarity between seed lots. As a result, further work was conducted with a single plant line derived from Toropi variant Toropi-6.4. A double haploid population with 168 lines derived from the cross Toropi-6.4 × Thatcher was phenotyped over multiple years and locations in Canada, New Zealand, and Kenya, with a total of seven field trials undertaken for leaf rust and nine for stripe rust. Genotyping with the 90K iSelect array, simple sequence repeat and Kompetitive allele-specific polymerase chain reaction markers resulted in a genetic map of 3,043 cM, containing 1,208 nonredundant markers. Significant quantitative trait loci (QTL) derived from Toropi-6.4 were identified in multiple environments on chromosomes 1B (QLr.crc-1BL/QYr.crc-1BL), 3B (QLr.crc-3BS), 4B (QYr.crc-4BL), 5A (QLr.crc-5AL and QYr.crc-5AL), and 5D (QLr.crc-5DS). The QTL QLr.crc-1BL/QYr.crc-1BL colocated with the multi-rust resistance locus Lr46/Yr29, while the QTL QLr.crc-5DS located to the Lr78 locus previously found in a wheat backcross population derived from Toropi. Comparisons of QTL combinations showed QLr.crc-1BL to contribute a significantly enhanced leaf rust resistance when combined with QLr.crc-5AL or QLr.crc-5DS, more so than when QLr.crc-5AL and QLr.crc-5DS were combined. A strong additive effect was also seen when the stripe rust resistance QTL QYr.crc-1BL and QYr.crc-5AL were combined.

Leaf rust and powdery mildew are two important foliar diseases in wheat. A recombinant inbred line (RIL) population, obtained by crossing two bread wheat cultivars (‘Victo’ and ‘Spada’), was evaluated for resistance to the two pathogens at seedling stage. Upon developing a genetic map of 8726 SNP loci, linkage analysis identified three resistance Quantitative Trait Loci (QTLs), with ‘Victo’ contributing the resistant alleles to all loci. One major QTL (QPm.gb-7A) was detected in response to Blumeria graminis on chromosome 7A, which explained 90% of phenotypic variation (PV). The co-positional relationship with known powdery mildew (Pm) resistance loci suggested that a new source of resistance was identified in T. aestivum. Two QTLs were detected in response to Puccinia triticina: a major gene on chromosome 5D (QLr.gb-5D), explaining a total PV of about 59%, and a minor QTL on chromosome 2B (QLr.gb-2B). A positional relationship was observed between the QLr.gb-5D with the known Lr1 gene, but polymorphisms were found between the cloned Lr1 and the corresponding ‘Victo’ allele, suggesting that QLr.gb-5D could represent a new functional Lr1 allele. Lastly, upon anchoring the QTL on the T. aestivum reference genome, candidate genes were hypothesized on the basis of gene annotation and in silico gene expression analysis.

In previous studies, we reported quantitative trait loci (QTL) associated with the heading, flowering, and maturity time in four hard red spring wheat recombinant inbred line (RIL) populations but the results are scattered in population-specific genetic maps, which is challenging to exploit efficiently in breeding. Here, we mapped and characterized QTL associated with these three earliness traits using the International Wheat Genome Sequencing Consortium (IWGSC) RefSeq v2.0 physical map. Our data consisted of (i) 6526 single nucleotide polymorphisms (SNPs) and two traits evaluated at five conventionally managed environments in the ‘Cutler’ × ‘AC Barrie’ population; (ii) 3158 SNPs and two traits evaluated across three organic and seven conventional managements in the ‘Attila’ × ‘CDC Go’ population; (iii) 5731 SilicoDArT and SNP markers and the three traits evaluated at four conventional and organic management systems in the ‘Peace’ × ‘Carberry’ population; and (iv) 1058 SNPs and two traits evaluated across two conventionally and organically managed environments in the ‘Peace’ × ‘CDC Stanley’ population. Using composite interval mapping, the phenotypic data across all environments, and the IWGSC RefSeq v2.0 physical maps, we identified a total of 44 QTL associated with days to heading (11), flowering (10), and maturity (23). Fifteen of the 44 QTL were common to both conventional and organic management systems, and the remaining QTL were specific to either the conventional (21) or organic (8) management systems. Some QTL harbor known genes, including the Vrn-A1, Vrn-B1, Rht-A1, and Rht-B1 that regulate photoperiodism, flowering time, and plant height in wheat, which lays a solid basis for cloning and further characterization.

The major aluminum (Al) tolerance gene in wheat ALMT1 confers. An Al-activated efflux of malate from root apices. We determined the genomic structure of the ALMT1 gene and found it consists of 6 exons interrupted by 5 introns. Sequencing a range of wheat genotypes identified 3 alleles for ALMT1, 1 of which was identical to the ALMT1 gene from an Aegilops tauschii accession. The ALMT1 gene was mapped to chromosome 4DL using 'Chinese Spring' deletion lines, and loss of ALMT1 coincided with the loss of both Al tolerance and Al-activated malate efflux. Aluminium tolerance in each of 5 different doubled-haploid populations was found to be conditioned by a single major gene. When ALMT1 was polymorphic between the parental lines, QTL and linkage analyses indicated that ALMT1 mapped to chromosome 4DL and cosegregated with Al tolerance. In 2 populations examined, Al tolerance also segregated with a greater capacity for Al-activated malate efflux. Aluminium tolerance was not associated with a particular coding allele for ALMT1, but was significantly correlated with the relative level of ALMT1 expression. These findings suggest that the Al tolerance in a diverse range of wheat genotypes is primarily conditioned by ALMT1.Key words: aluminum, tolerance, genetic marker, Triticum aestivum, QTL, deletion mapping.

Endosperm texture has a tremendous impact on the end-use quality of wheat (Triticum aestivum L.). Cultivars of barley (Hordeum vulgare L.), a close relative of wheat, also vary measurably in grain hardness. However, in contrast to wheat, little is known about the genetic control of barley grain hardness. Puroindolines are endosperm-specific proteins found in wheat and its relatives. In wheat, puroindoline sequence variation controls the majority of wheat grain texture variation. Hordoindolines, the puroindoline homologs of barley, have been identified and mapped. Recently, substantial allelic variation was found for hordoindolines among commercial barley cultivars. Our objective was to determine the influence of hordoindoline allelic variation upon grain hardness and dry matter digestibility in the 'Steptoe' × 'Morex' mapping population. This population is segregating for hordoindoline allele type, which was measured by a HinA/HinB/Gsp composite marker. One-hundred and fifty lines of the 'Steptoe' × 'Morex' population were grown in a replicated field trial. Grain hardness was estimated by near-infrared reflectance (NIR) and measured using the single kernel characterization system (SKCS). Variation attributable to the HinA/HinB/Gsp locus averaged 5.7 SKCS hardness units (SKCS U). QTL analysis revealed the presence of several areas of the genome associated with grain hardness. The largest QTL mapped to the HinA/HinB/Gsp region on the short arm of chomosome 7 (5H). This QTL explains 22% of the SKCS hardness difference observed in this study. The results indicate that the Hardness locus is present in barley and implicates the hordoindolines in endosperm texture control.Key words: puroindolines, grain hardness, digestibility.

Yellow rust (YR) epidemics have affected wheat productivity worldwide. YR resistance (Yr) is eminent in wheat; however, it is continuously invaded by evolving YR pathogen Puccinia striiformis (Pst.). Understanding the Yr genes’ diversity among the available germplasm is paramount to developing YR-resistant cultivars. In this study, 14 wheat genotypes were screened for their relative resistance index (RRI) and Yr genes/QTL via linked microsatellite markers. RRI screening categorized the studied genotypes into susceptible (<5; 4.44 ± 0.75), moderate (5–7; 6.11 ± 0.64), and resistant (>7; 8.45 ± 0.25) bulks (p < 0.001). Genetic analysis using 19 polymorphic microsatellite markers revealed 256 alleles, which were divergent among the three resistance bulks. Markers Xbarc7 and Xgwm429 showed the highest allelic diversity in comparison to Xbarc181, Xwmc419, SCAR1400, and Xgwm130. Resistant bulk showed associated alleles at Yr18 gene-linked markers Xgwm295, cssfr6, and csLV34. Other RRI-associated alleles at markers Xbarc7 and Xbarc101 showed weak and moderate linkages, respectively, with the Yr5 gene; whereas, a moderate association was noted for the Yr15 gene-linked marker Xgwm11. Marker Xwe173 linked with the Yr26 gene showed associated alleles among the susceptible bulk. Cross combinations of the parental lines forming recombinant inbred lines (RILs) demonstrated net higher RRI implying favorable allelic recombination. These results support reports and field observations on novel Pst. races that triggered Yr26, Yr5, and Yr15 busts in recent past. This study further implies that pyramiding all stage resistance genes (Yr5, Yr10, Yr15, and Yr26) with adult plant resistance genes (Yr18 and Yr62) should provide sustained YR resistance. The associated alleles at Yr genes-linked markers provide a basis for marker-assisted YR resistance breeding in wheat.

The photosynthesis of wheat glumes makes important contributions to the yield. Stomata play a crucial role in regulating photosynthesis and transpiration in plants. However, the genetic base of wheat glume stomata is not fully understood. In this study, stomatal length (SL), stomatal width (SW), stomatal density (SD), potential conductance index (PCI) of stomata, stomatal area (SA), and stomatal relative area (SRA) were measured in different parts of wheat glumes from a doubled haploid (DH) population and their parents. Quantitative trait loci (QTLs) of these traits were anchored on a high-density genetic linkage map of the DH population. A total of 61 QTLs for stoma-related traits were mapped onto 16 chromosomes, and each one accounted for 3.63 to 19.02% of the phenotypic variations. Two QTL hotspots were detected in two marker intervals, AX-109400932∼AX-110985652 and AX-108972184∼AX-108752564, on chromosome 6A. Five possibly candidate genes (TraesCS6A02G105400, TraesCS6A02G106400, TraesCS6A02G115100, TraesCS6A02G115400, and TraesCS6A02G116200) for stoma-related traits of wheat glumes were screened out , according to their predicted expression levels in wheat glumes or spikes. The expression of these genes may be induced by a variety of abiotic stresses. These findings provide insights for cloning and functional characterization of stoma-related candidate genes in wheat glumes.

Since 1984, the ‘Chilero’ spring wheat line developed by CIMMYT has proven to be highly resistant to leaf rust and stripe rust. Amid efforts to understand the basis of resistance of this line, a recombinant inbred line (RIL) population derived from a cross between Avocet and Chilero was studied. The parents and RILs were characterized in field trials for leaf rust and stripe rust in three locations in Mexico between 2012 and 2015 and genotyped with DArT-array, DArT-GBS, and SSR markers. A total of 6,168 polymorphic markers were used to construct genetic linkage maps. Inclusive composite interval mapping detected four colocated resistance loci to both rust diseases and two stripe rust resistant loci in the Avocet × Chilero population. Among these, the quantitative trait locus (QTL) on chromosome 1BL was identified as a pleotropic adult plant resistance gene Lr46/Yr29, whereas QLr.cim-5DS/QYr.cim-5DS was a newly discovered colocated resistance locus to both rust diseases in Chilero. Additionally, one new stripe rust resistance locus on chromosome 7BL was mapped in the current population. Avocet also contributed two minor colocated resistance QTLs situated on chromosomes 1DL and 4BS. The flanking SNP markers can be converted to breeder friendly Kompetitive Allele Specific PCR (KASP) markers for wheat breeding programs.

La fusariose de l’épi est une maladie fongique qui touche toutes les cultures de céréales à paille à travers le monde entrainant des baisses de rendements et de la qualité des grains. La fusariose pose également un problème pour la sécurité alimentaire lié à la contamination des grains infectés par des mycotoxines. Le développement de variétés résistantes est considéré comme la méthode la plus efficace et la plus durable pour réduire les dommages causés par la maladie et pour limiter la contamination par les mycotoxines. L’amélioration de la résistance à la fusariose chez le blé dur (Triticum durum Desf.) demeure un défi du fait de son extrême sensibilité à la maladie et de la faible variabilité génétique disponible pour ce caractère. L’objectif principal de cette thèse a été d’évaluer l’effet de Fhb1, le QTL majeur de résistance à la fusariose chez le blé tendre (Triticum aestivumL.), au sein de fonds génétiques de blé dur élite. Pour cela, trois populations de cartographie, comprenant chacune environ 100 F7-RIL (lignées pures recombinantes ou « recombinant inbred lines »), ont été développées à partir de croisements entre la lignée expérimentale de blé dur DBC-480, portant une introgression de Fhb1, et les cultivars de blé dur Karur, Durobonus et SZD1029K. Les lignées ont été évaluées au champ, sur trois saisons, pour leur résistance globale à la fusariose après inoculation en spray de Fusarium culmorum. Des notations morphologiques (date de floraison, hauteur des plantes) ont également été réalisées afin d'évaluer leur influence sur l'infestation. Les lignées ont été génotypées à l’aide de marqueurs SSR et de marqueurs GBS (génotypage par séquençage ou « genotyping-by-sequencing ») développés par DArTseq. L’analyse de liaison a permis d’identifier des QTL de résistance sur les bras des chromosomes 2BL, 3BS, 4AL, 4BS, 5AL et 6AS. DBC-480 contribuait à l’allèle de résistance à tous ces loci. Le QTL sur 3BS a été détecté au sein des trois populations centré sur l’intervalle de Fhb1, confirmant, pour la première fois, son introgression dans le blé dur. L’évaluation de la résistance à la propagation après inoculation ponctuelle, réalisé au sein d’une des trois populations, a également permis de valider l’effet de Fhb1 sur la résistance de type 2 chez le blé dur. La hauteur des plantes influe fortement sur la résistance globale à la fusariose et, en particulier, l’allèle de nanisme Rht-B1b est associé à une plus grande sensibilité à la maladie dans les trois populations. Cependant, l’effet négatif de Rht-B1b sur la résistance est largement compensé dans les lignées possédant Fhb1. Des lignées semi-naines avec un meilleur niveau de résistance ont été sélectionnées et favoriseront le développement de cultivars de blé dur résistants à la fusariose
Fusarium head blight (FHB) is a devastating disease affecting small-grain cereals worldwide causing yield and quality losses. FHB affects food safety due to the contamination of infected grains by mycotoxins. Host plant resistance is considered the most efficient and sustainable approach to contain FHB and mycotoxin contaminations. In durum wheat (Triticum durum Desf.) breeding for FHB resistance remains a challenge due to its extreme susceptibility and to lack of genetic variation available in the primary durum wheat gene pool. The primary goal of this thesis was to evaluate the effect of Fhb1, the major common wheat (Triticum aestivum L.) FHB resistance QTL, in elite durum wheat background. Three F7-RIL (recombinant inbred lines) mapping populations of about 100 lines were developed from crosses between the durum wheat experimental line DBC-480, harboring Fhb1, and the durum wheat cultivars Karur, Durobonus and SZD1029K. The RILs were tested under field conditions by artificial spray inoculation with Fusarium culmorum in three seasons. Morphological traits (flowering date, height) were also recorded to assess their influence on FHB infestation. Genotyping of the lines was performed with SSR and genotyping-by-sequencing (GBS) DArTseq markers. QTL analysis identified genomic regions associated with FHB resistance on chromosome arms 2BL, 3BS, 4AL, 4BS, 5AL and 6AS. DBC-480 contributed the resistant allele at all loci. Fhb1 was detected in all three populations, demonstrating for the first time its successful deployment in durum wheat. The effect of Fhb1 on FHB resistance in durum wheat was further verified by evaluating type 2 resistance in one of the three populations. Plant height had a strong influence in modulating FHB severity. Although the semi-dwarf allele Rht-B1b was associated with increased FHB susceptibility, its negative effect was efficiently counterbalanced in lines carrying Fhb1. Semi-dwarf lines with enhanced levels of resistance were selected and will assist the development of FHB resistant cultivars

L’étude des rendements en blé a mis en évidence une stagnation apparue dans les années 1990, notamment en France, et principalement lié aux stress hydrique et thermique. Dans ce contexte, améliorer la tolérance du blé européen à ces stress est de première importance. Cette étude avait pour but d’étudier le déterminisme génétique de la tolérance à ces stress chez le blé. Pour ce faire, trois populations de blé tendre du CIMMYT combinant des caractères d’adaptation à ces stress ont été cultivées en conditions irriguée, sèche et stress thermique irriguée plusieurs années. Des caractères physiologiques et agronomiques ont été mesurés sur un réseau de 15 essais. Une méthodologie de caractérisation environnementale a été développée et a permis l’identification de six scenarii de stress au sein du réseau. Une covariable environnementale représentative de chacun a été extraite. L’utilisation des modèles de régression factorielles a permis la décomposition de l’interaction génotype x environnement ainsi que la mise en évidence d’une sensibilité différentielle au stress dans le germplasm. Une recherche de QTL multi-environnementale a conduit à la détection de régions génomiques contrôlant les caractères physiologiques et agronomiques ainsi que leurs interactions avec l’environnement. De la caractérisation environnementale à la détection de QTL, cette étude a abouti au développement d’un outil pour les sélectionneurs permettant l’évaluation du potentiel des génotypes face à une gamme d’environnement, mais aussi à l’identification de régions génomiques impliquées dans le contrôle de la tolérance aux stress hydrique et thermique chez le blé tendre. Ceci pourrait améliorer la tolérance à ces stress au sein du germplasm européen
A stagnation of wheat yield was reported in France and other countries worldwide since the 1990’s, which incriminated mainly drought and heat stress. Improving the European wheat tolerance to them is of first importance. This study aimed to investigate the genetic determinism of the tolerance to such stresses. Three CIMMYT bread wheat populations combining complementary heat and drought adaptive habits were grown in Northern Mexico under irrigated, drought and heat-irrigated treatments from 2011 to 2013. The trial network comprised 15 trials and both physiological and agronomic traits were scored. First, an environmental characterization methodology was developed and resulted in the identification of six main environmental scenarios in the network. A representative environmental covariate was extracted from each of them. Then, a factorial regression model leaded to the dissection of the genotype-by-environment interaction and highlighted differential stress sensitivity of the germplasm. Finally, a multi-environmental QTL detection resulted in the discovery of genomic regions involved in the control of both physiological and agronomic traits and the study of their sensitivity to the environment. From the environmental characterization to the QTL detection, this study resulted in the development of a tool for breeders which may enable the evaluation of the potential of any genotypes in front of a range of environment, but also the identification of genomic regions involved in the control of the tolerance to drought and heat stress in bread wheat. This may help in improving the tolerance of the European bread wheat germplasm to drought and heat stress

Genetic diversity of domesticated wheats has been significantly reduced compared to that of their wild progenitors, through a long selection procedure for those phenotypic traits which led the wild plants to better suit the human needs. Tetraploid wheat landraces were largely cultivated until the first decades of the twentieth century, being progressively abandoned from the early 1970s and replaced with improved, genetically uniform semidwarf cultivars as a consequence of the Green Revolution. Nevertheless, since the current climate change is affecting grain yields worldwide and threatening food securety, sources for specific adaption to drough and heat are urgently needed. In this context, addressing the research towards the study of the level and the structure of genetic diversity in tetraploid wheats, linked to the dection of specific chromosomic traits of interest, has become very important. The relatively high level of genetic variation in modern crops could be obtained through the genetic drift and introgressions between or among the domesticated crops and their close wild relatives. In particular, landraces, characterized by a wide variability in terms of morphological, phenological and quality traits, provide a large source of genetic variability. Many researches have showed their specific adaptation to local environmental conditions according to their place of origin, and, very recently, their ability to form mycorrhizal symbiosis. Positive advances have been reported regarding the mutualistic relationship between the plant and the mycorrhizal fungus, revealing better performance for the host in terms of nutrient uptake and protection from salinity, lack of water, and excess phytotoxic elements. Mycorrhiza studies and the recent progress in research in this sector have shown a possible solution for environmental sustainability: AMF represent a valid alternative to overcome the loss of biological fertility of soils, reduce chemical inputs, and alleviate the effects of biotic and abiotic stresses. However, the actual role of the single wheat genotype in establishing this type of association is still poorly investigated. In this work, the genetic diversity and population genetic structure of a collection of 265 accessions of eight tetraploid Triticum turgidum L. subspecies were investigated using 35,143 single nucleotide polymorphisms (SNPs) screened with a 35K Axiom® array. Neighbor joining algorithm, discriminant analysis of principal components (DAPC) and Bayesian model-based clustering algorithm implemented in STRUCTURE software revealed clusters in accordance to the taxonomic classification, reflecting the evolutionary history and the phylogenetic relationships among Triticum turgidum L. subspecies. Starting from these results, 130 accessions have been inoculated with the AMF species Funneliformis mosseae (F. mosseae) and Rhizoglomus irregulare (R. irregulare), and a genome wide association study (GWAS) was conducted to identify genetic markers in linkage with chromosome regions involved in this symbiosis. 5 Six clusters of genetically related accessions were identified, showing a different mycorrhizal colonization among them. GWAS revealed four significant quantitative trait nucleotides (QTNs) involved in mycorrhizal symbiosis, located on chromosome 1A, 2A, 2B and 6A. The results of this work enrich future breeding activities aimed at developing new grains on the basis of genetic diversity on low or high susceptibility to mycorrhization, and, possibly, maximizing the symbiotic effects.

Fructans are polysaccharides that are made up mainly of fructose. They are non-digestible carbohydrates and act as prebiotics to selectively promote the growth of colonic bifidobacteria, thereby improving human gut health. Fructans are present in the grain of wheat (Triticum aestivum L.), a staple food crop. Until now, there has been no research on genetic improvement of the concentration of fructans in wheat grain, partly because it has been difficult to accurately measure. One aim of this research project was to develop a simple and effective method to measure the fructan concentration in wheat grain. This was achieved by modifying a method that involves extraction of fructans from wheat grain followed by enzymatic hydrolysis to break down fructans into monosaccharides and quantification by anion-exchange liquid chromatography coupled with pulsed amperometric detection. The modified procedure is reliable and allows the handling of large numbers of flour samples at a relatively low cost, and can therefore be useful for assessing large numbers of wheat breeding lines. Using this method, grain samples taken from a diverse set of 117 wheat cultivars and breeding lines, including parents of mapping populations, were analysed for grain fructan concentration. There was significant genotypic variation among these materials, with grain fructan concentration ranging from 0.3 to 2.3% of grain dry weight. There was no evidence of strong genotype-byenvironment interaction; the fructan concentrations of the same genotypes were positively correlated over different environments in Australia. Genetic mapping was carried out to detect and map loci affecting grain fructan concentration in wheat using a doubled haploid population derived from a cross between Berkut (high fructan) and Krichauff (low fructan). Grain samples were obtained from two field sites in South Australia and one in Kazakhstan. Fructan concentration varied widely within the population (0.6-2.6% of grain dry weight), with heritability estimated as h² = 0.71. A linkage map of 528 molecular markers covering 21 wheat chromosomes was used for locating quantitative trait loci (QTL). Genetic mapping identified two major QTLs on chromosomes 6D and 7A, with the (high fructan concentration) alleles contributed from Berkut, contributing to a 30-40% increase in wheat grain fructan compared to the Krichauff alleles. Effects of these chromosome regions were validated in additional environments and in another mapping population, Sokoll/Krichauff, with the favourable alleles contributed from Sokoll. The major QTL on chromosome 7A was in the same region with a reported fructosyltransferase orthologue (AB029888), while the major QTL on chromosome 6D seemed to be co-located with a reported gene encoding for a fructan-degrading enzyme 1-exohydrolase (1-FEHw2). It is concluded that grain fructan concentration of wheat can be improved by breeding and that molecular markers could be used to select effectively for favourable alleles in two regions of the wheat genome.
Thesis (Ph.D.) - University of Adelaide, School of Agriculture, Food and Wine, 2009

Fructans are polysaccharides that are made up mainly of fructose. They are non-digestible carbohydrates and act as prebiotics to selectively promote the growth of colonic bifidobacteria, thereby improving human gut health. Fructans are present in the grain of wheat (Triticum aestivum L.), a staple food crop. Until now, there has been no research on genetic improvement of the concentration of fructans in wheat grain, partly because it has been difficult to accurately measure. One aim of this research project was to develop a simple and effective method to measure the fructan concentration in wheat grain. This was achieved by modifying a method that involves extraction of fructans from wheat grain followed by enzymatic hydrolysis to break down fructans into monosaccharides and quantification by anion-exchange liquid chromatography coupled with pulsed amperometric detection. The modified procedure is reliable and allows the handling of large numbers of flour samples at a relatively low cost, and can therefore be useful for assessing large numbers of wheat breeding lines. Using this method, grain samples taken from a diverse set of 117 wheat cultivars and breeding lines, including parents of mapping populations, were analysed for grain fructan concentration. There was significant genotypic variation among these materials, with grain fructan concentration ranging from 0.3 to 2.3% of grain dry weight. There was no evidence of strong genotype-byenvironment interaction; the fructan concentrations of the same genotypes were positively correlated over different environments in Australia. Genetic mapping was carried out to detect and map loci affecting grain fructan concentration in wheat using a doubled haploid population derived from a cross between Berkut (high fructan) and Krichauff (low fructan). Grain samples were obtained from two field sites in South Australia and one in Kazakhstan. Fructan concentration varied widely within the population (0.6-2.6% of grain dry weight), with heritability estimated as h² = 0.71. A linkage map of 528 molecular markers covering 21 wheat chromosomes was used for locating quantitative trait loci (QTL). Genetic mapping identified two major QTLs on chromosomes 6D and 7A, with the (high fructan concentration) alleles contributed from Berkut, contributing to a 30-40% increase in wheat grain fructan compared to the Krichauff alleles. Effects of these chromosome regions were validated in additional environments and in another mapping population, Sokoll/Krichauff, with the favourable alleles contributed from Sokoll. The major QTL on chromosome 7A was in the same region with a reported fructosyltransferase orthologue (AB029888), while the major QTL on chromosome 6D seemed to be co-located with a reported gene encoding for a fructan-degrading enzyme 1-exohydrolase (1-FEHw2). It is concluded that grain fructan concentration of wheat can be improved by breeding and that molecular markers could be used to select effectively for favourable alleles in two regions of the wheat genome.
Thesis (Ph.D.) - University of Adelaide, School of Agriculture, Food and Wine, 2009

The primary goals of this project were: (1) development of a genetically characterized association panel of wild emmer for high resolution analysis of the genetic basis of complex traits; (2) characterization and mapping of genes and QTL for seedling and adult plant resistance to stripe rust in wild emmer populations; (3) characterization of LD patterns along wild emmer chromosomes; (4) elucidation of the multi-locus genetic structure of wild emmer populations and its correlation with geo-climatic variables at the collection sites. Introduction In recent years, Stripe (yellow) rust (Yr) caused by Pucciniastriiformis f. sp. tritici(PST) has become a major threat to wheat crops in many parts of the world. New races have overcome most of the known resistances. It is essential, therefore, that the search for new genes will continue, followed by their mapping by molecular markers and introgression into the elite varieties by marker-assisted selection (MAS). The reservoir of genes for disease and pest resistance in wild emmer wheat (Triticumdicoccoides) is an important resource that must be made available to wheat breeders. The majority of resistance genes that were introgressed so far in cultivated wheat are resistance (R) genes. These genes, though confering near-immunity from the seedling stage, are often overcome by the pathogen in a short period after being deployed over vast production areas. On the other hand, adult-plant resistance (APR) is usually more durable since it is, in many cases, polygenic and confers partial resistance that may put less selective pressure on the pathogen. In this project, we have screened a collection of 480 wild emmer accessions originating from Israel for APR and seedling resistance to PST. Seedling resistance was tested against one Israeli and 3 North American PST isolates. APR was tested on accessions that did not have seedling resistance. The APR screen was conducted in two fields in Israel and in one field in the USA over 3 years for a total of 11 replicates. We have found about 20 accessions that have moderate stripe rust APR with infection type (IT<5), and about 20 additional accessions that have novel seedling resistance (IT<3). We have genotyped the collection using genotyping by sequencing (GBS) and the 90K SNP chip array. GBS yielded a total 341K SNP that were filtered to 150K informative SNP. The 90K assay resulted in 11K informative SNP. We have conducted a genome-wide association scan (GWAS) and found one significant locus on 6BL ( -log p >5). Two novel loci were found for seedling resistance. Further investigation of the 6BL locus and the effect of Yr36 showed that the 6BL locus and the Yr36 have additive effect and that the presence of favorable alleles of both loci results in reduction of 2 grades in the IT score. To identify alleles conferring adaption to extreme climatic conditions, we have associated the patterns of genomic variation in wild emmer with historic climate data from the accessions’ collection sites. The analysis of population stratification revealed four genetically distinct groups of wild emmer accessions coinciding with their geographic distribution. Partitioning of genomic variance showed that geographic location and climate together explain 43% of SNPs among emmer accessions with 19% of SNPs affected by climatic factors. The top three bioclimatic factors driving SNP distribution were temperature seasonality, precipitation seasonality, and isothermality. Association mapping approaches revealed 57 SNPs associated with these bio-climatic variables. Out of 21 unique genomic regions controlling heading date variation, 10 (~50%) overlapped with SNPs showing significant association with at least one of the three bioclimatic variables. This result suggests that a substantial part of the genomic variation associated with local adaptation in wild emmer is driven by selection acting on loci regulating flowering. Conclusions: Wild emmer can serve as a good source for novel APR and seedling R genes for stripe rust resistance. APR for stripe rust is a complex trait conferred by several loci that may have an additive effect. GWAS is feasible in the wild emmer population, however, its detection power is limited. A panel of wild emmer tagged with more than 150K SNP is available for further GWAS of important traits. The insights gained by the bioclimatic-gentic associations should be taken into consideration when planning conservation strategies.