A new wild emmer wheat panel allows to map new loci associated with resistance to stem rust at seedling stage.

Wheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is a major wheat disease worldwide. A collection of 283 wild emmer wheat [Triticum turgidum L. subsp. dicoccoides (Körn. ex Asch. & Graebn.) Thell] accessions, representative of the entire Fertile Crescent region where wild emmer naturally occurs, was assembled, genotyped, and characterized for population structure, genetic diversity, and rate of linkage disequilibrium (LD) decay. Then, the collection was employed for mapping Pgt resistance genes, as a proof of concept of the effectiveness of genome-wide association studies in wild emmer. The collection was evaluated in controlled conditions for reaction to six common Pgt pathotypes (TPMKC, TTTTF, JRCQC, TRTTF, TTKSK/Ug99, and TKTTF). Most resistant accessions originated from the Southern Levant wild emmer lineage, with some showing a resistance reaction toward three to six tested races. Association analysis was conducted considering a 12K polymorphic single-nucleotide polymorphisms dataset, kinship relatedness between accessions, and population structure. Eleven significant marker-trait associations (MTA) were identified across the genome, which explained from 17% to up to 49% of phenotypic variance with an average 1.5 additive effect (based on the 1-9 scoring scale). The identified loci were either effective against single or multiple races. Some MTAs colocalized with known Pgt resistance genes, while others represent novel resistance loci useful for durum and bread wheat prebreeding. Candidate genes with an annotated function related to plant response to pathogens were identified at the regions linked to the resistance and defined according to the estimated small LD (about 126 kb), as typical of wild species.

Population structure, allelic variation at Rht-B1 and Ppd-A1 loci and its effects on agronomic traits in Argentinian durum wheat.

Exploring the genetic variability in yield and yield-related traits is essential to continue improving genetic gains. Fifty-nine Argentinian durum wheat cultivars were analyzed for important agronomic traits in three field experiments. The collection was genotyped with 3565 genome-wide SNPs and functional markers in order to determine the allelic variation at Rht-B1 and Ppd-A1 genes. Population structure analyses revealed the presence of three main groups, composed by old, modern and genotypes with European or CIMMYT ancestry. The photoperiod sensitivity Ppd-A1b allele showed higher frequency (75%) than the insensitivity one Ppd-A1a (GS105). The semi-dwarfism Rht-B1b and the Ppd-A1a (GS105) alleles were associated with increases in harvest index and decreases in plant height, grain protein content and earlier heading date, although only the varieties carrying the Rht-B1 variants showed differences in grain yield. Out of the two main yield components, grain number per plant was affected by allelic variants at Rht-B1 and Ppd-A1 loci, while no differences were observed in thousand kernel weight. The increases in grain number per spike associated with Rht-B1b were attributed to a higher grain number per spikelet, whereas Ppd-A1a (GS105) was associated with higher grain number per spikelet, but also with lower spikelets per spike.

Allelic Variation at Glutenin Loci (Glu-1, Glu-2 and Glu-3) in a Worldwide Durum Wheat Collection and Its Effect on Quality Attributes.

Durum wheat grains (Triticum turgidum L. ssp. durum) are the main source for the production of pasta, bread and a variety of products consumed worldwide. The quality of pasta is mainly defined by the rheological properties of gluten, an elastic network in wheat endosperms formed of gliadins and glutenins. In this study, the allelic variation at five glutenin loci was analysed in 196 durum wheat genotypes. Two loci (Glu-A1 and Glu-B1), encoding for high-molecular-weight glutenin subunits (HMW-GS), and three loci (Glu-B2, Glu-A3 and Glu-B3), encoding for low molecular weight glutenin subunits (LMW-GS), were assessed by SDS-PAGE. The SDS-sedimentation test was used and the grain protein content was evaluated. A total of 32 glutenin subunits and 41 glutenin haplotypes were identified. Four novel alleles were detected. Fifteen haplotypes represented 85.7% of glutenin loci variability. Some haplotypes carrying the 7 + 15 and 7 + 22 banding patterns at Glu-B1 showed a high gluten strength similar to those that carried the 7 + 8 or 6 + 8 alleles. A decreasing trend in grain protein content was observed over the last 85 years. Allelic frequencies at the three main loci (Glu-B1, Glu-A3 and Glu-B3) changed over the 1915-2020 period. Gluten strength increased from 1970 to 2020 coinciding with the allelic changes observed. These results offer valuable information for glutenin haplotype-based selection for use in breeding programs.

Linkage disequilibrium patterns, population structure and diversity analysis in a worldwide durum wheat collection including Argentinian genotypes.

BACKGROUND: Durum wheat (Triticum turgidum L. ssp. durum Desf. Husn) is the main staple crop used to make pasta products worldwide. Under the current climate change scenarios, genetic variability within a crop plays a crucial role in the successful release of new varieties with high yields and wide crop adaptation. In this study we evaluated a durum wheat collection consisting of 197 genotypes that mainly comprised a historical set of Argentinian germplasm but also included worldwide accessions. RESULTS: We assessed the genetic diversity, population structure and linkage disequilibrium (LD) patterns in this collection using a 35 K SNP array. The level of polymorphism was considered, taking account of the frequent and rare allelic variants. A total of 1547 polymorphic SNPs was located within annotated genes. Genetic diversity in the germplasm collection increased slightly from 1915 to 2010. However, a reduction in genetic diversity using SNPs with rare allelic variants was observed after 1979. However, larger numbers of rare private alleles were observed in the 2000-2009 period, indicating that a high reservoir of rare alleles is still present among the recent germplasm in a very low frequency. The percentage of pairwise loci in LD in the durum genome was low (13.4%) in our collection. Overall LD and the high (r2 > 0.7) or complete (r2 = 1) LD presented different patterns in the chromosomes. The LD increased over three main breeding periods (1915-1979, 1980-1999 and 2000-2020). CONCLUSIONS: Our results suggest that breeding and selection have impacted differently on the A and B genomes, particularly on chromosome 6A and 2A. The collection was structured in five sub-populations and modern Argentinian accessions (cluster Q4) which were clearly differentiated. Our study contributes to the understanding of the complexity of Argentinian durum wheat germplasm and to derive future breeding strategies enhancing the use of genetic diversity in a more efficient and targeted way.

The Global Durum Wheat Panel (GDP): An International Platform to Identify and Exchange Beneficial Alleles.

Representative, broad and diverse collections are a primary resource to dissect genetic diversity and meet pre-breeding and breeding goals through the identification of beneficial alleles for target traits. From 2,500 tetraploid wheat accessions obtained through an international collaborative effort, a Global Durum wheat Panel (GDP) of 1,011 genotypes was assembled that captured 94-97% of the original diversity. The GDP consists of a wide representation of Triticum turgidum ssp. durum modern germplasm and landraces, along with a selection of emmer and primitive tetraploid wheats to maximize diversity. GDP accessions were genotyped using the wheat iSelect 90K SNP array. Among modern durum accessions, breeding programs from Italy, France and Central Asia provided the highest level of genetic diversity, with only a moderate decrease in genetic diversity observed across nearly 50 years of breeding (1970-2018). Further, the breeding programs from Europe had the largest sets of unique alleles. LD was lower in the landraces (0.4 Mbp) than in modern germplasm (1.8 Mbp) at r 2 = 0.5. ADMIXTURE analysis of modern germplasm defined a minimum of 13 distinct genetic clusters (k), which could be traced to the breeding program of origin. Chromosome regions putatively subjected to strong selection pressure were identified from fixation index (F st ) and diversity reduction index (DRI) metrics in pairwise comparisons among decades of release and breeding programs. Clusters of putative selection sweeps (PSW) were identified as co-localized with major loci controlling phenology (Ppd and Vrn), plant height (Rht) and quality (gliadins and glutenins), underlining the role of the corresponding genes as driving elements in modern breeding. Public seed availability and deep genetic characterization of the GDP make this collection a unique and ideal resource to identify and map useful genetic diversity at loci of interest to any breeding program.

Genetic diversity and linkage disequilibrium using SNP (KASP) and AFLP markers in a worldwide durum wheat (Triticum turgidum L. var durum) collection.

The aim of this work was to analyze the genetic diversity and linkage disequilibrium in a collection of 168 durum wheat accessions (Triticum turgidum L. var. durum) of different origins. Our collection was mainly composed of released and unreleased Argentinian germplasm, with additional genotypes from Italy, Chile, France, CIMMYT, Cyprus, USA and WANA region. To this end, the entire collection was characterized with 85 Single Nucleotide Polymorphism (SNP) markers obtained by Kompetitive Allele Specific PCR (KASP), giving a heterozygosity (He) mean value of 0.183 and a coefficient of genetic differentiation (Gst) value of 0.139. A subset of 119 accessions was characterized with six Amplified Fragment Length Polymorphism (AFLP) primer combinations. A total of 181 polymorphic markers (125 AFLP and 56 SNP) amplified across this subset revealed He measures of 0.352 and 0.182, respectively. Of these, 134 were selected to estimate the genome-wide linkage disequilibrium obtaining low significant values (r2 = 0.11) in the subset, indicating its suitability for future genome-wide association studies (GWAS). The structure analysis conducted in the entire collection with SNP detected two subpopulations. However, the structure analysis conducted with AFLP markers in the subset of 119 accessions proved to have greater degree of resolution and detect six subpopulations. The information provided by both marker types was complementary and showed a strong association between old Argentinian and Italian germplasm and a contribution of CIMMYT germplasm to modern Argentinian, Chilean and Cypriot accessions. The influence of Mediterranean germplasm, mainly from Italy, on part of the modern Argentinian cultivars or breeding lines was also clearly evidenced. Although our analysis yields conclusive results and useful information for association mapping studies, further analyses are needed to refine the number of subpopulations present in the germplasm collection analyzed.

Biocontrol of Fusarium graminearum sensu stricto, Reduction of Deoxynivalenol Accumulation and Phytohormone Induction by Two Selected Antagonists.

Fusarium head blight (FHB) is a devastating disease that causes extensive yield and quality losses to wheat and other small cereal grains worldwide. Species within the Fusarium graminearum complex are the main pathogens associated with the disease, F. graminearum sensu stricto being the main pathogen in Argentina. Biocontrol can be used as part of an integrated pest management strategy. Phytohormones play a key role in the plant defense system and their production can be induced by antagonistic microorganisms. The aims of this study were to evaluate the effect of the inoculation of Bacillus velezensis RC 218, F. graminearum and their co-inoculation on the production of salicylic acid (SA) and jasmonic acid (JA) in wheat spikes at different periods of time under greenhouse conditions, and to evaluate the effect of B. velezensis RC 218 and Streptomyces albidoflavus RC 87B on FHB disease incidence, severity and deoxynivalenol accumulation on Triticum turgidum L. var. durum under field conditions. Under greenhouse conditions the production of JA was induced after F. graminearum inoculation at 48 and 72 h, but JA levels were reduced in the co-inoculated treatments. No differences in JA or SA levels were observed between the B. velezensis treatment and the water control. In the spikes inoculated with F. graminearum, SA production was induced early (12 h), as it was shown for initial FHB basal resistance, while JA was induced at a later stage (48 h), revealing different defense strategies at different stages of infection by the hemibiotrophic pathogen F. graminearum. Both B. velezensis RC 218 and S. albidoflavus RC 87B effectively reduced FHB incidence (up to 30%), severity (up to 25%) and deoxynivalenol accumulation (up to 51%) on durum wheat under field conditions.

Mapping of main and epistatic effect QTLs associated to grain protein and gluten strength using a RIL population of durum wheat.

Quality, specifically protein content and gluten strength are among the main objectives of a durum wheat breeding program. The aim of this work was to validate quantitative trait loci (QTLs) associated with grain protein content (GPC) and gluten strength measured by SDS sedimentation volume (SV) and to find additional QTLs expressed in Argentinean environments. Also, epistatic QTL and QTL x environmental interactions were analyzed. A mapping population of 93 RILs derived from the cross UC1113 x Kofa showing extreme values in gluten quality was used. Phenotypic data were collected along six environments (three locations, two years). Main effect QTLs associated with GPC were found in equivalent positions in two environments on chromosomes 3BS (R(2)=21.0-21.6%) and 7BL (R(2)=12.1-13%), and in one environment on chromosomes 1BS, 2AL, 2BS, 3BL, 4AL, 5AS, 5BL and 7AS. The most important and stable QTL affecting SV was located on chromosome 1BL (Glu-B1) consistently detected over the six environments (R(2)=20.9- 54.2%). Additional QTLs were found in three environments on chromosomes 6AL (R(2)=6.4-12.5%), and in two environments on chromosomes 6BL (R(2)=11.5-12.1%), 7AS (R(2)=8.2-10.2%) and 4BS (R(2)=11-16.4%). In addition, pleiotropic effects were found affecting grain yield, test weight, thousand-kernel-weight and days to heading in some of these QTLs. Epistatic QTLs and QTL x environment interactions were found for both quality traits, mostly for GPC. The flanking markers of the QTLs detected in this work could be efficient tools to select superior genotypes for the mentioned traits.