Genomewide association study reveals novel quantitative trait loci associated with resistance towards Septoria tritici blotch in North European winter wheat

Fungal diseases are a major constraint for wheat production. Effective disease resistance is essential for ensuring a high production quality and yield. One of the most severe fungal diseases of wheat is Septoria tritici blotch (STB), which influences wheat production across the world. In this study, genomewide association mapping was used to identify new chromosomal regions on the wheat genome conferring effective resistance towards STB. A winter wheat population of 164 North European varieties and breeding lines was genotyped with 15K single nucleotide polymorphism (SNP) wheat array. The varieties were evaluated for STB in field trials at three locations in Denmark and across 3 years. The association analysis revealed four quantitative trait loci, on chromosomes 1B, 2A, 5D and 7A, highly associated with STB resistance. By comparing varieties containing several quantitative trait loci (QTL) with varieties containing none of the found QTL, a significant difference was found in the mean disease score. This indicates that an effective resistance can be obtained by pyramiding several QTL.     

Cation and total macroelement concentrations influence Ca uptake in micropropagated potato plantlets

Ca uptake by micropropagated ‘Norland’ potato plantlets was investigated in relation to cation and macroelement salt concentrations in modified Murashige and Skoog (1962) basal media. Increased Ca:specific cations, by lowering NH₄, Mg, or K, had little effect on the Ca content of plantlets grown on these media. Increased Ca:total cation ratio, by raising Ca from 3 to 6 mM in the medium, promoted Ca content of micropropagated plantlets. At equivalent Ca:total cation ratio, macroelement dilution increased the osmotic potential of the medium, and had an even greater impact on plantlet shoot Ca content. Root pressure, which is sensitive to variations in salt concentration, was probably the primary Ca delivery method to developing tissues since transpiration was limited in vitro. In vitro systems appear ideal for studying some aspects of nutrient relationships of plants.     

Changes in allelic frequency over time in European bread wheat (Triticum aestivum L.) varieties revealed using DArT and SSR markers

A collection of 189 bread wheat landraces and cultivars, primarily of European origin, released between 1886 and 2009, was analyzed using two DNA marker systems. A set of 76 SSR markers and ~7,000 DArT markers distributed across the wheat genome were employed in these analyses. All of the SSR markers were found to be polymorphic, whereas only 2,532 of the ~7,000 DArT markers were polymorphic. A Mantel test between the genetic distances calculated based on the SSR and DArT data showed a strong positive correlation between the two marker types, with a Pearson’s value (r) of 0.66. We assessed the genetic diversity and allelic frequencies among the accessions based on spring- versus winter-wheat type as well as between landraces and cultivars. We also analyzed the changes in genetic diversity and allelic frequencies in these samples over time. We observed separation based on both vernalization type and release date. Interestingly, we detected a decrease in genetic diversity in wheat accessions released over the period from 1960 to 1980. However, our results also showed that modern plant breeding have succeeded in maintaining genetic diversity in modern wheat cultivars. Studying allelic frequencies using SSR and DArT markers over time revealed changes in allelic frequencies for a number of markers that are known to be linked to important traits, which should be useful for genomic screening efforts. Monitoring changes in the frequency of molecular DNA markers over time in wheat cultivars may yield insight into alleles linked to important traits that have been the subject of positive or negative selection in the past and that may be useful for marker-assisted breeding programs in the future.     

Genetic diversity,population structure and linkage disequilibrium in Nordic spring barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L. subsp. <Emphasis Type="Italic">vulgare</Emphasis>)

Genetic diversity, population structure and genome-wide linkage disequilibrium (LD) was estimated in Nordic spring barley (Hordeum vulgare L. subsp. vulgare) by genotyping 180 breeding lines with 48 SSR markers and 7842 high-confidence SNPs using the Illumina Infinium 9K assay. In total 6208 SNPs were polymorphic and selected for further statistical analysis. A Mantel test revealed a strong positive correlation with a Pearson’s correlation coefficient (r) of 0.86, between the estimates of genetic distances based on SSR and SNP data. Population structure analysis identified two groups with a clear ancestry and one group with an admixed ancestry. The groups were primarily separated based on row-type and geographical origin. Average LD for the whole population decayed below a critical level of r² = 0.20 within a range of 0–4 cM. To avoid confounding effects of the strong population structure, LD decay for the different groups was analysed separately and ranged from 0 to 12 cM. A slower LD decay was found within the two-rowed lines compared to the six-rowed lines and the two-rowed lines originating from the northern part, which could be the result of strong selection for malting quality and yield in the southern part. No large difference in genetic diversity was observed between population sub-groups, but differences at certain chromosomal regions were evident.     

The Identification of Two New Races of Pyrenophora tritici-repentis from the Host Center of Diversity Confirms a One-to-One Relationship in Tan Spot of Wheat

ABSTRACT Pyrenophora tritici-repentis, causal agent of tan spot, induces necrosis and chlorosis in its wheat host. The tan spot system conforms to the toxin model and three host-specific toxins have been identified (Ptr ToxA, Ptr ToxB, and putative Ptr ToxC). Processing of a collection of isolates, obtained in the Fertile Crescent and Caucasus regions, yielded two new virulence patterns. Isolate Az35-5 combined the virulences of races 2 and 5 and was classified in the new race 7. Isolates TS93-71B and TS93-71F had a virulence pattern that combined those of races 2, 3, and 5 and were grouped in the new race 8. Southern analysis revealed that all three isolates possessed copies of the ToxA and ToxB genes, the first time the genes were found in a common background. The production of Ptr ToxA and Ptr ToxB by the isolates was confirmed by western blotting. Virulence patterns suggested that TS93-71B and TS93-71F may also produce Ptr ToxC, even though it was not present at detectable levels in culture filtrates. The identification of races 7 and 8 complete the theoretical maximum number of races that can be differentiated by three loci in the host (2(3) = 8), assuming a one-to-one relationship. It appears that the wheat/P. tritici-repentis system is a mirror image of the classical gene-for-gene relationship.     

Enhanced calcium uptake in micropropagated potato plantlets by macroelement dilution

Ca deficiency disorders are common in micropropagated plantlets but little is known regarding the uptake of Ca in vitro. Ca uptake by in vitra-grown potato (Solanum tuberosum L. cvs. Bintje and Norland) plantlets was investigated when Ca levels were maintained but the total macroelement salts or just NH₄, the major competing cation, were reduced in Murashige and Skoog (MS) medium. This increased the ratio of Ca:cations or Ca:NH₄ as well as the osmotic potential of the medium. The increased Ca-cations improved Ca uptake of shoot tips, leaves, and stems while increased Ca:NH₄ had no effect. MS medium macroelement dilution may improve Ca uptake by micropropagated plantlets and prevent Ca deficiency disorders.     

High genetic diversity revealed in barley (Hordeum vulgare) collected from small-scale farmer’s fields in Eritrea

Remarkable morphological variation has been found within small Eritrean barley fields. Barley was collected from fields approximately 50 m² in size. Spike shape, type, and colour were observed to vary both between and within fields. A set of 39 Simple Sequence Repeats (SSR) markers were used to explore the genetic diversity of the Eritrean barley collected from small-scale farmer’s fields. Significant genetic diversity was found within the barley fields. Out of 240 spikes collected from 24 fields (10 spikes per field), only two spikes from geographically distant fields were genetically similar. Based on the SSR data, individual farmers’ fields were found to possess 97.3% of the genetic variation present in the Eritrean barley. We discuss a strategy to improve the barley yield in Eritrea, and to facilitate the in situ conservation of barley genetic diversity. Gunter Backes and Jihad Orabi contributed equally to this work.     

High levels of genetic and genotypic diversity in field populations of the barley pathogen Ramularia collo-cygni

The ascomycete pathogen Ramularia collo-cygni causes Ramularia leaf spot (RLS) on barley. Although R. collo-cygni is considerd an emerging disease of barley, little is known about genetic diversity or population genetic structure of this pathogen. We applied a set of polymorphic AFLP (Amplified Fragment Length Polymorphism) markers to investigate population genetic structure in two Northern European populations of R. collo-cygni. The distribution of AFLP alleles revealed low levels of population subdivision and high levels of genetic diversity at both locations. Our analyses included 87 isolates and of these 84 showed a unique genotype pattern. The genetic structure of populations in Scotland and Denmark is highly similar and we find no evidence of population sub-division. An analysis of molecular variance was used to show that 86 % of the variance is attributable to within field genetic variance. In spite of the high levels of genetic and genotypic diversity in the R. collo-cygni populations, we find significant evidence of linkage disequilibrium among the AFLP alleles using a multilocus analysis. We propose that the high levels of genotypic diversity and the lack of population differentiation result from considerable levels of gene flow between populations most likely mediated by seed borne dispersal of inoculum.     

Araguspongine C Induces Autophagic Death in Breast Cancer Cells through Suppression of c-Met and HER2 Receptor Tyrosine Kinase Signaling

Receptor tyrosine kinases are key regulators of cellular growth and proliferation. Dysregulations of receptor tyrosine kinases in cancer cells may promote tumorigenesis by multiple mechanisms including enhanced cell survival and inhibition of cell death. Araguspongines represent a group of macrocyclic oxaquinolizidine alkaloids isolated from the marine sponge Xestospongia species. This study evaluated the anticancer activity of the known oxaquinolizidine alkaloids araguspongines A, C, K and L, and xestospongin B against breast cancer cells. Araguspongine C inhibited the proliferation of multiple breast cancer cell lines in vitro in a dose-dependent manner. Interestingly, araguspongine C-induced autophagic cell death in HER2-overexpressing BT-474 breast cancer cells was characterized by vacuole formation and upregulation of autophagy markers including LC3A/B, Atg3, Atg7, and Atg16L. Araguspongine C-induced autophagy was associated with suppression of c-Met and HER2 receptor tyrosine kinase activation. Further in-silico docking studies and cell-free Z-LYTE assays indicated the potential of direct interaction between araguspongine C and the receptor tyrosine kinases c-Met and HER2 at their kinase domains. Remarkably, araguspongine C treatment resulted in the suppression of PI3K/Akt/mTOR signaling cascade in breast cancer cells undergoing autophagy. Induction of autophagic death in BT-474 cells was also associated with decreased levels of inositol 1,4,5-trisphosphate receptor upon treatment with effective concentration of araguspongine C. In conclusion, results of this study are the first to reveal the potential of araguspongine C as an inhibitor to receptor tyrosine kinases resulting in the induction of autophagic cell death in breast cancer cells.     

Population structure and frequency differences of CYP51 mutations in <Emphasis Type="Italic">Zymoseptoria tritici</Emphasis> populations in the Nordic and Baltic regions

Septoria tritici blotch caused by the fungus Zymoseptoria tritici (formerly Mycosphaerella graminicola) is one of the most yield-reducing diseases worldwide. Effective disease management involves the use of resistant cultivars and application of fungicides. In this study, the population structure and genetic diversity of 183 Z. tritici isolates from Denmark, Sweden, Finland and the Baltic countries were analysed by molecular markers. In population structure analysis, isolates from Denmark and Sweden were grouped together, whereas isolates from the Baltics and Finland were grouped together. Analysis of genetic diversity and ?-values confirmed the division of Nordic and Baltic regions. Danish isolates sampled from different regions and different varieties were not genetically different. However, significant genetic differences were detected between isolates sampled from different years in Denmark and for isolates sampled from specific cultivars in different years. Additionally, the frequency of several known point mutations in the gene cyp51, conferring decreased sensitivity to DMI fungicides, was investigated. Several of the examined mutations were detected at a lower frequency in Baltic isolates compared to Danish and Swedish isolates. Analysis of the Danish population revealed a significant increase in specific mutations over the years. Lastly, some mutations were significantly more frequent in isolates derived from certain varieties. By using different resistance sources in breeding programmes and application of a wide range of fungicides, a sustainable and efficient disease management can be obtained.