Identification of Ideal Allele Combinations for the Adaptation of Spring Barley to Northern Latitudes.

The northwards expansion of barley production requires adaptation to longer days, lower temperatures and stronger winds during the growing season. We have screened 169 lines of the current barley breeding gene pool in the Nordic region with regards to heading, maturity, height, and lodging under different environmental conditions in nineteen field trials over 3 years at eight locations in northern and central Europe. Through a genome-wide association scan we have linked phenotypic differences observed in multi-environment field trials (MET) to single nucleotide polymorphisms (SNP). We have identified an allele combination, only occurring among a few Icelandic lines, that affects heat sum to maturity and requires 214 growing degree days (GDD) less heat sum to maturity than the most common allele combination in the Nordic spring barley gene pool. This allele combination is beneficial in a cold environment, where autumn frost can destroy a late maturing harvest. Despite decades of intense breeding efforts relying heavily on the same germplasm, our results show that there still exists considerable variation within the current breeding gene pool and we identify ideal allele combinations for regional adaptation, which can facilitate the expansion of cereal cultivation even further northwards.

Exposure to Ultraviolet (UV-C) Radiation Increases Germination Rate of Maize (Zea maize L.) and Sugar Beet (Beta vulgaris) Seeds.

This study investigated the effect of ultraviolet (UV-C) radiation on the germination percentage, germination rate, radicle length, and plumule length of maize and sugar beet seeds. The experiment was implemented in six replicates of 30 seeds per replicate and in sterilized petri dishes under laboratory conditions. Treatments included UV-C (254 nm) radiation exposure durations of 0 min (control), 30 min, 2 h, 4 h, 8 h, and 12 h. The UV-C radiation treatments did not significantly affect the germination percentage of the seeds (p < 0.05). However, the seeds germination rate was significantly affected by the UV-C radiation treatments. The treatments of 8 h and 12 h exposure duration led to the highest seed germination rates in maize and sugar beet, respectively. Lowest seed germination rates belonged to the controls. The radicle length of maize seeds was significantly affected by the UV-C radiation treatments, but the treatments did not significantly affect the radicle length of sugar beet seeds. The 12 h exposure to UV-C radiation treatment resulted in the largest radicle in maize, which was 2.08 cm larger than the radicle of the control seeds. The UV-C radiation treatments had a statistically significant effect on the plumule length of maize and sugar beet seeds. The treatment 8 h UV-C exposure duration led to the largest plumule in maize and sugar beet, which were 0.32 cm and 0.83 cm larger than the plumule of the control seeds, respectively. Breaking down the seed coat and increasing the temperature by UV-C radiation are potential reasons for the observed positive effects.

Mapping of common bunt resistance gene Bt9 in wheat.

KEY MESSAGE: The Bt9 resistance locus was mapped and shown to be distinct from the Bt10 locus. New markers linked to Bt9 have been identified and may be used to breed for resistance towards the seed-borne disease. Increasing organic wheat production in Denmark, and in other wheat-producing areas, in conjunction with legal requirements for organic seed production, may potentially lead to a rise in common bunt occurrence. As systemic pesticides are not used in organic farming, organic wheat production systems may benefit from genetic resistances. However, little is known about the underlying genetic mechanisms and locations of the resistance factors for common bunt resistance in wheat. A double haploid (DH) population segregating for common bunt resistance was used to identify the chromosomal location of common bunt resistance gene Bt9. DH lines were phenotyped in three environments and genotyped with DArTseq and SSR markers. The total length of the resulting linkage map was 2882 cM distributed across all 21 wheat chromosomes. Bt9 was mapped to the distal end of chromosome 6DL. Since wheat common bunt resistance gene Bt10 is also located on chromosome 6D, the possibility of their co-location was investigated. A comparison of marker sequences linked to Bt9 and Bt10 on physical maps of chromosome 6D confirmed that Bt9 and Bt10 are two distinct resistance factors located at the distal (6DL) and proximal (6DS) end, respectively, of chromosome 6D. Five new SSR markers Xgpw4005-1, Xgpw7433, Xwmc773, Xgpw7303 and Xgpw362 and many SNP and PAV markers flanking the Bt9 resistance locus were identified and they may be used in the future for marker-assisted selection.

Genetic diversity and population structure analysis of European hexaploid bread wheat (Triticum aestivum L.) varieties.

Progress in plant breeding is facilitated by accurate information about genetic structure and diversity. Here, Diversity Array Technology (DArT) was used to characterize a population of 94 bread wheat (Triticum aestivum L.) varieties of mainly European origin. In total, 1,849 of 7,000 tested markers were polymorphic and could be used for population structure analysis. Two major subgroups of wheat varieties, GrI and GrII, were identified using the program STRUCTURE, and confirmed by principal component analysis (PCA). These subgroups were largely separated according to origin; GrI comprised varieties from Southern and Eastern Europe, whereas GrII contained mostly modern varieties from Western and Northern Europe. A large proportion of the markers contributing most to the genetic separation of the subgroups were located on chromosome 2D near the Reduced height 8 (Rht8) locus, and PCR-based genotyping suggested that breeding for the Rht8 allele had a major impact on subgroup separation. Consistently, analysis of linkage disequilibrium (LD) suggested that different selective pressures had acted on chromosome 2D in the two subgroups. Our data provides an overview of the allele composition of bread wheat varieties anchored to DArT markers, which will facilitate targeted combination of alleles following DArT-based QTL studies. In addition, the genetic diversity and distance data combined with specific Rht8 genotypes can now be used by breeders to guide selection of crossing parents.

Genome-wide association study of resistant starch (RS) phenotypes in a barley variety collection.

Barley is primarily grown for feed and malt, but in some regions of the world it is also considered to be a staple food. Some barley types such as high-amylose barley have also gained importance as health-promoting foods. Starch that is not readily digested in the upper mammalian gastrointestinal system, or resistant starch (RS), is considered to be valuable because it prevents some diet-related diseases such as colon cancer. RS was quantified in a diverse collection of 209 spring barley varieties released in Europe during the past 100 years. The RS content varied from <1% to >15% in the collection, with 13 varieties having high RS content (>11%) and 15 varieties below 1%. Combined with genome-wide association scanning (GWAS), SNP markers and candidate genes controlling the RS content in grains were identified. This identified 40 SNP markers with a LOD score above 2, located on chromosomes 2H, 3H, 5H, and 6H, respectively. Among these SNPs, 10 genes with a known role in starch biosynthesis were associated on the basis of synteny conservation to the rice genome. The ß-glucan content was quantified in 61 varieties selected to represent extreme as well as medium RS values. The ß-glucan amount in the 15 varieties with RS <1% ranged from 1.7 to 3.2%, ranged from 1.76 to 2.54% in the 13 varieties with RS >11%, and ranged from 1.95 to 2.82% for those with 1%< RS < 11%. No statistically significant correlation between RS content and ß-glucan content was found. This association analysis of commercial varieties revealed a large variation in RS content and identified a number of SNP markers that can be explored for selection and further dissection of the pathway and control of RS phenotype.

Integration of the barley genetic and seed proteome maps for chromosome 1H, 2H, 3H, 5H and 7H.

Two-dimensional gel electrophoresis was used to screen spring barley cultivars for differences in seed protein profiles. In parallel, 72 microsatellite (simple sequence repeat (SSR)) markers and 11 malting quality parameters were analysed for each cultivar. Over 60 protein spots displayed cultivar variation, including peroxidases, serpins and proteins with unknown functions. Cultivars were clustered based on the spot variation matrix. Cultivars with superior malting quality grouped together, indicating malting quality to be more closely correlated with seed proteomes than with SSR profiles. Mass spectrometry showed that some spot variations were caused by amino acid differences encoded by single nucleotide polymorphisms (SNPs). Coding SNPs were validated by mass spectrometry, expressed sequence tag and 2D gel data. Coding SNPs can alter function of affected proteins and may thus represent a link between cultivar traits, proteome and genome. Proteome analysis of doubled haploid lines derived from a cross between a malting (Scarlett) and a feed cultivar (Meltan) enabled genetic localisation of protein phenotypes represented by 48 spot variations, involving e.g. peroxidases, serpins, alpha-amylase/trypsin inhibitors, peroxiredoxin and a small heat shock protein, in relation to markers on the chromosome map.

The Horn of Africa as a centre of barley diversification and a potential domestication site.

According to a widely accepted theory on barley domestication, wild barley (Hordeum vulgare ssp. spontaneum) from the Fertile Crescent is the progenitor of all cultivated barley (H. vulgare ssp. vulgare). To determine whether barley has undergone one or more domestication events, barley accessions from three continents have been studied (a) using 38 nuclear SSR (nuSSRs) markers, (b) using five chloroplast SSR (cpSSR) markers yielding 5 polymorphic loci and (c) by detecting the differences in a 468 bp fragment from the non-coding region of chloroplast DNA. A clear separation was found between Eritrean/Ethiopian barley and barley from West Asia and North Africa (WANA) as well as from Europe. The data from chloroplast DNA clearly indicate that the wild barley (H. vulgare ssp. spontaneum) as it is found today in the "Fertile Crescent" might not be the progenitor of the barley cultivated in Eritrea (and Ethiopia). Consequently, an independent domestication might have taken place at the Horn of Africa.