Analysis and mapping quantitative trait loci for histidine content in barley (Hordeum vulgare L.) using microsatellite markers

Mining the gene of histidine content in barley grain helps with the breeding of functional barley varieties. The study constructed a recombinant inbred lines (RILs) containing 193 families derived from the cross between Ziguangmangluoerling (ZGMLEL) (♀) and Schooner No.3 (♂). The histidine (HIS) content in the grain of the mapping population and its parents were determined by an automatic amino acid analyzer. The HIS content of ZGMLEL was 0.53 mg/g. The grain HIS content of Schooner No. 3 was 0.21 mg/g, and the grain HIS content of population ranged from 0.23 to 0.54. Genetic linkage maps, including those of seven chromosomes of barley, were constructed by using 180 pairs of simple sequence repeat (SSR) markers, with a total genetic distance of 2671.03 cM and average marker spacing of 14.84 cM. Quantitative trait locus (QTL) IciMappingV3.3 was used to analyze QTL of HIS content in barley grains, and three QTLs were detected. Mapping results showed that the three loci were located on chromosomes 2H, 4H, and 7H, respectively. The major QTL with a contribution rate of 10.11% was located on barley chromosome 4H (HVBAMMGB84-BMAG0808). The additive effect is positive (0.025). Thus, it comes from the high-value parent ZGMLEL. Another major QTL with a contribution rate of 13.75% was located on barley chromosome 7H (GBM1303-GMS056). The minor QTL with a contribution rate of 6.01% was located on chromosome 2H (Scssr03381-Scssr07759). The additive effects of 4H and 7H QTLs were negative (− 0.02 and − 0.033). So, they came from the male parent Schooner. The results provided a reference for further fine mapping, cloning, and transformation of HIS genes in barley grains.

Inheritance and Characterization of Rph27: A Third Race-Specific Resistance Gene in the Barley Cultivar Quinn

The barley cultivar Quinn was previously reported to carry two genes for resistance to Puccinia hordei, viz. Rph2 and Rph5. In this study, we characterized and mapped a third resistance gene (RphCRQ3) in cultivar Quinn. Multipathotype testing in the greenhouse on a panel of barley genotypes previously postulated to carry Rph2 revealed rare race specificity in four genotypes in response to P. hordei pathotype (pt.) 222 P+ (virulent on Rph2 and Rph5). This suggested either the presence of a race-specific allele variant of Rph2 or the presence of an independent uncharacterized leaf rust resistance locus. A test of allelism on 1,271 F2 Peruvian (Rph2)/Quinn (Rph2 + Rph5) derived seedlings with P. hordei pt. 220 P+ (avirulent on Rph2 and virulent on Rph5) revealed no susceptible segregants. To determine whether the race-specific resistance in Quinn was due to an allele of Rph2 on chromosome 5H or a third uncharacterized resistance gene, we tested the Peruvian/Quinn F3 population with 222 P+ and observed monogenic inheritance. Subsequent bulked segregant analysis indicated the presence of complete in-phase marker fixation near the telomere on the short arm of chromosome 4H, confirming the presence of a third resistance locus in Quinn in addition to Rph2 and Rph5. In accordance with the rules and numbering system of barley gene nomenclature, RphCRQ3 has been designated Rph27.

Physical organization of repetitive sequences and chromosome diversity of barley revealed by fluorescence in situ hybridization (FISH)

Fluorescence in situ hybridization (FISH) using oligonucleotides is a simple and convenient method for chromosome research. In this study, 34 of 46 previously developed oligonucleotides produced signals in barley. Together with two plasmid clones and one PCR-amplified cereal centromere repeat (CCS1) probe, 37 repetitive sequences were chromosomally located produced three types of signals covering different positions on the chromosomes. The centromeric and pericentric regions had a more complex genomic organization and sequence composition probably indicative of higher contents of heterochromatin. An efficient multi-plex probe containing eight oligonucleotides and a plasmid clone of 45S rDNA was developed. Thirty-three barley karyotypes were developed and compared. Among them, 11 irradiation-induced mutants of cultivar 08-49 showed no chromosomal variation, whereas 22 cultivar and landrace accessions contained 28 chromosomal polymorphisms. Chromosome 4H was the most variable and 6H was the least variable based on chromosome polymorphic information content (CPIC). Five polymorphic chromosomes (1H-2, 2H-1, 3H-3, 5H-2, and 6H-2) were dominant types, each occurring in more than 50% of accessions. The multi-plex probe should facilitate identification of further chromosomal polymorphisms in barley.

Redox-dependent interaction between thaumatin-like protein and β-glucan influences malting quality of barley

Barley is the cornerstone of the malting and brewing industry. It is known that 250 quantitative trait loci (QTLs) of the grain are associated with 19 malting-quality phenotypes. However, only a few of the contributing genetic components have been identified. One of these, on chromosome 4H, contains a major malting QTL, QTL2, located near the telomeric region that accounts, respectively, for 28.9% and 37.6% of the variation in the β-glucan and extract fractions of malt. In the current study, we dissected the QTL2 region using an expression- and microsynteny-based approach. From a set of 22 expressed sequence tags expressed in seeds at the malting stage, we identified a candidate gene,TLP8(thaumatin-like protein 8), which was differentially expressed and influenced malting quality. Transcript abundance and protein profiles ofTLP8were studied in different malt and feed varieties using quantitative PCR, immunoblotting, and enzyme-linked immunosorbent assay (ELISA). The experiments demonstrated that TLP8 binds to insoluble (1, 3, 1, 4)-β-D glucan in grain extracts, thereby facilitating the removal of this undesirable polysaccharide during malting. Further, the binding of TLP8 to β-glucan was dependent on redox. These findings represent a stride forward in our understanding of the malting process and provide a foundation for future improvements in the final beer-making process.

Mapping barley genes to chromosome arms by transcript profiling of wheat–barley ditelosomic chromosome addition lines

Wheat–barley disomic and ditelosomic chromosome addition lines have been used as genetic tools for a range of applications since their development in the 1980s. In the present study, we used the Affymetrix Barley1 GeneChip for comparative transcript analysis of the barley cultivar Betzes, the wheat cultivar Chinese Spring, and Chinese Spring – Betzes ditelosomic chromosome addition lines to physically map barley genes to their respective chromosome arm locations. We mapped 1257 barley genes to chromosome arms 1HS, 2HS, 2HL, 3HS, 3HL, 4HS, 4HL, 5HS, 5HL, 7HS, and 7HL based on their transcript levels in the ditelosomic addition lines. The number of genes assigned to individual chromosome arms ranged from 24 to 197. We validated the physical locations of the genes through comparison with our previous chromosome-based physical mapping, comparative in silico mapping with rice and wheat, and single feature polymorphism (SFP) analysis. We found our physical mapping of barley genes to chromosome arms to be consistent with our previous physical mapping to whole chromosomes. In silico comparative mapping of barley genes assigned to chromosome arms revealed that the average genomic synteny to wheat and rice chromosome arms was 63.2% and 65.5%, respectively. In the 1257 mapped genes, we identified SFPs in 924 genes between the appropriate ditelosomic line and Chinese Spring that supported physical map placements. We also identified a single small rearrangement event between rice chromosome 9 and barley chromosome 4H that accounts for the loss of synteny for several genes.