Genetic analyses of BaMMV/BaYMV resistance in barley accession HOR4224 result in the identification of an allele of the translation initiation factor 4e (Hv-eIF4E) exclusively effective against Barley mild mosaic virus (BaMMV).

KEY MESSAGE: Based on a strategy combining extensive segregation analyses and tests for allelism with allele-specific re-sequencing an Hv-eIF4E allele exclusively effective against BaMMV was identified and closely linked markers for BaYMV resistance were developed. Soil-borne barley yellow mosaic disease is one of the most important diseases of winter barley. In extensive screenings for resistance, accession 'HOR4224' being resistant to three strains of Barley mild mosaic virus (BaMMV-ASL1, BaMMV-Sil, and BaMMV-Teik) and two strains of Barley yellow mosaic virus (BaYMV-1 and BaYMV-2) was identified. Analyses using Bmac29, being to some extent diagnostic for the rym4/5 locus, gave hint to the presence of the susceptibility-encoding allele at this locus. Therefore, 107 DH lines derived from the cross 'HOR4224' × 'HOR10714' (susceptible) were screened for resistance. Genetic analyses revealed an independent inheritance of resistance to BaMMV and BaYMV ([Formula: see text] = 5.58) both encoded by a single gene (BaMMV [Formula: see text] = 0.477; BaYMV [Formula: see text] = 0.770). Although Bmac29 indicated the susceptibility-encoding allele, BaMMV resistance of 'HOR4224' co-localized with rym4/rym5. The BaYMV resistance was mapped to chromosome 5H in the region of rym3. Sequencing of full length cDNA of the Hv-eIF4E gene displayed an already sequenced allele described to be efficient against BaMMV and BaYMV. However, the F1 progenies of crosses involving 'HOR4224' and rym4/rym5 donors were all resistant to BaMMV but susceptible to BaYMV. Therefore, this is the first report of an allele at the rym4/rym5 locus exclusively efficient against BaMMV. Changes in the specificity are due to one non-synonymous amino acid substitution (I118K). Results obtained elucidate that combining extensive segregation analyses and tests for allelism involving different strains of BaMMV/BaYMV in combination with allele-specific re-sequencing is an efficient strategy for gene and allele detection in complex pathosystems.

The ability of a bymovirus to overcome the rym4-mediated resistance in barley correlates with a codon change in the VPg coding region on RNA1.

The genome difference(s) that enable the European pathotype 2 isolates of Barley yellow mosaic virus (BaYMV-2) to infect barley genotypes with the rym4 resistance gene were investigated. Stable deletions of different sizes occurred in RNA2 of laboratory isolates of the common pathotype (BaYMV-1) and BaYMV-2. After mechanical inoculation of susceptible or rym4 genotypes with a mixture of both isolates, immunocapture-RT-PCR with RNA2-specific primers flanking stable deletion regions was used to detect and distinguish the two pathotypes. Individual leaves contained RNA2 of either or both isolates, showing that RNA2 of BaYMV-1 can replicate and move systemically in rym4 plants when co-inoculated with BaYMV-2. In contrast, sequences of RNA1-specific RT-PCR fragments showed that in resistant plants these were always BaYMV-2, suggesting that the pathogenicity determinant was on RNA1. The complete ORFs of RNA1 of three BaYMV-1 and four BaYMV-2 isolates from the UK and Germany were sequenced, and the RNA2 sequences of one BaYMV-1 and two BaYMV-2 isolates from the UK were also determined. All sequences were very similar to one another and to the published German BaYMV-1 isolate. The only consistent amino acid difference between the BaYMV-1 and BaYMV-2 isolates was in the RNA1-encoded polyproteins and this was confirmed by sequencing the relevant region of eight further German isolates. All BaYMV-1 isolates had lysine at aa 1307, whereas BaYMV-2 isolates had asparagine (or, in one isolate, histidine). The polymorphism occurred in the central region of VPg, which has been shown to be required for pathogenicity on genotypes carrying recessive resistance genes in several potyvirus/dicotyledonous plant pathosystems.