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1.
Plant Mol Biol ; 98(3): 249-260, 2018 Oct.
Article in English | MEDLINE | ID: mdl-30244408

ABSTRACT

KEY MESSAGE: We have isolated a novel powdery mildew resistance gene in wheat that was originally introgressed from rye. Further analysis revealed evolutionary divergent history of wheat and rye orthologous resistance genes. Wheat production is under constant threat from a number of fungal pathogens, among them is wheat powdery mildew (Blumeria graminis f. sp. tritici). Deployment of resistance genes is the most economical and sustainable method for mildew control. However, domestication and selective breeding have narrowed genetic diversity of modern wheat germplasm, and breeders have relied on wheat relatives for enriching its gene pool through introgression. Translocations where the 1RS chromosome arm was introgressed from rye to wheat have improved yield and resistance against various pathogens. Here, we isolated the Pm17 mildew resistance gene located on the 1RS introgression in wheat cultivar 'Amigo' and found that it is an allele or a close paralog of the Pm8 gene isolated earlier from 'Petkus' rye. Functional validation using transient and stable transformation confirmed the identity of Pm17. Analysis of Pm17 and Pm8 coding regions revealed an overall identity of 82.9% at the protein level, with the LRR domains being most divergent. Our analysis also showed that the two rye genes are much more diverse compared to the variants encoded by the Pm3 gene in wheat, which is orthologous to Pm17/Pm8 as concluded from highly conserved upstream sequences in all these genes. Thus, the evolutionary history of these orthologous loci differs in the cereal species rye and wheat and demonstrates that orthologous resistance genes can take different routes towards functionally active genes. These findings suggest that the isolation of Pm3/Pm8/Pm17 orthologs from other grass species, additional alleles from the rye germplasm as well as possibly synthetic variants will result in novel resistance genes useful in wheat breeding.


Subject(s)
Biological Evolution , Genetic Variation , Plant Proteins/metabolism , Secale/genetics , Triticum/genetics , Genetic Predisposition to Disease , Genetic Speciation , Plant Diseases/genetics , Plant Diseases/immunology , Plant Diseases/microbiology , Plant Proteins/genetics
2.
Plant J ; 79(6): 893-903, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24942051

ABSTRACT

The development of high-yielding varieties with broad-spectrum durable disease resistance is the ultimate goal of crop breeding. In plants, immune receptors of the nucleotide-binding-leucine-rich repeat (NB-LRR) class mediate race-specific resistance against pathogen attack. When employed in agriculture this type of resistance is often rapidly overcome by newly adapted pathogen races. The stacking of different resistance genes or alleles in F1 hybrids or in pyramided lines is a promising strategy for achieving more durable resistance. Here, we identify a molecular mechanism which can negatively interfere with the allele-pyramiding approach. We show that pairwise combinations of different alleles of the powdery mildew resistance gene Pm3 in F1 hybrids and stacked transgenic wheat lines can result in suppression of Pm3-based resistance. This effect is independent of the genetic background and solely dependent on the Pm3 alleles. Suppression occurs at the post-translational level, as levels of RNA and protein in the suppressed alleles are unaffected. Using a transient expression system in Nicotiana benthamiana, the LRR domain was identified as the domain conferring suppression. The results of this study suggest that the expression of closely related NB-LRR resistance genes or alleles in the same genotype can lead to dominant-negative interactions. These findings provide a molecular explanation for the frequently observed ineffectiveness of resistance genes introduced from the secondary gene pool into polyploid crop species and mark an important step in overcoming this limitation.


Subject(s)
Ascomycota/physiology , Disease Resistance , Plant Diseases/immunology , Plant Proteins/genetics , Proteins/genetics , Triticum/immunology , Alleles , Chimera , Crops, Agricultural , Gene Expression , Leucine , Leucine-Rich Repeat Proteins , Plant Diseases/microbiology , Plant Leaves/genetics , Plant Leaves/immunology , Plant Leaves/microbiology , Plant Proteins/metabolism , Plants, Genetically Modified , Polyploidy , Proteins/metabolism , Nicotiana/genetics , Nicotiana/immunology , Nicotiana/microbiology , Triticum/genetics , Triticum/microbiology
3.
Plant J ; 79(6): 904-13, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24942074

ABSTRACT

The powdery mildew resistance gene Pm8 derived from rye is located on a 1BL.1RS chromosome translocation in wheat. However, some wheat lines with this translocation do not show resistance to isolates of the wheat powdery mildew pathogen avirulent to Pm8 due to an unknown genetically dominant suppression mechanism. Here we show that lines with suppressed Pm8 activity contain an intact and expressed Pm8 gene. Therefore, the absence of Pm8 function in certain 1BL.1RS-containing wheat lines is not the result of gene loss or mutation but is based on suppression. The wheat gene Pm3, an ortholog of rye Pm8, suppressed Pm8-mediated powdery mildew resistance in lines containing Pm8 in a transient single-cell expression assay. This result was further confirmed in transgenic lines with combined Pm8 and Pm3 transgenes. Expression analysis revealed that suppression is not the result of gene silencing, either in wheat 1BL.1RS translocation lines carrying Pm8 or in transgenic genotypes with both Pm8 and Pm3 alleles. In addition, a similar abundance of the PM8 and PM3 proteins in single or double homozygous transgenic lines suggested that a post-translational mechanism is involved in suppression of Pm8. Co-expression of Pm8 and Pm3 genes in Nicotiana benthamiana leaves followed by co-immunoprecipitation analysis showed that the two proteins interact. Therefore, the formation of a heteromeric protein complex might result in inefficient or absent signal transmission for the defense reaction. These data provide a molecular explanation for the suppression of resistance genes in certain genetic backgrounds and suggest ways to circumvent it in future plant breeding.


Subject(s)
Ascomycota/physiology , Disease Resistance , Plant Diseases/immunology , Plant Proteins/genetics , Secale/genetics , Triticum/genetics , Alleles , Ascomycota/pathogenicity , Dimerization , Gene Expression , Genes, Reporter , Genotype , Immunoprecipitation , Inbreeding , Molecular Sequence Data , Plant Diseases/microbiology , Plant Leaves/genetics , Plant Proteins/metabolism , Plants, Genetically Modified , Protein Interaction Mapping , Protein Processing, Post-Translational , Nicotiana/genetics , Nicotiana/immunology , Nicotiana/microbiology , Transgenes , Translocation, Genetic , Triticum/microbiology
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