Mapping of powdery mildew resistance genes transferred to common wheat from wild emmer wheat revealed three functional Pm60 haplotypes

Powdery mildew(PM),caused by Blumeria graminis f.sp.tritici(Bgt),is one of the destructive wheat diseases worldwide.Wild emmer wheat(Triticum turgidum ssp.dicoccoides,WEW),a tetraploid progenitor of common wheat,is a valuable genetic resource for wheat disease resistance breeding programs.We developed three hexaploid pre-breeding lines with PM resistance genes derived from three WEW accessions.These resistant pre-breeding lines were crossed with susceptible common wheat accessions.Segregations in the F2populations were 3 resistant:1 susceptible,suggesting a single dominant allele in each resistant parent.Mapping of the resistance gene in each line indicated a single locus on the long arm of chromosome 7A,at the approximate location of previously cloned Pm60 from T.urartu.Sanger sequencing revealed three different Pm60 haplotypes(Hap 3,Hap 5,and Hap 6).Co-segregating diagnostic markers were developed for identification and selection of each haplotype.The resistance function of each haplotype was verified by the virus-induced gene silencing(VIGS).Common wheat lines carrying each of these Pm60 haplotypes were resistant to most Bgt isolates and differences in the response arrays suggested allelic variation in response.

Molecular mapping of YrTZ2,a stripe rust resistance gene in wild emmer accession TZ-2 and its comparative analyses with Aegilops tauschii

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici （Pst）, is a devastating disease that can cause severe yield losses. Identification and utilization of stripe rust resistance genes are essential for effective breeding against the disease. Wild emmer accession TZ-2, originally collected from Mount Hermon, Israel, confers near-immunity resistance against several prevailing Pst races in China. A set of 200 F6：7 recombinant inbred lines （RILs） derived from a cross between susceptible durum wheat cultivar Langdon and TZ-2 was used for stripe rust evaluation. Genetic analysis indicated that the stripe rust resistance of TZ-2 to Pst race CYR34 was controlled by a single dominant gene, temporarily designated YrTZ2. Through bulked segregant analysis （BSA） with SSR markers, YrTZ2 was located on chromosome arm 1BS flanked by Xwmc230 and Xgwm413 with genetic distance of 0.8 cM （distal） and 0.3 cM （proximal）, respectively. By applying wheat 90K iSelect SNP genotyping assay, 11 polymorphic loci （consisting of 250 SNP markers） closely linked to YrTZ2 were identified. YrTZ2 was further delimited into a 0.8-cM genetic interval between SNP marker IWB19368 and SSR marker Xgwm413, and cosegregated with SNP marker IWB28744 （co-segregated with 28 SNP）. Comparative genomics analyses revealed high level of collinearity between the YrTZ2 genomic region and the orthologous region of Aegilops tauschii 1DS. The genomic region between loci IWB19368 and IWB31649 harboring YrTZ2 is orthologous to a 24.5-Mb genomic region between AT1D0112 and AT1D0150, spanning 15 contigs on chromosome 1DS. The genetic and comparative maps of YrTZ2 rovide a framework for map-based cloning and marker-assisted selection of YrTZ2.

Functional characterization of powdery mildew resistance gene MIIW172,a new Pm60 allele and its allelic variation in wild emmer wheat

Wild emmer wheat(Triticum dicoccoides,WEW)is an immediate progenitor of both the cultivated tetraploid and hexaploid wheats and it harbors rich genetic diversity against powdery mildew caused by Blumeria graminis f.sp.tritici(Bgt).A powdery mildew resistance gene Ml I^(W172)originated from WEW accession I^(W172)(G-797-M)is fine mapped in a 0.048 centimorgan(c M)genetic interval on 7 AL,corresponding to a genomic region spanning 233 kb,1 Mb and 800 kb in Chinese Spring,WEW Zavitan,and T.urartu G1812,respectively.Ml I^(W172)encodes a typical NLR protein NLRI^(W172)and physically locates in an NBS-LRR gene cluster.NLRI^(W172)is subsequently identified as a new allele of Pm60,and its function is validated by EMS mutagenesis and transgenic complementation.Haplotype analysis of the Pm60 alleles reveals diversifications in sequence variation in the locus and presence and absence variations(PAV)in WEW populations.Four common single nucleotide variations(SNV)are detected between the Pm60 alleles from WEW and T.urartu,indicative of speciation divergence between the two different wheat progenitors.The newly identified Pm60 alleles and haplotypes in WEW are anticipated to be valuable for breeding powdery mildew resistance wheat cultivars via marker-assisted selection.