Additional file 2: Table S1. of Wheat WCBP1 encodes a putative copper-binding protein involved in stripe rust resistance and inhibition of leaf senescence

. Detailed information on 112 ESTs. A BLAST search was performed using BLASTX in NCBI. In silico mapping was performed using the IWGSC draft wheat sequence database. Table S2. BLAST searches performed for the forward subtraction clones found in the stripe rust-resistant genotype L693. Table S3. The primers used to amplify ESTs. Primer pairs were only designed for 62 out of the 112 ESTs because the remaining ESTs were too short to be used for primer design. Table S4. Primers were designed according to the sequence of the contig (3837062) carrying the polymorphic amplicon, which was located on wheat chromosome 1B ( http://www.wheatgenome.org/ ). These amplicons were used to determine the linkage relationship between the markers and the stripe rust resistance gene in L693. Table S5. Results of the amplification of 5 WCBP1-linked genetic markers in an F2 population derived from crosses between L661 and L693 and between L693 and L661. The genotypes (RR, Rr, rr) of the F2 individuals were determined according to the phenotypes of the corresponding F2:3 families; "A" and "B" for the individual F2 populations were derived from crosses between L661 and L693 and between L693 and L661, respectively. R represents the same amplicon as the resistant parent L693; S represents the same amplicon as the susceptible parent L661; H represents the heterozygous condition. The yield components of the F2 plants, including the number of grains per ear (N s ), the total grain weight per ear (T w ) and the 1000-grain weight (S w ), are provided. ‘\’ represents a missing yield component in the F2 plants. Table S6. The sequences of seven primer pairs designed according to the sequence of a cloned candidate gene in silico; these primers were used for PCR-based gene cloning from genomic DNA. Table S7. Primers for q-PCR analysis. (ZIP 94 kb

Additional file 1: Figure S1. of Wheat WCBP1 encodes a putative copper-binding protein involved in stripe rust resistance and inhibition of leaf senescence

Length distributions of the wheat leaf ESTs (A) and the assembled uniESTs (B), including contigs (C) and singletons (D). Figure S2. Primers and the amplified region of genomic DNA. To clone the WCBP1 gene, primers were designed according to the Chinese Spring draft genome sequence. Full-length amplification was performed using the 1B-1 forward primer and the 1B-6 reverse primer. Figure S3. Comparison of WCBP1 gene sequences. WCBP1 encodes a heavy metal copper-binding protein with two copper sensing domains. The four sequences were cloned using PCR with primers designed according to the polymorphic fragment. L693 is the stripe rust-resistant line; L661 is the stripe rust-susceptible line; YU25 is the resistant parent of L693 and L661; MY11 is the susceptible parent of L693 and L661. Figure S4. Quality map of the sequences flanking the SNP site. A represents L693; B represents YU25; C represents L661; D represents MY11, which showed that the SNP existed among the different genotypes. Figure S5. GISH analysis of mitotic metaphase chromosomes of TAI 7047, L661, L693 and YU25 using genomic DNA of Th. Intermedium as a probe (green). A TAI 7047, B L661, C L693, D YU25. Chromosomes were counterstained with DAPI (blue). Figure S6. Results demonstrating the high quality and good replication of q-PCR. (A), the melt curve of the q-PCR product during the amplification of WCBP2. (B), The amplification curve shows the consistent crossing points of the three technological replicates of the WCBP2 gene and the reference gene GAPDH. This chart shows L661 at 0 h. (ZIP 837 kb