Fig 1.
FISH pattern of CD-3 and the leaf response to stripe rust.
(a) Chromosomes of CD-3 stained with DAPI (blue), Oligo-pTa535 (red) and Oligo-pSc119.2 (green), (b) chromosomes stained by DAPI (blue) and Oligo-pHv62-1(green), (c) chromosomes stained with propidium iodide, PI (red) and oligo-(GAA)7, (d) the FISH karyotype of CD-3, (e) The leaf response to stripe rust of D. villosum, CD-3 and CS.
Fig 2.
PCR products of PLUG markers in wheat-D. villosum alien introgression lines.
(a)-(e) PCR amplification generated by markers TNAC1248, TNAC1294, TNAC1301, TNAC1277 and TNAC1267, respectively; M, Trans2k plus DNA markers (Trangene, Beijing, China); 1, common wheat CS “Chinese Spring”; 2, D. villosum, 3–4, CS-D. villosum 1V -2V additional lines; 5, CD-3; 6–9, CS-D. villosum 4V -7V additional lines. The asterisks indicated the D. villosum-specific bands and the arrows showed the 3D-specific band which was absent in CD-3.
Table 1.
The PLUG primers belonging to Triticeae homoeologous Group 3 used in the study.
Fig 3.
The phenotypic response to stripe rust of D. villosum, CD-3, CS and 3 plants of F2 (CD-3/CS).
(a-c) The response to stripe rust of D.villosum, CD-3 and CS at adult stage. (d) The response to stripe rust of CS, D.villosum, CD-3 and F2 (CD-3/CS) plants at seedling stage. 2–2 was the F2 plant without 3V chromosome, 3–5 was the F2 plant holding one 3V chromosome, and 19–2 was the F2 plant having two 3Vchromosomes. (e-j) The FISH pattern of 2-2(e, f), 3-5(g-h) and 19-2(i-j). Chromosomes were stained by DAPI(blue), Oligo-pHv62-1(white), Oligo-pTa535 (red) and Oligo-pSc119.2 (green). The arrows showed 3V chromosomes.
Fig 4.
PCR products of primer pairs TaGS5-P6 and TaGS5-P7 in CS, CD-3 and D. villosum. M, trans2k plus DNA Marker; 1–3, the bands amplified by TaGS5-P6 in CS, CD-3 and D. villosum, respectively; 4–5, the bands amplified by TaGS5-P7 in CS, CD-3 and D. villosum.
The white arrows showed the D. villosum-specific bands.
Table 2.
Primers used for the identification of GS5 gene.
Fig 5.
Sequence alignment of DvGS5-1 and TaGS5-1.
Arrows showed binding sites of SCAR primer pair DG-3VF and DG-3VR.
Fig 6.
PCR bands generated by DG-3VF and DG-3VR.
1, D. villosum; 2–3, CS -D. villosum 1V-2V additional lines; 4, CD-3; 5–8, CS-D. villosum 4V-7V additional lines; 9–13, common wheat cultivars CS, Chuanmai49, Chuanmai50, Chuanmai60, and rye cultivar JZHM. White arrows indicated the 3V chromosome-specific marker DvGS5-1443.