PMC

Specific chromosomes and fluorescent bands are indicated by numbers and arrows, respectively. Metaphase chromosomes of: (a) D. villosum Bomarzo accession and (b) D. villosum ‘Greek’ accession, both after ND-FISH (GAA)₇−FITC labeling (green fluorescence) and DAPI staining (blue fluorescence) are shown. All D. villosum Bomarzo chromosomes display a characterizing banding pattern while D. villosum ‘Greek’ accession chromosomes 3 V and 4 V differ in the GAA hybridization pattern, with respect to the Bomarzo accession. (c) Identification of chromosome 1 V in metaphase chromosomes of D. villosum Bomarzo after double target FISH with pTa71-Cy3 (red fluorescence) and (GAA)₇−FITC. (d) Identification of chromosomes 4 V with double target FISH labeling with pSc119.2-Cy3 and (GAA)₇−FITC oligonucleotide to D. villosum Bomarzo metaphase spreads. The pSc119.2 probe hybridized to the 4VS arm only. (e) Metaphase chromosomes of D. villosum Bomarzo after double FISH with pHv62-Cy3 and (GAA)₇−FITC oligonucleotide; chromosome 7 V has been identified by pHv62 labeling. (f) Metaphase chromosomes of T. aestivum CS-6V D. villosum addition line after double FISH labeling of pHv62-Cy3 and (GAA)₇-FITC oligonucleotide. The D. villosum 6 V chromosome added to T. aestivum CS standard complement has been identified by pHv62, a V genome specific probe.

FISH analysis using the modified ONPM#4 multiplex probe and chromosome-specific molecular markers in progeny of the newly developed durum ‘Zhongyin 1286’–D. villosum amphiploid. (A) Karyotypes of spontaneous durum wheat–D. villosum amphiploids of ‘Zhongyin1286’/GRA2712 F₁. Probe colours are as in . Red boxes indicate wheat or D. villosum chromosome variation; 20WN38-2 is durum wheat–D. villosum Amphiploid #1; 20WN88 are 20WN89 are new amphiploids. (B) PCR analysis of D. villosum chromosome-specific markers in spontaneous durum wheat–D. villosum amphiploids. M, DL2000; 1, ‘Chinese Spring’; 2, durum wheat–D. villosum Amphiploid #1; 3, 20WN88-1; 4, 20WN88-2; 5, 20WN88-3; 6, 20WN88-4; 7, 20WN88-5; 8, 20WN89-1; 9, 20WN89-2; 10, 20WN89-3. Arrows indicate D. villosum chromosome-specific markers.

Screening of six wheat–D. villosum 6V#3L recombinants by combining molecular markers and GISH analyses. (A) Electrophoresis patterns of proximal 6V#3L-specific markers BE422631/HaeIII; (B) Electrophoresis patterns of distal 6V#3L-specific markers 6L11/MboI; (C) GISH analyses of wheat–D. villosum 6V#3L recombinants. D. villosum chromosomal segments were in green, while wheat chromosomes were in blue counterstained by DAPI. Lanes: M, 100 bp DNA Ladder; 1, CS; 2, CS–D. villosum 6V#3 disomic addition line TA7682; 3, CS–D. villosum T6AL·6V#3S translocation line TA5618; 4, CS–D. villosum T6AS·6V#3L translocation line TA5617 (Sr52 stock); 5–10, the newly developed CS–D. villosum 6V#3L translocation lines 1386, 1382, 1385, 1381, 1380, and 1392. Polymorphic bands of 6V#3L markers are indicated by arrows.

a, Wild A. Villosum; b, A. Villosum in Artificial cultivating; c, A. Villosum in Natural fostering