Table 1.
Wheat cytogenetic stocks used in this study.
Table 2.
Wheat genome specific EST markers used in this study.
Fig 1.
Detection of CENH3 and CRWs on mitotic chromosome of Chinese Spring (CS) wheat and the derived 4D telosomic stocks; a: the insert shows chromosome 4D probed with CENH3 (green) and CRWs (red), straightening was performed using the ‘straighten-curved-objects’ command of the Image J software, the signals are clearly separated from each other; Relative CENH3 signal intensity of b: t4DS; c: t4DL; f. dDt4D; Chromosome 4D with d: CENH3 (white arrow) and e: CRWs (red arrow) signals.
Fig 2.
Probing of t1DS telosomes present in the Dt1DS stock with a: CENH3, b: CRWs, c: pAs1, d: 1S-3 (red dots) and pAs1 (green), e: 1S-1 (red dots) and pAs1 (green); Probing of 1DS telosomes present in the dDt1DS stock with f: CENH3, g: pAs1, h: 1S-1 (not detected), and i: 1S-3 (red dots) and CRWs (faint green signals).
Simultaneous detection of CENH3, CRWs and pAs1 on dDt1DS also provided in S3 Fig. Ideograms depicting localization of each probe on telosomes, Dt1DS and dDt1DS (j-k, and S3 Fig). Possible scenario of the origin of the chromosomal rearrangements observed in the dDt1DS: (1) chromatin breakage, (2) loss of original centromere, (3) de novo formation of a centromere, (4) paracentric inversion, (5) pericentric inversion (s).
Fig 3.
Immuno-FISH based karyotype of D-genome chromosomes of wheat and their derived telosomes using CENH3 (white), CRWs (red) and pAs1 (green) as probes.
CRWs (red signals) co-localized with CENH3 (white signals) in most of the chromosome except dDt1DS, 4D, Dt4DS, dDt4DS, Dt5DL and dDt5DL. The centromeric regions of chromosome or chromosome arm were seen as pinkish red colors because the CRWs (red signals) are abundant in centromeric region and much brighter than CENH3 signals except in the above mentioned telosomes. The dDt1DS stock contained multiple chromosome rearrangement including inversion, deletion and centromere shift. Note that the CRWs were not detected in Dt4DS and dDt4DS, instead the pAs1 signal was overlapped with the CENH3 signal in these telosomes. A very faint pAs1 FISH site was detected in the terminal region of dDt6DS, indicating a terminal deletion. Short arm and long arm telosomes present in the ditelosomic stocks are represented as (DtS) and (DtL), respectively and short arm and long arm telosomes present in the double ditelosomic stocks are represented as (dDtS) and (dDtL), respectively.
Fig 4.
Localization of CENH3 (arrow in a), CRWs (arrow in b) and pAs1 (arrow in c) on Dt6DS; localization of CENH3 (d), CRWs (e) and pAs1 (f) on dDt6D, arrows and arrowhead indicate the 6DS and 6DL telosomes, respectively. Two color detection of single gene probe, 6S-2 and pAs1 on Dt6DS (g) and dDt6DS (h); the hybridization signal for single copy probe 6S-2 (red dots) is indicated by arrows and the pAs1 was labeled with green colors by arrowheads.
Fig 5.
Two color FISH mapping of CRWs (red) and GAAn (green) repeats on mitotic metaphase chromosome of CS containing 28 (24 telosomes from B-genome telosomes and 4 telosomes from t4AS and t4AL).
The signals for CRWs were not detected on two pairs of t1BS and t6BS (arrowheads) (a). Instead GAAn repeats were presented in their centromeric region. Arrows indicate the t4AS (a). Arrows and arrowheads indicate the t1BS and t6BS, respectively (b).