Figure 1.
The isolation of alien chromosomes in TAi-27 by a micromanipulator.
a Mitosis metaphase chromosomes of a root tip cell before chromosome isolation (Arrows show the smallest chromosome). b Metaphase chromosomes after isolation of the chromosome indicated by arrows in (a) by a glass needle. Bar = 10 µm.
Figure 2.
GISH and chromosome painting of mitotic metaphase chromosomes of TAi-27 and Th. intermedium.
a Identification of alien chromosomes in TAi-27 by GISH with the probe, Chroma Tide Alexa Fluor 488-5-dUTP-labeled genomic DNA of Th. Intermedium (blocked with genomic DNA of “Chinese Spring”). b Identification of alien chromosomes in TAi-27 by chromosome painting with the probe, Chroma Tide Alexa Fluor 488-5-dUTP-labeled DOP-PCR products of the microdissected chromosome in TAi-27. c Chromosome painting of Th. intermedium chromosomes using Chroma Tide Alexa Fluor 488-5-dUTP-labeled DOP-PCR products of the microdissected chromosome in TAi-27 as the probe. The alien chromosomes in TAi-27 are indicated by arrows and enlarged on the right corner in (a) and (b), respectively. Bar = 10 µm.
Figure 3.
Sequential FISH and GISH on the root-tip cells at mitotic metaphase in Th. intermedium.
a The hybridization results using the DOP-PCR products of the smallest alien chromosome of TAi-27 (green) as the probe. b The hybridization results using genomic DNA from Ps. spicata (St genome, red) as the probe. C The merged figure of (a) and (b). Bar = 10 µm.
Figure 4.
FISH patterns on the root tip cells at mitotic metaphase in Th. intermedium using (a) the DOP-PCR products of the smallest alien chromosome of TAi-27 (green) and (b) pDbH12 (Js genome specific probe, red) as the probe.
C The merged figure of (a) and (b). Bar = 10 µm.
Figure 5.
The karyotype of chromosomes of Th. intermedium from Figure 4c.
Figure 6.
Ideogram showing how the pair of chromosomes from Th. intermedium changed into the smallest submetacentric chromosome in TAi-27.
St chromosome breakages occurred, and only the centromeric area remained. A new submetacentric chromosome was subsequently formed.