PMID- 14608406
OWN - NLM
STAT- MEDLINE
DCOM- 20040708
LR  - 20161020
IS  - 0831-2796 (Print)
IS  - 0831-2796 (Linking)
VI  - 46
IP  - 5
DP  - 2003 Oct
TI  - Analysis and sorting of rye (Secale cereale L.) chromosomes using flow cytometry.
PG  - 893-905
AB  - Procedures for chromosome analysis and sorting using flow cytometry (flow 
      cytogenetics) were developed for rye (Secale cereale L.). Suspensions of intact 
      chromosomes were prepared by mechanical homogenization of synchronized root tips 
      after mild fixation with formaldehyde. Histograms of relative fluorescence 
      intensity obtained after the analysis of DAPI-stained chromosomes (flow 
      karyotypes) were characterized and the chromosome content of the DNA peaks was 
      determined. Chromosome 1R could be discriminated on a flow karyotype of S. 
      cereale 'Imperial'. The remaining rye chromosomes (2R-7R) could be discriminated 
      and sorted from individual wheat-rye addition lines. The analysis of lines with 
      reconstructed karyotypes demonstrated a possibility of sorting translocation 
      chromosomes. Supernumerary B chromosomes could be sorted from an experimental rye 
      population and from S. cereale 'Adams'. Flow-sorted chromosomes were identified 
      by fluorescence in situ hybridization (FISH) with probes for various DNA repeats. 
      Large numbers of chromosomes of a single type sorted onto microscopic slides 
      facilitated detection of rarely occurring chromosome variants by FISH with 
      specific probes. PCR with chromosome-specific primers confirmed the identity of 
      sorted fractions and indicated suitability of sorted chromosomes for physical 
      mapping. The possibility to sort large numbers of chromosomes opens a way for the 
      construction of large-insert chromosome-specific DNA libraries in rye.
FAU - Kubalakova, M
AU  - Kubalakova M
AD  - Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental 
      Botany, Olomouc, Czech Republic.
FAU - Valarik, M
AU  - Valarik M
FAU - Barto, J
AU  - Barto J
FAU - Vrana, J
AU  - Vrana J
FAU - Cihalikova, J
AU  - Cihalikova J
FAU - Molnar-Lang, M
AU  - Molnar-Lang M
FAU - Dolezel, J
AU  - Dolezel J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Canada
TA  - Genome
JT  - Genome
JID - 8704544
SB  - IM
MH  - Cell Separation/methods
MH  - Chromosomes, Plant/*genetics
MH  - Flow Cytometry/*methods
MH  - In Situ Hybridization, Fluorescence
MH  - Karyotyping
MH  - Microsatellite Repeats
MH  - Physical Chromosome Mapping
MH  - Polymerase Chain Reaction
MH  - Secale/*genetics
MH  - Translocation, Genetic
EDAT- 2003/11/11 05:00
MHDA- 2004/07/09 05:00
CRDT- 2003/11/11 05:00
PHST- 2003/11/11 05:00 [pubmed]
PHST- 2004/07/09 05:00 [medline]
PHST- 2003/11/11 05:00 [entrez]
AID - g03-054 [pii]
AID - 10.1139/g03-054 [doi]
PST - ppublish
SO  - Genome. 2003 Oct;46(5):893-905. doi: 10.1139/g03-054.