Development of flow cytogenetics and physical genome mapping in chickpea (Cicer arietinum L.)
Language English Country Netherlands Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Cell Cycle MeSH
- Chromosomes, Plant genetics MeSH
- Cicer genetics MeSH
- Cytogenetics MeSH
- DNA, Plant genetics metabolism MeSH
- Physical Chromosome Mapping methods MeSH
- Genetic Linkage MeSH
- Genome, Plant * MeSH
- In Situ Hybridization, Fluorescence MeSH
- Indoles MeSH
- Karyotyping MeSH
- Plant Roots genetics MeSH
- Metaphase MeSH
- Microsatellite Repeats MeSH
- Sequence Tagged Sites MeSH
- Mitosis MeSH
- Polymerase Chain Reaction MeSH
- Flow Cytometry methods MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DAPI MeSH Browser
- DNA, Plant MeSH
- Indoles MeSH
Procedures for flow cytometric analysis and sorting of mitotic chromosomes (flow cytogenetics) have been developed for chickpea (Cicer arietinum). Suspensions of intact chromosomes were prepared from root tips treated to achieve a high degree of metaphase synchrony. The optimal protocol consisted of a treatment of roots with 2 mmol/L hydroxyurea for 18 h, a 4.5-h recovery in hydroxyurea-free medium, 2 h incubation with 10 micromol/L oryzalin, and ice-water treatment overnight. This procedure resulted in an average metaphase index of 47%. Synchronized root tips were fixed in 2% formaldehyde for 20 min, and chromosome suspensions prepared by mechanical homogenization of fixed root tips. More than 4 x 10(5) morphologically intact chromosomes could be isolated from 15 root tips. Flow cytometric analysis of DAPI-stained chromosomes resulted in histograms of relative fluorescence intensity (flow karyotypes) containing eight peaks, representing individual chromosomes and/or groups of chromosomes with a similar relative DNA content. Five peaks could be assigned to individual chromosomes (A, B, C, G, H). The parity of sorted chromosome fractions was high, and chromosomes B and H could be sorted with 100% purity. PCR on flow-sorted chromosome fractions with primers for sequence-tagged microsatellite site (STMS) markers permitted assignment of the genetic linkage group LG8 to the smallest chickpea chromosome H. This study extends the number of legume species for which flow cytogenetics is available, and demonstrates the potential of flow cytogenetics for genome mapping in chickpea.
See more in PubMed
Planta. 1992 Aug;188(1):93-8 PubMed
Methods Cell Sci. 2001;23(1-3):71-82 PubMed
Mol Gen Genet. 1994 Mar;242(5):551-8 PubMed
Chromosoma. 2000 Nov;109(7):482-9 PubMed
World J Microbiol Biotechnol. 1995 Jul;11(4):438-48 PubMed
Chromosome Res. 1999;7(6):431-44 PubMed
Methods Cell Biol. 2001;64:3-31 PubMed
Genetics. 2001 Jul;158(3):1269-77 PubMed
Mol Gen Genet. 1999 Aug;262(1):90-101 PubMed
Cytometry. 2000 Oct 1;41(2):102-8 PubMed
Theor Appl Genet. 1993 Feb;85(6-7):665-72 PubMed
Theor Appl Genet. 1994 Mar;87(8):893-9 PubMed
Theor Appl Genet. 1996 May;92(6):744-51 PubMed
Chromosome Res. 2002;10(1):63-71 PubMed
Biotechniques. 1998 Jul;25(1):32-4, 36, 38 PubMed
Chromosome Res. 1993 Jul;1(2):107-15 PubMed
Cell. 1988 Apr 8;53(1):127-36 PubMed
Mol Cells. 1999 Aug 31;9(4):436-9 PubMed
Genome. 1996 Apr;39(2):258-65 PubMed
Chromosome Res. 1996 Nov;4(7):531-9 PubMed
Genome. 1999 Apr;42(2):210-7 PubMed
Genome. 1997 Oct;40(5):633-8 PubMed
Genome. 1994 Aug;37(4):713-6 PubMed
Genetics. 2000 Dec;156(4):2033-41 PubMed
Genome. 1994 Aug;37(4):656-63 PubMed
Chromosome Res. 1998 Feb;6(2):97-104 PubMed
Methods Cell Sci. 1999;21(2-3):95-107 PubMed
Science. 2000 Jun 2;288(5471):1602-3 PubMed
Genome. 1996 Aug;39(4):697-703 PubMed
Theor Appl Genet. 1998 Mar;96(3-4):348-53 PubMed
Chromosome analysis and sorting
Chromosomes in the flow to simplify genome analysis
Coupling amplified DNA from flow-sorted chromosomes to high-density SNP mapping in barley
Chromosome sorting in tetraploid wheat and its potential for genome analysis
Flow cytogenetics and plant genome mapping
