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Biologia Plantarum

, Volume 60, Issue 1, pp 25–36 | Cite as

Precise karyotyping of carrot mitotic chromosomes using multicolour-FISH with repetitive DNA

  • A. Nowicka
  • E. Grzebelus
  • D. Grzebelus
Original Papers

Abstract

Carrot (Daucus carota L.) chromosomes are small and uniform in shape and length. Here, mitotic chromosomes were subjected to multicolour fluorescence in situ hybridization (mFISH) with probes derived from conserved plant repetitive DNA (18-25S and 5S rDNA, telomeres), a carrot-specific centromeric repeat (Cent-Dc), carrot-specific repetitive elements (DCREs), and miniature inverted-repeat transposable elements (MITEs). A set of major chromosomal landmarks comprising rDNA and telomeric and centromeric sequences in combination with chromosomal measurements enabled discrimination of carrot chromosomes. In addition, reproducible and unique FISH patterns generated by three carrot genome-specific repeats (DCRE22, DCRE16, and DCRE9) and two transposon families (DcSto and Krak) in combination with telomeric and centromeric reference probes allowed identification of chromosome pairs and construction of detailed carrot karyotypes. Hybridization patterns for DCREs were observed as pericentromeric and interstitial dotted tracks (DCRE22), signals in pericentromeric regions (DCRE16), or scattered signals (DCRE9) along chromosomes similar to those observed for both MITE families.

Additional key words

Daucus carota repetitive elements fluorescence in situ hybridization miniature inverted-repeat transposable elements 

Abbreviations

BAC

bacterial artificial chromosome

DAPI

4′,6-diamidino-2-phenylindole

DCRE

Daucus carota repetitive element

FISH

fluorescence in situ hybridization

FITC

fluorescein isothiocyanate

mFISH

multicolour fluorescence in situ hybridization

MITE

miniature inverted-repeat transposable element

NOR

nucleolar organizer region

SSC

saline-sodium citrate

TE

transposable element

TIR

terminal inverted repeat

TRS

telomeric repeat sequence

TSD

target site duplications

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Supplementary material

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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  1. 1.Department of Cell Biology, Franciszek Gorski Institute of Plant PhysiologyPolish Academy of SciencesKrakowPoland
  2. 2.Department of Genetics, Plant Breeding and Seed Science, Institute of Plant Biology and Biotechnology, Faculty of Biotechnology and HorticultureUniversity of Agriculture in KrakowKrakowPoland

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