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.
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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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Acknowledgments: This research was financed by the Ministry of Science and Higher Education of the Republic of Poland.
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Nowicka, A., Grzebelus, E. & Grzebelus, D. Precise karyotyping of carrot mitotic chromosomes using multicolour-FISH with repetitive DNA. Biol Plant 60, 25–36 (2016). https://doi.org/10.1007/s10535-015-0558-2
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DOI: https://doi.org/10.1007/s10535-015-0558-2