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. 2004 Sep 14;101(37):13554-9.
doi: 10.1073/pnas.0403659101. Epub 2004 Sep 1.

Chromosome Painting Using Repetitive DNA Sequences as Probes for Somatic Chromosome Identification in Maize

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Free PMC article

Chromosome Painting Using Repetitive DNA Sequences as Probes for Somatic Chromosome Identification in Maize

Akio Kato et al. Proc Natl Acad Sci U S A. .
Free PMC article

Abstract

Study of the maize (Zea mays L.) somatic chromosomes (2n = 20) has been difficult because of a lack of distinguishing characteristics. To identify all maize chromosomes, a multicolor fluorescence in situ hybridization procedure was developed. The procedure uses tandemly repeated DNA sequences to generate a distinctive banding pattern for each of the 10 chromosomes. Fluorescence in situ hybridization screening trials of nonsubtracted or subtracted PCR libraries resulted in the isolation of microsatellite 1-26-2, subtelomeric 4-12-1, and 5S rRNA 2-3-3 clones. These three probes, plus centromeric satellite 4 (Cent4), centromeric satellite C (CentC), knob, nucleolus-organizing region (NOR), pMTY9ER telomere-associated sequence, and tandemly repeated DNA sequence 1 (TR-1) were used as a mixture for hybridization to root-tip chromosomes. All 10 chromosomes were identified by the banding and color patterns in the 14 examined lines. There was significant quantitative variation among lines for the knob, microsatellite, TR-1, and CentC signals. The same probe mixture identifies meiotic pachytene, late prophase I, and metaphase I chromosomes. The procedure could facilitate the study of chromosomal structure and behavior and be adapted for other plant species.

Figures

Fig. 1.
Fig. 1.
Probe localization on maize Oh43 root-tip chromosomes. (a) Microsatellite 1-26-2 clone (red), CentC (green), TR-1 (white), and knob 180-bp (blue) signals. (b) The 5S 2-3-3 clone (yellow), Cent4 (red), NOR-173 clone (green), and knob180-bp (blue) signals. (c) pMTY9ER telomere-associated sequence (red), subtelomeric 4-12-1 clone (green), and knob 180-bp (blue) signals. (d) The signals of all nine probes. (Scale bar, 10 μm.)
Fig. 2.
Fig. 2.
Somatic-chromosome identification in four maize inbred lines probed with the FISH mixture described in the text. (Scale bar, 10 μm.)
Fig. 3.
Fig. 3.
FISH signals on maize Oh43 meiotic cells. (Upper) Late prophase I. All 10 chromosome pairs are identifiable. (Lower) Metaphase I. Chromosomes are identifiable from the signal combinations. (Scale bar, 10 μm.)
Fig. 4.
Fig. 4.
Somatic chromosome karyotyping of 14 maize lines probed with the FISH mixture. Knob 180-bp repeat (blue), 5S 2-3-3 (yellow, 2L), NOR-173 clone (green, 6S), CentC (green), subtelomeric 4-12-1 clone (green), Cent4 (red, 4C), microsatellite 1-26-2 clone (red), pMTY9ER telomere-associated sequence (red), and TR-1 (white). (Scale bar, 10 μm.)

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