Chromosome Research

, Volume 21, Issue 1, pp 75–85 | Cite as

An assessment of karyotype restructuring in the neoallotetraploid Tragopogon miscellus (Asteraceae)

  • Michael Chester
  • Malorie J. Lipman
  • Joseph P. Gallagher
  • Pamela S. Soltis
  • Douglas E. Soltis
Article

Abstract

Tragopogon miscellus and Tragopogon mirus are two rare examples of allopolyploids that have formed recently in nature. Molecular cytogenetic studies have revealed chromosome copy number variation and intergenomic translocations in both allotetraploids. Due to a lack of interstitial chromosome markers, there remained the possibility of additional karyotype restructuring in these neopolyploids, via intrachromosomal and intragenomic rearrangements. To address this issue, we searched for additional high-copy tandem repeats in genomic sequences of the diploid progenitor species—Tragopogon dubius, Tragopogon pratensis and Tragopogon porrifolius—for application to the chromosomes of the allotetraploids. Eight novel repeats were localised by fluorescence in situ hybridisation (FISH) in the diploids; one of these repeats, TTR3, provided interstitial coverage. TTR3 was included in a cocktail with other previously characterised probes, producing better-resolved karyotypes for the three diploids. The cocktail was then used to test a hypothesis of karyotype restructuring in the recent allotetraploid T. miscellus by comparing repeat distributions to its diploid progenitors, T. dubius and T. pratensis. Five individuals of T. miscellus were selected from across the range of karyotypic variation previously observed in natural populations. FISH signal distributions mostly matched those observed in the diploid progenitors, with the exception of several losses or gains of signal at chromosomal subtermini and previously noted intergenomic translocations. Thus, in T. miscellus, we find most changes restricted to the subterminal regions, and although some were recurrent, none of the changes were common to all individuals analysed. We consider these findings in relation to the gene loss reported previously for T. miscellus.

Keywords

polyploidy translocation homeologous substitution FISH 

Abbreviations

Cy3

Cyanine 3

Cy5

Cyanine 5

DAPI

4′,6-Diamidino-2-phenylindole

dsDNA

Double-stranded deoxyribonucleic acid

FISH

Fluorescence in situ hybridisation

GISH

Genomic in situ hybridisation

NGS

Next-generation sequencing

rDNA

Ribosomal DNA

TGP7

Tragopogon porrifolius subtelomeric repeat-7

TPRMBO

Tragopogon pratensis tandemly repetitive sequence MboI-rich

TR

Tandem repeat

TTR

Tragopogon tandem repeat

Supplementary material

10577_2013_9339_Fig4_ESM.jpg (16 kb)
Fig. S1

Bar chart showing the percentage abundance of the 40 most common tandem repeats. (JPEG 16 kb)

10577_2013_9339_MOESM1_ESM.tif (564 kb)
High resolution image(TIFF 563 kb)
10577_2013_9339_Fig5_ESM.jpg (44 kb)
Fig. S2

FISH applied to mitotic chromosomes of T. dubius, T. porrifolius and T. pratensis with probes to a dub198; b pra001; c TTR4; d TTR6; e 5S rDNA. In c, arrows indicate hybridisation to minor 35S rDNA sites, on the short arm of chromosome DPo. Scale bar 5 μm. (JPEG 43 kb)

10577_2013_9339_MOESM2_ESM.tif (634 kb)
High resolution image(TIFF 634 kb)
10577_2013_9339_Fig6_ESM.jpg (6 kb)
Fig. S3

FISH reprobing with 5S rDNA probe (magenta) merged with the same metaphase chromosomes as shown in Fig. 2d of T. porrifolius. Arrows indicate 5S rDNA array positions. Scale bar 5 μm. (JPEG 5 kb)

10577_2013_9339_MOESM3_ESM.tif (139 kb)
High resolution image(TIFF 139 kb)
10577_2013_9339_MOESM4_ESM.doc (62 kb)
Table S1Repeat monomer consensus sequences (DOC 62 kb)
10577_2013_9339_MOESM5_ESM.doc (32 kb)
Table S2PCR primer sequences and cloned probe sequences (DOC 32 kb)
10577_2013_9339_MOESM6_ESM.doc (46 kb)
Table S3Complementary oligonucleotide probe template sequences (DOC 46 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Michael Chester
    • 1
  • Malorie J. Lipman
    • 1
  • Joseph P. Gallagher
    • 1
    • 2
  • Pamela S. Soltis
    • 3
  • Douglas E. Soltis
    • 1
    • 3
  1. 1.Department of BiologyUniversity of FloridaGainesvilleUSA
  2. 2.Department of Ecology, Evolution, and Organismal BiologyIowa State UniversityAmesUSA
  3. 3.Florida Museum of Natural HistoryUniversity of FloridaGainesvilleUSA

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