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Theoretical and Applied Genetics

, Volume 107, Issue 6, pp 988–996 | Cite as

Molecular cytogenetic analysis of genome structure in Lupinus angustifolius and Lupinus cosentinii

  • Inga Hajdera
  • Dorota Siwinska
  • Robert Hasterok
  • Jolanta Maluszynska
Article
First Online:

Abstract

Molecular cytogenetic analysis of Lupinus angustifolius and Lupinus cosentinii was performed using flow cytometry, fluorescence in situ hybridisation (FISH) and differential chromosome staining. Genome size was determined as 2.07 pg for L. angustifolius and 1.54 pg for L. cosentinii. Analysis of nuclear DNA amount in cells during plant development has shown endopolyploidisation in different organs. The highest level of endopolyploidy was in cotyledons and reached 32C in L. angustifolius and 64C in L. cosentinii. Both of the investigated Lupinus species belong to the polysomatic type of plants. Double FISH with rDNA probes provided chromosomal landmarks for 10 out of 40 chromosomes for L. angustifolius and 8 out of 32 chromosomes for L. cosentinii. In L. angustifolius, the number and localisation of 25S rDNA hybridisation signals precisely corresponded to the chromomycin A3 (CMA+) bands, while in L. cosentinii both 25S and 5S rDNA loci overlapped with CMA+ bands. Silver staining revealed that only 45S rRNA genes located in secondary constriction regions were transcriptionally active. FISH with Arabidopsis-type telomeric arrays revealed the presence of signals at termini of all chromosomes. Despite the application of different DNA probes for FISH and different chromosome staining, a relatively small proportion of chromosomes in the Lupinus karyotypes can be distinguished. Identification of all chromosomes requires the use of more chromosome-specific markers.

Keywords

FISH Flow cytometry Lupinus rDNA Telomere 
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Notes

Acknowledgements

The authors thank Prof. Andrzej Legocki (Institute of Bioorganic Chemistry, Poznan, Poland) and Prof. Ewa Sawicka-Sienkiewicz (University of Agriculture, Wroclaw, Poland) for providing seeds of Lupinus species, Dr. Jaroslav Dolezel (Institute of Experimental Botany, Olomouc, Czech Republic) for valuable advice on flow cytometry measurement and providing seeds for DNA standards and Dr. Tim Langdon (University of Wales, Aberystwyth, UK) for providing telomeric sequences. The experiments conducted conformed to accepted practices in Poland.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Inga Hajdera
    • 1
  • Dorota Siwinska
    • 1
  • Robert Hasterok
    • 1
  • Jolanta Maluszynska
    • 1
  1. 1.Department of Plant Anatomy and Cytology, University of Silesia, Jagiellonska 28, 40-032 Katowice, Poland

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