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Chromosome Research

, 17:755 | Cite as

A novel, simple and rapid nondenaturing FISH (ND-FISH) technique for the detection of plant telomeres. Potential used and possible target structures detected

  • Ángeles CuadradoEmail author
  • Hieronim Golczyk
  • Nicolás Jouve
Article

Abstract

We report a new technique—nondenaturing FISH (ND-FISH)—for the rapid detection of plant telomeres without the need for prior denaturation of the chromosomes. In its development, two modified, synthetic oligonucleotides, 21 nt in length, fluorescently labelled at their 5′ and 3′ ends and complementary to either the cytidine-rich (C3TA3) or guanosine-rich (T3AG3) telomeric DNA strands, were used as probes. The high binding affinity of these probes and the short hybridization time required allows the visualization of plant telomeres in less than an hour. In tests, both probes gave strong signals visualized as double spots at both chromosome ends; this was true of both the mitotic and meiotic chromosomes of barley, wheat, rye, maize, Brachypodium distachyon and Rhoeo spathacea. They were also able to detect telomere motifs at certain intercalary sites in the chromosomes of R. spathacea. To investigate the nature of the target structures detected, the chromosomes were treated with RNase A and single strand-specific nuclease S1 before ND-FISH experiments. Signal formation was resistant to standard enzymatic treatment, but sensitive when much higher enzyme concentrations were used. The results are discussed in relation to current knowledge of telomere structure.

Keywords

plant telomere ND-FISH 

Abbreviations

dsDNA

double stranded DNA

ND-FISH

nondenaturing FISH

ssDNA

single-stranded DNA

Notes

Acknowledgements

This study was supported by grants from the Spanish Ministry of Education and Science (AGL2006-09018-C02) and the Polish State Committee for Scientific Research (N301 116 32/4008). The authors thank Adrian Burton for linguistic assistance.

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Ángeles Cuadrado
    • 1
    Email author
  • Hieronim Golczyk
    • 2
  • Nicolás Jouve
    • 1
  1. 1.Department of Cell Biology and GeneticsUniversity of Alcalá de HenaresMadridSpain
  2. 2.Department of Plant Cytology and Embryology, Institute of BotanyJagiellonian UniversityKrakówPoland

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