Biologia Plantarum

, Volume 52, Issue 3, pp 445–452

Localization of BAC clones on mitotic chromosomes of Musa acuminata using fluorescence in situ hybridization

Original Papers

DOI: 10.1007/s10535-008-0089-1

Cite this article as:
Hřibová, E., Doleželová, M. & Doležel, J. Biol Plant (2008) 52: 445. doi:10.1007/s10535-008-0089-1

Abstract

A bacterial artificial chromosome (BAC) library of banana (Musa acuminata) was used to select BAC clones that carry low amounts of repetitive DNA sequences and could be suitable as probes for fluorescence in situ hybridization (FISH) on mitotic metaphase chromosomes. Out of eighty randomly selected BAC clones, only one clone gave a single-locus signal on chromosomes of M. acuminata cv. Calcutta 4. The clone localized on a chromosome pair that carries a cluster of 5S rRNA genes. The remaining BAC clones gave dispersed FISH signals throughout the genome and/or failed to produce any signal. In order to avoid the excessive hybridization of repetitive DNA sequences, we subcloned nineteen BAC clones and selected their ‘low-copy’ subclones. Out of them, one subclone gave specific signal in secondary constriction on one chromosome pair; three subclones were localized into centromeric and peri-centromeric regions of all chromosomes. Other subclones were either localized throughout the banana genome or their use did not result in visible FISH signals. The nucleotide sequence analysis revealed that subclones, which localized on different regions of all chromosomes, contained short fragments of various repetitive DNA sequences. The chromosome-specific BAC clone identified in this work increases the number of useful cytogenetic markers for Musa.

Additional key words

cytogenetic mappingchromosome structurerepetitive DNAribosomal DNAsubcloning

Abbreviations

BAC

bacterial artificial chromosome

FISH

fluorescence in situ hybridization

DAPI

4′,6-diamidino-2-phenylindole

FITC

fluorescein isothiocyanate

PFGE

pulsed field gel electrophoresis

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  1. 1.Laboratory of Molecular Cytogenetics and CytometryInstitute of Experimental BotanyOlomoucCzech Republic
  2. 2.Department of Cell Biology and GeneticsPalacký UniversityOlomoucCzech Republic