Short Communication. A robust method for chromosome quantification and ploidy determination in woody species

Gamage, Harshi K. and Schmidt, Susanne (2009) Short Communication. A robust method for chromosome quantification and ploidy determination in woody species. Australian Journal of Botany, 57 2: 87-93. doi:10.1071/BT09006


Author Gamage, Harshi K.
Schmidt, Susanne
Title Short Communication. A robust method for chromosome quantification and ploidy determination in woody species
Journal name Australian Journal of Botany   Check publisher's open access policy
ISSN 0067-1924
Publication date 2009-05
Sub-type Article (original research)
DOI 10.1071/BT09006
Open Access Status
Volume 57
Issue 2
Start page 87
End page 93
Total pages 7
Editor B. Hill
S. Farrer
Place of publication Australia
Publisher C S I R O Publishing
Collection year 2010
Language eng
Subject C1
060702 Plant Cell and Molecular Biology
829899 Environmentally Sustainable Plant Production not elsewhere classified
960806 Forest and Woodlands Flora, Fauna and Biodiversity
070508 Tree Nutrition and Physiology
Formatted abstract
Accurate determination of ploidy level of putative polyploid plants is essential for tree breeding and other applications. Methods for ploidy determination include quantification of chromosome numbers in root-tip cells via light microscopy and indirect assessment via anatomical and morphological traits. Flow cytometry is potentially a high-throughput method to quantify nuclear DNA content; however, it does not allow determining chromosome numbers and interfering compounds often prevent its use. Microscopy-based quantification of chromosomes in active root-tip cells remains the most unambiguous method for ploidy determination, although root tips are difficult to obtain from field-grown plants, and light microscopy can result in insufficient resolution in species with many and small chromosomes. Here, we present a robust technique that uses 2, 4-diamidino-2-phenylindole (DAPI) dye and 1000-fold magnification fluorescence microscopy for quantification of chromosomes in root and shoot tips of woody angiosperms and gymnosperms, and overcomes the reported difficulties. Rather than the conventional tip squashing, spreading tips on glass slides resulted in very good chromosome separation in diverse species, with up to 56 chromosomes and a chromosome size of 2–20 μm. Chromosome counts were performed in diploid Agathis robusta, Elaeocarpus angustifolius, Eucalyptus robusta, Paulownia tomentosa, Pongamia pinnata and Toona ciliata, and di- and tetraploid Acacia crassicarpa and Citrus species.
Q-Index Code C1
Q-Index Status Confirmed Code

 
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Created: Thu, 03 Sep 2009, 08:10:45 EST by Mr Andrew Martlew on behalf of School of Biological Sciences