Annals of Botany 87: 719-728, 2001
© 2001 Annals of Botany Company
Plant Genome Size Measurement with DNA Image Cytometry
University of Ljubljana, Biotechnical Faculty, Department of Biology, Ve
na pot 111, 1000, Ljubljana, Slovenia Institute of Botany, University of Vienna, Rennweg 14, A-1030, Vienna, Austria
Received: 16 October 2000 ; Returned for revision: 12 December 2000 . Accepted: 4 February 2001
To test the reliability of DNA image cytometry for the measurement of nuclear DNA content in plant material, we conducted independent experiments in two laboratories using different image analysis instruments for densitometric measurement of nuclear DNA amount in Feulgen-stained squash preparations of root tips. The 2C nuclear DNA content of the nine species studied spanned a 100-fold range (approx. 0.3–33 pg). The estimates of nuclear DNA content measured with image cytometry methods were comparable to values obtained previously using both photometric cytometry and flow cytometry. Image cytometry methods showed little variation among repeated experiments within each laboratory or among different operators using the same instrument. Furthermore, the interphase-peak method (measurement of several hundred interphase nuclei per slide) was comparable to the classical prophase/telophase approach (measurement of ten early prophase and ten late telophase nuclei per slide). Hence, DNA image cytometry gives accurate and reproducible results and may be used as an alternative to photometric cytometry in plant nuclear DNA content measurements. In the present study, we propose that two standards for quality control of nuclear DNA content measurement are used in plant DNA image cytometry: (1) the coefficient of variation of the peak should be lower than 6%, and (2) the 4C/2C ratio should be between 1.9 and 2.1.Copyright 2001 Annals of Botany Company
DNA image cytometry, nuclear DNA content, genome size, quality control standards for slides, prophase/telophase method, interphase-peak method
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