Plant Genome Size Measurement with DNA Image Cytometry |
| |
Affiliation: | 1. Department of Biology, Biotechnical faculty, University of Ljubljana, SI-1000 Ljubljana, Slovenia;2. Department of Molecular and Biomedical Sciences, Jožef Stefan Institute, SI-1000 Ljubljana, Slovenia;1. Department of Biotechnology, K L University, Vaddesweram, Guntur, Andhra Pradesh, India;2. Institute of Biotechnology, Prof. Jaya Shankar Telangana State Agricultural University, Hyderabad, India |
| |
Abstract: | 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. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|