Reliable flow cytometric estimation of nuclear DNA content in coffee trees
Flow cytometry gave high resolution of genome size in two coffee taxa (Coffea liberica dewevrei and C. pseudozanguebariae). Propidium iodide (PI) and Petunia hybrida were used as dye and internal standard, respectively. Proportionality between the DNA content and the digitized fluorescence signal was checked. Five main results were evident. First, optimal experimental conditions were established for peak location estimation (mean or mode), staining time (at least 2 minutes), high voltage (557 V) for the photomultiplier tube (PMT), and PI concentration (333 ug/ml). Second, a parameterization of the effects of high voltage and PI concentration were deduced from curve fitting. Third, two biases in DNA content estimation were recorded for high voltage and PI concentration, and were minimized. Fourth, the genome sizes of C. liberica dewevrei and C. pseudozanguebariae were estimated with accuracy 2C=1.421 más menos 0.005 pg and 2c=1.129 más menos 0.005 pg, respectively. Fifth, between-genotype variance was emphasized within each taxon.
| Main Authors: | , , , , , |
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| Format: | biblioteca |
| Published: |
Montpellier (Francia)
1994
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| Subjects: | COFFEA LIBERICA, COFFEA EXCELSA, COFFEA ZANGUEBARIAE, ADN, ESTIMACION, |
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