Reliable flow cytometric estimation of nuclear DNA content in coffee trees

✍ Scribed by Philippe Barre; Michel Noirot; Jacques Louarn; Christophe Duperray; Serge Hamon

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 659 KB
• Volume: 24
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/(sici)1097-0320(19960501)24:1<32::aid-cyto4>3.0.co;2-k

✦ Synopsis

Flow cytometry gave high resolution of genome size in two coffee taxa (Coffea Iiberica 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 pg/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 minimiz ed. Fourth, the genome sizes of C. liberica dewevrei and C. pseudozanguebariae were estimatedwith accuracy 2C = 1.421 f 0.005 pg and 2C = 1.129 k 0.005 pg, respectively. Fifth, between-genotype variance was emphasized within each taxon. 0 19% Wiley-Liis, Inc.