Genome size estimation: a new methodology

Recently, within the cytogenetic analysis, the evolutionary relations implied in the content of nuclear DNA in plants and animals have received a great attention. The first detailed measurements of the nuclear DNA content were made in the early 40's, several years before Watson and Crick proposed the molecular structure of the DNA. In the following years Hewson Swift developed the concept of "C-value" in reference to the haploid phase of DNA in plants. Later Mirsky and Ris carried out the first systematic study of genomic size in animals, including representatives of the five super classes of vertebrates as well as of some invertebrates. From these preliminary results it became evident that the DNA content varies enormously between the species and that this variation does not bear relation to the intuitive notion from the complexity of the organism. Later, this observation was reaffirmed in the following years as the studies increased on genomic size, thus denominating to this characteristic of the organisms like the "Paradox of the C-value". Few years later along with the no-codification discovery of DNA the paradox was solved, nevertheless, numerous questions remain until nowadays unfinished, taking to denominate this type of studies like the "C-value enigma". In this study, we reported a new method for genome size estimation by quantification of fluorescence fading. We measured the fluorescence intensity each 1600 milliseconds in DAPI-stained nuclei. The estimation of the area under the graph (integral fading) during fading period was related with the genome size.

[1]  J. Macas,et al.  Flow cytogenetics and plant genome mapping , 2004, Chromosome Research.

[2]  Laboratorio de Gen Genome-size variation in bivalve molluscs determined by flow cytometry , 1996 .

[3]  B. Bertino,et al.  A comparative study of DNA content as measured by flow cytometry and image analysis in 1864 specimens. , 1994, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[4]  H. Sawada,et al.  Quantitative Comparison of Anti-Fading Mounting Media for Confocal Laser Scanning Microscopy , 2001, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[5]  J L Haynes,et al.  Principles of flow cytometry. , 1988, Cytometry. Supplement : the journal of the Society for Analytical Cytology.

[6]  A. Komaru,et al.  Androgenetic Reproduction in a Freshwater Diploid Clam Corbicula fluminea (Bivalvia: Corbiculidae) , 2003, Zoological science.

[7]  T. Gregory,et al.  Coincidence, coevolution, or causation? DNA content, cellsize, and the C‐value enigma , 2001, Biological reviews of the Cambridge Philosophical Society.

[8]  W. Allsbrook,et al.  Flow cytometric DNA analysis. , 1990, Clinical laboratory science : journal of the American Society for Medical Technology.

[9]  D. Morse,et al.  Hydrogen peroxide induces spawning in mollusks, with activation of prostaglandin endoperoxide synthetase. , 1977, Science.

[10]  G. D. Johnson,et al.  A simple method of reducing the fading of immunofluorescence during microscopy. , 1981, Journal of immunological methods.

[11]  E J Holborow,et al.  Fading of immunofluorescence during microscopy: a study of the phenomenon and its remedy. , 1982, Journal of immunological methods.

[12]  A. Mirsky,et al.  THE DESOXYRIBONUCLEIC ACID CONTENT OF ANIMAL CELLS AND ITS EVOLUTIONARY SIGNIFICANCE , 1951, The Journal of general physiology.

[13]  T. Gregory Genome size and developmental parameters in the homeothermic vertebrates. , 2002, Genome.

[14]  P. Hebert,et al.  From Pixels to Picograms , 2002, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[15]  T. Hirschfeld Fluorescence background discrimination by prebleaching. , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[16]  T. Gregory Genome size and developmental complexity , 2002, Genetica.

[17]  T. Gregory Genome size estimates for two important freshwater molluscs, the zebra mussel (Dreissena polymorpha) and the schistosomiasis vector snail (Biomphalaria glabrata). , 2003, Genome.

[18]  Schulte Ek Standardization of the Feulgen reaction for absorption DNA image cytometry: a review. , 1991 .

[19]  A. Komaru,et al.  Detection of induced triploidy at different ages for larvae of the Japanese pearl oyster, Pinctada fucata martensii, by microfluorometry with DAPI staining , 1989 .