The determination of genome size in male and female germ cells of Drosophila melanogaster by DNA-feulgen cytophotometry

Mulligan, Pamela K.; Rasch, Ellen M.

doi:10.1007/bf00493241

Cited by 37 publications

(18 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is remarkable that B chromosomes in L. migratoria do contain as much DNA as the C-value in Tribolium albax (0.16 pg) [Alvarez-Fuster et al, 2009] or Drosophila melanogaster (0.18 pg) [Rasch et al, 1971;Mulligan and Rasch, 1980;Adams et al, 2000]. Likewise, B chromosomes in E. plorans contain DNA amounts comparable to the C-value in the beetle Tenebrio molitor (0.52 pg) [Juan and Petitpierre, 1989], or the worm Lumbricus terrestris (0.60 pg) .…”

Section: Resultsmentioning

confidence: 99%

DNA Amount of X and B Chromosomes in the Grasshoppers Eyprepocnemis plorans and Locusta migratoria

Ruiz‐Ruano

Ruiz-Estévez

Rodriguez-Perez

et al. 2011

Cytogenet Genome Res

View full text Add to dashboard Cite

We analyzed the DNA amount in X and B chromosomes of 2 XX/X0 grasshopper species (Eyprepocnemis plorans and Locusta migratoria), by means of Feulgen image analysis densitometry (FIAD), using previous estimates in L. migratoria as standard (5.89 pg). We first analyzed spermatids of 0B males and found a bimodal distribution of integrated optical densities (IODs), suggesting that one peak corresponded to +X and the other to –X spermatids. The difference between the 2 peaks corresponded to the X chromosome DNA amount, which was 1.28 pg in E. plorans and 0.80 pg in L. migratoria. In addition, the +X peak in E. plorans gave an estimate of the C-value in this species (10.39 pg). We next analyzed diplotene cells from 1B males in E. plorans and +B males in L. migratoria (a species where Bs are mitotically unstable and no integer B number can be defined for an individual) and measured B chromosome IOD relative to X chromosome IOD, within the same cell, taking advantage of the similar degree of condensation for both positively heteropycnotic chromosomes at this meiotic stage. From this proportion, we estimated the DNA amount for 3 different B chromosome variants found in individuals from 3 E. plorans Spanish populations (0.54 pg for B1 from Saladares, 0.51 pg for B2 from Salobreña and 0.64 for B24 from Torrox). Likewise, we estimated the DNA amount of the B chromosome in L. migratoria to be 0.15 pg. To automate measurements, we wrote a GPL3 licensed Python program (pyFIA). We discuss the utility of the present approach for estimating X and B chromosome DNA amount in a variety of situations, and the meaning of the DNA amount estimates for X and B chromosomes in these 2 species.

show abstract

Section: Resultsmentioning

confidence: 99%

DNA Amount of X and B Chromosomes in the Grasshoppers Eyprepocnemis plorans and Locusta migratoria

Ruiz‐Ruano

Ruiz-Estévez

Rodriguez-Perez

et al. 2011

Cytogenet Genome Res

View full text Add to dashboard Cite

show abstract

“…Fluorescent flow cytometry can accurately estimate genome size: As genome size estimates were previously available for D. melanogaster (Laird 1971;Rasch et al 1971;Kavenoff and Zimm 1973;Mulligan and Rasch 1980;Celniker et al 2002;Bennett et al 2003) and D. virilis (Kavenoff and Zimm 1973;Laird 1973), we assessed the ability of PI and DAPI flow cytometry to accurately reproduce the previously described genome size differences for these two species. For example, previous estimates described the D. virilis genome to be much larger than D. melanogaster and to have a higher heterochromatin content (Gall et al 1971;Schweber 1974).…”

Section: Resultsmentioning

confidence: 99%

“…(Laird 1971(Laird , 1973Rasch et al 1971;Kavenoff and Zimm 1973;Mulligan and Rasch 1980;Celniker et al 2002;Hoskins et al 2002;Bennett et al 2003). Two best-fit curves (one for PI and another for DAPI) were obtained, which then were used to convert 2C measurements into megabase values.…”

Section: Methodsmentioning

confidence: 99%

Analysis of Drosophila Species Genome Size and Satellite DNA Content Reveals Significant Differences Among Strains as Well as Between Species

et al. 2007

View full text Add to dashboard Cite

The size of eukaryotic genomes can vary by several orders of magnitude, yet genome size does not correlate with the number of genes nor with the size or complexity of the organism. Although ''whole''-genome sequences, such as those now available for 12 Drosophila species, provide information about euchromatic DNA content, they cannot give an accurate estimate of genome sizes that include heterochromatin or repetitive DNA content. Moreover, genome sequences typically represent only one strain or isolate of a single species that does not reflect intraspecies variation. To more accurately estimate whole-genome DNA content and compare these estimates to newly assembled genomes, we used flow cytometry to measure the 2C genome values, relative to Drosophila melanogaster. We estimated genome sizes for the 12 sequenced Drosophila species as well as 91 different strains of 38 species of Drosophilidae. Significant differences in intra-and interspecific 2C genome values exist within the Drosophilidae. Furthermore, by measuring polyploid 16C ovarian follicle cell underreplication we estimated the amount of satellite DNA in each of these species. We found a strong correlation between genome size and amount of satellite underreplication. Addition and loss of heterochromatin satellite repeat elements appear to have made major contributions to the large differences in genome size observed in the Drosophilidae.

show abstract

“…), invertebrate hemocytes and sperm do appear suitable for comparison with chicken erythrocytes. The classic example of this is the remarkably accurate estimate of the genome size of Drosophila melanogaster by Ellen Rasch, using sperm and hemocyte nuclei against a chicken erythrocyte standard, made 30 years before genome sequencing confirmed its size with certainty (Rasch et al 1971;Mulligan and Rasch 1980;Adams et al 2000).…”

Section: Additional Sources Of Errormentioning

confidence: 99%

From Pixels to Picograms

Hardie

Gregory

Hebert

2002

J Histochem Cytochem.

228

View full text Add to dashboard Cite

S U M M A R YThe study of genome size variation is important from a number of practical and theoretical perspectives. For example, the long-standing "C-value enigma" relating to the more than 200,000-fold range in eukaryotic genome sizes is best studied from a broad comparative standpoint. Genome size data are also required in detailed analyses of genome structure and evolution. The choice of future genome sequencing projects will be dependent on knowledge regarding the sizes of genomes to be sequenced, and so on. To date, genome size data have been acquired primarily by Feulgen microdensitometry or flow cytometry. Each has several advantages but also important limitations. In this review, we provide a practical guide to the new technique of Feulgen image analysis densitometry. The review is designed for those interested in genome size measurements but not extensively experienced in histochemistry, densitometry, or microscopy. Therefore, relevant historical and technical background information is included. For easy reference, we provide recipes for required reagents, guidelines for cell staining, and a checklist of steps for successful image analysis. We hope that the accuracy, rapidity, and cost-effectiveness of Feulgen image analysis demonstrated here will stimulate further surveys of genome sizes in a variety of taxa.

show abstract

The determination of genome size in male and female germ cells of Drosophila melanogaster by DNA-feulgen cytophotometry

Abstract: The amounts of DNA in haploid and diploid cells of Drosophila melanogaster have been determined by DNA-Feulgen cytophotometry, using Xenopus laevis erythrocyte nuclei as a reference standard. The haploid male genome is estimated to be 0.18 pg DNA and the haploid female genome, 0.20 pg DNA.

Cited by 37 publications

References 27 publications

DNA Amount of X and B Chromosomes in the Grasshoppers <i>Eyprepocnemis plorans</i> and <i>Locusta migratoria</i>

DNA Amount of X and B Chromosomes in the Grasshoppers <i>Eyprepocnemis plorans</i> and <i>Locusta migratoria</i>

Analysis of Drosophila Species Genome Size and Satellite DNA Content Reveals Significant Differences Among Strains as Well as Between Species

From Pixels to Picograms

Contact Info

Product

Resources

About