High-resolution flow cytometry was used to analyze nuclear DNA content in dioecious plants Melandrium album and M. rubrum. Very low coefficients of variation (0.53-0.70%) of G0/1 peaks were obtained after the analysis of nuclei isolated by chopping leaf tissues. In both species, females have lower DNA content than males due to sex chromosome heteromorphism. This made it possible to discriminate peaks belonging to female and male nuclei on the same histogram and permitted unambiguous sex identification in both species. The results indicated the possibility of diagnosing sex also in other heterogametic dioecious plants. The assay described here might be interesting in any area where rapid sex identification is required at an early stage of plant development.

https://onlinelibrary.wiley.com/doi/pdf/10.1002/cyto.990190203

Sex determination in dioecious plants Melandrium album and M. rubrum using high-resolution flow cytometry

The recent development of simple, DNA-based methods for the determination of an individual's sex will make possible large-scale studies of sex allocation and the consequence of gender in birds. Birds provide ideal systems for studying these questions in vertebrates, as so much is known about their biology and determinants of fitness. Until recently, however, little quantitative work has been possible because of the difficulty in determining gender in most cases. Recent studies suggest that biased sex allocation be more widespread in birds than has been realized.

New tools for sex identification and the study of sex allocation in birds

We measured genome size (nuclear DNA content) by fluorescence flow cytometry in 55 species of birds representing 12 different orders. Similar studies were performed in approximately 100 species by laboratories using absorption cytophotometry of Feulgen-stained nuclei. Although there have been apparent discrepancies in the assigned values for the species used as a reference, the values obtained in the different laboratories are generally in agreement. When the data are standardized in relation to a diploid (2C) value of 2.5 picograms (pg) of DNA for the domestic chicken (Gallus gallus domesticus), the mean for DNA content in 135 species representing 17 orders is 2.82 +/- 0.33 (SD) pg with a range of 2.0-3.8 pg. Thus the genome size of birds is the most conservative of any vertebrate class and, all values considered, is smaller and more uniform in size than previous estimates would indicate. This could be explained by a previously unexplored hypothesis: that the genome of birds has evolved from a small ancestral genome that was reduced before emergence of the protoavian.

On the Evolution of Genome Size of Birds

This is the first of a series of units discussing the application of cytometry to plant material. Techniques commonly used for mammalian nuclei evaluation need considerable modification to be successful with plant material. David Galbraith and his colleagues bring together many years of knowledge in plant cytometry. Their unit provides detailed protocols on measuring DNA content, ploidy, and cell cycle status of plant tissue using both conventional laser based instruments as well as arc lamp cytometers. This unit provides an excellent starting point for those interested in doing cytometry with plants.

Analysis of Nuclear DNA Content and Ploidy in Higher Plants

The nuclear DNA content in 10 species of chondrostean fishes was measured by flow cytometry. The sterlet Acipenser ruthenus blood cells were used as an internal standard. The sterlet DNA content was calculated on the basis of comparison with the Xenopus laevis blood cells, 2C = 6.30 pg. In the tetraploid A. ruthenus and A. stellatus the DNA content comprises 3.74 pg/nucleus and is practically invariant; in Huso dauricus it is almost the same, 3.74–3.81 pg; and in A. nudiventris it is a little higher, 3.88–4.04 pg. In the oldest chondrostean, Pseudoscaphirhynchus kaufmanni, the nuclear DNA content is slightly lower, 2C = 3.46–3.48 pg, and in the American paddlefish Polyodon spathula it is lower still, 3.17 pg. In two octoploid sturgeons, A. baeri and A. gueldenstaedti, the DNA content is twice as high as that of the sterlet, 8.29–8.31 and 7.86–7.88 pg, respectively; a very similar amount, 8.24–8.42 pg, was determined in the hybrid Huso huso × A. ruthenus. In the Sakhalin sturgeon, A. medirostris (=A. mikadoi), the DNA content is two times higher than in the octoploids, 13.93–14.73 pg; therefore its ploidy may be 16n and the number of chromosomes could be 500. © 1993 Wiley-Liss, Inc.

https://onlinelibrary.wiley.com/doi/pdf/10.1002/cyto.990140406

DNA content in eurasian sturgeon species determined by flow cytometry

A rapid method to identify sex in birds is described. The method requires microliter volumes of blood, and, under appropriate conditions, results can be available within an hour of sample collection. Samples can be stored at 4 C or–20 C with out sacrificing the ability to discriminate sex differences in DNA content. The assay will find utility in laboratory, field, and applied studies, in other classes of vertebrates, and in studies on the dynamics of genome size within and among populations.

Rapid identification of sex in birds by flow cytometry.

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Rapid identification of sex in birds by flow cytometry.