Estimating percentage constitutive heterochromatin by flow cytometry

Rayburn, A. Lane; Auger, Julie; McMurphy, L. M.

doi:10.1016/0014-4827(92)90165-5

Cited by 44 publications

(23 citation statements)

References 12 publications

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“…One fluorochrome particularly susceptible to chromatin structure is PI, the dye used in the present study. Upon comparing the potential binding of two fluorochromes with nuclei with differing amounts of compact heterochromatin, Rayburn et al (1992) found that PI fluorescence was influenced by the amount of heterochromatin. They hypothesized that DNA binding of PI was reduced in heterochromatin resulting in an underestimate of total DNA.…”

Section: Discussionmentioning

confidence: 99%

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Metamorphosis inXenopus laevisis not associated with large-scale nuclear DNA content variation

Freeman

Rayburn

2004

Journal of Experimental Biology

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presence of nuclei of variable fluorescence intensity within the unfixed nuclei. Upon optimum fixation, which has been speculated to result in more homogeneous chromatin conformation and to reduce staining artifacts, the nuclei were observed to have less fluorescence intensity variation. The differential fluorescence observed in this study is consistent with the hypothesis that large-scale intraindividual DNA variation is not associated with amphibian metamorphosis.

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Section: Discussionmentioning

confidence: 99%

“…PI is a base-pairintercalating dye. Rayburn et al (1992) observed that PI fluorescence did not reflect reported DNA content differences among maize lines differing with respect to the amount of heterochromatin each contained. The authors hypothesized that compacted heterochromatin was not as accessible to the PI dye.…”

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confidence: 88%

Metamorphosis inXenopus laevisis not associated with large-scale nuclear DNA content variation

Freeman

Rayburn

2004

Journal of Experimental Biology

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“…About 1 cm 2 of fresh leaf tissue from a putative hybrid and 1-cm-long maize stem segment were finely chopped and combined for each isolation. Isolation and staining procedures followed were those indicated previously by Rayburn et al (1992). Flow cytometry was conducted on a Coulter EPICS XL-MCL four-color flow cytometer (Beckman Coulter) equipped with an argon ion laser operated at 15 mW, with an excitation wavelength of 488 nm.…”

Section: Dna Content Analysismentioning

confidence: 99%

Amaranthus hybridus can be pollinated frequently by A. tuberculatus under field conditions

et al. 2004

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Recent studies have confirmed that weedy Amaranthus species are capable of interspecific hybridization, and such hybridization may foster the evolution of herbicide resistance. However, the extent to which hybridization among these species occurs in nature is unknown. The purpose of this study was to determine the frequency under field conditions at which A. hybridus, a monoecious and predominantly selfpollinated species, would be pollinated by A. tuberculatus, a dioecious species. To do this, parents carrying different alleles at the ALS locus, which encodes a herbicide target site, were used. Male A. tuberculatus parents were homozygous for a dominant herbicide-insensitive allele, while A. hybridus parents were homozygous for a sensitive form. Hybrid progeny therefore could be detected via herbicide selection. Mean hybridization frequencies between 0.4 and 2.3% were obtained, depending on the proximity between parents (P ¼ 0.02). The robustness of the hybrid selection assay was verified using a molecular marker and DNA content analyses. Using these techniques, more than 99% of the progeny that survived the herbicide were confirmed to be hybrids. Frequencies obtained in this study were many times higher than the generally expected rate of mutation. Therefore, even minimal fertility in hybrid progeny would support the view that hybridization could play a role in adaptive evolution of weedy Amaranthus species. Heredity (2005) 94, 64-70.

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“…PI, being an intercalating dye, does not easily bind to tightly coiled DNA found in heterochromatin (2,13,16). Subtracting the amount of heterochromatin from each chromosome and running the correlation between euchromatin per chromosome and nuclear DNA content, however, does not improve the correlation, r = 0.63.…”

Section: Discussionmentioning

confidence: 99%