Reference standards for determination of DNA content of plant nuclei

J. Spencer Johnston, Michael D. Bennett, A. Lane Rayburn, David W. Galbraith, H. James Price

Research output: Contribution to journalArticle

200 Scopus citations

Abstract

Flow cytometry was used to compare 14 potential reference standards for plant DNA content determination. Both chicken and plant internal standards were used, as were propidium iodide (PI) and 4'-6-diamidino-2-phenylindole (DAPI) as fluorochromes. Means and standard errors of the means are presented for the 14 potential reference standards, and the means are compared to those obtained by Feulgen densitometry. Five species are recommended as an initial set of international standards for future plant DNA content determinations: Sorghum bicolor cv. Pioneer 8695 (2C = 1.74 pg), Pisum sativum cv. Minerva Maple (2C = 9.56 pg), Hordeum vulgare cv. Sultan (2C = 11.12 pg), Vicia faba (2C = 26.66 pg), and Allium cepa cv. Ailsa Craig (2C = 33.55 pg). It is recommended that the reference standard of choice be one with 2C and 4C nuclear DNA content peaks similar to, but not overlapping, the 2C and 4C peaks of the target species. We recommend PI as the fluorochrome of choice for flow cytometric determination of plant DNA content. DAPI should be used only if the estimated DNA value is corroborated by using a second stain that has no bias for AT- or GC-rich sequences within genomes.

Original languageEnglish (US)
Pages (from-to)609-613
Number of pages5
JournalAmerican journal of botany
Volume86
Issue number5
DOIs
StatePublished - Jan 1 1999

Keywords

  • DAPI
  • DNA content
  • Flow cytometry
  • Propidium iodide
  • Reference standards

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Genetics
  • Plant Science

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