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

197 Citations (Scopus)

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
StatePublished - 1999

Fingerprint

Plant DNA
reference standards
DNA
fluorescent dyes
Propidium
iodide
Fluorescent Dyes
densitometry
GC Rich Sequence
Acer
Allium cepa
Vicia faba
Sorghum bicolor
Sorghum
Onions
nuclear genome
Pisum sativum
Hordeum vulgare
Densitometry
Peas

Keywords

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

ASJC Scopus subject areas

  • Plant Science

Cite this

Johnston, J. S., Bennett, M. D., Rayburn, A. L., Galbraith, D. W., & Price, H. J. (1999). Reference standards for determination of DNA content of plant nuclei. American Journal of Botany, 86(5), 609-613.

Reference standards for determination of DNA content of plant nuclei. / Johnston, J. Spencer; Bennett, Michael D.; Rayburn, A. Lane; Galbraith, David W; Price, H. James.

In: American Journal of Botany, Vol. 86, No. 5, 1999, p. 609-613.

Research output: Contribution to journalArticle

Johnston, JS, Bennett, MD, Rayburn, AL, Galbraith, DW & Price, HJ 1999, 'Reference standards for determination of DNA content of plant nuclei', American Journal of Botany, vol. 86, no. 5, pp. 609-613.
Johnston, J. Spencer ; Bennett, Michael D. ; Rayburn, A. Lane ; Galbraith, David W ; Price, H. James. / Reference standards for determination of DNA content of plant nuclei. In: American Journal of Botany. 1999 ; Vol. 86, No. 5. pp. 609-613.
@article{8db2adf9d59447c18e0e0f1e68c53965,
title = "Reference standards for determination of DNA content of plant nuclei",
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.",
keywords = "DAPI, DNA content, Flow cytometry, Propidium iodide, Reference standards",
author = "Johnston, {J. Spencer} and Bennett, {Michael D.} and Rayburn, {A. Lane} and Galbraith, {David W} and Price, {H. James}",
year = "1999",
language = "English (US)",
volume = "86",
pages = "609--613",
journal = "American Journal of Botany",
issn = "0002-9122",
publisher = "Botanical Society of America Inc.",
number = "5",

}

TY - JOUR

T1 - Reference standards for determination of DNA content of plant nuclei

AU - Johnston, J. Spencer

AU - Bennett, Michael D.

AU - Rayburn, A. Lane

AU - Galbraith, David W

AU - Price, H. James

PY - 1999

Y1 - 1999

N2 - 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.

AB - 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.

KW - DAPI

KW - DNA content

KW - Flow cytometry

KW - Propidium iodide

KW - Reference standards

UR - http://www.scopus.com/inward/record.url?scp=0033302291&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033302291&partnerID=8YFLogxK

M3 - Article

C2 - 10330063

AN - SCOPUS:0033302291

VL - 86

SP - 609

EP - 613

JO - American Journal of Botany

JF - American Journal of Botany

SN - 0002-9122

IS - 5

ER -