Reference standards for determination of DNA content of plant nuclei
Open Access
- 1 May 1999
- journal article
- Published by Wiley in American Journal of Botany
- Vol. 86 (5), 609-613
- https://doi.org/10.2307/2656569
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
Funding Information
- National Science Foundation
This publication has 26 references indexed in Scilit:
- Heterochromatin study demonstrating the non‐linearity of fluorometry useful for calculating genomic base compositionCytometry, 1993
- Nuclear DNA content in F1 hybrids of maizeHeredity, 1993
- Nuclease-induced DNA structural changes assessed by flow cytometry with the intercalating dye propidium iodideCytometry, 1991
- Detection of Intraspecific DNA Content Variation inZea maysL. by Flow CytometryJournal of Experimental Botany, 1989
- A Comparison of Internal Standards for Plant CytophotometryStain Technology, 1986
- Nuclear DNA content in the genera Zea and Sorghum. Intergeneric, interspecific and intraspecific variationHeredity, 1985
- Rapid Flow Cytometric Analysis of the Cell Cycle in Intact Plant TissuesScience, 1983
- Detection of Intraspecific Variation in Nuclear DNA Content in Microseris douglasiiBotanical Gazette, 1980
- DNA sequence organization in the pea genomeBiochemistry, 1978
- Relationships between mitotic cycle duration, S period duration and the average rate of DNA synthesis in the root meristem cells of several plantsExperimental Cell Research, 1965