Reference standards for flow cytometric estimation of absolute
nuclear DNA content in plants

J 2022

Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants

TEMSCH, Eva M., Petr KOUTECKÝ, Tomáš URFUS, Petr ŠMARDA, Jaroslav DOLEŽEL et. al.

Základní údaje

Originální název

Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants

Autoři

TEMSCH, Eva M. (40 Rakousko, garant), Petr KOUTECKÝ (203 Česká republika), Tomáš URFUS (203 Česká republika), Petr ŠMARDA (203 Česká republika, domácí) a Jaroslav DOLEŽEL (203 Česká republika)

Vydání

Cytometry: Part A, HOBOKEN, NJ, Wiley, 2022, 1552-4922

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10611 Plant sciences, botany

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Odkazy

Odkaz na článek na stránkách časopisu

Impakt faktor

Impact factor: 3.700

Kód RIV

RIV/00216224:14310/22:00125113

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1002/cyto.a.24495

UT WoS

000685868000001

Klíčová slova anglicky

best practices; C-value; flow cytometry; GC content; genome size; plant sciences; plant standard species; standardization

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 23. 1. 2023 12:12, Mgr. Marie Šípková, DiS.

Anotace

V originále

The estimation of nuclear DNA content has been by far the most popular application of flow cytometry in plants. Because flow cytometry measures relative fluorescence intensities of nuclei stained by a DNA fluorochrome, ploidy determination, and estimation of the nuclear DNA content in absolute units both require comparison to a reference standard of known DNA content. This implies that the quality of the results obtained depends on the standard selection and use. Internal standardization, when the nuclei of an unknown sample and the reference standard are isolated, stained, and measured simultaneously, is mandatory for precise measurements. As DNA peaks representing G(1)/G(0) nuclei of the sample and standard appear on the same histogram of fluorescence intensity, the quotient of their position on the fluorescence intensity axis provides the quotient of DNA amounts. For the estimation of DNA amounts in absolute units, a number of well-established standards are now available to cover the range of known plant genome sizes. Since there are different standards in use, the standard and the genome size assigned to it has always to be reported. When none of the established standards fits, the introduction of a new standard species is needed. For this purpose, the regression line approach or simultaneous analysis of the candidate standard with several established standards should be prioritized. Moreover, the newly selected standard organism has to fulfill a number of requirements: it should be easy to identify and maintain, taxonomically unambiguous, globally available, with known genome size stability, lacking problematic metabolites, suitable for isolation of sufficient amounts of nuclei, and enabling measurements with low coefficients of variation of DNA peaks, hence suitable for the preparation of high quality samples.

Návaznosti

GA19-13231S, projekt VaV

Název: Růstové strategie trvalek: od buněk k celým rostlinám

Investor: Grantová agentura ČR, Růstové strategie trvalek: od buněk k celým rostlinám

GA19-18545S, projekt VaV

Název: Eko-geografická limitace rostlinných polyploidů: experimentální testování nových hypotéz souvisejících s velikostí buněk

Investor: Grantová agentura ČR, Eko-geografická limitace rostlinných polyploidů: experimentální testování nových hypotéz souvisejících s velikostí buněk

GA20-15989S, projekt VaV

Název: Evoluce velikosti genomu - centromerický drajv v nové roli (Akronym: Centrogenomtah)

Investor: Grantová agentura ČR, Evolution of genome size - a new role for the centromere drive

Citovat

TEMSCH, Eva M., Petr KOUTECKÝ, Tomáš URFUS, Petr ŠMARDA a Jaroslav DOLEŽEL. Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants. Cytometry: Part A. HOBOKEN, NJ: Wiley, 2022, roč. 101, č. 9, s. 710-724. ISSN 1552-4922. Dostupné z: https://dx.doi.org/10.1002/cyto.a.24495.

@article{1814077,
   author = {Temsch, Eva M. and Koutecký, Petr and Urfus, Tomáš and Šmarda, Petr and Doležel, Jaroslav},
   article_location = {HOBOKEN, NJ},
   article_number = {9},
   doi = {http://dx.doi.org/10.1002/cyto.a.24495},
   keywords = {best practices; C-value; flow cytometry; GC content; genome size; plant sciences; plant standard species; standardization},
   language = {eng},
   issn = {1552-4922},
   journal = {Cytometry: Part A},
   title = {Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants},
   url = {https://onlinelibrary.wiley.com/doi/10.1002/cyto.a.24495},
   volume = {101},
   year = {2022}
}

TY  - JOUR
ID  - 1814077
AU  - Temsch, Eva M. - Koutecký, Petr - Urfus, Tomáš - Šmarda, Petr - Doležel, Jaroslav
PY  - 2022
TI  - Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants
JF  - Cytometry: Part A
VL  - 101
IS  - 9
SP  - 710-724
EP  - 710-724
PB  - Wiley
SN  - 15524922
KW  - best practices
KW  - C-value
KW  - flow cytometry
KW  - GC content
KW  - genome size
KW  - plant sciences
KW  - plant standard species
KW  - standardization
UR  - https://onlinelibrary.wiley.com/doi/10.1002/cyto.a.24495
N2  - The estimation of nuclear DNA content has been by far the most popular application of flow cytometry in plants. Because flow cytometry measures relative fluorescence intensities of nuclei stained by a DNA fluorochrome, ploidy determination, and estimation of the nuclear DNA content in absolute units both require comparison to a reference standard of known DNA content. This implies that the quality of the results obtained depends on the standard selection and use. Internal standardization, when the nuclei of an unknown sample and the reference standard are isolated, stained, and measured simultaneously, is mandatory for precise measurements. As DNA peaks representing G(1)/G(0) nuclei of the sample and standard appear on the same histogram of fluorescence intensity, the quotient of their position on the fluorescence intensity axis provides the quotient of DNA amounts. For the estimation of DNA amounts in absolute units, a number of well-established standards are now available to cover the range of known plant genome sizes. Since there are different standards in use, the standard and the genome size assigned to it has always to be reported. When none of the established standards fits, the introduction of a new standard species is needed. For this purpose, the regression line approach or simultaneous analysis of the candidate standard with several established standards should be prioritized. Moreover, the newly selected standard organism has to fulfill a number of requirements: it should be easy to identify and maintain, taxonomically unambiguous, globally available, with known genome size stability, lacking problematic metabolites, suitable for isolation of sufficient amounts of nuclei, and enabling measurements with low coefficients of variation of DNA peaks, hence suitable for the preparation of high quality samples.
ER  -

TEMSCH, Eva M., Petr KOUTECKÝ, Tomáš URFUS, Petr ŠMARDA a Jaroslav DOLEŽEL. Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants. \textit{Cytometry: Part A}. HOBOKEN, NJ: Wiley, 2022, roč.~101, č.~9, s.~710-724. ISSN~1552-4922. Dostupné z: https://dx.doi.org/10.1002/cyto.a.24495.

Podrobný výpis o publikaci