Cytochemistry and C-values: The Less-well-known World of Nuclear DNA Amounts. Export

@article{citeulike:1950124, abstract = {Background In the plant sciences there are two widely applied technologies for measuring nuclear DNA content: Feulgen absorbance cytophotometry and flow cytometry (FCM). While FCM is, with good reasons, increasingly popular among plant scientists, absorbance-cytophotometric techniques lose ground. This results in a narrowing of the methodological repertoire, which is neither desirable nor beneficial. Both approaches have their advantages, but static cytophotometry seems to pose more instrumental difficulties and material-based problems than FCM, so that Feulgen-based data in the literature are often less reliable than one would expect. Scope The purpose of this article is to present a selective overview of the field of nuclear DNA content measurement, and C-values in particular, with a focus on the technical difficulties imposed by the characteristics of the biological material and with some comments on the photometrical aspects of the work. For over 20 years it has been known that plant polyphenols cause problems in Feulgen DNA cytophotometry, since they act as major staining inhibitors leading to unreliable results. However, little information is available about the chemical classes of plant metabolites capable of DNA staining interference and the mechanisms of their inhibition. Plant slimes are another source of concern. Conclusions In FCM research to uncover the effects of secondary metabolites on measurement results has begun only recently. In particular, the analysis of intraspecific genome size variation demands a stringent methodology which accounts for inhibitors. FCM tests for inhibitory effects of endogenous metabolites should become obligatory. The use of dry seeds for harvesting embryo and endosperm nuclei for FCM and Feulgen densitometry may often provide a means of circumventing staining inhibitors. The importance of internal standardization is highlighted. Our goal is a better understanding of phytochemical/cytochemical interactions in plant DNA photometry for the benefit of an ever-growing list of plant genome sizes.}, address = {Department of Systematic and Evolutionary Botany, Faculty of Life Sciences, University of Vienna, Rennweg 14, A 1030 Vienna, Austria.}, author = {Greilhuber, J}, citeulike-article-id = {1950124}, citeulike-linkout-0 = {http://dx.doi.org/10.1093/aob/mcm250}, citeulike-linkout-1 = {http://aob.oxfordjournals.org/cgi/content/abstract/101/6/791}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/17951594}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=17951594}, day = {18}, doi = {10.1093/aob/mcm250}, issn = {0305-7364}, journal = {Ann Bot (Lond)}, keywords = {flow\_cytometry, methods}, month = {October}, posted-at = {2008-06-30 11:45:12}, priority = {2}, title = {Cytochemistry and C-values: The Less-well-known World of Nuclear DNA Amounts.}, url = {http://dx.doi.org/10.1093/aob/mcm250}, year = {2007} }

TY - JOUR ID - citeulike:1950124 L3 - citeulike-article-id:1950124 N2 - Background In the plant sciences there are two widely applied technologies for measuring nuclear DNA content: Feulgen absorbance cytophotometry and flow cytometry (FCM). While FCM is, with good reasons, increasingly popular among plant scientists, absorbance-cytophotometric techniques lose ground. This results in a narrowing of the methodological repertoire, which is neither desirable nor beneficial. Both approaches have their advantages, but static cytophotometry seems to pose more instrumental difficulties and material-based problems than FCM, so that Feulgen-based data in the literature are often less reliable than one would expect. Scope The purpose of this article is to present a selective overview of the field of nuclear DNA content measurement, and C-values in particular, with a focus on the technical difficulties imposed by the characteristics of the biological material and with some comments on the photometrical aspects of the work. For over 20 years it has been known that plant polyphenols cause problems in Feulgen DNA cytophotometry, since they act as major staining inhibitors leading to unreliable results. However, little information is available about the chemical classes of plant metabolites capable of DNA staining interference and the mechanisms of their inhibition. Plant slimes are another source of concern. Conclusions In FCM research to uncover the effects of secondary metabolites on measurement results has begun only recently. In particular, the analysis of intraspecific genome size variation demands a stringent methodology which accounts for inhibitors. FCM tests for inhibitory effects of endogenous metabolites should become obligatory. The use of dry seeds for harvesting embryo and endosperm nuclei for FCM and Feulgen densitometry may often provide a means of circumventing staining inhibitors. The importance of internal standardization is highlighted. Our goal is a better understanding of phytochemical/cytochemical interactions in plant DNA photometry for the benefit of an ever-growing list of plant genome sizes. JF - Ann Bot (Lond) SN - 0305-7364 AD - Department of Systematic and Evolutionary Botany, Faculty of Life Sciences, University of Vienna, Rennweg 14, A 1030 Vienna, Austria. TI - Cytochemistry and C-values: The Less-well-known World of Nuclear DNA Amounts. KW - flow_cytometry KW - methods AU - Greilhuber, J PY - 2007/10/18/ UR - http://dx.doi.org/10.1093/aob/mcm250 ER -