Apoplastic fluid was extracted from maize (Zea mays L) roots using two procedures: collection from the surface of intact plant roots by filter paper strips (AF) or vacuum infiltration and/or centrifugation from excised root segments (AWF). The content of cytoplasmic marker (glucose-6-phosphate, G-6-P) and antioxidative components (enzymes, organic acids, phenolics, sugars, ROS) were compared in the extracts. The results obtained demonstrate that AF was completely free of G-6-P, as opposed to AWF where the cytoplasmic constituent was detected even at mildest centrifugation (200 x g). Isoelectric focusing of POD and SOD shows the presence of cytoplasmic isoforms in AWF, and HPLC of sugars and phenolics a much more complex composition of AWF, due to cytoplasmic contamination. Organic acid composition differed in the two extracts, much higher concentrations of malic acid being registered in AF, while oxalic acid due to intracellular contamination being present only in AWF. EPR spectroscopy... of DEPMPO spin trap in the extracts showed persistent generation of hydroxyl radical adduct in AF. The results obtained argue in favor of the filter strip method for the root apoplastic fluid extraction, avoiding the problems of cytoplasmic contamination and dilution and enabling concentration measurements in minute regions of the root.
Keywords:
Spin trapping electron paramagnetic resonance (EPR) / Isoelectric focusing / Infiltration/centrifugation / HPLC / Filter paper strips / Apoplastic fluid
TY - JOUR
AU - Dragišić Maksimović, Jelena
AU - Živanović, Branka D.
AU - Maksimović, Vuk
AU - Mojović, Miloš
AU - Nikolic, Miroslav
AU - Vučinić, Željko
PY - 2014
UR - http://rimsi.imsi.bg.ac.rs/handle/123456789/751
AB - Apoplastic fluid was extracted from maize (Zea mays L) roots using two procedures: collection from the surface of intact plant roots by filter paper strips (AF) or vacuum infiltration and/or centrifugation from excised root segments (AWF). The content of cytoplasmic marker (glucose-6-phosphate, G-6-P) and antioxidative components (enzymes, organic acids, phenolics, sugars, ROS) were compared in the extracts. The results obtained demonstrate that AF was completely free of G-6-P, as opposed to AWF where the cytoplasmic constituent was detected even at mildest centrifugation (200 x g). Isoelectric focusing of POD and SOD shows the presence of cytoplasmic isoforms in AWF, and HPLC of sugars and phenolics a much more complex composition of AWF, due to cytoplasmic contamination. Organic acid composition differed in the two extracts, much higher concentrations of malic acid being registered in AF, while oxalic acid due to intracellular contamination being present only in AWF. EPR spectroscopy of DEPMPO spin trap in the extracts showed persistent generation of hydroxyl radical adduct in AF. The results obtained argue in favor of the filter strip method for the root apoplastic fluid extraction, avoiding the problems of cytoplasmic contamination and dilution and enabling concentration measurements in minute regions of the root.
PB - Elsevier Ireland Ltd, Clare
T2 - Plant Science
T1 - Filter strip as a method of choice for apoplastic fluid extraction from maize roots
EP - 58
SP - 49
VL - 223
DO - 10.1016/j.plantsci.2014.03.009
ER -
@article{
author = "Dragišić Maksimović, Jelena and Živanović, Branka D. and Maksimović, Vuk and Mojović, Miloš and Nikolic, Miroslav and Vučinić, Željko",
year = "2014",
abstract = "Apoplastic fluid was extracted from maize (Zea mays L) roots using two procedures: collection from the surface of intact plant roots by filter paper strips (AF) or vacuum infiltration and/or centrifugation from excised root segments (AWF). The content of cytoplasmic marker (glucose-6-phosphate, G-6-P) and antioxidative components (enzymes, organic acids, phenolics, sugars, ROS) were compared in the extracts. The results obtained demonstrate that AF was completely free of G-6-P, as opposed to AWF where the cytoplasmic constituent was detected even at mildest centrifugation (200 x g). Isoelectric focusing of POD and SOD shows the presence of cytoplasmic isoforms in AWF, and HPLC of sugars and phenolics a much more complex composition of AWF, due to cytoplasmic contamination. Organic acid composition differed in the two extracts, much higher concentrations of malic acid being registered in AF, while oxalic acid due to intracellular contamination being present only in AWF. EPR spectroscopy of DEPMPO spin trap in the extracts showed persistent generation of hydroxyl radical adduct in AF. The results obtained argue in favor of the filter strip method for the root apoplastic fluid extraction, avoiding the problems of cytoplasmic contamination and dilution and enabling concentration measurements in minute regions of the root.",
publisher = "Elsevier Ireland Ltd, Clare",
journal = "Plant Science",
title = "Filter strip as a method of choice for apoplastic fluid extraction from maize roots",
pages = "58-49",
volume = "223",
doi = "10.1016/j.plantsci.2014.03.009"
}
Dragišić Maksimović, J., Živanović, Branka D., Maksimović, V., Mojović, M., Nikolic, M.,& Vučinić, Ž.. (2014). Filter strip as a method of choice for apoplastic fluid extraction from maize roots. in Plant Science
Elsevier Ireland Ltd, Clare., 223, 49-58.
https://doi.org/10.1016/j.plantsci.2014.03.009
Dragišić Maksimović J, Živanović, Branka D., Maksimović V, Mojović M, Nikolic M, Vučinić Ž. Filter strip as a method of choice for apoplastic fluid extraction from maize roots. in Plant Science. 2014;223:49-58.
doi:10.1016/j.plantsci.2014.03.009 .
Dragišić Maksimović, Jelena, Živanović, Branka D., Maksimović, Vuk, Mojović, Miloš, Nikolic, Miroslav, Vučinić, Željko, "Filter strip as a method of choice for apoplastic fluid extraction from maize roots" in Plant Science, 223 (2014):49-58,
https://doi.org/10.1016/j.plantsci.2014.03.009 . .
