TY  - JOUR
AB  - BACKGROUND: A screening method for elicitor and priming agents does not only allow detecting new bioactive substances, it can also be used to understand structure–function relationships of known agents by testing different derivatives of them. This can not only provide new lead compounds for the development of novel, more environment-benign, bio-based agro-chemicals, it may eventually also lead to a better understanding of defense mechanisms in plants. Reactive oxygen species (ROS) are sensitive indicators of these mechanisms but current assay formats are not suitable for multiplex screening, in particularly not in the case of monocot systems. RESULTS: Here we describe continuous monitoring of ROS in 96-well microtiter plates using the chemiluminescent probe L012, a luminol derivative producing chemiluminescence when oxidised by ROS like hydrogen peroxide, superoxide, or hydroxyl radical that can thus be used as an indicator for these ROS. We were able to measure ROS in both monocot (Oryza sativa) and dicot (Medicago truncatula) cell suspension cultures and record dose dependencies for the carbohydrate elicitors and priming agents ulvan and chitosan at low substrate concentrations (0.3–2.5 µg/ml). The method was optimized in terms of cell density, L012 concentration, and pre-incubation time. In contrast to the single peak observed using a cuvette luminometer, the improved method revealed a double burst in both cell systems during the 90-min measuring period, probably due to the detection of multiple ROS rather than only H2O2. CONCLUSION: We provide a medium throughput screening method for monocot and dicot suspension-cultured cells that enables direct comparison of monocot and dicot plant systems regarding their reaction to different signaling molecules.
AU  - Melcher, Rebecca
AU  - Moerschbacher, Bruno
DA  - 2016-01-26
DO  - 10.1186/s13007-016-0110-1
KW  - Oxidative burst
KW  - Monocots
KW  - Dicots
KW  - Microtiter plate
KW  - L-012
KW  - Plant defense
KW  - Ulvan
KW  - Chitosan
LA  - eng
N1  - Plant Methods 12 (2016) 5, 1-11
N1  - Finanziert durch den Open-Access-Publikationsfonds 2015/2016 der Westfälischen Wilhelms-Universität Münster (WWU Münster).
PY  - 2016-01-26
SN  - 1746-4811
TI  - An improved microtiter plate assay to monitor the oxidative burst in monocot and dicot plant cell suspension cultures
UR  - https://nbn-resolving.org/urn:nbn:de:hbz:6-95279639567
Y2  - 2024-11-21T22:54:52
ER  -