A sensitive chemiluminescence method for quantification of the oxidative burst in grapevine cells and rice roots.

Roots are prominent plant-microbe interaction sites and of great biological relevance for many studies. The root response is of interest when searching for potential systemic resistance inducers. Screening of elicitors often focuses on the oxidative burst, the rapid and transient production of Reactive Oxygen Species (ROS). However, to our knowledge, no high-throughput, sensitive methods have been developed for the quantification of ROS released by roots. Here, we report on the development of an L-012-based chemiluminescence bioassay to quantitatively determine the oxidative burst following elicitation events in roots. Rice and grapevine were used as monocot and dicot models. We demonstrate that chitosan, a recognized elicitor in rice cells, was able to elicit ROS production in rice roots. Chitosan also triggered a strong oxidative burst in grapevine cell suspension cultures, while grapevine roots were not responsive. Although this method is broadly applicable, the L-012 probe requires careful consideration of solvents and plant species. Insufficient extracellular ROS, quenching, and the interference of solvents with the probe can undermine the assay sensitivity.

Responses to Drought Stress Modulate the Susceptibility to Plasmopara viticola in Vitis vinifera Self-Rooted Cuttings.

Climate change will increase the occurrence of plants being simultaneously subjected to drought and pathogen stress. Drought can alter the way in which plants respond to pathogens. This research addresses how grapevine responds to the concurrent challenge of drought stress and Plasmopara viticola, the causal agent of downy mildew, and how one stress affects the other. Self-rooted cuttings of the drought-tolerant grapevine cultivar Xynisteri and the drought-sensitive cultivar Chardonnay were exposed to full or deficit irrigation (40% of full irrigation) and artificially inoculated with P. viticola in vitro or in planta. Leaves were sampled at an early infection stage to determine the influence of the single and combined stresses on oxidative parameters, chlorophyll, and phytohormones. Under full irrigation, Xynisteri was more susceptible to P. viticola than the drought-sensitive cultivar Chardonnay. Drought stress increased the susceptibility of grapevine leaves inoculated in vitro, but both cultivars showed resistance against P. viticola when inoculated in planta. Abscisic acid, rather than jasmonic acid and salicylic acid, seemed to play a prominent role in this resistance. The irrigation-dependent susceptibility observed in this study indicates that the practices used to mitigate the effects of climate change may have a profound impact on plant pathogens.