Dear all,
I am wondering if there is anyway to develop a positive control for ROS generation study in plants. In principle, I have higher signal in stressed plants compared to non-stressed but our concern is to validate if the signal really associated with ROS. So, I am just wondering how can I generate ROS in healthy plants and then compare the strength of signal with both stressed and non-stressed plant parts. Do you have any suggestion(s)?
Thanks in advance.
Muhammad
Reactive oxygen, Nitrogen and sulfur are in continuous generations which antagonized by antioxidant to a level by which Acquired Systematic Resistance. However, under stress conditions, performance of electron transition on cellular membrane of mitochondria, Chloroplast and peroxysomes inhibited and electron hits compounds creating oxidants.
If you want to urge Oxidant accumulation you have to expose your plants to slight stress for instance salt Osmosity less than 50 micromole NaCl these treatments called Adaptation or hardening. Reducing or mitigating the adversity of ROS required the application of antioxidants . Thanks
There are several ways, you can use pathogen elicitor, mechanical damage. Also there are regiones with high ROS production like root tips; superoxide and hyrogen peroxide, differentially produced according to the region of the root.
Hello.
The way you pose the question is a little difficult.
You want to induce ROS but not to stress the plants and in order to validate if the signal is really associated with ROS? The way you pose your hypothesis may cause trouble in the way you will interpret your results.
Are you going to measure ROS or something you think is related to ROS induction? You should distinguish between what is stress related (even if stress tolerated), and what is a variable metabolic response and thus anticipated in non stressful conditions.
I think the most "natural" way to induce ROS is to increase light or leaf heat. That is easily measured, controlled, changed and has immediate and measurable impact in comparison to changes in the soil conditions. It will induce ROS and the related metabolic changes are well described in the literature, so that you can interpret the results safely.
Make sure you measure the time you implement the stress factor you choose or what ever it is to induce ROS. I wrote about this in Alexou 2013 and in Alexou and Dimitrakopoulos 2014 both in Tree Physiology.
I hope this helps, best regards.
Maria
Abiotic or biotic stress in plants trigger ROS generation . One may, however, use methyl viologen (paraquat) that generates superoxide and eventually hydrogen peroxide as positive control. More interesting would be to use inhibitors of cell wall bound NADH-peroxidase (salicylhydroxamic acid, sodium azide) or plasma membrane bound NADPH-oxidase (diphenylene iodonium chloride, imidazole) that inhibit generation of hydrogen peroxide or superoxide, respectively, as negative control that would help to check if the signal ROS-specific.
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