Recognition of N-acetylchitooligosaccharide elicitor by rice protoplasts

The response by rice protoplasts to N-acetylchitooligosaccharide elicitor was examined by monitoring the production of reactive oxygen species (ROS), and the expression of the two early-responsive genes, EL2 and EL3. Freshly prepared rice protoplasts produced a high level of ROS in the absence of the elicitor, and did not show further increase of the ROS generation in response to N-acetylchitooligosaccharide elicitor. By incubating protoplasts for 1 d, the background level decreased and the induction of ROS production and the induction of mRNAs for the two genes were observed. The structural requirements of N-acetylchitooligosaccharides for elicitor-activity, as well as the effects of inhibitors of protein kinase (K-252a), protein phosphatase (calyculin A) and protein synthesis (cycloheximide) on the ROS production and gene expression were very similar to those observed in suspension-cultured rice cells, indicating that rice protoplasts retain the machinery for the recognition of, and initial signaling from, N-acetylchitooligosaccharide elicitor.