Cell-specific visualization of jasmonates in wounded tomato and Arabidopsis leaves using jasmonate-specific...

Jasmonates are well-characterized signals in the development of plants and their response to abiotic and biotic stresses, such as touch and wounding by herbivores. A gap in our knowledge on jasmonate-induced processes, however, is the cellular localization of jasmonates. Here, a novel antibody-based approach was developed to visualize jasmonates in cross-sections of plant material. Antibodies raised in rabbits against BSA-coupled jasmonic acid (JA) are specific for JA, its methyl ester and isoleucine conjugate. They do not bind to 12-oxophytodienoic acid, 12-hydoxy-JA or coronatine. These antibodies were used in combination with newly established fixation and embedding methods. Jasmonates were rapidly and uniformly distributed within all cells near the site of damage of a mechanically wounded tomato (Solanum lycopersicum) leaf. Leaf tissue distally located to the wound site exhibited identical distribution, but had a lower signal intensity. The occurrence of jasmonates in all cell types of a wounded leaf was accompanied by transcript accumulation of early JA-induced genes visualized by in situ hybridization. With these new antibodies, a powerful tool is available to detect cell-specifically the occurrence of jasmonates in any jasmonate-dependent stress response or developmental process of plants.