Leibniz-Institut für Pflanzenbiochemie, D-06120 Halle, Germany.
Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK), D-06466 Gatersleben, Germany.
New Phytol. 2011 Jun;190(4):1069-1080. doi: 10.1111/j.1469-8137.2010.03638.x. Epub 2011 Jan 25.
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.
茉莉酸是植物发育和对非生物及生物胁迫（如食草动物的触碰和伤害）反应中特征明显的信号分子。然而，我们对茉莉酸诱导过程的了解还存在一个空白，即茉莉酸在细胞内的定位。本文开发了一种基于抗体的新方法，用于对植物材料的横切片中的茉莉酸进行可视化。用 BSA 偶联的茉莉酸（JA）免疫兔子产生的抗体特异性识别 JA、其甲酯和异亮氨酸轭合物。它们不与 12-氧代-植二烯酸、12-羟基-JA 或冠菌素结合。这些抗体与新建立的固定和包埋方法结合使用。在机械损伤的番茄（Solanum lycopersicum）叶片中，靠近损伤部位的所有细胞内，茉莉酸都迅速且均匀分布。位于伤口部位远端的叶片组织表现出相同的分布，但信号强度较低。在受伤叶片的所有细胞类型中都存在茉莉酸，同时通过原位杂交可视化观察到早期 JA 诱导基因的转录积累。有了这些新的抗体，就可以在任何依赖茉莉酸的应激反应或植物发育过程中，特异性地检测细胞内茉莉酸的发生。
