Tubulin marker line of grapevine suspension cells as a tool to follow early stress responses

Abstract

Plant microtubules (MTs), in addition to their role in cell division and cell expansion, respond to various stress signals. To understand the biological function of this early response requires non-destructive strategies for visualization in cellular models that are highly responsive to stress signals. We have therefore generated a transgenic tubulin marker line for a cell line from the grapevine Vitis rupestris that readily responds to stress factors of defense-related and abiotic stresses based on a fusion of the green fluorescent protein with Arabidopsis β-tubulin 6. By a combination of spinning-disk confocal microscopy with quantitative image analysis, we could detect early and specific responses of MTs to defense-related and abiotic stress factors in vivo. We observed that Harpin Z (HrpZ), a bacterial elicitor that can trigger programmed cell death, rapidly eliminated radial MTs, followed by a slower depletion of the cortical array. Jasmonic acid (JA), in contrast, induced bundling of cortical MTs. Auxin reduced the thickness of cortical MTs. This effect followed a characteristic bell-shaped dose-dependency and could revert JA-induced bundling. Impeded cell expansion as a consequence of stress treatment or superoptimal auxin was linked with the appearance of intranuclear tubulin speckles. The early and stimulus-specific responses of MTs are discussed with respect to a function in processing or decoding of stress signals.