Age, wound size and position of injury – dependent vascular regeneration assay in growing leaves

Abstract

Background Recurring damage to aerial organs of plants necessitates their prompt repair, particularly their vasculature. While vascular regeneration assay in aerial plant parts such as stem and inflorescence stalk are well established, those on leaf vasculature remained unexplored. Recently we established a new vascular regeneration assay in growing leaf and discovered the underlying molecular mechanism. Results Here we describe the detailed stepwise method of incision and the regeneration assay used for studying the leaf vascular regeneration. By using a combination of micro−surgical perturbations, brightfield microscopy and other experimental approaches, our new findings show that the regeneration efficiency decreases with aging of the leaf, and increases with the nearness of the wound towards the proximal end of the leaf. Conclusion This easy−to−master vascular regeneration assay is an efficient and rapid method to study the mechanism of vascular regeneration in growing leaves. It can be readily adapted for other plant species and can be combined with cellular and molecular biology techniques.