Atomic force microscopic view of the fine topography on the tobacco stigma surface during its response to pollination

During compatible pollination in tobacco, an extracellular matrix (ECM) is secreted from the stigma surface; however, it is unknown whether the pattern of secretion across the stigma depends on the pollen source. In fact, technical limitations have prevented clear observation of ECM secretion. Here, we report the detailed topographic changes on the stigma surface that accompanies intraspecies and interspecies pollination in tobacco using contact mode atomic force microscopy (AFM). Our results, which show the dynamics and time course of ECM secretion after pollination, indicate that a certain pattern of secretion already exists on the stigma prior to pollination. Intraspecies induced a two-step response, characterized by topographical changes on the stigma surface several hours after pollina-tion, which was distinct from the pattern of ECM secretion induced by interspecies pollination. This difference was confirmed by root-mean-square analysis, which assessed the roughness of the stigma surface. Our findings indicate that compatible pollination not only induces ECM secretion from the stigma, but also results in a specific distribution of the ECM. Thus, this study demonstrates the pow-erful potential of AFM in studying the pollen-stigma interaction.