Charophytes represent a diverse assemblage of extant green algae that are the sister lineage to land plants. 500-600+ million years ago, a charophyte progenitor successfully colonized land and subsequently gave rise to land plants. Charophytes have diverse but relatively simple body plans that make them highly attractive organisms for many areas of biological research. At the cellular level, many charophytes have been used for deciphering cytoskeletal networks and their dynamics, membrane trafficking, extracellular matrix secretion and cell division mechanisms. Some charophytes live in challenging habitats and have become excellent models for elucidating the cellular and molecular effects of various abiotic stressors on plant cells. Recent sequencing of several charophyte genomes has also opened doors for the dissection of biosynthetic and signaling pathways. While we are only in an infancy stage in elucidating the cell biology of charophytes, the future application of novel analytical methodologies in charophyte studies that include a broader survey of inclusive taxa will enhance our understanding of plant evolution and cell dynamics.

Background and Aims Endosidins are a group of small molecular weight compounds, first identified by “chemical biology” screening assays, that have been used to target specific components of the endomembrane system. In this study, we employed multiple microscopy-based screening techniques to elucidate effects of Endosidin 5 (ES5) on the Golgi Apparatus and the secretion of extracellular matrix (ECM) components, Penium margaritaceum. These effects were compared with those caused by treatments with brefeldin A and concanamycin A. Penium margaritaceum’s extensive Golgi Apparatus and endomembrane system make it an outstanding model organism for screening changes to the endomembrane system. Here we detail changes to the Golgi Apparatus and secretion of ECM material caused by Endosidin 5. Methods Changes to extracellular polymeric substance (EPS) secretion and cell wall expansion were screened using fluorescence microscopy. Confocal laser scanning microscopy, and transmission electron microscopy were used to assess changes to the Golgi Apparatus, the cell wall and the vesicular network. Electron tomography was also performed in order to detail the changes to the Golgi Apparatus. Key Results While other endosidins were able to impact EPS secretion and cell wall expansion only ES5 completely inhibited EPS secretion and cell wall expansion over 24 h. Short treatments of ES5 resulted in displacement of the Golgi bodies from their typical linear alignment. The number of cisternae decreased per Golgi stack and trans face cisternae in-curled to form distinct elongate circular profiles. Longer treatment resulted in a transformation of the Golgi body to an irregular aggregate of cisternae. These alterations could be reversed by removal of ES5 and returning cells to culture. Conclusions ES5 alters secretion of ECM material in Penium by affecting the Golgi apparatus and does so in a markedly different way than other endomembrane inhibitors such as Brefeldin A and Concanamycin A.