Regeneration and transient gene expression in protoplasts of Draparnaldia (chlorophytes), an emerging model for comparative analyses with basal streptophytes

Abstract

Background Green plants comprise two lineages: (1) the streptophytes that colonised land and (2) the chlorophytes that have adaptations to land but remained mostly aquatic. To better understand what made streptophytes so successful, we are currently establishing the chlorophyte alga Draparnaldia sp. (Chaetophorales, Chlorophyceae) as a model for comparative analyses between these two lineages. However, establishing Draparnaldia as a valuable model requires that it can be transformed. Thus, our goal is to develop a transformation protocol for this alga. Results We have established the first transformation protocol for Draparnaldia . This protocol is based on protoplast transformation by electroporation. It includes instructions on protoplast isolation, regeneration and transient transfection. It also provides a list of the effective selective agents for future Draparnaldia transformations. Conclusions Our protocol opens a way for Draparnaldia functional genomics analyses. Moreover, it also provides an important base for establishment of stable transformation. Electronic supplementary material The online version of this article (10.1186/s13007-019-0460-6) contains supplementary material, which is available to authorized users.

Rice MEL2 regulates the timing of meiotic transition as a component of cytoplasmic RNA granules

“…Our analysis revealed that MEL2-like genes are widely conserved in land plants and charophyte algae with diverse domain structures. Green plants comprise two lineages, the streptophytes that colonized land and the chlorophytes that have adaptations for land but remained mostly aquatic [52]. All land plants are thought to have evolved from aquatic charophyte algae, implying that the prototype MEL2 evolved after the divergence from chlorophytes.…”

Protoplast Preparation for Algal Single-Cell Omics Sequencing

“…One common separation method is membrane filtration, taking advantage of shape/size differences between protoplasts and undigested cells. For example, polyester (PET) mesh filters of 40 μm, 15 μm, and 10 μm are used in this procedure [ 66 ]. The undigested cells still having cell walls are left on the mesh surface while the protoplasts pass through the mesh successfully.…”

“…The process was repeated three times to harvest protoplasts. The protoplast generation was confirmed by OD 750 measurement and M2R staining [ 66 ].…”