Characterization of Programmable Transcription Activators in the Model Monocot Setaria viridis Via Protoplast Transfection.

Recent advances in DNA synthesis and assembly allow for genetic constructs to be designed and constructed in high throughput. Characterizing large numbers of variant genetic designs is not feasible with low-throughput and time-consuming plant transformation workflows. Protoplast transformation offers a rapid, high-throughput compatible alternative for testing genetic constructs in plant-relevant molecular environments.

Osmotic stress induces phosphorylation of histone H3 at threonine 3 in pericentromeric regions of Arabidopsis thaliana.

Histone phosphorylation plays key roles in stress-induced transcriptional reprogramming in metazoans but its function(s) in land plants has remained relatively unexplored. Here we report that an Arabidopsis mutant defective in At3g03940 and At5g18190, encoding closely related Ser/Thr protein kinases, shows pleiotropic phenotypes including dwarfism and hypersensitivity to osmotic/salt stress. The double mutant has reduced global levels of phosphorylated histone H3 threonine 3 (H3T3ph), which are not enhanced, unlike the response in the wild type, by drought-like treatments.

Intron-regulated expression of SUVH3, an Arabidopsis Su(var)3-9 homologue.

SU(VAR)3-9 proteins are key regulators of heterochromatin structure and function in plants, mammals, Drosophila, and yeast. In contrast to animals and fungi, plants contain numerous Su(var)3-9 homologues (SUVH), the members of which form a discrete subfamily. The SU(VAR)3-9 and SUVH proteins associate with heterochromatin and possess histone methyltransferase activity, indicating that they participate in the organization of transcriptionally repressive chromatin.