Biology subjects

Piepers, M.

Publications and source records attributed to Piepers, M..

3 recordsLinked to original sources

bioRxiv2025

Mechanics and growth coordination define SOSEKI-based polarity fields

The formation of organs requires the coordinated growth of cells and tissues relative to the main body axes. Plant growth is typically anisotropic and mechanically coupled through contiguous cell walls, yet how these physical patterns link to cell and tissue polarity remains unclear. Here, we use SOSEKI (SOK) proteins--previously thought to report global polarity fields--as markers to dissect how polarity arises during lateral root (LR) organogenesis. Live imaging revealed that SOK polarisation does not follow a uniform global field but instead responds to local differences in growth and mechanical state between neighbouring tissues. SOKs accumulate at interfaces separating domains of distinct growth behaviour and dissipate under compressive stress. After perturbing tissue interfaces or the mechanical continuity of the tissue, the coherence of the polarity was disrupted, indicating that stable axis establishment requires mechanical coupling across the cell wall network. Our results suggest that SOK polarisation is mechanoresponsive, linking tissue mechanics to polarity and axis establishment.

plant biology↗

bioRxiv2023

GreenGate 2.0: backwards compatible addons for assembly of complex transcriptional units and their stacking with GreenGate

Molecular cloning is a crucial technique in genetic engineering that enables the precise design of synthetic transcriptional units (TUs) and the manipulation of genomes. GreenGate and several other modular molecular cloning systems were developed about ten years ago and are widely used in plant research. All these systems define grammars for assembling transcriptional units from building blocks, cloned as Level 0 modules flanked by four-base pair overhangs and recognition sites for a particular Type IIs endonuclease. Modules are efficiently assembled into Level 1 TUs in a hierarchical assembly process, and Level 2 multigene constructs are assembled by stacking Level 1 TUs. GreenGate is highly popular but has three main limitations. First, using ad-hoc overhangs added by PCR and classical restriction/ligation prevents the efficient use of a one-pot, one-step reaction to generate entry clones and domesticate internal sites; second, a Level 1 TU is assembled from a maximum of six modules, which may be limiting for applications such as multiplex genome editing; third, the generation of Level 2 assemblies is sequential and inefficient. GreenGate 2.0 (GG2.0) expands GreenGate features. It introduces additional overhangs, allowing for the combination of up to 12 Level 0 modules in a Level 1 TU. It includes a Universal Entry Generator plasmid (pUEG) to streamline the generation of Level 0 modules. GG2.0 introduces GreenBraid, a convenient method for stacking transcriptional units iteratively for multigene assemblies. Importantly, GG2.0 is backwards compatible with most existing GreenGate modules. Additionally, GG2.0 includes Level 0 modules for multiplex expression of guide RNAs for CRISPR/Cas9 genome editing and pre-assembled Level 1 vectors for dexamethasone-inducible gene expression and ubiquitous expression of plasma membrane and nuclear fluorescent markers. GG2.0 streamlines and increases the versatility of assembling complex transcriptional units and their combination.

plant biology↗

bioRxiv2020

Suspensor-derived somatic embryogenesis in Arabidopsis

In many flowering plants, including Arabidopsis thaliana, asymmetric division of the zygote generates apical and basal cells with different fates. The apical cell continues to produce the embryo while the basal cell undergoes a restricted number of anticlinal divisions leading to a suspensor of 6-9 quiescent cells that remain extra-embryonic and eventually senesce. In some genetic backgrounds, or upon ablation of the embryo, suspensor cells can however undergo periclinal cell divisions and eventually form a second, twin seedling. Likewise, embryogenesis can be induced from somatic cells by various genes, but the relation to suspensor-derived embryos is unclear. Here, we addressed the nature of the suspensor to embryo fate transformation, and its genetic triggers. We expressed most known embryogenesis-inducing transcriptional regulators and receptor-like kinases specifically in suspensor cells. Among these, only RKD1 and WUS could induce a heritable twin seedling phenotype. We next analyzed morphology and fate marker expression in embryos in which suspensor division were activated by different triggers to address the developmental paths towards reprogramming. Our results show that reprogramming of Arabidopsis suspensor cells towards embryonic identity is a specific cellular response that is triggered by defined regulators, follows a conserved developmental trajectory and shares similarity to the process of somatic embryogenesis from post-embryonic tissues.

plant biology↗