Biology subjects

Bascom, C. S.

Publications and source records attributed to Bascom, C. S..

3 recordsLinked to original sources

bioRxiv2024

The role of auxin-mediated gene activation in the bryophyte, Physcomitrium patens

Perception and response to the hormone auxin is critical to plant growth and development. Expression of auxin-response genes is tightly regulated via known mechanisms of both activation and repression. Across the plant lineage, auxin-response gene induction is performed by AUXIN-REPSONSE FACTOR (ARF) activating transcription factors. Conversely, AUXIN/INDOLE ACETIC ACID proteins repress expression. Studies of gain-of-function constitutive-repression lines and ARF loss-of-function mutants have advanced the field. Yet, there is a need for a comparative study of aberrant auxin-signaling mutants to understand the developmental consequences of constitutive repression versus the absence of auxin-mediated gene induction. Using CRISPR/Cas9 gene-editing tools, we mutated each activating ARF gene in the model bryophyte, Physcomitrium patens. The resulting septuple loss-of-function mutant line (arfasept) has severe developmental phenotypes and a diminished ability to respond to exogenous auxin. However, phenotypic analysis revealed that the arfasept line is not as severely affected as the constitutive-repression lines. Expression analysis of several auxin-response genes demonstrate that auxin-mediated gene induction is abolished in both arfasept and constitutive-repression lines but that basal expression levels are higher in the arfaseptlines. Our results suggest that the expression of auxin-regulated genes important for developmental progression is maintained, albeit at reduced levels, in the absence of ARFs. HighlightResearchers used CRIPSR/Cas9 to produce a septuple Auxin Response Factor mutant in the model bryophyte Physcomitrium patens revealing the consequences of the complete loss of auxin-mediated gene activation on development.

plant biology↗

bioRxiv2022

A Clade-D Auxin Response Factor is a Major Regulator of Auxin Signaling in Physcomitrium patens.

Auxin Response Factors (ARFs) are a family of transcription factors that are responsible for regulating gene expression in response to changes in auxin level. The analysis of ARF sequence and activity indicates that there are two major groups-activators and repressors. One clade of ARFs, clade-D, is sister to clade-A activating ARFs, but are unique in that they lack a DNA binding domain. Clade-D ARFs are present in lycophytes and bryophytes but absent in pteridophytes and spermatophytes. The transcriptional activity of clade-D ARFs, as well as how they regulate gene expression, is not well understood. Here, we report that clade-D ARFs are transcriptional activators in the model bryophyte P. patens and have a major role in the development of this species. {Delta}arfd1,d2 protonemata exhibit a delay in filament branching, as well as a delay in a key cell differentiation event. Additionally, leafy gametophore development in {Delta}arfd1,d2 lines lag behind wild-type. We present evidence that ARFd1 interacts with activating, but not repressing, ARFs. An ARFd1 hypomorph, arfd1T653L, cannot multimerize. Therefore, we propose a model by which clade-D ARFs enhance gene expression by oligomerizing to DNA-bound archetypal ARFs.

plant biology↗

bioRxiv2020

Regulation of Vacuole Morphology by PIEZO Channels in Spreading Earth Moss

The perception of mechanical force is a fundamental property of most, if not all cells. PIEZO channels are plasma membrane-embedded mechanosensitive calcium channels that play diverse and essential roles in mechanobiological processes in animals1,2. PIEZO channel homologs are found in plants3,4, but their role(s) in the green lineage are almost completely unknown. Plants and animals diverged approximately 1.5 billion years ago, independently evolved multicellularity, and have vastly different cellular mechanics5. Here, we investigate PIEZO channel function in the moss Physcomitrium patens, a representative of one of the first land plant lineages. PpPIEZO1 and PpPIEZO2 were redundantly required for normal growth, size, and shape of tip-growing caulonema cells. Both were localized to vacuolar membranes and facilitated the release of calcium into the cytosol in response to hypoosmotic shock. Loss-of-function ({Delta}Pppiezo1/2) and gain-of-function (PpPIEZO2-R2508K and -R2508H) mutants revealed a role for moss PIEZO homologs in regulating vacuole morphology. Our work here shows that plant and animal PIEZO homologs have diverged in both subcellular localization and in function, likely co-opted to serve different needs in each lineage. The plant homologs of PIEZO channels thus provide a compelling lens through which to study plant mechanobiology and the evolution of mechanoperceptive strategies in multicellular eukaryotes.

plant biology↗