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Roeder, A. H. K.

Publications and source records attributed to Roeder, A. H. K..

24 records · Page 2Linked to original sources

The ratio of auxin to cytokinin controls leaf development and meristem initiation in Physcomitrium patens

Crosstalk between auxin and cytokinin contributes to widespread developmental processes, including root and shoot meristem maintenance, phyllotaxy, and vascular patterning. Although auxin and cytokinin are potent regulators of plant development, our understanding of crosstalk between these hormones is limited to few model systems. The moss Physcomitrium patens (formerly Physcomitrella patens) is a powerful system for studying plant hormone function. Auxin and cytokinin play similar roles in regulating moss gametophore (upright shoot) architecture, as they do in flowering plant shoots. However, auxin-cytokinin crosstalk is poorly understood in moss. Here we find that the ratio of auxin to cytokinin is an important determinant of development in P. patens, especially during leaf development and branch stem cell initiation. Addition of high levels of auxin to P. patens gametophores blocks leaf outgrowth. However, simultaneous addition of high levels of both auxin and cytokinin partially restores leaf outgrowth, suggesting that the ratio of these hormones is the overriding factor. Likewise, during branch initiation and outgrowth, chemical inhibition of auxin synthesis phenocopies cytokinin application. Finally, cytokinin insensitive mutants resemble plants with altered auxin signaling and are hypersensitive to auxin. In summary, our results suggest that the ratio between auxin and cytokinin signaling is the basis for developmental decisions in the moss gametophore.

plant biology↗

An optimized pipeline for live imaging whole Arabidopsis leaves at cellular resolution

Live imaging is the gold standard for determining how cellular development gives rise to organs. However, tracking all individual cells across whole organs over large developmental time windows is extremely challenging. In this work, we provide a comparably simple method for confocal live imaging of Arabidopsis thaliana first leaves across early development. Our imaging method works for both wild-type leaves and the complex curved leaves of the jaw-1D mutant. We find that dissecting the cotyledons, affixing a coverslip above the samples and mounting samples with perfluorodecalin yields optimal imaging series for robust cellular and organ level analysis. We provide details of our complementary image processing steps in MorphGraphX software for segmenting cells, tracking the cell lineages, and measuring a suite of cellular growth properties. We also provide MorphoGraphX image processing scripts that we developed to automate analysis of segmented images and data presentation. Our imaging techniques and processing steps combine into a robust imaging pipeline. With this pipeline we are able to examine important nuances in the cellular growth and differentiation of jaw-D versus WT leaves that have not been demonstrated before. Our pipeline is a practical starting place for researchers new to live imaging plant leaves, but also to anyone interested in improving the throughput and reliability of their live imaging process.

plant biology↗

Dynamic growth re-orientation orchestrates flatness in the Arabidopsis leaf

The growth and division of cells in plant leaves is highly dynamic in time and space, even though cells cannot move relative to their neighbors. Thus, organ shape must emerge from carefully coordinated growth, especially in leaves that remain relatively flat as they grow. Here we explored the phenotype of the jagged and wavy (jaw-D) mutant in Arabidopsis thaliana, in which the leaves do not remain flat. It has previously been shown that the jaw-D mutant phenotype is caused by the overexpression of miR319, which represses TCP transcription factors, thus delaying maturation of the leaf. We analyzed cell dynamics in wild type and jaw-D by performing time lapse live imaging of developing leaves. We found that the progression of maturation from the tip of the leaf downward was delayed in jaw-D relative to wild type based on several markers of maturation, in agreement with the role of TCP transcription factors in promoting maturation. We further found that these changes in maturation were accompanied by differences in the coordination of growth across the leaf, particularly across the medial-lateral axis, causing growth conflicts that prevent the leaf from remaining flat. Although leaf flatness is often framed as a problem that requires the local synchronization of growth on the abaxial vs adaxial sides of the leaf, our results based on the jaw-D phenotype suggest that wild-type plants also need to coordinate growth more globally across the leaf blade to maintain flatness.

plant biology↗

A giant cell enhancer achieves cell-type specificity through activation via TCP and repression by Dof transcription factors

Proper pattern formation relies on the tight coordination of cell fate specification and cell cycle regulation in growing tissues. How this can be organized at enhancers that activate gene expression necessary for differentiation is not well understood. One such example is the patterning of the Arabidopsis thaliana sepal epidermis where giant cell fate specification is associated with the endoreduplication cell cycle. Previously, we identified an enhancer region capable of driving giant cell-specific expression. In this study, we use the giant cell enhancer as a model to understand the regulatory logic that promotes cell-type specific expression. Our dissection of the enhancer revealed that giant cell specificity is achieved primarily through the combination of two elements: an activator and a repressor. TCP transcription factors are involved in activation of non-specific expression throughout the epidermis with higher expression in endoreduplicated giant cells than small cells. Dof transcription factors act via the second element to repress activity of the enhancer and limit expression to giant cells. Thus, we find that cell-type specific expression emerges from the combined activities of two broadly acting enhancer elements.

plant biology↗

Timing of organ initiation is crucial for robust organ size

AbstractOrgans precisely regulate their size and shape to ensure proper function1-6. The contribution of organ initiation timing to final organ size and shape is often masked by compensatory adjustments to growth later in development7-9. Here we show that DEVELOPMENT RELATED MYB-LIKE1 (DRMY1) is required for both proper organ initiation timing and growth leading to robust sepal size in Arabidopsis. Within each drmy1 flower, the initiation of some sepals is variably delayed. Late-initiating sepals in drmy1 mutants remain smaller throughout development resulting in variability in sepal size. DRMY1 focuses the spatiotemporal signaling patterns of the plant hormones auxin and cytokinin, which jointly control the timing of sepal initiation. Contrary to expectation, our findings demonstrate that timing of organ initiation contributes to robust organ size throughout development.

plant biology↗

Cells From The Same Lineage Switch From Reduction To Enhancement Of Size Variability In Arabidopsis Sepals

Organs form with remarkably consistent sizes and shapes during development, whereas a high variability in size and growth is observed at cell level. Given this contrast, it is unclear how such consistency at organ scale can emerge from cellular behavior. We examine the growth of cell lineages, or groups of cells that are the progeny of a single mother cell. At early stages of the lineage, we find that initially smaller lineages grow faster than the larger ones reducing variability in lineage size, a phenomenon we refer to as size uniformization. In contrast at later stages of the lineage, size variability is enhanced when initially larger cell lineages grow faster than the smaller ones. Our results imply that the cell lineage changes its growth pattern at a tipping point. Finally, we found that the growth heterogeneity of individual cells within a lineage is correlated with fast growth of the lineage. Consequently, fast growing lineages show greater cell growth heterogeneity, leading to uniformization in lineage size. Thus, cellular variability in growth contributes toward decreasing variability of cell lineages throughout the sepal.

developmental biology↗