The shoot apical meristem (SAM) is a reservoir of stem cells that gives rise to all post-embryonic aboveground plant organs. The size of the SAM remains stable over time due to a precise balance of stem cell replenishment versus cell incorporation into organ primordia. The WUSCHEL (WUS)/CLAVATA (CLV) negative feedback loop is central to SAM size regulation. Its correct function depends on accurate spatial expression of WUS and CLV3. A signaling pathway, consisting of ERECTA family (ERf) receptors and EPIDERMAL PATTERNING FACTOR LIKE (EPFL) ligands, restricts SAM width and promotes leaf initiation. While ERf receptors are expressed throughout the SAM, EPFL ligands are expressed in its periphery. Our genetic analysis demonstrated that ERfs and CLV3 synergistically regulate the size of the SAM, and wus is epistatic to erfs. Furthermore, activation of ERf signaling with exogenous EPFLs resulted in a rapid decrease of CLV3 and WUS expression. ERf-EPFL signaling inhibits expression of WUS and CLV3 in the periphery of the SAM, confining them to the center. These findings establish the molecular mechanism for stem cell positioning along the radial axis.

2 Summary statementERf signaling restricts the width of the shoot apical meristem, a structure which generates aboveground plant organs, by inhibiting expression of two principal regulators, CLV3 and WUS, at its periphery. Abstract: The shoot apical meristem (SAM) is a reservoir of stem cells that gives rise to all post-embryonic aboveground plant organs. The size of the SAM remains stable over time due to a precise balance of stem cell replenishment versus cell incorporation into organ primordia. The WUSCHEL (WUS)/CLAVATA (CLV) negative feedback loop is central to SAM size regulation. Its correct function depends on accurate spatial expression of WUS and CLV3. A signaling pathway, consisting of ERECTA family (ERf) receptors and EPIDERMAL PATTERNING FACTOR LIKE (EPFL) ligands, restricts SAM width and promotes leaf initiation. While ERf receptors are expressed throughout the SAM, EPFL ligands are expressed in its periphery. Our genetic analysis demonstrated that ERfs and CLV3 synergistically regulate the size of the SAM, and wus is epistatic to erfs. Furthermore, activation of ERf signaling withexogenous EPFLs resulted in a rapid decrease of CLV3 and WUS expression. ERf-EPFL signaling inhibits expression of WUS and CLV3 in the periphery of the SAM, confining them to the center. These findings establish the molecular mechanism for stem cell positioning along the radial axis.

Leaves and flowers are produced by the shoot apical meristem (SAM) at a certain distance from its center, a process that requires the hormone auxin. The amount of auxin and the pattern of its distribution in the initiation zone determine the size and spatial arrangement of organ primordia. Auxin gradients in the SAM are formed by PIN-FORMED (PIN) auxin efflux carriers whose polar localization in the plasma membrane depends on the protein kinase PINOID (PID). Previous work determined that ERECTA (ER) family genes (ERfs) control initiation of leaves. ERfs are plasma membrane receptors that enable cell-to-cell communication by sensing extracellular small proteins from the EPIDERMAL PATTERNING FACTOR/EPF-LIKE (EPF/EPFL) family. Here, we investigated whether ERfs regulate initiation of organs by altering auxin distribution or signaling in Arabidopsis (Arabidopsis thaliana). Genetic and pharmacological data suggested that ERfs do not regulate organogenesis through PINs while transcriptomics data showed ERfs do not alter primary transcriptional responses to auxin. Our results indicated that in the absence of ERf signaling the peripheral zone cells inefficiently initiate leaves in response to auxin signals and that increased accumulation of auxin in the er erecta-like1 (erl1) erl2 SAM can partially rescue organ initiation defects. We propose that both auxin and ERfs are essential for leaf initiation and that they have common downstream targets. Genetic data also indicated that the role of PID in initiation of cotyledons and leaves cannot be attributed solely to regulation of PIN polarity, and PID is likely to have other functions in addition to regulation of auxin distribution.

In-line graphene characterization to determine quality, area coverage fraction, and layer number on transparent substrates is critical to large-scale commercial graphene production. Many applications, including biosensors and imbedded diagnostics, flexible electronics, and transparent electrodes, require uniform graphene transfer from its native chemical vapor deposition foil to transparent films. To enable high-volume production of these devices, graphene layer number, quality, and area coverage must be mapped at high spatial resolution to enable growth and transfer process optimization. To this end, we present a spatially resolved optical transmission technique combined with statistical analysis of the measurements to determine graphene layer number on different transparent substrates, including polymer films and glass. This method can be automated and does not require user-inputted threshold values. Our method can effectively map >1 cm2 areas at 10 micron resolution and is not limited by type of substrate or thickness assuming the substrate is transparent. We corroborate these experimental results with simulated data and present guidelines to reasonably assess graphene quality, layer number, and feature size as functions of the experimental parameters.

Leaves and flowers are produced by the shoot apical meristem (SAM) at a certain distance from its center, a process that requires the hormone auxin. The amount of auxin and the pattern of its distribution in the initiation zone determine the size and spatial arrangement of organ primordia. Auxin gradients in the SAM are formed by PIN-FORMED (PIN) auxin efflux carriers whose polar localization in the plasma membrane depends on the protein kinase PINOID (PID). Previous work determined that ERECTA family genes (ERfs) control initiation of leaves. ERfs are plasma membrane receptors that enable cell-to-cell communications by sensing extracellular small proteins from Epidermal Patterning Factor/EPF-like (EPF/EPFL) family. Here, we investigate whether ERfs regulate initiation of organs by altering auxin distribution or signaling. Genetic and pharmacological data suggest that ERfs do not regulate organogenesis through PINs while transcriptomics data show ERfs do not alter primary transcriptional responses to auxin. Our results indicate that in the absence of ERf signaling, the peripheral zone cells inefficiently initiate leaves in response to auxin signals and that increased accumulation of auxin in the er erl1 erl2 SAM can partially rescue organ initiation defects. We propose that both auxin and ERfs are essential for leaf initiation, and that they have common downstream targets. Genetic data also indicate that the role of PID in initiation of cotyledons and leaves cannot be attributed solely to regulation of PIN polarity, and PID is likely to have other functions in addition to regulation of auxin distribution.