Plant Stem Cell Niches Research Articles

Transcription Factor MdPLT1 Involved Adventitious Root Initiation in Apple Rootstocks

The induction of adventitious roots is a key factor restricting the vegetative propagation of apple dwarf rootstocks. PLETHORA (PLT) transcription factors are involved in the regulation of plant stem cell niche and adventitious root development. In this study, we identified the PLT1 gene in apples by bioinformatics and analyzed its evolutionary relationship. The MdPLT1 gene was cloned from M9-T337 to verify its subcellular localization and analyze its function in transgenic tobacco. The MdPLT1 protein contained two conserved AP2 domains which may be similar to those of poplar the PtrPLT2a and PtrPLT2b with 85% support. The CDS sequence of the MdPLT1 gene was 1638 bp, encoding 545 amino acids. The transcription factor MdPLT1 was localized in the nucleus. The number of adventitious roots of tobacco plants overexpressing MdPLT1 significantly increased. In the adventitious roots of MdPLT1-overexpressed plants, the expression levels of genes related to the NtPINs family and the NtYUCCAs family were significantly increased. The results showed that MdPLT1 positively regulated adventitious root formation. This study provided a theoretical basis for the establishment of the fast vegetative propagation of apple dwarf rootstocks.

The plant stem-cell niche and pluripotency: 15 years of an epigenetic perspective.

Pluripotent stem-cells are slowly dividing cells giving rise to daughter cells that can either differentiate to new tissues and organs, or remain stem-cells. In plants, stem-cells are located in specific niches of the shoot and root apical meristems (SAMs and RAMs). After ablation of stem-cell niches, pluripotent meristematic cells can establish new stem-cells, whereas the removal of the whole meristem destructs the regeneration process. In tissue cultures, after detached plant organs are transferred to rooting or callus induction medium (G5 or CIM), vasculature-associated pluripotent cells (VPCs) immediately start proliferation to form adventitious roots or callus, respectively, while other cell types of the organ explants basically play no part in the process. Hence, in contrast to the widely-held assumption that all plant cells have the ability to reproduce a complete organism, only few cell types are pluripotent in practice, raising the question how pluripotent stem-cells differ from differentiated cells. It is now clear that, in addition to gene regulatory networks of pluripotency factors and phytohormone signaling, epigenetics play a crucial role in initiation, maintenance and determination of plant stem-cells. Although, more and more epigenetic regulators have been shown to control plant stem-cell fate, only a few studies demonstrate how they are recruited and how they change the chromatin structure and transcriptional regulation of pluripotency factors. Here, we highlight recent breakthroughs but also revisited classical studies of epigenetic regulation and chromatin dynamics of plant stem-cells and their pluripotent precursor-cells, and point out open questions and future directions.

Calcium signaling in coordinating plant development, circadian oscillations and environmental stress responses in plants

Calcium (Ca 2+ ) is an intracellular second messenger involved in many signal transduction pathways in plants. The perception of environmental and internal cues leads to alterations in the cytosolic Ca 2+ signatures which translate into changes in gene expression and cellular functions. Herein, we review the novel roles of Ca 2+ signalling in coordinating plant stem cell niche homeostasis for root and shoot development. We also summarize the regulatory networks between Ca 2+ signalling and other plant signalling pathways which commence into activation or repression of genes involved in regulating vegetative to reproductive phase transition and light-mediated plant growth responses. The possible functions of Ca 2+ in the input pathway as a second messenger, external stimuli and the output of the clock regulating the rhythmicity of the plant circadian system are being reviewed. Additionally, we have also highlighted the new insights into the roles of Ca 2+ signalling components in the perception and regulation of abiotic/ biotic stress responses in plants. • Ca 2+ signalling integrates several signalling pathways in response to various environmental stimuli in plants. • Nuclear Ca 2+ signalling correlates cell activity, plant development, developmental phase transition, and hormonal homeostasis. • Ca 2+ signalling plays crucial roles in coordinating phosphorylation-mediated photoacclimation processes in plants. • Cytosolic-free Ca 2+ oscillations regulate circadian clocks in plants through Ca 2+ dependent calcium binding proteins. • Spatiotemporal regulation of Ca 2+ binding proteins couple biotic/abiotic stress responses contributing towards plant adaptation.

Cell Fate Decisions Within the Vascular Cambium-Initiating Wood and Bast Formation.

Precise coordination of cell fate decisions is a hallmark of multicellular organisms. Especially in tissues with non-stereotypic anatomies, dynamic communication between developing cells is vital for ensuring functional tissue organization. Radial plant growth is driven by a plant stem cell niche known as vascular cambium, usually strictly producing secondary xylem (wood) inward and secondary phloem (bast) outward, two important structures serving as much-needed CO2 depositories and building materials. Because of its bidirectional nature and its developmental plasticity, the vascular cambium serves as an instructive paradigm for investigating principles of tissue patterning. Although genes and hormones involved in xylem and phloem formation have been identified, we have a yet incomplete picture of the initial steps of cell fate transitions of stem cell daughters into xylem and phloem progenitors. In this mini-review perspective, we describe two possible scenarios of cell fate decisions based on the current knowledge about gene regulatory networks and how cellular environments are established. In addition, we point out further possible research directions.

A single-cell analysis of the Arabidopsis vegetative shoot apex.

The shoot apical meristem allows for reiterative formation of new aerial structures throughout the life cycle of a plant. We use single-cell RNA sequencing to define the cellular taxonomy of the Arabidopsis vegetative shoot apex at the transcriptome level. We find that the shoot apex is composed of highly heterogeneous cells, which can be partitioned into 7 broad populations with 23 transcriptionally distinct cell clusters. We delineate cell-cycle continuums and developmental trajectories of epidermal cells, vascular tissue, and leaf mesophyll cells and infer transcription factors and gene expression signatures associated with cell fate decisions. Integrative analysis of shoot and root apical cell populations further reveals common and distinct features of epidermal and vascular tissues. Our results, thus, offer a valuable resource for investigating the basic principles underlying cell division and differentiation in plants at single-cell resolution.

The Arabidopsis Shoot Apex Cell Atlas

Plant stem cell niches are embedded within an organized group of dividing cells known as the meristem. The shoot apical meristem allows for reiterative development and the formation of new aerial structures throughout the life cycle of a plant. As in animals, plant stem cells possess the capacity to self-renew and at the same time to differentiate into lateral organs through transit-amplifying cells. Here, we use single-cell RNA sequencing to define the cellular taxonomy of the Arabidopsis vegetative shoot apex at the transcriptome level. We find that the shoot apex is composed of highly heterogeneous cells, which can be partitioned into 7 broad populations with 23 transcriptionally distinct cell clusters. We delineate cell cycle continuums and developmental trajectories of epidermal cells, vascular tissue and leaf mesophyll cells, and infer transcription factors and gene expression signatures associated with cell fate decisions. Moreover, panoramic integration analysis of shoot and root apical meristem cell populations reveals common vascular cell types and uncovers shoot-specific parenchymatous bundle sheath cell cluster. Our results thus provide a molecular definition of the cell populations making up the shoot apex, and offer an unprecedented opportunity to investigate conserved principles underlying cell division and differentiation in multicellular organisms at single-cell resolution.

A system-level mechanistic explanation for asymmetric stem cell fates: Arabidopsis thaliana root niche as a study system

Asymmetric divisions maintain long-term stem cell populations while producing new cells that proliferate and then differentiate. Recent reports in animal systems show that divisions of stem cells can be uncoupled from their progeny differentiation, and the outcome of a division could be influenced by microenvironmental signals. But the underlying system-level mechanisms, and whether this dynamics also occur in plant stem cell niches (SCN), remain elusive. This article presents a cell fate regulatory network model that contributes to understanding such mechanism and identify critical cues for cell fate transitions in the root SCN. Novel computational and experimental results show that the transcriptional regulator SHR is critical for the most frequent asymmetric division previously described for quiescent centre stem cells. A multi-scale model of the root tip that simulated each cell’s intracellular regulatory network, and the dynamics of SHR intercellular transport as a cell-cell coupling mechanism, was developed. It revealed that quiescent centre cell divisions produce two identical cells, that may acquire different fates depending on the feedback between SHR’s availability and the state of the regulatory network. Novel experimental data presented here validates our model, which in turn, constitutes the first proposed systemic mechanism for uncoupled SCN cell division and differentiation.

3DCellAtlas Meristem: a tool for the global cellular annotation of shoot apical meristems

Modern imaging approaches enable the acquisition of 3D and 4D datasets capturing plant organ development at cellular resolution. Computational analyses of these data enable the digitization and analysis of individual cells. In order to fully harness the information encoded within these datasets, annotation of the cell types within organs may be performed. This enables data points to be placed within the context of their position and identity, and for equivalent cell types to be compared between samples. The shoot apical meristem (SAM) in plants is the apical stem cell niche from which all above ground organs are derived. We developed 3DCellAtlas Meristem which enables the complete cellular annotation of all cells within the SAM with up to 96% accuracy across all cell types in Arabidopsis and 99% accuracy in tomato SAMs. Successive layers of cells are identified along with the central stem cells, boundary regions, and layers within developing primordia. Geometric analyses provide insight into the morphogenetic process that occurs during these developmental processes. Coupling these digital analyses with reporter expression will enable multidimensional analyses to be performed at single cell resolution. This provides a rapid and robust means to perform comprehensive cellular annotation of plant SAMs and digital single cell analyses, including cell geometry and gene expression. This fills a key gap in our ability to analyse and understand complex multicellular biology in the apical plant stem cell niche and paves the way for digital cellular atlases and analyses.

Calcium signals are necessary to establish auxin transporter polarity in a plant stem cell niche

In plants mechanical signals pattern morphogenesis through the polar transport of the hormone auxin and through regulation of interphase microtubule (MT) orientation. To date, the mechanisms by which such signals induce changes in cell polarity remain unknown. Through a combination of time-lapse imaging, and chemical and mechanical perturbations, we show that mechanical stimulation of the SAM causes transient changes in cytoplasmic calcium ion concentration (Ca2+) and that transient Ca2+ response is required for downstream changes in PIN-FORMED 1 (PIN1) polarity. We also find that dynamic changes in Ca2+ occur during development of the SAM and this Ca2+ response is required for changes in PIN1 polarity, though not sufficient. In contrast, we find that Ca2+ is not necessary for the response of MTs to mechanical perturbations revealing that Ca2+ specifically acts downstream of mechanics to regulate PIN1 polarity response.

Control of stem cell activity in the carpel margin meristem (CMM) in Arabidopsis.

Overview of the current understanding of the molecular mechanisms that regulate meristem activity in the CMM compared to the SAM. Meristems are undifferentiated cells responsible for post-embryonic plant development. The meristems are able to form new organs continuously by carefully balancing between stem cell proliferation and cell differentiation. The plant stem cell niche in each meristem harbors the stem cells that are important to maintain each meristem. The shoot apical meristem (SAM) produces all above-parts of a plant and the molecular mechanisms active in the SAM are actively studied since many years, and models are available. During the reproductive phase of the plant, the inflorescence meristem gives rise to floral meristems, which give rise to the flowers. During floral development, the gynoecium forms that contains a new meristem inside, called the carpel margin meristem (CMM). In Arabidopsis, the gynoecium consists out of two fused carpels, where the CMM forms along the fused carpel margins. In this review, we focus on the molecular mechanisms taking place in the CMM, and we discuss similarities and differences found in the SAM.

Direct conversion of root primordium into shoot meristem relies on timing of stem cell niche development.

To understand how the identity of an organ can be switched, we studied the transformation of lateral root primordia (LRP) into shoot meristems in Arabidopsis root segments. In this system, the cytokinin-induced conversion does not involve the formation of callus-like structures. Detailed analysis showed that the conversion sequence starts with a mitotic pause and is concomitant with the differential expression of regulators of root and shoot development. The conversion requires the presence of apical stem cells, and only LRP at stages VI or VII can be switched. It is engaged as soon as cell divisions resume because their position and orientation differ in the converting organ compared with the undisturbed emerging LRP. By alternating auxin and cytokinin treatments, we showed that the root and shoot organogenetic programs are remarkably plastic, as the status of the same plant stem cell niche can be reversed repeatedly within a set developmental window. Thus, the networks at play in the meristem of a root can morph in the span of a couple of cell division cycles into those of a shoot, and back, through transdifferentiation.

MOL1 is required for cambium homeostasis in Arabidopsis.

SummaryPlants maintain pools of pluripotent stem cells which allow them to constantly produce new tissues and organs. Stem cell homeostasis in shoot and root tips depends on negative regulation by ligand–receptor pairs of the CLE peptide and leucine‐rich repeat receptor‐like kinase (LRR‐RLK) families. However, regulation of the cambium, the stem cell niche required for lateral growth of shoots and roots, is poorly characterized. Here we show that the LRR‐RLK MOL1 is necessary for cambium homeostasis in Arabidopsis thaliana. By employing promoter reporter lines, we reveal that MOL1 is active in a domain that is distinct from the domain of the positively acting CLE41/PXY signaling module. In particular, we show that MOL1 acts in an opposing manner to the CLE41/PXY module and that changing the domain or level of MOL1 expression both result in disturbed cambium organization. Underlining discrete roles of MOL1 and PXY, both LRR‐RLKs are not able to replace each other when their expression domains are interchanged. Furthermore, MOL1 but not PXY is able to rescue CLV1 deficiency in the shoot apical meristem. By identifying genes mis‐expressed in mol1 mutants, we demonstrate that MOL1 represses genes associated with stress‐related ethylene and jasmonic acid hormone signaling pathways which have known roles in coordinating lateral growth of the Arabidopsis stem. Our findings provide evidence that common regulatory mechanisms in different plant stem cell niches are adapted to specific niche anatomies and emphasize the importance of a complex spatial organization of intercellular signaling cascades for a strictly bidirectional tissue production.

An epidermis-driven mechanism positions and scales stem cell niches in plants

How molecular patterning scales to organ size is highly debated in developmental biology. We explore this question for the characteristic gene expression domains of the plant stem cell niche residing in the shoot apical meristem. We show that a combination of signals originating from the epidermal cell layer can correctly pattern the key gene expression domains and notably leads to adaptive scaling of these domains to the size of the tissue. Using live imaging, we experimentally confirm this prediction. The identified mechanism is also sufficient to explain de novo stem cell niches in emerging flowers. Our findings suggest that the deformation of the tissue transposes meristem geometry into an instructive scaling and positional input for the apical plant stem cell niche.

Plant stem cells: unity and diversity

Stem cells are undifferentiated cells of multicellular organisms that can divide, self-renew and differentiate. Despite the differences of properties, general principles of the existence of stem cells can be distinguished in all multicellular organisms. In plants, stem cells are found in meristems – the structures that ensure the continuous growth of plant and provide material for the formation of various specialized tissues. There are numerous types of meristems: shoot and root apical meristems, lateral meristems (procambium, cambium, pericycle), as well as the so-called irregular meristems, developing under certain conditions (callus, meristems of symbiotic nodules, spontaneous and pathogen-induced tumors, etc.). For each of meristems, specific mechanisms of regulation, which are based on the interaction of plant hormones and the major groups of transcription factors, were identified. The activity of meristems is based on two opposite processes: proliferation and self-renewal of stem cells in the central part of the meristem and differentiation of specialized cells in the periphery. WOX-CLAVATA systems are a regulatory component conservative for different meristems and providing consistency of the composition of the meristem, as well as the balance of stem cell proliferation and differentiation. In this review, we will consider the similarities and differences between the principles of organization of stem cell niches in plants and animals, as well as in a variety of meristems of higher plants; special attention will be paid to the role of WOX-CLAVATA systems in maintaining meristems and their interaction with other meristem regulators.

Asymmetric cell divisions constructing Arabidopsis stem cell niches: the emerging role of protein phosphatases.

Plant stem cell niches (SCNs) can be maintained in time through asymmetric cell divisions (ACDs) that allow the production of new cell types while constantly renewing the pools of stem cells (SCs). ACDs in plants require the asymmetric distribution of molecular components inside the cells as well as external asymmetric positional information. These two types of asymmetric information are controlled by inter- and intracellular signalling events. Phosphorylation of proteins is a major intermediate step in these signalling events, serving either as an activator or repressor of signalling, via fast auto- and trans-phosphorylation mechanisms. Whereas protein kinases, which phosphorylate proteins on serine, threonine or tyrosine residues, have been thoroughly studied, less attention has been given to protein phosphatases, which de-phosphorylate their protein targets on these same residues. Phosphatases modulate the activity of signalling pathways by balancing the action of kinases, and are therefore critical in the regulation of ACDs in plants. In this review, we first present the different types of ACDs that operate during Arabidopsis embryonic and post-embryonic development and participate in the construction and maintenance of its root and shoot SCNs; we then give a brief description of the main protein phosphatases so far described in the Arabidopsis genome; and finally discuss their functions toward the regulation of the ACDs introduced in the first part of the paper.

Organizer-Derived WOX5 Signal Maintains Root Columella Stem Cells through Chromatin-Mediated Repression of CDF4 Expression

Stem cells in plants and animals are maintained pluripotent by signals from adjacent niche cells. In plants, WUSCHEL HOMEOBOX (WOX) transcription factors are central regulators of stem cell maintenance in different meristem types, yet their molecular mode of action has remained elusive. Here we show that in the Arabidopsis root meristem, the WOX5 protein moves from the root niche organizer, the quiescent center, into the columella stem cells, where it directly represses the transcription factor gene CDF4. This creates a gradient of CDF4 transcription, which promotes differentiation opposite to the WOX5 gradient, allowing stem cell daughter cells to exit the stem cell state. We further show that WOX5 represses CDF4 transcription by recruiting TPL/TPR co-repressors and the histone deacetylase HDA19, which consequently induces histone deacetylation at the CDF4 regulatory region. Our results show that chromatin-mediated repression of differentiation programs is a common strategy in plant and animal stem cell niches.

Plant and animal stem cells: similar yet different.

The astonishingly long lives of plants and their regeneration capacity depend on the activity of plant stem cells. As in animals, stem cells reside in stem cell niches, which produce signals that regulate the balance between self-renewal and the generation of daughter cells that differentiate into new tissues. Plant stem cell niches are located within the meristems, which are organized structures that are responsible for most post-embryonic development. The continuous organ production that is characteristic of plant growth requires a robust regulatory network to keep the balance between pluripotent stem cells and differentiating progeny. Components of this network have now been elucidated and provide a unique opportunity for comparing strategies that were developed in the animal and plant kingdoms, which underlie the logic of stem cell behaviour.

A SCARECROW-RETINOBLASTOMA Protein Network Controls Protective Quiescence in the Arabidopsis Root Stem Cell Organizer

Author SummaryIn the plant Arabidposis thaliana, root meristems (in the growing tip of the root) contain slowly dividing cells that act as an organizing center for the root stem cells that surround them. This centre is called the quiescent centre (QC). In this study, we show that the slow rate of division in the QC is regulated by the interaction between two proteins: Retinoblastoma homolog (RBR) and SCARECROW (SCR), a transcription factor that controls stem cell maintenance. RBR and SCR regulate quiescence in the QC by repressing an asymmetric cell division that generates short-term stem cells. Here we genetically manipulate the cells in the QC to alter their quiescence by regulating the RBR/SCR interaction to demonstrate that quiescence is not needed for the organizing capacity of the QC but instead provides cells with a higher resistance to genotoxic stress, allowing stem cells in the QC to survive even if more rapidly cycling stem cells are damaged. A role for mitotic quiescence has been reported in animal stem cells, in which Rb has been implicated. These findings indicate that it might serve a similar role in plant stem cells.

Transcriptome Study Outlines Ontogeny of the Maize Shoot Apical Meristem

There are three primary stem cell niches in land plants: the shoot apical meristem (SAM), root apical meristem, and floral meristem, giving rise to the shoot, root, and floral organs, respectively. These meristems carry out the opposing functions of stem cell maintenance and organogenesis,

Combined in silico/in vivo analysis of mechanisms providing for root apical meristem self-organization and maintenance

Background and AimsThe root apical meristem (RAM) is the plant stem cell niche which provides for the formation and continuous development of the root. Auxin is the main regulator of RAM functioning, and auxin maxima coincide with the sites of RAM initiation and maintenance. Auxin gradients are formed due to local auxin biosynthesis and polar auxin transport. The PIN family of auxin transporters plays a critical role in polar auxin transport, and two mechanisms of auxin maximum formation in the RAM based on PIN-mediated auxin transport have been proposed to date: the reverse fountain and the reflected flow mechanisms.MethodsThe two mechanisms are combined here in in silico studies of auxin distribution in intact roots and roots cut into two pieces in the proximal meristem region. In parallel, corresponding experiments were performed in vivo using DR5::GFP Arabidopsis plants.Key ResultsThe reverse fountain and the reflected flow mechanism naturally cooperate for RAM patterning and maintenance in intact root. Regeneration of the RAM in decapitated roots is provided by the reflected flow mechanism. In the excised root tips local auxin biosynthesis either alone or in cooperation with the reverse fountain enables RAM maintenance.ConclusionsThe efficiency of a dual-mechanism model in guiding biological experiments on RAM regeneration and maintenance is demonstrated. The model also allows estimation of the concentrations of auxin and PINs in root cells during development and under various treatments. The dual-mechanism model proposed here can be a powerful tool for the study of several different aspects of auxin function in root.