Postembryonic Growth Research Articles

Root stem cells: how to establish and maintain the eternal youth

Differently from animals, most of the plant organs are generated during post-embryonic development. This depends on meristems, regions located at the distal sides of the plants. In the meristems a set of self-renewal stem cells divides asymmetrically providing new cells to the growing organs. During embryogenesis, the acquisition of stem cell identity by two different sets of cells located at the basal and apical pole of the embryo, guarantees the generation of the primary meristems: the shoot apical meristem (SAM) and the root apical meristem (RAM). During post-embryonic growth these sets of stem cells are maintained and give rise to all organs of the mature plant, sustaining its growth. Here we review the state of the art on the actual knowledge on root stem cells. In particular, we focus on those mechanisms that permit stem cell fate acquisition and that allow their fate maintenance during growth, and how interspecific variability of stem cells activity provides differences in species-specific anatomical features.

Dissecting mechanisms in root growth from the transition zone perspective.

The root of the plant Arabidopsis thaliana is a dynamic structure in which cells continuously divide and differentiate to sustain its postembryonic undetermined growth. Cells at different developmental stages are organized in distinguished zones whose position and activities are maintained constant during root growth. In this review, we will discuss the latest discoveries on the regulatory networks involved in root zonation and, in particular, in the mechanisms involved in maintaining the position of the transition zone, a root developmental boundary. Developmental boundaries physically divide cells with different functions and identities. The transition zone separates dividing cells from differentiating cells in two functional domains, preserving their identity during root growth and thus controlling root development.

A Reservoir of Pluripotent Phloem Cells Safeguards the Linear Developmental Trajectory of Protophloem Sieve Elements.

Plant cells can change their identity based on positional information, a mechanism that confers developmental plasticity to plants. This ability, common to distinct multicellular organisms, is particularly relevant for plant phloem cells. Protophloem sieve elements (PSEs), one type of phloem conductive cells, act as the main organizers of the phloem pole, which comprises four distinct cell files organized in a conserved pattern. Here, we report how Arabidopsis roots generate a reservoir of meristematic phloem cells competent to swap their cell identities. Although PSE misspecification induces cell identity hybridism, the activity of RECEPTOR LIKE PROTEIN KINASE 2 (RPK2) by perceiving CLE45 peptide contributes to restrict PSE identity to the PSE position. By maintaining a spatiotemporal window when PSE and PSE-adjacent cells' identities are interchangeable, CLE45 signaling endows phloem cells with the competence to re-pattern a functional phloem pole when protophloem fails to form.

YUCCA4 overexpression modulates auxin biosynthesis and transport and influences plant growth and development via crosstalk with abscisic acid in Arabidopsis thaliana.

Auxin regulates a plethora of events during plant growth and development, acting in concert with other phytohormones. YUCCA genes encode flavin monooxygenases that function in tryptophan-dependent auxin biosynthesis. To understand the contribution of the YUCCA4 (YUC4) gene on auxin homeostasis, plant growth and interaction with abscisic acid (ABA) signaling, 35S::YUC4 seedlings were generated, which showed elongated hypocotyls with hyponastic leaves and changes in root system architecture that correlate with enhanced auxin responsive gene expression. Differential expression of PIN1, 2, 3 and 7 auxin transporters was detected in roots of YUC4 overexpressing seedlings compared to the wild-type: PIN1 was down-regulated whereas PIN2, PIN3 and PIN7 were up-regulated. Noteworthy, 35S::YUC4 lines showed enhanced sensitivity to ABA on seed germination and post-embryonic root growth, involving ABI4 transcription factor. The auxin reporter genes DR5::GUS, DR5::GFP and BA3::GUS further revealed that abscisic acid impairs auxin responses in 35S::YUC4 seedlings. Our results indicate that YUC4 overexpression influences several aspects of auxin homeostasis and reveal the critical roles of ABI4 during auxin-ABA interaction in germination and primary root growth.

Chromium Differentially Affects Hydrogen Peroxide Distribution in Primary and Adventitious Roots of Arabidopsis thaliana L.

The post-embryonic growth of the Arabidopsis thaliana root system can be modified by different types of stress, such as sublethal concentrations of metals, which may induce the production of reactive oxygen species (ROS). In this study, the effects of different concentrations of potassium chromate (K₂CrO4) on the distribution and relative quantity of hydrogen peroxide (H2O2) were determined in primary and adventitious roots in A. thaliana HyPer line seedlings. This line has a biosensor that specifically reports H2O2 levels within tissues as fluorescence. Primary root growth was inhibited at 100 μM Cr (VI); in contrast, adventitious root formation was induced over the main root growth axis. These structures proliferated from 100-160 μM Cr (VI), and much higher concentrations (180-200 μM) of K₂CrO4 were required to affect their growth. The H2O2 distributions were observed in the columella and lateral root cap of primary roots of plants grown in medium lacking dichromate, but following the development of toxicity symptoms, H2O2 changed its distribution to the meristem and differentiation zones. Conversely, adventitious roots had comparable H2O2 distribution patterns in untreated plants and those exposed to Cr (VI) supplementation. Thus, differential H2O2 distribution correlates with the resistance of adventitious roots, but not primary roots, to dichromate and underlies cell reorganization at the apex to support growth.

Integration of behavioural tests and transcriptome sequencing of C. elegans reveals how the nematode responds to peanut shell biochar amendment

Biochars have drawn wide attention as adsorbents, carbon sequesters and soil re-mediators. However, these substances are ambiguous regarding their effects on the motility, phenotypic changes and potential adaptative mechanisms of soil organisms. This study investigated how peanut shell biochar (PBC) affects the C. elegans model via a one-choice selection test and RNA-seq analysis. The results showed that C. elegans were able to select either PBC or a water control, and a clear preference for PBC was observed after 48 h of exposure, with the chemotaxis index (CI) reaching approximately 1.0. The nematode preferences for PBC vs sterile PBC/graphite were not significant, which demonstrated that initial microorganisms and appearances were not the reasons for the worms' selection, but the selection behaviour was instead determined by volatile odours. The treatments also showed that biochar amendment significantly decreased the body length, brood size and superoxide dismutase (SOD) activity of C. elegans to 960.20 ± 15.23 μm, 173.22 ± 4.56, 165.81 ± 3.82 U/mL SOD, respectively. Then, a possible molecular mechanism of PBC-induced developmental and reproductive effects on C. elegans was explored. Differential gene expression analysis was performed, and 1625 genes (1425 up- and 225 downregulated genes) were regulated in response to PBC treatment. The top 20 regulated genes were col genes (col-129; col-81; col-139; col-71), bli-6, perm-4 and his-24, which indicated that cuticle collagen synthesis, eggshell formation and/or heterochromatin in postembryonic growth may be disrupted following exposure to PBC. Therefore, our study suggested that quality standards be used to test nematode preferences and responses to biochar amendment, with the aim of safe application in soils, seedling substrates or fertilizers.

The Evolution of Insect Metamorphosis.

The evolution of insect metamorphosis is one of the most important sagas in animal history, transforming small, obscure soil arthropods into a dominant terrestrial group that has profoundly shaped the evolution of terrestrial life. The evolution of flight initiated the trajectory towards metamorphosis, favoring enhanced differences between juvenile and adult stages. The initial step modified postembryonic development, resulting in the nymph-adult differences characteristic of hemimetabolous species. The second step was to complete metamorphosis, holometaboly, and occurred by profoundly altering embryogenesis to produce a larval stage, the nymph becoming the pupa to accommodate the deferred development needed to make the adult. These changing life history patterns were intimately linked to two hormonal systems, the ecdysteroids and the juvenile hormones (JH), which function in both embryonic and postembryonic domains and control the stage-specifying genes Krüppel homolog 1 (Kr-h1), broad and E93. The ecdysteroids induce and direct molting through the ecdysone receptor (EcR), a nuclear hormone receptor with numerous targets including a conserved transcription factor network, the 'Ashburner cascade', which translates features of the ecdysteroid peak into the different phases of the molt. With the evolution of metamorphosis, ecdysteroids acquired a metamorphic function that exploited the repressor capacity of the unliganded EcR, making it a hormone-controlled gateway for the tissue development preceding metamorphosis. JH directs ecdysteroid action, controlling Kr-h1 expression which in turn regulates the other stage-specifying genes. JH appears in basal insect groups as their embryos shift from growth and patterning to differentiation. As a major portion of embryogenesis was deferred to postembryonic life with the evolution of holometaboly, JH also acquired a potent role in regulating postembryonic growth and development. Details of its involvement in broad expression and E93 suppression have been modified as life cycles became more complex and likely underlie some of the changes seen in the shift from incomplete to complete metamorphosis.

A high-resolution gene expression atlas links dedicated meristem genes to key architectural traits.

The shoot apical meristem (SAM) orchestrates the balance between stem cell proliferation and organ initiation essential for postembryonic shoot growth. Meristems show a striking diversity in shape and size. How this morphological diversity relates to variation in plant architecture and the molecular circuitries driving it are unclear. By generating a high-resolution gene expression atlas of the vegetative maize shoot apex, we show here that distinct sets of genes govern the regulation and identity of stem cells in maize versus Arabidopsis. Cell identities in the maize SAM reflect the combinatorial activity of transcription factors (TFs) that drive the preferential, differential expression of individual members within gene families functioning in a plethora of cellular processes. Subfunctionalization thus emerges as a fundamental feature underlying cell identity. Moreover, we show that adult plant characters are, to a significant degree, regulated by gene circuitries acting in the SAM, with natural variation modulating agronomically important architectural traits enriched specifically near dynamically expressed SAM genes and the TFs that regulate them. Besides unique mechanisms of maize stem cell regulation, our atlas thus identifies key new targets for crop improvement.

Two Wheat Cultivars with Contrasting Post-Embryonic Root Biomass Differ in Shoot Re-Growth after Defoliation: Implications for Breeding Grazing Resilient Forages.

The ability of forages to quickly resume aboveground growth after grazing is a trait that enables farmers to better manage their livestock for maximum profitability. Leaf removal impairs root growth. As a consequence of a deficient root system, shoot re-growth is inhibited leading to poor pasture performance. Despite the importance of roots for forage productivity, they have not been considered as breeding targets for improving grazing resilience due in large part to the lack of knowledge on the relationship between roots and aboveground biomass re-growth. Winter wheat (Triticum aestivum) is extensively used as forage source in temperate climates worldwide. Here, we investigated the impact of leaf clipping on specific root traits, and how these influence shoot re-growth in two winter wheat cultivars (i.e., Duster and Cheyenne) with contrasting root and shoot biomass. We found that root growth angle and post-embryonic root growth in both cultivars are strongly influenced by defoliation. We discovered that Duster, which had less post-embryonic roots before defoliation, reestablished its root system faster after leaf cutting compared with Cheyenne, which had a more extensive pre-defoliation post-embryonic root system. Rapid resumption of root growth in Duster after leaf clipping was associated with faster aboveground biomass re-growth even after shoot overcutting. Taken together, our results suggest that lower investments in the production of post-embryonic roots presents an important ideotype to consider when breeding for shoot re-growth vigor in dual purpose wheat.

Plant Cell Polarity: Creating Diversity from Inside the Box.

Cell polarity in plants operates across a broad range of spatial and temporal scales to control processes from acute cell growth to systemic hormone distribution. Similar to other eukaryotes, plants generate polarity at both the subcellular and tissue levels, often through polarization of membrane-associated protein complexes. However, likely due to the constraints imposed by the cell wall and their extremely plastic development, plants possess novel polarity molecules and mechanisms highly tuned to environmental inputs. Considerable progress has been made in identifying key plant polarity regulators, but detailed molecular understanding of polarity mechanisms remains incomplete in plants. Here, we emphasize the striking similarities in the conceptual frameworks that generate polarity in both animals and plants. To this end, we highlight how novel, plant-specific proteins engage in common themes of positive feedback, dynamic intracellular trafficking, and posttranslational regulation to establish polarity axes in development. We end with a discussion of how environmental signals control intrinsic polarity to impact postembryonic organogenesis and growth.

Evolution, Initiation, and Diversity in Early Plant Embryogenesis.

During embryogenesis in plants, cell identities are specified de novo, starting from a single cell. By combining imaging, genomic profiling, and genetics, principles of early plant development have been unraveled in the dicotyledonous plant Arabidopsis. A central emerging question, however, is how well zygotic embryogenesis in Arabidopsis reflects homologous processes in other plant species, including early diverging, non-flowering, and non-seed plants. Here, we consider plant embryogenesis with an emphasis on its evolutionary history, the diverse modes of its initiation, and the concepts in pattern formation among morphologically distinct plant groups. Furthermore, we explore challenges and future directions in plant embryogenesis research.

Staging and normal table of postembryonic development of the clownfish (Amphiprion ocellaris).

BackgroundThe clownfish Amphiprion ocellaris is one of the rare coral reef fish species that can be reared in aquaria. With relatively short embryonic and larval development, it could be used as a model species to study the impact of global changes such as temperature rise or anthropogenic threats (eg, pollution) on the postembryonic development at molecular and endocrinological levels. Establishing a developmental table allows us to standardize sampling for the scientific community willing to conduct experiments on this species on different areas: ecology, evolution, and developmental biology.ResultsHere, we describe the postembryonic developmental stages for the clownfish A. ocellaris from hatching to juvenile stages (30 days posthatching). We quantitatively followed the postembryonic growth and described qualitative traits: head, paired and unpaired fins, notochord flexion, and pigmentation changes. The occurrence of these changes over time allowed us to define seven stages, for which we provide precise descriptions.ConclusionsOur work gives an easy system to determine A. ocellaris postembryonic stages allowing, thus, to develop this species as a model species for coral reef fishes. In light of global warming, the access to the full postembryonic development stages of coral reef fish is important to determine stressors that can affect such processes.

Ascorbic acid stimulates the in vitro myoblast proliferation and migration of pacu (Piaractus mesopotamicus)

The postembryonic growth of skeletal muscle in teleost fish involves myoblast proliferation, migration and differentiation, encompassing the main events of embryonic myogenesis. Ascorbic acid plays important cellular and biochemical roles as an antioxidant and contributes to the proper collagen biosynthesis necessary for the structure of connective and bone tissues. However, whether ascorbic acid can directly influence the mechanisms of fish myogenesis and skeletal muscle growth remains unclear. The aim of our work was to evaluate the effects of ascorbic acid supplementation on the in vitro myoblast proliferation and migration of pacu (Piaractus mesopotamicus). To provide insight into the potential antioxidant role of ascorbic acid, we also treated myoblasts in vitro with menadione, which is a powerful oxidant. Our results show that ascorbic acid-supplemented myoblasts exhibit increased proliferation and migration and are protected against the oxidative stress caused by menadione. In addition, ascorbic acid increased the activity of the antioxidant enzyme superoxide dismutase and the expression of myog and mtor, which are molecular markers related to skeletal muscle myogenesis and protein synthesis, respectively. This work reveals a direct influence of ascorbic acid on the mechanisms of pacu myogenesis and highlights the potential use of ascorbic acid for stimulating fish skeletal muscle growth.

Tempo-Spatial Pattern of Stepharine Accumulation in Stephania Glabra Morphogenic Tissues.

Alkaloids attract great attention due to their valuable therapeutic properties. Stepharine, an aporphine alkaloid of Stephania glabra plants, exhibits anti-aging, anti-hypertensive, and anti-viral effects. The distribution of aporphine alkaloids in cell cultures, as well as whole plants is unknown, which hampers the development of bioengineering strategies toward enhancing their production. The spatial distribution of stepharine in cell culture models, plantlets, and mature micropropagated plants was investigated at the cellular and organ levels. Stepharine biosynthesis was found to be highly spatially and temporally regulated during plant development. We proposed that self-intoxication is the most likely reason for the failure of the induction of alkaloid biosynthesis in cell cultures. During somatic embryo development, the toxic load of alkaloids inside the cells increased. Only specialized cell sites such as vascular tissues with companion cells (VT cells), laticifers, and parenchymal cells with inclusions (PI cells) can tolerate the accumulation of alkaloids, and thus circumvent this restriction. S. glabra plants have adapted to toxic pressure by forming an additional transport secretory (laticifer) system and depository PI cells. Postembryonic growth restricts specialized cell site formation during organ development. Future bioengineering strategies should include cultures enriched in the specific cells identified in this study.

In vivo action of RNA G-quadruplex in phloem development

Phloem network integrates cellular energy status into post-embryonic growth, and development by tight regulation of carbon allocation. Phloem development involves complicated coordination of cell fate determination, cell division, and terminal differentiation into sieve elements (SEs), functional conduit. All of these processes must be tightly coordinated, for optimization of systemic connection between source supplies and sink demands throughout plant life cycle, that has substantial impact on crop productivity. Despite its pivotal role, surprisingly, regulatory mechanisms underlying phloem development have just begun to be explored, and we recently identified a novel translational regulatory network involving RNA G-quadruplex and a zinc-finger protein, JULGI, for phloem development. From this perspective, we further discuss the role of RNA G-quadruplex on post-transcriptional control of phloem regulators, as a potential interface integrating spatial information for asymmetric cell division, and phloem development.

Anamorphic development and extended parental care in a 520 million-year-old stem-group euarthropod from China

BackgroundExtended parental care is a complex reproductive strategy in which progenitors actively look after their offspring up to – or beyond – the first juvenile stage in order to maximize their fitness. Although the euarthropod fossil record has produced several examples of brood-care, the appearance of extended parental care within this phylum remains poorly constrained given the scarcity of developmental data for Palaeozoic stem-group representatives that would link juvenile and adult forms in an ontogenetic sequence.ResultsHere, we describe the post-embryonic growth of Fuxianhuia protensa from the early Cambrian Chengjiang Lagerstätte in South China. Our data demonstrate anamorphic post-embryonic development for F. protensa, in which new tergites were sequentially added from a posterior growth zone, the number of tergites varies from eight to 30. The growth of F. protensa is typified by the alternation between segment addition, followed by the depletion of the anteriormost abdominal segment into the thoracic region. The transformation of abdominal into thoracic tergite is demarcated by the development of laterally tergopleurae, and biramous walking legs. The new ontogeny data leads to the recognition of the rare Chengjiang euarthropod Pisinnocaris subconigera as a junior synonym of Fuxianhuia. Comparisons between different species of Fuxianhuia and with other genera within Fuxianhuiida suggest that heterochrony played a prominent role in the morphological diversification of fuxianhuiids. Functional analogy with the flexible trunk ontogeny of Cambrian and Silurian olenimorphic trilobites suggests an adaptation to sporadic low oxygen conditions in Chengjiang deposits for F. protensa. Finally, understanding the growth of F. protensa allows for the interpretation of an exceptional life assemblage consisting of a sexually mature adult alongside four ontogenetically coeval juveniles, which constitutes the oldest occurrence of extended parental care by prolonged cohabitation in the panarthropod fossil record.ConclusionsOur findings constitute the most detailed characterization of the post-embryonic development in a soft-bodied upper stem-group euarthropod available to date. The new ontogeny data illuminates the systematics, trunk segmentation and palaeoecology of F. protensa, offers insights on the macroevolutionary processes involved in the diversification of this clade, and contributes towards an improved understanding of complex post-embryonic reproductive ecology in Cambrian euarthropods.

Ion microprobe–measured stable isotope evidence for ammonite habitat and life mode during early ontogeny

AbstractAmmonites have disparate adult morphologies indicative of diverse ecological niches, but ammonite hatchlings are small (~1 mm diameter), which raises questions about the similarity of egg incubation and hatchling life mode in ammonites. ModernNautilusis sometimes used as a model organism for understanding ammonites, but despite their outward similarities, the groups are only distantly related. Trends in ammonite diversity and extinction vulnerability in the fossil record contrast starkly with those of nautilids, and embryonic shells from Late Cretaceous ammonites are two orders of magnitude smaller than nautilid embryonic shells. To investigate possible environmental changes experienced by ammonite hatchlings, we used secondary ion mass spectrometry to analyze the oxygen and carbon isotope composition of the embryonic shells and early postembryonic whorls of five juveniles ofHoploscaphites comprimusobtained from a single concretion in the Fox Hills Formation of South Dakota. Co-occurring bivalves and diagenetic calcite were also analyzed to provide a benthic baseline for comparison. The oxygen isotope ratios of embryonic shells are more like those of benthic bivalves, suggesting that ammonite eggs were laid on the bottom. Ammonite shell immediately after hatching has more negative δ18O, suggesting movement to more shallow water that is potentially warmer and/or fresher. After approximately one whorl of postembryonic growth, the values of δ18O become more positive in three of the five individuals, suggesting that these animals transitioned to a more demersal mode of life. Two other individuals transition to even lower δ18O values that could suggest movement to nearshore brackish water. These data suggest that ammonites, like many modern coleoids, may have spawned at different times of the year. Because scaphites were one of the short-term Cretaceous–Paleogene extinction survivors, it is possible that this characteristic allowed them to develop a broader geographic range and, consequently, a greater resistance to extinction.

SCARECROW and SHORTROOT control the auxin/cytokinin balance necessary for embryonic stem cell niche specification

ABSTRACTThe root apical meristem is established during embryogenesis, when its organizer, the quiescent center, is specified and the stem cell niche is positioned. The SCARECROW-SHORTROOT heterodimer is essential for quiescent center specification and maintenance. As continuous post-embryonic root growth relies upon the SCARECROW-mediated control of the cytokinin/auxin balance, we investigated the role of SCARECROW and SHORTROOT in controlling cytokinin signaling during embryonic quiescent center specification. We found that from embryogenesis onward both SCARECROW and SHORTROOT antagonize cytokinin signaling, thus repressing the expression of the auxin biosynthetic enzyme ANTRANILATHE SYNTHASE BETA 1. This mechanism prevents detrimental and premature high auxin levels in the QC allowing the establishment of a functional embryonic root pole.

Potential Regenerative Capacity of physiological serum, L-arginine and IGF-1 on Skeletal Muscle

In post–mitotic tissues such as skeletal muscles there are no significant cell replacements throughout life; the cells sustain local damage through its pool of specified stem cells. Muscle satellite cells are mononuclear cells that remain in a quiescent state till be activated; when they proliferate and fuse with muscle fibers to donate myonuclei, a process necessary for post-embryonic growth, hypertrophy and tissue repair in this post-mitotic tissue.Modern trends in using (autologous) biological agents ininitiation of regenerative process encourage us to evaluate the efficacy of some of these agents.In this study muscle damage was induced by cardiotoxin; followed by administration of biological agents; autologous physiological serum, L-arginine and IGF-1as a stimulating factors of myogenic satellite cells to initiate the myogenic regenerative process.Our study refers to variable degrees of efficacy in stimulation of myogenic stem cells differentiation and the outcome of the regenerative process. Although the physiological serum had marked ability to stimulate MSC but high number of newly formed myotubules are split. L-arginine despite had delayed regenerating time, but more organization of newly formed myotubules was retained. IGF-1 had rapid effect but associated with considerable number of newly formed split myotubules.We could conclude that for perfect regenerative myogenic tissue not main just formation muscle fibers, but theses fibers must be organized and functional. Healthy functional regenerative process need synchronization of several agents should be considered

Suppression of DELLA signaling induces procambial cell formation in culture.

The post-embryonic growth of plants requires the activities of apical meristems and lateral meristems. In the meristems, self-proliferation and differentiation of stem cells is tightly modulated by plant hormone signaling networks and specific transcription factors. Despite extensive studies on stem cell maintenance in plants, the mechanism by which stem cells are initially established is largely unknown. Vascular stem cells consisting of procambial/cambial cells give rise to xylem and phloem cells. In this study, we analyzed the establishment of procambial cells using the invitro culture system VISUAL, in which mesophyll cells rapidly differentiate into xylem tracheary elements and phloem sieve elements via procambial cells. We found that procambial cell formation in VISUAL is initiated by light, which can be replaced by application of gibberellin (GA). Gibberellin was able to promote procambial cell formation through degradation of DELLA, whereas light did not elevate the endogenous GA content. Indeed, light in combination with bikinin reduced the accumulation of DELLA protein in VISUAL. Consistently, overexpression of a constitutively active DELLA protein repressed vascular cell differentiation even under light. These combined results suggest that DELLA signaling suppresses procambial cell formation during vascular development in VISUAL.