Glypican 4 regulates planar cell polarity of endoderm cells by controlling the localization of Cadherin 2

Non-canonical Wnt/Planar Cell Polarity (Wnt/PCP) signaling has been implicated in endoderm morphogenesis. However, the underlying cellular and molecular mechanisms of this process are unclear. We found that during convergence and extension (C&E) in zebrafish, gut endodermal cells are polarized mediolaterally, with GFP-Vangl2 enriched at the anterior edges. Endoderm cell polarity is lost, and intercalation is impaired, in the absence of glypican 4 (gpc4), a heparan-sulfate proteoglycan that promotes Wnt/PCP signaling, suggesting that this signaling is required for endodermal cell polarity. Live imaging revealed that endoderm C&E is accomplished by polarized cell protrusions and junction remodeling, which are impaired in gpc4-deficient endodermal cells. Furthermore, in the absence of gpc4, Cadherin 2 expression on the endodermal cell surface is increased due to impaired Rab5c-mediated endocytosis, which partially accounts for the endodermal defects in these mutants.These findings indicate that Gpc4 regulates endodermal planar cell polarity during endoderm C&E by influencing localization of Cadherin 2. Thus, our study uncovers a new mechanism by which Gpc4 regulates planar cell polarity and reveals the role of Wnt/PCP signaling in endoderm morphogenesis.

Overexpression of OsCASP1 Improves Calcium Tolerance in Rice

The Casparian strip domain protein 1 (OsCASP1) is necessary for the formation of the Casparian strip (CS) in the rice endodermis. It also controls Ca2+ transport to the stele. Here, we demonstrated that OsCASP1 overexpression enhanced Ca tolerance in rice. Under normal conditions, OsCASP1-overexpressed lines showed similar concentrations of essential metals in the roots and shoots compared to the wild type, while under high Ca conditions, Ca in the roots, shoots, and xylem sap of the OsCASP1-overexpressed lines was significantly decreased. This did not apply to other essential metals. Ca-inhibited growth was significantly alleviated in the OsCASP1-overexpressed lines. Furthermore, OsCASP1 overexpression resulted in earlier formation of both the CS and functional apoplastic barrier in the endodermis but did not induce ectopic CS formation in non-endodermal cell layers and affect suberin accumulation in the endodermis. These results indicate that the overexpression of OsCASP1 promotes CS formation in endodermal cells and inhibits Ca2+ transport by the apoplastic pathway, restricting Ca accumulation in the roots and shoots under high Ca conditions. Taken together, the results suggest that OsCASP1 overexpression is an effective way to improve rice adaptation to high Ca environments.

Polarly localized receptor-like kinases PXC2 and IRK act redundantly during Arabidopsis root development in the radial axis

SUMMARYIn plants, coordination of cell division and differentiation is critical for tissue patterning and organ development. Directional cell signaling and cell polarity have been proposed to participate in coordination of these developmental processes. For instance, a leucine-rich repeat receptor-like kinase (LRR-RLK) named INFLORESCENCE AND ROOT APICES KINASE (IRK) functions to restrict stele area and inhibit longitudinal anticlinal divisions (LADs) in the endodermis where it is polarly localized. The LRR-RLK most closely related to IRK is PXY/TDR CORRELATED 2 (PXC2) and we find that PXC2 shows similar polarized accumulation as IRK in root cell types. To further understand how these proteins operate in directional cell-cell signaling and root development we explored PXC2 function. pxc2 roots have an increase in stele area, indicating that PXC2 also functions to restrict stele size. Additionally, compared to either single mutant, irk pxc2 roots have an enhanced phenotype with further increases in endodermal LADs and stele area indicating redundant activities of these receptors. The double mutant also exhibits abnormal root growth, suggesting broader functions of PXC2 and IRK in the root. However, PXC2 is not functionally equivalent to IRK, as endodermal misexpression of PXC2 did not fully rescue irk. We propose that PXC2 is at least partially redundant to IRK with a more predominant role for IRK in repression of endodermal LADs. Our results are consistent with the hypothesis that repression of specific endodermal cell divisions and stele area through a PXC2/IRK-mediated directional signaling pathway is required for coordinated root growth and development.SIGNIFICANCE STATEMENTIn the root, coordination of growth and developmental processes is critical for organ function and directional cell-to-cell signaling and cell polarity are implicated in these processes. Our studies indicate that laterally polar transmembrane receptor kinases, PXC2 and IRK, have redundant functions in restriction of specific endodermal cell divisions and stele size, and are important for gravitropic root growth.

Timed mesodermal FGF and BMP govern the multi-step thyroid specification

SummaryThe thyroid plays an essential role in homeostasis and development, but the extrinsic regulation of its embryonic development remains poorly understood. Recently, we have identified the FGF and BMP pathways to be crucial for thyroid specification and have confirmed the hypothesis that the cardiac mesoderm provides the FGF and BMP ligands to regulate this process. However, it is not clear how these ligands control thyroid specification. To study the molecular mechanisms underlying early thyroid development, we combined a pharmacological approach in zebrafish embryos with genetic models, to modulate the activity of the FGF and BMP pathways at different embryonic stages. We first characterized the expression of the transcription factors pax2a and nkx2.4b - the two main early thyroid markers - in the anterior foregut endoderm and observed that pax2a was expressed from 18 hours post fertilization (hpf) and marked a large endodermal cell population while nkx2.4b was expressed from 24 hpf and marked only a subset of the pax2a-positive endodermal cells. Interestingly, the activity profiles of FGF and BMP coincided with the detection of pax2a and nkx2.4b expression, respectively. Brief modulations of the FGF and/or BMP pathways support the hypothesis that the FGF pathway regulates the expression of pax2a and the BMP pathway regulates the expression of nkx2.4b. Furthermore, inhibition of the BMP pathway during early segmentation has dramatic effects on thyroid specification, probably via the FGF pathway. Together with our previous observations, we propose here, an updated model of early thyroid development in which the foregut endoderm receives several synchronized waves of FGF and BMP signals from the cardiac mesoderm, which result in sequential activation of pax2a and nkx2.4b gene expression and subsequent thyroid specification.

A Nodal/Eph signalling relay drives the transition from apical constriction to apico-basal shortening in ascidian endoderm invagination

ABSTRACTGastrulation is the first major morphogenetic event during animal embryogenesis. Ascidian gastrulation starts with the invagination of 10 endodermal precursor cells between the 64- and late 112-cell stages. This process occurs in the absence of endodermal cell division and in two steps, driven by myosin-dependent contractions of the acto-myosin network. First, endoderm precursors constrict their apex. Second, they shorten apico-basally, while retaining small apical surfaces, thereby causing invagination. The mechanisms that prevent endoderm cell division, trigger the transition between step 1 and step 2, and drive apico-basal shortening have remained elusive. Here, we demonstrate a conserved role for Nodal and Eph signalling during invagination in two distantly related ascidian species, Phallusia mammillata and Ciona intestinalis. Specifically, we show that the transition to step 2 is triggered by Nodal relayed by Eph signalling. In addition, our results indicate that Eph signalling lengthens the endodermal cell cycle, independently of Nodal. Finally, we find that both Nodal and Eph signals are dispensable for endoderm fate specification. These results illustrate commonalities as well as differences in the action of Nodal during ascidian and vertebrate gastrulation.

Silicification of Root Tissues

Silicon (Si) is not considered an essential element, however, its tissue concentration can exceed that of many essential elements in several evolutionary distant plant species. Roots take up Si using Si transporters and then translocate it to aboveground organs. In some plant species, root tissues are also places where a high accumulation of Si can be found. Three basic modes of Si deposition in roots have been identified so far: (1) impregnation of endodermal cell walls (e.g., in cereals, such as Triticum (wheat)); (2) formation of Si-aggregates associated with endodermal cell walls (in the Andropogoneae family, which includes Sorghum and Saccharum (sugarcane)); (3) formation of Si aggregates in “stegmata” cells, which form a sheath around sclerenchyma fibers e.g., in some palm species (Phoenix (date palm)). In addition to these three major and most studied modes of Si deposition in roots, there are also less-known locations, such as deposits in xylem cells and intercellular deposits. In our research, the ontogenesis of individual root cells that accumulate Si is discussed. The documented and expected roles of Si deposition in the root is outlined mostly as a reaction of plants to abiotic and biotic stresses.

VIT1-dependent Fe distribution in seeds is conserved in dicots

One third of the people suffer from iron (Fe) Fe deficiency. An underlying factor for this malnutrition is insufficient Fe intake from the diet. A major part of the human diet includes seeds of staple crops, which contain Fe that is poorly bioavailable. One reason for the low bioavailability is these seeds store Fe in cellular compartments that also contain antinutrients, such as phytate. Thus, several studies focused on decreasing phytate concentrations. As an alternative approach to increase bioavailable Fe, Fe reserves might be directed to cellular compartments such as plastids that are free of phytate. Previous studies indicated that Fe reserves can be relocalized inside the seed to the desired compartment by genetic modification, provided that a suitable iron transporter protein is used. However, to the best of our knowledge, a Fe transporter localizing to plastids have not been identified in seeds to date. To discover novel Fe transporters, we screened Fe patterns in seeds of distinct plant lineages, hypothesizing Fe hyperaccumulating sites would indicate Fe transporter presence. To this end, metal localizations in seeds of more than twenty species were investigated using histochemical or X-ray based techniques. Results showed that in Rosids, the largest clade of eudicots, Fe reserves were primarily confined in the embryo part of the seeds. Furthermore, inside the embryos, Fe was enriched in the endodermal cell layer, a well-known feature that is mediated by vacuolar Fe transporter, VIT1 in model plantArabidopsis thaliana. This enrichment was well conserved in and beyond Rosid species. Finally, a few seeds showed novel Fe patterns, includingCarica papayawhich concentrated large Fe reserves exclusively in plastids called amyloplasts. Generally, Fe stored in amyloplast is considered bioavailable. Taken together, this study suggests dicot seeds store Fe mainly in the embryo, with a VIT1-dependent enrichment in its endodermal cell layer and indicateCarica papayapossess a strong Fe transporter at the plastid membrane. Once it is identified that might be useful in biofortification, as a novel tool to shift Fe to compartments where it is more bioavailable.

Toddler signaling regulates mesodermal cell migration downstream of Nodal signaling

Toddler/Apela/Elabela is a conserved secreted peptide that regulates mesendoderm development during zebrafish gastrulation. Two non-exclusive models have been proposed to explain Toddler function. The ‘specification model’ postulates that Toddler signaling enhances Nodal signaling to properly specify endoderm, whereas the ‘migration model’ posits that Toddler signaling regulates mesendodermal cell migration downstream of Nodal signaling. Here, we test key predictions of both models. We find that in toddler mutants Nodal signaling is initially normal and increasing endoderm specification does not rescue mesendodermal cell migration. Mesodermal cell migration defects in toddler mutants result from a decrease in animal pole-directed migration and are independent of endoderm. Conversely, endodermal cell migration defects are dependent on a Cxcr4a-regulated tether of the endoderm to mesoderm. These results suggest that Toddler signaling regulates mesodermal cell migration downstream of Nodal signaling and indirectly affects endodermal cell migration via Cxcr4a-signaling.

The odd roots of Campylocentrum (Angraeciinae-Orchidaceae): an anatomical study of its morphologically variable roots

Although some anatomical studies have been performed in Angraecinae, knowledge about the anatomy of the genus Campylocentrum is as yet incipient. The aim of this study is to anatomically characterize the structure of the different kinds of roots in the genus. Roots from 12 species were analyzed, including all the morphological variation in the genus (smooth and granulose surface). The leafless species are characterized by endovelamen, exodermal and endodermal cell walls thicker than in the leafy species. The species with terete leaves can be split in two groups: one constituting C. poeppigii, whose roots have a granulose surface produced by numerous unicellular, absorbent hairs; the second formed by six species from the Atlantic Forest. In this second group, the same granulose root appearance is produced by tufts of epivelamen in addition to the unicellular, absorbent root hairs. The other species in the genus, with conduplicate leaves, do not present a pattern for grouping. Some of them, such as C. serranum and C. micranthum, share a similar structure with the leafless species, but with thinner exodermal and endodermal cell walls. Other species, such as C. crassirhizum and C. jamaicense, are the only ones in the genus with ○-thickened cells in the exodermis.