Co-option of the limb patterning program in cephalopod eye development

Background Across the Metazoa, similar genetic programs are found in the development of analogous, independently evolved, morphological features. The functional significance of this reuse and the underlying mechanisms of co-option remain unclear. Cephalopods have evolved a highly acute visual system with a cup-shaped retina and a novel refractive lens in the anterior, important for a number of sophisticated behaviors including predation, mating, and camouflage. Almost nothing is known about the molecular-genetics of lens development in the cephalopod. Results Here we identify the co-option of the canonical bilaterian limb patterning program during cephalopod lens development, a functionally unrelated structure. We show radial expression of transcription factors SP6-9/sp1, Dlx/dll, Pbx/exd, Meis/hth, and a Prdl homolog in the squid Doryteuthis pealeii, similar to expression required in Drosophila limb development. We assess the role of Wnt signaling in the cephalopod lens, a positive regulator in the developing Drosophila limb, and find the regulatory relationship reversed, with ectopic Wnt signaling leading to lens loss. Conclusion This regulatory divergence suggests that duplication of SP6-9 in cephalopods may mediate the co-option of the limb patterning program. Thus, our study suggests that this program could perform a more universal developmental function in radial patterning and highlights how canonical genetic programs are repurposed in novel structures.

Root Patterning: Tuning SHORT ROOT Function Creates Diversity in Form

Roots have a fundamental role in plant growth and adaptation to different environments. Diversity in root morphology and architecture enables plants to acquire water and nutrients in contrasting substrate conditions, resist biotic and abiotic stress, and develop symbiotic associations. At its most fundamental level, morphology is determined by discrete changes in tissue patterning. Differences in the number and arrangement of the cell layers in the root can change tissue structure, as well as root length and girth, affecting important productivity traits. Therefore, understanding the molecular mechanisms controlling variation in developmental patterning is an important goal in biology. The ground tissue (GT) system is an ideal model to study the genetic basis of morphological diversity because it displays great interspecific variability in cell layer number. In addition, the genetic circuit controlling GT patterning in Arabidopsis thaliana has been well described, although little is known about species with more complex root anatomies. In this review, we will describe the Arabidopsis model for root radial patterning and present recent progress in elucidating the genetic circuitry controlling GT patterning in monocots and the legume Medicago truncatula (Mt), species that develop roots with more complex anatomies and multilayered cortex.

Co-option of the Limb Patterning Program in Cephalopod Lens Development

Across the Metazoa, similar genetic programs are found in the development of analogous, independently evolved, morphological features. The functional significance of this reuse and the underlying mechanisms of co-option remain unclear. Here we identify the co-option of the canonical bilaterian limb pattering program redeployed during cephalopod lens development, a functionally unrelated structure. We show radial expression of transcription factorsSP6-9/sp1, Dlx/dll, Pbx/exd, Meis/hth, and aPrdlhomolog in the squidDoryteuthis pealeii, similar to expression required inDrosophilalimb development. We assess the role of Wnt signaling in the cephalopod lens, a positive regulator in the developing limb, and find the regulatory relationship reversed, with ectopic Wnt signaling leading to lens loss. This regulatory divergence suggests that duplication of SP6-9 in cephalopods may mediate this co-option. These results suggest that the limb network does not exclusively pattern appendage outgrowth but is performing a more universal developmental function: radial patterning.

A mechanism coordinating root elongation, endodermal differentiation, redox homeostasis and response

Root growth relies on both cell division and elongation, which occur in the meristem and elongation zones respectively. SCARECROW (SCR) is a GRAS family gene essential for root growth and radial patterning in the Arabidopsis root. Previous studies showed that SCR promotes root growth by suppressing cytokinin response in the meristem, but there is also evidence that SCR expressed beyond the meristem is required as well for root growth. Here we report that SCR promotes root growth by promoting cell elongation through suppression of oxidative stress response and maintenance of redox homeostasis in the elongation zone. In the scr root, a higher level of hydrogen peroxide was detected, which can be attributed to down-regulation of peroxidase gene 3. When stress response was blocked or redox status was ameliorated by the aba2 or upb1 mutation, the scr mutant produced a significantly longer root with longer cells and a larger and mitotically more active meristem, even though the stem cell and radial patterning defects still persisted. We showed that WRKY15, an oxidative responsive gene, was a direct target of SCR down-regulated in the scr mutant, which suggests that SCR has an active role in suppressing oxidative stress response. Since hydrogen peroxide and peroxidases are essential for endodermal differentiation, these results suggest that SCR plays a central role in coordinating cell elongation, endodermal differentiation, redox homeostasis, and oxidative stress response in plant root.One sentence summaryThis study reveals a novel mechanism of root growth regulation, which involves a previously unrecognized role of SCR in regulating cell elongation, endodermal differentiation, and redox homeostasis.

ROOT PATTERNING AND REGENERATION ARE MEDIATED BY THE QUIESCENT CENTER AND INVOLVE BLUEJAY, JACKDAW AND SCARECROW REGULATION OF VASCULATURE FACTORS

ABSTRACTStem cells are central to plant development. During root postembryonic development stem cells generating tissues are patterned in layers around a stem cell organizer or quiescent center (QC). How stem cell lineages are initially patterned is unclear. Here, we describe a role for BLUEJAY (BLJ), JACKDAW (JKD) and SCARECROW (SCR) transcription factors in patterning of cell lineages during growth and in patterning reestablishment during regeneration through regulation of number of QC cells and their regenerative capacities. In blj jkd scr mutants, QC cells are progressively lost which results in loss of tissue layers. Upon laser ablation blj jkd scr is impaired in QC division and specification resulting in severe impairment in pattern regeneration. Beyond direct regulation of QC activity by these transcription factors, reduced levels of SHORT-ROOT (SHR) and of PIN auxin transporters were observed in the vasculature of blj jkd scr, leading to strong reduction in the auxin response in the QC. We narrowed down non-cell-autonomous regulation of vascularly expressed genes in blj jkd scr to C-REPEAT BINDING FACTOR 3 (CBF3). cbf3 mutant shows reduced levels of SHR in the vasculature, and in addition QC disorganization and downregulation of the QC regulator WUSCHEL-RELATED HOMEODOMAIN 5 (WOX5). CBF3 gene is primarily expressed in the ground tissue downstream of BLJ, JKD and SCR, while CBF3 protein may move. Targeted-expression of CBF3 to the ground tissue of blj jkd scr recovers radial patterning and regeneration. We propose that BLJ, JKD and SCR regulate QC-mediated patterning, and that part of this regulation involves CBF3.

β-Catenin is required for radial cell patterning and identity in the developing mouse cochlea

Development of multicellular organs requires the coordination of cell differentiation and patterning. Critical for sound detection, the mammalian organ of Corti contains functional units arranged tonotopically along the cochlear turns. Each unit consists of sensory hair cells intercalated by nonsensory supporting cells, both specified and radially patterned with exquisite precision during embryonic development. However, how cell identity and radial patterning are jointly controlled is poorly understood. Here we show that β-catenin is required for specification of hair cell and supporting cell subtypes and radial patterning of the cochlea in vivo. In 2 mouse models of conditional β-catenin deletion, early specification of Myosin7-expressing hair cells and Prox1-positive supporting cells was preserved. While β-catenin-deficient cochleae expressed FGF8 and FGFR3, both of which are essential for pillar cell specification, the radial patterning of organ of Corti was disrupted, revealed by aberrant expression of cadherins and the pillar cell markers P75 and Lgr6. Moreover, β-catenin ablation caused duplication of FGF8-positive inner hair cells and reduction of outer hair cells without affecting the overall hair cell density. In contrast, in another transgenic model with suppressed transcriptional activity of β-catenin but preserved cell adhesion function, both specification and radial patterning of the organ of Corti were intact. Our study reveals specific functions of β-catenin in governing cell identity and patterning mediated through cell adhesion in the developing cochlea.

Paralogues of the PXY and ER receptor kinases enforce radial patterning in plant vascular tissue

Plant cell walls do not allow cells to migrate, thus plant growth and development is entirely the consequence of changes to cell division and cell elongation. Where tissues are arranged in concentric rings, expansion of inner tissue, such as that which occurs during vascular development, must be coordinated with cell division and/or expansion of the outer tissue layers, endodermis, cortex, and epidermis, in order for tissue integrity to be maintained. Little is known of how coordination between cell layers occurs, but non-cell autonomous signalling could provide an explanation. Endodermis-derived EPIDERMAL PATTERNING FACTOR-LIKE (EPFL) ligands have been shown to signal to the ERECTA (ER) receptor kinase present in the phloem. ER interacts with PHLOEM INTERCALLATED WITH XYLEM (PXY), a receptor present in the procambium. The PXY ligand, TRACHEARY ELEMENT DIFFERENTIATION INHIBITORY FACTOR (TDIF) is derived from CLE41 which is expressed in the phloem. These factors therefore represent a mechanism by which intertissue signalling could occur to control radial expansion between vascular and non-vascular tissue in plant stems. Here we show that ER regulates expression of PXY paralogues, PXL1 and PXL2, and that in turn PXY, PXL1 and PXL2 together with ER, regulate the expression of ERL1 and ERL2, genes paralogous to ER. PXY, PXL1, PXL2 and ER also regulate the expression of ER-ligands. Genetic analysis of these six receptor kinase genes demonstrated that they are required to control organisation, proliferation and cell size across multiple tissue layers. Taken together, our experiments demonstrate that ER signalling attenuates PXL expression in the stem, thus influencing vascular expansion and patterning. We anticipate that similar regulatory relationships, where tissue growth is controlled via cell signals moving across different tissue layers, will coordinate tissue layer expansion throughout the plant body.

Elongator is required for root stem cell maintenance by regulating SHORT ROOT transcription

SHORTROOT (SHR) is essential for stem cell maintenance and radial patterning in Arabidopsis thaliana roots, but how its expression is regulated is still unknown. Here, we report that Elongator regulates the transcription of SHR. The depletion of Elongator drastically reduced SHR expression and led to defective root stem cell maintenance and radial patterning. The importance of the nuclear localization of Elongator for its functioning, together with the insensitivity of the elp1 mutant to the transcription elongation inhibitor 6-azauracil and the direct interaction of the ELP4 subunit with the C-terminal domain of RNA polymerase II (RNAPII CTD), support the notion that Elongator plays important roles in transcription elongation. Indeed, we found that ELP3 associates with the pre-mRNA of SHR and that mutation of Elongator reduces the enrichment of RNAPII on the SHR gene body. Moreover, Elongator interacted in vivo with SUPPRESSOR OF Ty4 (SPT4), a well-established transcription elongation factor that was recruited to the SHR locus. Together, these results demonstrate that Elongator acts in concert with SPT4 to regulate the transcription of SHR.

Epidermal expression of a sterol biosynthesis gene regulates root growth by a non-cell autonomous mechanism in Arabidopsis

The epidermis has been hypothesized to play a signalling role during plant development. One class of mutants showing defects in signal transduction and radial patterning are those in sterol biosynthesis. The expectation is that sterol biosynthesis is a constitutive cell-autonomous process for the maintenance of basic cellular functions. The HYDRA1 (HYD1) gene of Arabidopsis encodes an essential sterol Δ8-Δ7 isomerase, and although hyd1 mutant seedlings are defective in radial patterning of several tissues, we show that the HYD1 gene is expressed primarily in the root epidermis. Cell type-specific transgenic activation of HYD1 transcription reveals that HYD1 expression in the epidermis of hyd1 null mutants is sufficient to rescue root patterning and growth. Unexpectedly, expression of HYD1 in the vascular tissues and root meristem, though not endodermis or pericycle, also leads to phenotypic rescue. Phenotypic rescue is associated with rescued patterning of the PIN1 and PIN2 auxin efflux carriers. The importance of the epidermis is in part due to its role as a site for tissue-specific sterol biosynthesis, and auxin is a candidate for a non-cell autonomous signal.