Cell differentiation in the embryonic periderm and in scaffolding epithelia of skin appendages.

Terminal differentiation of epithelial cells is critical for the barrier function of the skin, the growth of skin appendages, such as hair and nails, and the development of the skin of amniotes. Here, we present the hypothesis that the differentiation of cells in the embryonic periderm shares characteristic features with the differentiation of epithelial cells that support the morphogenesis of cornified skin appendages during postnatal life. The periderm prevents aberrant fusion of adjacent epithelial sites during early skin development. It is shed off when keratinocytes of the epidermis form the cornified layer, the stratum corneum. A similar role is played by epithelia that ensheath cornifying skin appendages until they disintegrate to allow the separation of the mature part of the skin appendage from the adjacent tissue. These epithelia, exemplified by the inner root sheath of hair follicles and the epithelia close to the free edge of nails or claws, are referred to as scaffolding epithelia. The periderm and scaffolding epithelia are similar with regard to their transient functions in separating tissues and the conserved expression of trichohyalin and trichohyalin-like genes in mammals and birds. Thus, we propose that parts of the peridermal differentiation program were coopted to a new postnatal function during the evolution of cornified skin appendages in amniotes.

Rhytidome- and cork-type barks of holm oak, cork oak and their hybrids highlight processes leading to cork formation.

BACKGROUND: The periderm is basic for land plants due to its protective role during radial growth, which is achieved by the polymers deposited in the cell walls. In most trees, like holm oak, the first periderm is frequently replaced by subsequent internal periderms yielding a heterogeneous outer bark made of a mixture of periderms and phloem tissues, known as rhytidome. Exceptionally, cork oak forms a persistent or long-lived periderm which results in a homogeneous outer bark of thick phellem cell layers known as cork. Cork oak and holm oak distribution ranges overlap to a great extent, and they often share stands, where they can hybridize and produce offspring showing a rhytidome-type bark. RESULTS: Here we use the outer bark of cork oak, holm oak, and their natural hybrids to analyse the chemical composition, the anatomy and the transcriptome, and further understand the mechanisms underlying periderm development. We also include a unique natural hybrid individual corresponding to a backcross with cork oak that, interestingly, shows a cork-type bark. The inclusion of hybrid samples showing rhytidome-type and cork-type barks is valuable to approach cork and rhytidome development, allowing an accurate identification of candidate genes and processes. The present study underscores that abiotic stress and cell death are enhanced in rhytidome-type barks whereas lipid metabolism and cell cycle are enriched in cork-type barks. Development-related DEGs showing the highest expression, highlight cell division, cell expansion, and cell differentiation as key processes leading to cork or rhytidome-type barks. CONCLUSION: Transcriptome results, in agreement with anatomical and chemical analyses, show that rhytidome and cork-type barks are active in periderm development, and suberin and lignin deposition. Development and cell wall-related DEGs suggest that cell division and expansion are upregulated in cork-type barks whereas cell differentiation is enhanced in rhytidome-type barks.

Neural crest and periderm-specific requirements of Irf6 during neural tube and craniofacial development.

IRF6 is a key genetic determinant of syndromic and non-syndromic cleft lip and palate. The ability to interrogate post-embryonic requirements of Irf6 has been hindered, as global Irf6 ablation in the mouse causes neonatal lethality. Prior work analyzing Irf6 in mouse models defined its role in the embryonic surface epithelium and periderm where it is required to regulate cell proliferation and differentiation. Several reports have also described Irf6 gene expression in other cell types, such as muscle, and neuroectoderm. However, analysis of a functional role in non-epithelial cell lineages has been incomplete due to the severity and lethality of the Irf6 knockout model and the paucity of work with a conditional Irf6 allele. Here we describe the generation and characterization of a new Irf6 floxed mouse model and analysis of Irf6 ablation in periderm and neural crest lineages. This work found that loss of Irf6 in periderm recapitulates a mild Irf6 null phenotype, suggesting that Irf6-mediated signaling in periderm plays a crucial role in regulating embryonic development. Further, conditional ablation of Irf6 in neural crest cells resulted in an anterior neural tube defect of variable penetrance. The generation of this conditional Irf6 allele allows for new insights into craniofacial development and new exploration into the post-natal role of Irf6.

Plant periderm as a continuum in structural organisation: a tracheophyte-wide survey and hypotheses on evolution.

Periderm is a well-known structural feature with vital roles in protection of inner plant tissues and wound healing. Despite its importance to plant survival, knowledge of periderm occurrences outside the seed plants is limited and the evolutionary origins of periderm remain poorly explored. Here, we review the current knowledge of the taxonomic distribution of periderm in its two main forms - canonical periderm (periderm formed as a typical ontogenetic stage) and wound periderm (periderm produced as a self-repair mechanism) - with a focus on major plant lineages, living and extinct. We supplement the published occurrences with data based on our own observations and experiments. This updated body of data reveals that the distribution of wound periderm is more widespread taxonomically than previously recognized and some living and extinct groups are capable of producing wound periderm, despite canonical periderm being absent from their normal developmental program. A critical review of canonical and wound periderms in extant and fossil lineages indicates that not all periderms are created equal. Their organisation is widely variable and the differences can be characterised in terms of variations in three structural features: (i) the consistency in orientation of periclinal walls within individual files of periderm cells; (ii) the lateral coordination of periclinal walls between adjacent cell files; and (iii) whether a cambial layer and conspicuous layering of inward and outward derivatives can be distinguished. Using a new system of scoring periderm structure based on these criteria, we characterise the level of organisation of canonical and wound periderms in different lineages. Looking at periderms through the lens provided by their level of organisation reveals that the traditional image of periderm as a single generalised feature, is best viewed as a continuum of structural configurations that are all predicated by the same basic process (periclinal divisions), but can fall anywhere between very loosely organized (diffuse periclinal growth) to very tightly coordinated (organized periclinal growth). Overall, wound periderms in both seed plants and seed-free plants have lower degrees of organisation than canonical periderms, which may be due to their initiation in response to inherently disruptive traumatic events. Wound and canonical periderms of seed plants have higher degrees of organisation than those of seed-free plants, possibly due to co-option of the programs responsible for organizing their vascular cambial growth. Given the importance of wound periderm to plant survival, its widespread taxonomic distribution, and its early occurrence in the fossil record, we hypothesise that wound periderm may have had a single origin in euphyllophytes and canonical periderm may have originated separately in different lineages by co-option of the basic regulatory toolkit of wound periderm formation. In one evolutionary scenario, wound periderm regulators activated initially by tissue tearing due to tensional stresses elicited by woody growth underwent heterochronic change that switched their activation trigger from tissue tearing to the tensional stresses that precede it, with corresponding changes in the signalling that triggered the regulatory cascade of periderm development from tearing-induced signals to signalling induced by tension in cells.

Death and Dying: Grapevine Survival, Cold Hardiness, and BLUPs and Winter BLUEs in North Dakota Vineyards.

A total of fourteen diverse, interspecific hybrid grapevines (Vitis spp.) were evaluated for their adaptability to North Dakota winter conditions using differential thermal analysis (DTA) of low-temperature exotherms (LTE) and bud cross-sectional assessment of survival techniques. This research was conducted in two vineyard locations in eastern North Dakota. This work demonstrates the use of DTA for monitoring and selecting cultivars capable of withstanding sub-zero temperatures. These results were assessed for quantitative genetic traits. High heritability was observed for bud LTE traits and may thus be a useful target for cold hardiness breeding programs; however, it is necessary to ensure that variance is reduced when pooling multiple sample events. After DTA sampling, grapevines were assessed for survival of primary and secondary dormant buds using cross-sectional visual evaluation of death. 'Valiant' had the greatest primary bud survival (68%), followed by 'Frontenac gris', 'Crimson Pearl', and 'King of the North'. These varieties are among those with potential for production in eastern North Dakota's environment. The newly evaluated relationships between traits and the heritability of DTA results provide valuable tools to grapevine breeders for the development of cold-tolerant genotypes for future climatic challenges.

Development-Associated Genes of the Epidermal Differentiation Complex (EDC).

The epidermal differentiation complex (EDC) is a cluster of genes that encode protein components of the outermost layers of the epidermis in mammals, reptiles and birds. The development of the stratified epidermis from a single-layered ectoderm involves an embryo-specific superficial cell layer, the periderm. An additional layer, the subperiderm, develops in crocodilians and over scutate scales of birds. Here, we review the expression of EDC genes during embryonic development. Several EDC genes are expressed predominantly or exclusively in embryo-specific cell layers, whereas others are confined to the epidermal layers that are maintained in postnatal skin. The S100 fused-type proteins scaffoldin and trichohyalin are expressed in the avian and mammalian periderm, respectively. Scaffoldin forms the so-called periderm granules, which are histological markers of the periderm in birds. Epidermal differentiation cysteine-rich protein (EDCRP) and epidermal differentiation protein containing DPCC motifs (EDDM) are expressed in the avian subperiderm where they are supposed to undergo cross-linking via disulfide bonds. Furthermore, a histidine-rich epidermal differentiation protein and feather-type corneous beta-proteins, also known as beta-keratins, are expressed in the subperiderm. The accumulating evidence for roles of EDC genes in the development of the epidermis has implications on the evolutionary diversification of the skin in amniotes.

Bark structure is coordinated with xylem hydraulic properties in branches of five Cupressaceae species.

The properties of bark and xylem contribute to tree growth and survival under drought and other types of stress conditions. However, little is known about the functional coordination of the xylem and bark despite the influence of selection on both structures in response to drought. To this end, we examined relationships between proportions of bark components (i.e. thicknesses of tissues outside the vascular cambium) and xylem transport properties in juvenile branches of five Cupressaceae species, focusing on transport efficiency and safety from hydraulic failure via drought-induced embolism. Both xylem efficiency and safety were correlated with multiple bark traits, suggesting that xylem transport and bark properties are coordinated. Specifically, xylem transport efficiency was greater in species with thicker secondary phloem, greater phloem-to-xylem thickness ratio and phloem-to-xylem cell number ratio. In contrast, species with thicker bark, living cortex and dead bark tissues were more resistant to embolism. Thicker phellem layers were associated with lower embolism resistance. Results of this study point to an important connection between xylem transport efficiency and phloem characteristics, which are shaped by the activity of vascular cambium. The link between bark and embolism resistance affirms the importance of both tissues to drought tolerance.

Climatic drivers of cork growth depend on site aridity.

Cork is one of the main non-timber forest products in the world. Most of its production is concentrated in the Iberian Peninsula, a climate change hotspot. Climate warming may lead to increased aridification and reduce cork production in that region. However, we still lack assessments of climate-cork relationships across ample geographical and climatic gradients explicitly considering site aridity. We quantified cork growth by measuring cork ring width and related it to climate variables and a drought index using dendrochronology. Four cork oak (Quercus suber) forests located from north eastern Spain to south western Morocco (31.5-41.5° N) and subjected to different aridity levels were sampled. Warm conditions in spring to early summer, when cork is formed, reduced cork width, whereas high precipitation in winter and spring enhanced it. The response of cork to increased water availability in summer peaked (r = 0.89, p = 0.00002) in the most arid and continental site considering 14-month long droughts. A severe drought caused a disproportionate loss of cork production in this site, where for every five-fold decrease in the drought index, the cork-width index declined by a factor of thirteen. Therefore, site aridity determines the responses of cork growth to the soil water availability resulting from accumulated precipitation during winter and spring previous to cork growth and until summer. In general, this cumulative water balance, which is very dependent on temperature and evapotranspiration rate, is critical for cork production, especially in continental, dry sites. The precipitation during the hydrological year can be used as a proxy of cork production in similar sites. Assessments of climate-cork relationships in the western Mediterranean basin could be used as analogues to forecast the impacts of aridification on future cork production.

Cork cellular and chemical features underlying bark environmental protection in the miombo species Parinari curatellifolia.

Parinari curatellifolia is an important evergreen tree from the Miombo woodland of south-central and eastern Africa. The bark is corky, suggesting an increased protection against the ecosystem high temperatures and drought conditions as well as against wild fires. The cork in the bark rhytidome of P. curatellifolia was analyzed here for the first time with a focus on chemical and cellular features. P. curatellifolia cork has the cellular characteristics of cork tissues, with typical honeycomb structure in the tangential section and a brick-wall layer in the transverse and radial sections, without intercellular voids. Chemically P. curatellifolia cork has 8.4 % extractives, 33.9 % suberin, 31.9 % lignin and 25.2 % polysaccharides of the cork. The hemicelluloses are mostly xylans, with a substantial proportion of arabinose and galactose. Suberin showed a proportion of long chain lipids to glycerol (LCLip:Gly, mass ratio) of 8.5, and the long chain monomeric composition included a similar proportion of α,ω-diacids and ω-hydroxy acids (35.4 % and 31.5 % of long chain monomers) with a substantial proportion of monoacids (19.4 % of long chain monomers). Lignin is a guaiacyl-syringyl lignin with S/G of 0.32 and H:G:S of 1:14.1:4.5. The rhytidome composition and the cellular and chemical features of its cork are in line with environment-targeted protective features namely as a transpiration and insulation barrier, and as an increased fire protection.

Time course of changes in the transcriptome during russet induction in apple fruit.

BACKGROUND: Russeting is a major problem in many fruit crops. Russeting is caused by environmental factors such as wounding or moisture exposure of the fruit surface. Despite extensive research, the molecular sequence that triggers russet initiation remains unclear. Here, we present high-resolution transcriptomic data by controlled russet induction at very early stages of fruit development. During Phase I, a patch of the fruit surface is exposed to surface moisture. For Phase II, moisture exposure is terminated, and the formerly exposed surface remains dry. We targeted differentially expressed transcripts as soon as 24 h after russet induction. RESULTS: During moisture exposure (Phase I) of 'Pinova' apple, transcripts associated with the cell cycle, cell wall, and cuticle synthesis (SHN3) decrease, while those related to abiotic stress increase. NAC35 and MYB17 were the earliest induced genes during Phase I. They are therefore linked to the initial processes of cuticle microcracking. After moisture removal (Phase II), the expression of genes related to meristematic activity increased (WOX4 within 24 h, MYB84 within 48 h). Genes related to lignin synthesis (MYB52) and suberin synthesis (MYB93, WRKY56) were upregulated within 3 d after moisture removal. WOX4 and AP2B3 are the earliest differentially expressed genes induced in Phase II. They are therefore linked to early events in periderm formation. The expression profiles were consistent between two different seasons and mirrored differences in russet susceptibility in a comparison of cultivars. Furthermore, expression profiles during Phase II of moisture induction were largely identical to those following wounding. CONCLUSIONS: The combination of a unique controlled russet induction technique with high-resolution transcriptomic data allowed for the very first time to analyse the formation of cuticular microcracks and periderm in apple fruit immediately after the onset of triggering factors. This data provides valuable insights into the spatial-temporal dynamics of russeting, including the synthesis of cuticles, dedifferentiation of cells, and impregnation of cell walls with suberin and lignin.

Priorities for Bark Anatomical Research: Study Venues and Open Questions.

The bark fulfils several essential functions in vascular plants and yields a wealth of raw materials, but the understanding of bark structure and function strongly lags behind our knowledge with respect to other plant tissues. The recent technological advances in sampling and preparation of barks for anatomical studies, along with the establishment of an agreed bark terminology, paved the way for more bark anatomical research. Whilst datasets reveal bark's taxonomic and functional diversity in various ecosystems, a better understanding of the bark can advance the understanding of plants' physiological and environmental challenges and solutions. We propose a set of priorities for understanding and further developing bark anatomical studies, including periderm structure in woody plants, phloem phenology, methods in bark anatomy research, bark functional ecology, relationships between bark macroscopic appearance, and its microscopic structure and discuss how to achieve these ambitious goals.

Epigenetic Modifications Related to Potato Skin Russeting.

Potato tuber skin is a protective corky tissue consisting of suberized phellem cells. Smooth-skinned varieties are characterized by a clean, shiny appearance compared to the darker hue of russeted potatoes. The rough skin of russeted cultivars is a desired, genetically inherited characteristic; however, unwanted russeting of smooth-skinned cultivars often occurs under suboptimal growth conditions. The involvement of epigenetic modifiers in regulating the smooth skin russeting disorder was tested. We used smooth-skin commercial cultivars with and without the russeting disorder and three lines from a breeding population segregating for russeting. Anatomically, the russet skin showed similar characteristics, whether the cause was environmentally triggered or genetically determined. The old outer layers of the corky phellem remain attached to the newly formed phellem layers instead of being sloughed off. Global DNA methylation analysis indicated a significant reduction in the percentage of 5-methylcytosine in mature vs. immature skin and russet vs. smooth skin. This was true for both the smooth-skin commercial cultivars and the russeted lines. The expression level of selected DNA methyltransferases was reduced in accordance. DNA demethylase expression did not change between the skin types and age. Hence, the reduced DNA methylation in mature and russet skin is more likely to be achieved through passive DNA demethylation and loss of methyltransferase activity.

Molecular and spatial landmarks of early mouse skin development.

A wealth of specialized cell populations within the skin facilitates its hair-producing, protective, sensory, and thermoregulatory functions. How the vast cell-type diversity and tissue architecture develops is largely unexplored. Here, with single-cell transcriptomics, spatial cell-type assignment, and cell-lineage tracing, we deconstruct early embryonic mouse skin during the key transitions from seemingly uniform developmental precursor states to a multilayered, multilineage epithelium, and complex dermal identity. We identify the spatiotemporal emergence of hair-follicle-inducing, muscle-supportive, and fascia-forming fibroblasts. We also demonstrate the formation of the panniculus carnosus muscle (PCM), sprouting blood vessels without pericyte coverage, and the earliest residence of mast and dendritic immune cells in skin. Finally, we identify an unexpected epithelial heterogeneity within the early single-layered epidermis and a signaling-rich periderm layer. Overall, this cellular and molecular blueprint of early skin development-which can be explored at https://kasperlab.org/tools-establishes histological landmarks and highlights unprecedented dynamic interactions among skin cells.

Insights into the formation and diversification of a novel chiropteran wing membrane from embryonic development.

BACKGROUND: Through the evolution of novel wing structures, bats (Order Chiroptera) became the only mammalian group to achieve powered flight. This achievement preceded the massive adaptive radiation of bats into diverse ecological niches. We investigate some of the developmental processes that underlie the origin and subsequent diversification of one of the novel membranes of the bat wing: the plagiopatagium, which connects the fore- and hind limb in all bat species. RESULTS: Our results suggest that the plagiopatagium initially arises through novel outgrowths from the body flank that subsequently merge with the limbs to generate the wing airfoil. Our findings further suggest that this merging process, which is highly conserved across bats, occurs through modulation of the programs controlling the development of the periderm of the epidermal epithelium. Finally, our results suggest that the shape of the plagiopatagium begins to diversify in bats only after this merging has occurred. CONCLUSIONS: This study demonstrates how focusing on the evolution of cellular processes can inform an understanding of the developmental factors shaping the evolution of novel, highly adaptive structures.

Complex wound response mechanisms and phellogen evolution - insights from Early Devonian euphyllophytes.

We analyze the oldest fossil occurrences of wound-response periderm to characterize the development of wound responses in early tracheophytes. The origin of periderm production by a cambium (phellogen), an innovation with key roles in protection of inner plant tissues, is poorly explored; understanding periderm development in early tracheophytes can illuminate key aspects of this process. Anatomy of wound-response tissues is characterized in serial sections in a new Early Devonian (Emsian; c. 400 Ma) euphyllophyte from Quebec (Canada) - Nebuloxyla mikmaqiana sp. nov. - and compared to previously described euphyllophyte periderm from the same fossil locality to reconstruct periderm development. Characterizing development in these oldest periderm occurrences allows us to propose a model for the development of wound-response periderm in early tracheophytes: by phellogen activity that is poorly coordinated laterally but bifacial, producing secondary tissues initially outwardly and subsequently inwardly. The earliest occurrences of wound periderm pre-date the oldest known periderm produced systemically as a regular ontogenetic stage (canonical periderm), suggesting that periderm evolved initially as a wound-response mechanism. We hypothesize that canonical periderm evolved by exaptation of this wound sealing mechanism, whose deployment was triggered by tangential tensional stresses induced in the superficial tissues by vascular cambial growth from within.

Cutinized and suberized barriers in leaves and roots: Similarities and differences.

Anatomical, histochemical, chemical, and biosynthetic similarities and differences of cutinized and suberized plant cell walls are presented and reviewed in brief. Based on this, the functional properties of cutinized and suberized plant cell walls acting as transport barriers are compared and discussed in more detail. This is of general importance because fundamental misconceptions about relationships in plant-environment water relations are commonly encountered in the scientific literature. It will be shown here, that cuticles represent highly efficient apoplastic transport barriers significantly reducing the diffusion of water and dissolved compounds. The transport barrier of cuticles is mainly established by the deposition of cuticular waxes. Upon wax extraction, with the cutin polymer remaining, cuticular permeability for water and dissolved non-ionized and lipophilic solutes are increasing by 2-3 orders of magnitude, whereas polar and charged substances (e.g., nutrient ions) are only weakly affected (2- to 3-fold increases in permeability). Suberized apoplastic barriers without the deposition of wax are at least as permeable as the cutin polymer matrix without waxes and hardly offer any resistance to the free movement of water. Only upon the deposition of significant amounts of wax, as it is the case with suberized periderms exposed to the atmosphere, an efficient transport barrier for water can be established by suberized cell walls. Comparing the driving forces (gradients between water potentials inside leaves and roots and the surrounding environment) for water loss acting on leaves and roots, it is shown that leaves must have a genetically pre-defined highly efficient transpiration barrier fairly independent from rapidly changing environmental influences. Roots, in most conditions facing a soil environment with relative humidities very close to 100%, are orders of magnitude more permeable to water than leaf cuticles. Upon desiccation, the permanent wilting point of plants is defined as -1.5 MPa, which still corresponds to nearly 99% relative humidity in soil. Thus, the main reason for plant water stress leading to dehydration is the inability of root tissues to decrease their internal water potential to values more negative than -1.5 MPa and not the lack of a transport barrier for water in roots and leaves. Taken together, the commonly mentioned concepts that a drought-induced increase of cuticular wax or root suberin considerably strengthens the apoplastic leaf or root transport barriers and thus aids in water conservation appears highly questionable.

Solving the regulation puzzle of periderm development using advances in fruit skin.

Periderm protects enlarged organs of most dicots and gymnosperms as a barrier to water loss and disease invasion during their secondary growth. Its development undergoes a complex process with genetically controlled and environmental stress-induced characters. Different development of periderm makes the full and partial russet of fruit skin, which diverges in inheritance with qualitative and quantitative characters, respectively, in pear pome. In addition to its specific genetics, fruit periderm has similar development and structure as that of stem and other organs, making it an appropriate material for periderm research. Recently, progress in histochemical as well as transcriptome and proteome analyses, and quantitative trait locus (QTL) mapping have revealed the regulatory molecular mechanism in the periderm based on the identification of switch genes. In this review, we concentrate on the periderm development, propose the conservation of periderm regulation between fruit and other plant organs based on their morphological and molecular characteristics, and summarize a regulatory network with the elicitors and repressors for the tissue development. Spontaneous programmed-cell death (PCD) or environmental stress produces the original signal that triggers the development of periderm. Spatio-temporal specific PCD produced by PyPPCD1 gene and its homologs can play a key role in the coordinated regulation of cell death related tissue development.

Comparing anatomy, chemical composition, and water permeability of suberized organs in five plant species: wax makes the difference.

MAIN CONCLUSION: The efficiency of suberized plant/environment interfaces as transpiration barriers is not established by the suberin polymer but by the wax molecules sorbed to the suberin polymer. Suberized cell walls formed as barriers at the plant/soil or plant/atmosphere interface in various plant organs (soil-grown roots, aerial roots, tubers, and bark) were enzymatically isolated from five different plant species (Clivia miniata, Monstera deliciosa, Solanum tuberosum, Manihot esculenta, and Malus domestica). Anatomy, chemical composition and efficiency as transpiration barriers (water loss in m s-1) of the different suberized cell wall samples were quantified. Results clearly indicated that there was no correlation between barrier properties of the suberized interfaces and the number of suberized cell layers, the amount of soluble wax and the amounts of suberin. Suberized interfaces of C. miniata roots, M. esculenta roots, and M. domestica bark periderms formed poor or hardly any transpiration barrier. Permeances varying between 1.1 and 5.1 × 10-8 m s-1 were very close to the permeance of water (7.4 × 10-8 m s-1) evaporating from a water/atmosphere interface. Suberized interfaces of aerial roots of M. deliciosa and tubers of S. tuberosum formed reasonable transpiration barriers with permeances varying between 7.4 × 10-10 and 4.2 × 10-9 m s-1, which were similar to the upper range of permeances measured with isolated cuticles (about 10-9 m s-1). Upon wax extraction, permeances of M. deliciosa and S. tuberosum increased nearly tenfold, which proves the importance of wax establishing a transpiration barrier. Finally, highly opposite results obtained with M. esculenta and S. tuberosum periderms are discussed in relation to their agronomical importance for postharvest losses and tuber storage.

Chemical and Molecular Characterization of Wound-Induced Suberization in Poplar (Populus alba × P. tremula) Stem Bark.

Upon mechanical damage, plants produce wound responses to protect internal tissues from infections and desiccation. Suberin, a heteropolymer found on the inner face of primary cell walls, is deposited in specific tissues under normal development, enhanced under abiotic stress conditions and synthesized by any tissue upon mechanical damage. Wound-healing suberization of tree bark has been investigated at the anatomical level but very little is known about the molecular mechanisms underlying this important stress response. Here, we investigated a time course of wound-induced suberization in poplar bark. Microscopic changes showed that polyphenolics accumulate 3 days post wounding, with aliphatic suberin deposition observed 5 days post wounding. A wound periderm was formed 9 days post wounding. Chemical analyses of the suberin polyester accumulated during the wound-healing response indicated that suberin monomers increased from 0.25 to 7.98 mg/g DW for days 0 to 28, respectively. Monomer proportions varied across the wound-healing process, with an overall ratio of 2:1 (monomers:glycerol) found across the first 14 days post wounding, with this ratio increasing to 7:2 by day 28. The expression of selected candidate genes of poplar suberin metabolism was investigated using qRT-PCR. Genes queried belonging to lipid polyester and phenylpropanoid metabolism appeared to have redundant functions in native and wound-induced suberization. Our data show that, anatomically, the wounding response in poplar bark is similar to that described in periderms of other species. It also provides novel insight into this process at the chemical and molecular levels, which have not been previously studied in trees.

Mouse models in palate development and orofacial cleft research: Understanding the crucial role and regulation of epithelial integrity in facial and palate morphogenesis.

Cleft lip and cleft palate are common birth defects resulting from genetic and/or environmental perturbations of facial development in utero. Facial morphogenesis commences during early embryogenesis, with cranial neural crest cells interacting with the surface ectoderm to form initially partly separate facial primordia consisting of the medial and lateral nasal prominences, and paired maxillary and mandibular processes. As these facial primordia grow around the primitive oral cavity and merge toward the ventral midline, the surface ectoderm undergoes a critical differentiation step to form an outer layer of flattened and tightly connected periderm cells with a non-stick apical surface that prevents epithelial adhesion. Formation of the upper lip and palate requires spatiotemporally regulated inter-epithelial adhesions and subsequent dissolution of the intervening epithelial seam between the maxillary and medial/lateral nasal processes and between the palatal shelves. Proper regulation of epithelial integrity plays a paramount role during human facial development, as mutations in genes encoding epithelial adhesion molecules and their regulators have been associated with syndromic and non-syndromic orofacial clefts. In this chapter, we summarize mouse genetic studies that have been instrumental in unraveling the mechanisms regulating epithelial integrity and periderm differentiation during facial and palate development. Since proper epithelial integrity also plays crucial roles in wound healing and cancer, understanding the mechanisms regulating epithelial integrity during facial development have direct implications for improvement in clinical care of craniofacial patients.