- Changing Carrot Color: Insertions in DcMYB7 Alter the Regulation of Anthocyanin Biosynthesis and Modification. [Journal Article]
- PPPlant Physiol 2019 Jun 18
- The original domesticated carrots (Daucus carota L.) are thought to have been purple, accumulating large quantities of anthocyanins in their roots. A quantitative trait locus associated with anthocya…
The original domesticated carrots (Daucus carota L.) are thought to have been purple, accumulating large quantities of anthocyanins in their roots. A quantitative trait locus associated with anthocyanin pigmentation in purple carrot roots has been identified on chromosome 3 and includes two candidate genes, DcMYB6 and DcMYB7. Here, we characterized the functions of DcMYB6 and DcMYB7 in carrots. Overexpression of DcMYB7, but not DcMYB6, in the orange carrot 'Kurodagosun' led to anthocyanin accumulation in roots. Knockout of DcMYB7 in the solid purple (purple periderm, phloem and xylem) carrot 'Deep purple' using the CRISPR/Cas9 system resulted in carrots with yellow roots. DcMYB7 could activate expression of its DcbHLH3 partner, a homologue of the anthocyanin-related apple bHLH3, and structural genes in the anthocyanin biosynthetic pathway. We determined that the promoter sequence of DcMYB7 in non-purple carrots was interrupted either by DcMYB8, a non-functional tandem duplication of DcMYB7, or by two transposons, leading to the transcriptional inactivation of DcMYB7 in non-purple carrot roots. As a result, non-purple carrots fail to accumulate anthocyanins in their roots. Our study supports the hypothesis that another genetic factor suppresses DcMYB7 expression in the phloem and xylem of purple peridermal carrot root tissues. DcMYB7 also regulated the glycosylation and acylation of anthocyanins by directly activating DcUCGXT1 and DcSAT1. We revealed the genetic factors conditioning anthocyanin pigmentation in purple versus non-purple carrot roots. Our results also provide insights into the mechanisms underlying anthocyanin glycosylation and acylation.
- The molecular anatomy of mammalian upper lip and primary palate fusion at single cell resolution. [Journal Article]
- DDevelopment 2019 Jun 17; 146(12)
- The mammalian lip and primary palate form when coordinated growth and morphogenesis bring the nasal and maxillary processes into contact, and the epithelia co-mingle, remodel and clear from the fusio…
The mammalian lip and primary palate form when coordinated growth and morphogenesis bring the nasal and maxillary processes into contact, and the epithelia co-mingle, remodel and clear from the fusion site to allow mesenchyme continuity. Although several genes required for fusion have been identified, an integrated molecular and cellular description of the overall process is lacking. Here, we employ single cell RNA sequencing of the developing mouse face to identify ectodermal, mesenchymal and endothelial populations associated with patterning and fusion of the facial prominences. This analysis indicates that key cell populations at the fusion site exist within the periderm, basal epithelial cells and adjacent mesenchyme. We describe the expression profiles that make each population unique, and the signals that potentially integrate their behaviour. Overall, these data provide a comprehensive high-resolution description of the various cell populations participating in fusion of the lip and primary palate, as well as formation of the nasolacrimal groove, and they furnish a powerful resource for those investigating the molecular genetics of facial development and facial clefting that can be mined for crucial mechanistic information concerning this prevalent human birth defect.
- StMYB44 negatively regulates anthocyanin biosynthesis at high temperatures in tuber flesh of potato. [Journal Article]
- JEJ Exp Bot 2019 Apr 25
- High temperatures are known to reduce anthocyanin accumulation in a number of diverse plant species. In potato (Solanum tuberosum L.), high temperature significantly reduces tuber anthocyanin pigment…
High temperatures are known to reduce anthocyanin accumulation in a number of diverse plant species. In potato (Solanum tuberosum L.), high temperature significantly reduces tuber anthocyanin pigment content. However, the mechanism of anthocyanin biosynthesis in potato tuber under heat stress remains unknown. Here we show that high temperature causes reduction of anthocyanin biosynthesis in both potato tuber skin and flesh, with white areas forming between the vasculature and periderm. Heat stress reduced the expression of the R2R3 MYB transcription factor (TF) StAN1 and StbHLH1, members of the transcriptional complex responsible for coordinative regulation of the skin and flesh pigmentation, as well as anthocyanin biosynthetic pathway genes in white regions. However, the core phenylpropanoid pathway genes, lignin and chlorogenic acid (CGA) pathway genes were up-regulated in white areas, suggesting that suppression of the anthocyanin branch may result in re-routing phenylpropanoid flux into the CGA or lignin biosynthesis branches. Two R2R3 MYB TFs, StMYB44-1 and StMYB44-2, were highly expressed in white regions under high temperature. In transient assays, StMYB44 represses anthocyanin accumulation in leaves of N. tabacum and N. benthamiana by directly suppressing the activity of the dihydroflavonol reductase (DFR) promoter. StMYB44-1 showed stronger repressive capacity than StMYB44-2 with both predicted proteins containing the repression-associated EAR motif with some variation. StMYB44-1 conferred repression without a requirement for a bHLH partner, suggesting a different repression mechanism from reported anthocyanin repressors. We propose that temperature-induced reduction of anthocyanin accumulation in potato flesh is caused by down-regulation of the activating anthocyanin regulatory complex, by enhancing the expression of flesh-specific StMYB44 and alteration of phenylpropanoid flux.
- Identification of regulatory elements recapitulating early expression of L-plastin in the zebrafish enveloping layer and embryonic periderm. [Journal Article]
- GEGene Expr Patterns 2019; 32:53-66
- We have cloned and characterized an intronic fragment of zebrafish lymphocyte cytosolic protein 1 (lcp1, also called L-plastin) that drives expression to the zebrafish enveloping layer (EVL). L-plast…
We have cloned and characterized an intronic fragment of zebrafish lymphocyte cytosolic protein 1 (lcp1, also called L-plastin) that drives expression to the zebrafish enveloping layer (EVL). L-plastin is a calcium-dependent actin-bundling protein belonging to the plastin/fimbrin family of proteins, and is necessary for the proper migration and attachment of several adult cell types, including leukocytes and osteoclasts. However, in zebrafish lcp1 is abundantly expressed much earlier, during differentiation of the EVL. The cells of this epithelial layer migrate collectively, spreading vegetally over the yolk. L-plastin expression persists into the larval periderm, a transient epithelial tissue that forms the first larval skin. This finding establishes that L-plastin is activated in two different embryonic waves, with a distinct regulatory switch between the early EVL and the later leukocyte. To better study L-plastin expressing cells we attempted CRISPR/Cas9 homology-driven recombination (HDR) to insert a self-cleaving peptide (Cre-P2A-EGFP-CAAX) downstream of the native lcp1 promoter. This produced a stable zebrafish line expressing Cre recombinase in EVL nuclei and green fluorescence in EVL cell membranes. In vivo tracking of these labeled cells provided enhanced views of EVL migration behavior, membrane extensions, and mitotic events. Finally, we experimentally dissected key elements of the targeted lcp1 locus, discovering a ∼300 bp intronic sequence sufficient to drive EVL expression. The lcp1: Cre-P2A-EGFP-CAAX zebrafish should be useful for studying enveloping layer specification, gastrulation movements and periderm development in this widely used vertebrate model. In addition, the conserved regulatory sequences we have isolated predict that L-plastin orthologs may have a similar early expression pattern in other vertebrate embryos.
- Tissue-specific study across the stem reveals the chemistry and transcriptome dynamics of birch bark. [Journal Article]
- NPNew Phytol 2019; 222(4):1816-1831
- Tree bark is a highly specialized array of tissues that plays important roles in plant protection and development. Bark tissues develop from two lateral meristems; the phellogen (cork cambium) produc…
Tree bark is a highly specialized array of tissues that plays important roles in plant protection and development. Bark tissues develop from two lateral meristems; the phellogen (cork cambium) produces the outermost stem-environment barrier called the periderm, while the vascular cambium contributes with phloem tissues. Although bark is diverse in terms of tissues, functions and species, it remains understudied at higher resolution. We dissected the stem of silver birch (Betula pendula) into eight major tissue types, and characterized these by a combined transcriptomics and metabolomics approach. We further analyzed the varying bark types within the Betulaceae family. The two meristems had a distinct contribution to the stem transcriptomic landscape. Furthermore, inter- and intraspecies analyses illustrated the unique molecular profile of the phellem. We identified multiple tissue-specific metabolic pathways, such as the mevalonate/betulin biosynthesis pathway, that displayed differential evolution within the Betulaceae. A detailed analysis of suberin and betulin biosynthesis pathways identified a set of underlying regulators and highlighted the important role of local, small-scale gene duplication events in the evolution of metabolic pathways. This work reveals the transcriptome and metabolic diversity among bark tissues and provides insights to its development and evolution, as well as its biotechnological applications.
- Fortification and bioavailability of zinc in potato. [Journal Article]
- JSJ Sci Food Agric 2019; 99(7):3525-3529
- CONCLUSIONS: A substantial increase in Zn bioavailability was obtained by priming the tubers for 12 h in 10 mg mL-1 Zn. © 2019 Society of Chemical Industry.
- Distribution and chemical forms of cadmium in Coptis chinensis Franch. determined by laser ablation ICP-MS, cell fractionation, and sequential extraction. [Journal Article]
- EEEcotoxicol Environ Saf 2019 Apr 30; 171:894-903
- Coptis chinensis Franch., is a widely used medicinal plant in China. This plant is often contaminated by cadmium (Cd) and render health risk to human consumers. Understanding distribution of Cd and i…
Coptis chinensis Franch., is a widely used medicinal plant in China. This plant is often contaminated by cadmium (Cd) and render health risk to human consumers. Understanding distribution of Cd and its chemical forms is important to evaluate accumulation of the metal and its detoxification mechanisms in this plant. Since few studies have focused on this aspect, we used laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) to spatially locate Cd in rhizome cross-sections, and ICP-MS to analyze the Cd subcellular distribution and the chemical forms of Cd in different tissues. Rhizome bioimaging results showed that Cd was distributed predominantly within the periderm, cortex, pith, and root trace vascular bundle. The LA-ICP-MS results suggested that Ca2+ channels might be a pathway for Cd entry into the plant. Subcellular distribution data indicated that most of Cd was associated with the cell wall (41.8-77.1%) and the soluble fraction (14.4-52.7%) in all tissues. Analysis of chemical forms revealed that majority Cd existed in less mobile and less toxic forms in all tissues, and P could convert to insoluble phosphate with Cd to moderate Cd toxicity. The new understanding of Cd accumulation and detoxification might provide novel strategies for reducing the levels of Cd in C. chinensis Franch., thereby mitigating its potential transfer to humans and providing a theoretical basis for evaluating the Cd status in other medicinal plants. Further, our findings might provide a basis for establishing a reasonable Cd limit level of traditional Chinese medicinal materials.
- Temporal resistance of potato tubers: Antibacterial assays and metabolite profiling of wound-healing tissue extracts from contrasting cultivars. [Journal Article]
- PPhytochemistry 2019; 159:75-89
- Solanum tuberosum, commonly known as the potato, is a worldwide food staple. During harvest, storage, and distribution the crop is at risk of mechanical damage. Wounding of the tuber skin can also be…
Solanum tuberosum, commonly known as the potato, is a worldwide food staple. During harvest, storage, and distribution the crop is at risk of mechanical damage. Wounding of the tuber skin can also become a point of entry for bacterial and fungal pathogens, resulting in substantial agricultural losses. Building on the proposal that potato tubers produce metabolites to defend against microbial infection during early stages of wound healing before protective suberized periderm tissues have developed, we assessed extracts of wound tissues from four potato cultivars with differing skin morphologies (Norkotah Russet, Atlantic, Chipeta, and Yukon Gold). These assays were conducted at 0, 1, 2, 3 and 7 days post wounding against the plant pathogen Erwinia carotovora and a non-pathogenic Escherichia coli strain that served as a control. For each of the potato cultivars, only polar wound tissue extracts demonstrated antibacterial activity. The polar extracts from earlier wound-healing time points (days 0, 1 and 2) displayed notably higher antibacterial activity against both strains than the later wound-healing stages (days 3 and 7). These results support a burst of antibacterial activity at early time points. Parallel metabolite profiling of the extracts revealed differences in chemical composition at different wound-healing time points and allowed for identification of potential marker compounds according to healing stage for each of the cultivars. It was possible to monitor the transformations in the metabolite profiles that could account for the phenomenon of temporal resistance by looking at the relative quantities of various metabolite classes as a function of time.
- Russeting partially restores apple skin permeability to water vapour. [Journal Article]
- PPlanta 2019; 249(3):849-860
- CONCLUSIONS: The higher water loss of russeted fruit results from the higher permeance of the periderm of the russeted skin as compared to that of the intact cuticle and epidermis. Apple fruit surfaces are often in-parallel composites, comprising areas of intact cuticle (atop a healthy epidermis) adjacent to areas covered by periderm (so-called russet). The occurrence of non-russeting and russeting genotypes makes this species an ideal model to study the barrier properties of its composite skin. The objective was to quantify the water vapour permeances of non-russeted ([Formula: see text]) and russeted fruit skins ([Formula: see text]). Rates of water loss from whole fruit ([Formula: see text]) and excised epidermal skin segments (ES) or peridermal skin segments (PS) were quantified gravimetrically. The [Formula: see text] was larger in russeting than in non-russeting genotypes because [Formula: see text] exceeded [Formula: see text] by about twofold. Also, the [Formula: see text] of russeting genotypes was larger than that of non-russeting genotypes. Generally, [Formula: see text] was more variable than [Formula: see text]. These differences were consistent across seasons and genotypes. The lower [Formula: see text] as compared to [Formula: see text] resulted primarily from the higher wax content of the cuticle of the [Formula: see text]. For non-russeted genotypes, the value of [Formula: see text] was significantly related to the permeance determined on the same intact fruit ([Formula: see text]). Close relationships were also found between the [Formula: see text] calculated from [Formula: see text] determined on the same fruit and the measured [Formula: see text]. For russeting genotypes, the [Formula: see text] or [Formula: see text] were not correlated with [Formula: see text]. The [Formula: see text] calculated from [Formula: see text], [Formula: see text], [Formula: see text] and [Formula: see text] (all determined on an individual-fruit basis) was significantly correlated with the measured [Formula: see text]. Our results demonstrate that the periderm permeance exceeds the cuticle permeance and that permeances of non-russeted surfaces of russeting genotypes exceed those of non-russeting genotypes.
New Search Next
- Embryonic development of parakeratinized epithelium of the tongue in the domestic duck (Anas platyrhynchos f. domestica): LM, SEM, and TEM observations. [Journal Article]
- PProtoplasma 2019; 256(3):631-642
- The parakeratinized epithelium is a common and widespread type of keratinized epithelium in the oral cavity in adult birds. In contrast to orthokeratinized epithelium, which mostly covers mechanical …
The parakeratinized epithelium is a common and widespread type of keratinized epithelium in the oral cavity in adult birds. In contrast to orthokeratinized epithelium, which mostly covers mechanical papillae and the lingual nail, parakeratinized epithelium covers almost the entire dorsal surface of the tongue in birds. The characteristic feature of parakeratinized epithelium is the presence of nuclei in the keratinized layer. The present study aimed to investigate for the first time the micro- and ultrastructural changes of parakeratinized epithelium during embryonic development and to assess the readiness of the epithelium to serve protective functions during food transport to the esophagus. Three developmental stages were distinguished: embryonic, transformation, and pre-hatching stages. The embryonic stage lasts from the 9th to the 14th day of incubation and the epithelium is composed of undifferentiated epithelial cells. The transformation stage lasts from the 15th to the 22nd day of incubation and the epithelium undergoes transformation into stratified epithelium consisting of basal, intermediate, and superficial layers. The characteristic feature of this stage is formation of the periderm with osmophilic granules. The pre-hatching stage starts on the 23rd day, and the epithelium with a fully developed keratinized layer resembles that of the epithelium in adult animals. No periderm was observed on the epithelial surface. It was confirmed that at the time of hatching the parakeratinized epithelium is fully differentiated and ready to fulfill its function during food transport. The presence of periderm is a common feature characteristic for para- and orthokeratinized epithelium in the oral cavity of birds. However, the formation of the keratinized/cornified layer is different for these two types of keratinized epithelia.