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The root parasitic weed Striga hermonthica has a devastating effect on sorghum and other cereal crops in Sub-Saharan Africa. Available Striga management strategies are rarely sufficient or not widely accessible or affordable. Identification of soil- or plant-associated microorganisms that interfere in the Striga infection cycle holds potential for development of complementary biological control measures. Such inoculants should be preferably based on microbes native to the regions of their application. We developed a method to assess microbiome-based soil suppressiveness to Striga with a minimal amount of field-collected soil. We previously used this method to identify the mechanisms of microbe-mediated suppression of Striga infection and to test individual microbial strains. Here, we present protocols to assess the functional potential of the soil microbiome and individual bacterial taxa that adversely affect Striga parasitism in sorghum via three major known suppression mechanisms. These methods can be further extended to other Striga hosts and other root parasitic weeds. Key features • This protocol provides a detailed description of the methods used in Kawa et al. [1]. • This protocol is optimized to assess soil suppressiveness to Striga infection by using natural field-collected soil and the same soil sterilized by gamma-radiation. • This protocol is optimized to test bacterial (and not fungal) isolates. • This protocol can be easily extended to other host-parasite-microbiome systems.

Background: The main application of cork is the production of stoppers for wine bottles. Cork sometimes contains 2,4,6-trichloroanisole, a compound that, at a concentration of ng/L, produces an unpleasant musty odor that destroys the organoleptic properties of wine and results in enormous economic losses for wineries and cork industries. Cork can exhibit a defect known as yellow stain, which is associated with high levels of 2,4,6-trichloroanisole. We describe how the microbiota of cork and yellow stain define a novel mechanism that explains the formation of chlorophenols and chloroanisoles (including 2,4,6-trichloroanisole) from p-hydroxybenzoate produced during lignin and/or suberin breakdown. Results: Electron microscopy revealed that cork affected by yellow stain exhibited significant structural degradation. This deterioration was attributed to the presence of higher microbial populations compared to those found in standard cork. Cork microbiota is rich in filamentous fungi able to metabolize lignin. A metataxonomic analysis confirmed that yellow stain contained significantly greater populations of fungal species belonging to Absidia, Geomyces, Mortierella, Mucor, Penicillium, Pseudogymnoascus, Talaromyces, and Umbelopsis. It also contained significantly greater amounts of bacteria belonging to Enterobacterales, Streptosporangiales, Tepidisphaerales, Pseudomonas, and several members of Burkholderiaceae, particularly species of the Burkholderia-Caballeronia-Paraburkholderia group. The extraction of aromatic compounds from cork samples allowed the identification of several compounds typically observed following lignin depolymerization. Notably, p-hydroxybenzoic acid and phenol were detected. Two strains of the genus Streptomyces isolated from yellow stain were able to biotransform p-hydroxybenzoate into phenol in resting cell assays. Phenol could be efficiently chlorinated in vitro to produce 2,4,6-trichlorophenol by a fungal chloroperoxidase, an enzymatic activity commonly found in filamentous fungi isolated from cork. Finally, as has been widely demonstrated before, 2,4,6-trichlorophenol can be efficiently O-methylated to 2,4,6-trichloroanisole by many of fungi that inhabit cork. Conclusions: Chlorophenols and chloroanisoles can be produced de novo in cork from p-hydroxybenzoate generated by the microbial biodegradation of lignin and/or suberin through the participation of different types of microorganisms present in cork. The natural origin of these compounds, which are of great interest for the chlorine cycle and represent a new source of environmental contamination that differs from that caused by human activity, is described. BmZYZKVv9XTmg3g6suYKG9 Video Abstract [ABSTRACT FROM AUTHOR]

Suberin biosynthesis involves the coordinated regulation of both phenolic and aliphatic metabolisms. HXXXD/BAHD acyltransferases occupy a unique place in suberization, as they function to crosslink phenolic and aliphatic monomers during suberin assembly. To date, only one suberin-associated HXXXD/BAHD acyltransferase, StFHT, has been described in potatoes, whereas, in Arabidopsis, at least two are implicated in suberin biosynthesis. RNAseq data from wound-induced potato tubers undergoing suberization indicate that transcripts for 28 HXXXD/BAHD acyltransferase genes accumulate in response to wounding. In the present study, we generated RNAi knockdown lines for StFHT and another highly wound-induced HXXXD/BAHD acyltransferase, designated StHCT, and characterized their wound-induced suberin phenotype. StFHT-RNAi and StHCT-RNAi knockdown lines share the same aliphatic suberin phenotype of reduced esterified ferulic acid and ferulates, which is similar to the previously described StFHT-RNAi knockdown suberin phenotype. However, the phenolic suberin phenotype differed between the two knockdown genotypes, with StHCT-RNAi knockdown lines having proportionately more p-hydroxyphenyl-derived moieties than either StFHT-RNAi knockdown or empty vector control lines. Analysis of soluble polar metabolites revealed that StHCT catalyzes a step upstream from StFHT. Overall, our data support the involvement of more than one HXXXD/BAHD acyltransferase in potato suberin biosynthesis. [ABSTRACT FROM AUTHOR]

Abstract Background The main application of cork is the production of stoppers for wine bottles. Cork sometimes contains 2,4,6-trichloroanisole, a compound that, at a concentration of ng/L, produces an unpleasant musty odor that destroys the organoleptic properties of wine and results in enormous economic losses for wineries and cork industries. Cork can exhibit a defect known as yellow stain, which is associated with high levels of 2,4,6-trichloroanisole. We describe how the microbiota of cork and yellow stain define a novel mechanism that explains the formation of chlorophenols and chloroanisoles (including 2,4,6-trichloroanisole) from p-hydroxybenzoate produced during lignin and/or suberin breakdown. Results Electron microscopy revealed that cork affected by yellow stain exhibited significant structural degradation. This deterioration was attributed to the presence of higher microbial populations compared to those found in standard cork. Cork microbiota is rich in filamentous fungi able to metabolize lignin. A metataxonomic analysis confirmed that yellow stain contained significantly greater populations of fungal species belonging to Absidia, Geomyces, Mortierella, Mucor, Penicillium, Pseudogymnoascus, Talaromyces, and Umbelopsis. It also contained significantly greater amounts of bacteria belonging to Enterobacterales, Streptosporangiales, Tepidisphaerales, Pseudomonas, and several members of Burkholderiaceae, particularly species of the Burkholderia-Caballeronia-Paraburkholderia group. The extraction of aromatic compounds from cork samples allowed the identification of several compounds typically observed following lignin depolymerization. Notably, p-hydroxybenzoic acid and phenol were detected. Two strains of the genus Streptomyces isolated from yellow stain were able to biotransform p-hydroxybenzoate into phenol in resting cell assays. Phenol could be efficiently chlorinated in vitro to produce 2,4,6-trichlorophenol by a fungal chloroperoxidase, an enzymatic activity commonly found in filamentous fungi isolated from cork. Finally, as has been widely demonstrated before, 2,4,6-trichlorophenol can be efficiently O-methylated to 2,4,6-trichloroanisole by many of fungi that inhabit cork. Conclusions Chlorophenols and chloroanisoles can be produced de novo in cork from p-hydroxybenzoate generated by the microbial biodegradation of lignin and/or suberin through the participation of different types of microorganisms present in cork. The natural origin of these compounds, which are of great interest for the chlorine cycle and represent a new source of environmental contamination that differs from that caused by human activity, is described. Video Abstract

Climate change may alter soil microbial communities and soil organic matter (SOM) composition. Soil carbon (C) cycling takes place over multiple time scales; therefore, long-term studies are essential to better understand the factors influencing C storage and help predict responses to climate change. To investigate this further, soils that were heated by 5 °C above ambient soil temperatures for 18 years were collected from the Barre Woods Soil Warming Study at the Harvard Forest Long-term Ecological Research site. This site consists of large 30 × 30 m plots (control or heated) where entire root systems are exposed to sustained warming conditions. Measurements included soil C and nitrogen concentrations, microbial biomass, and SOM chemistry using gas chromatography–mass spectrometry and solid-state 13C nuclear magnetic resonance spectroscopy. These complementary techniques provide a holistic overview of all SOM components and a comprehensive understanding of SOM composition at the molecular-level. Our results showed that soil C concentrations were not significantly altered with warming; however, various molecular-level alterations to SOM chemistry were observed. We found evidence for both enhanced SOM decomposition and increased above-ground plant inputs with long-term warming. We also noted shifts in microbial community composition while microbial biomass remained largely unchanged. These findings suggest that prolonged warming induced increased availability of preferred substrates, leading to shifts in the microbial community and SOM biogeochemistry. The observed increase in gram-positive bacteria indicated changes in substrate availability as gram-positive bacteria are often associated with the decomposition of complex organic matter, while gram-negative bacteria preferentially break down simpler organic compounds altering SOM composition over time. Our results also highlight that additional plant inputs do not effectively offset chronic warming-induced SOM decomposition in temperate forests. [ABSTRACT FROM AUTHOR]

Bark, the outermost tissue, plays an important role in protecting trees from damage induced by living organisms and the surrounding environment. Bark differs from the xylem primarily by the presence of suberin in cork cell walls. However, few studies have examined the role of suberin and its interactions with other chemical components in the cork. Consequently, this study aimed to understand the distribution of chemical components, including suberin and lignin, and their respective roles in cork cell walls, using Cerasus jamasakura (Siebold ex Koidz.) H. Ohba. Suberin and lignin were gradually and selectively removed from thin strip specimens. Fourier transform infrared (FTIR) spectroscopy suggested that desuberinization removed both suberin and part of the other matrix substances within a few minutes of treatment, whereas delignification exclusively removed lignin. Further microscopic observation revealed that suberin present was mainly in the secondary wall of cork cells, whereas lignin was present in both the tertiary wall and compound middle lamella. In addition, the cell wall collapse of the cork was only found in desuberinized specimens, whereas delignified specimens only showed monotonic contraction. Taken together, these results suggest that the presence of suberin in the cork contributes to the shape stability of cork cell walls.

All-cellulose composites (ACCs) have been fabricated by using a variety of cellulosic sources, versatile technologies, and are sustainable alternatives for traditional composites. In this study, nonwoven-woven ACC laminates were created from wood-based Spinnova short fibers and Lyocell fabrics via partial dissolution and an NaOH-urea solvent system. The less-known wood-based Spinnova fiber is created for the textile industry, but it also has great potential for the composite industry. To identify the mechanical properties of ACCs—which greatly influence the range of material application—tensile, impact, and flexural tests were conducted. The mechanical properties indicated only moderate properties, which are influenced by high porosity and weak fiber bonding. Despite this, valuable information on the nonwoven-woven structured ACCs was obtained. To improve the ACC laminate’s ability to resist moisture, bio-based coatings (e.g., commercially available birch bark betulin and suberin acid mixture) were applied on the surface of ACCs and it successfully improved the wetting resistance. The results of contact angle analyses demonstrated that the highest contact angle of 128° was measured for betulin-coated laminates and the best stable hydrophobicity calculated a minute after the beginning of the experiment were observed at 109° for the uncommercial pressurized hot ethanol (PHE) extract of birch bark.

All-cellulose composites (ACCs) have been fabricated by using a variety of cellulosic sources, versatile technologies, and are sustainable alternatives for traditional composites. In this study, nonwoven-woven ACC laminates were created from wood-based Spinnova short fibers and Lyocell fabrics via partial dissolution and an NaOH-urea solvent system. The less-known wood-based Spinnova fiber is created for the textile industry, but it also has great potential for the composite industry. To identify the mechanical properties of ACCs--which greatly influence the range of material application--tensile, impact, and flexural tests were conducted. The mechanical properties indicated only moderate properties, which are influenced by high porosity and weak fiber bonding. Despite this, valuable information on the nonwoven-woven structured ACCs was obtained. To improve the ACC laminate's ability to resist moisture, biobased coatings (e.g., commercially available birch bark betulin and suberin acid mixture) were applied on the surface of ACCs and it successfully improved the wetting resistance. The results of contact angle analyses demonstrated that the highest contact angle of 128° was measured for betulin-coated laminates and the best stable hydrophobicity calculated a minute after the beginning of the experiment were observed at 109° for the uncommercial pressurized hot ethanol (PHE) extract of birch bark. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Lipid polymers such as cutin and suberin strengthen the diffusion barrier properties of the cell wall in specific cell types and are essential for water relations, mineral nutrition, and stress protection in plants. Land plant-specific glycerol-3-phosphate acyltransferases (GPATs) of different clades are central players in cutin and suberin monomer biosynthesis. Here, we show that the GPAT4/6/8 clade in Arabidopsis thaliana, which is known to mediate cutin formation, is also required for developmentally regulated root suberization, in addition to the established roles of GPAT5/7 in suberization. The GPAT5/7 clade is mainly required for abscisic acid-regulated suberization. In addition, the GPAT5/7 clade is crucial for the formation of the typical lamellated suberin ultrastructure observed by transmission electron microscopy, as distinct amorphous globular polyester structures were deposited in the apoplast of the gpat5 gpat7 double mutant, in contrast to the thinner but still lamellated suberin deposition in the gpat4 gpat6 gpat8 triple mutant. Site-directed mutagenesis revealed that the intrinsic phosphatase activity of GPAT4, GPAT6, and GPAT8, which leads to monoacylglycerol biosynthesis, contributes to suberin formation. GPAT5/7 lack an active phosphatase domain and the amorphous globular polyester structure observed in the gpat5 gpat7 double mutant was partially reverted by treatment with a phosphatase inhibitor or the expression of phosphatase-dead variants of GPAT4/6/8. Thus, GPATs that lack an active phosphatase domain synthetize lysophosphatidic acids that might play a role in the formation of the lamellated structure of suberin. GPATs with active and nonactive phosphatase domains appear to have nonredundant functions and must cooperate to achieve the efficient biosynthesis of correctly structured suberin. [ABSTRACT FROM AUTHOR]

The utilization of polyols derived from renewable sources presents an opportunity to enhance the sustainability of rigid polyurethane (PUR) foams, thereby contributing to the advancement of a circular bioeconomy. This study explores the development of PUR rigid foams exclusively using polyols sourced from second-generation renewable biomass feedstocks, specifically depolymerized birch bark suberin (suberinic acids) and tall oil fatty acids. The polyols achieved a total renewable material content as high as 74%, with a suberinic acid content of 37%. Response surface modeling was employed to determine the optimal bio-polyol, blowing agents, and catalyst content, hence, optimizing the bio-based foam formulations. In addition, response surface modeling was applied to rigid PUR foam formulations based on commercially available petroleum-based polyols for comparison. The results, including apparent density (~40–44 kg/m3), closed cell content (~95%), compression strength (>0.2 MPa, parallel to the foaming direction), and thermal conductivity (~0.019 W/(m·K)), demonstrated that the suberinic acids-based rigid PUR foam exhibited competitive qualities in comparison to petroleum-based polyols. Remarkably, the bio-based rigid PUR foams comprised up to 29% renewable materials. These findings highlight the potential of suberinic acid-tall oil polyols as effective candidates for developing rigid PUR foams, offering promising solutions for sustainable insulation applications. [ABSTRACT FROM AUTHOR]

Suberin biosynthesis involves the coordinated regulation of both phenolic and aliphatic metabolisms. HXXXD/BAHD acyltransferases occupy a unique place in suberization, as they function to crosslink phenolic and aliphatic monomers during suberin assembly. To date, only one suberin-associated HXXXD/BAHD acyltransferase, StFHT, has been described in potatoes, whereas, in Arabidopsis, at least two are implicated in suberin biosynthesis. RNAseq data from wound-induced potato tubers undergoing suberization indicate that transcripts for 28 HXXXD/BAHD acyltransferase genes accumulate in response to wounding. In the present study, we generated RNAi knockdown lines for StFHT and another highly wound-induced HXXXD/BAHD acyltransferase, designated StHCT, and characterized their wound-induced suberin phenotype. StFHT-RNAi and StHCT-RNAi knockdown lines share the same aliphatic suberin phenotype of reduced esterified ferulic acid and ferulates, which is similar to the previously described StFHT-RNAi knockdown suberin phenotype. However, the phenolic suberin phenotype differed between the two knockdown genotypes, with StHCT-RNAi knockdown lines having proportionately more p-hydroxyphenyl-derived moieties than either StFHT-RNAi knockdown or empty vector control lines. Analysis of soluble polar metabolites revealed that StHCT catalyzes a step upstream from StFHT. Overall, our data support the involvement of more than one HXXXD/BAHD acyltransferase in potato suberin biosynthesis.

Mangrove plants demonstrate an impressive ability to tolerate environmental pollutants, but excessive levels of cadmium (Cd) can impede their growth. Few studies have focused on the effects of apoplast barriers on heavy metal tolerance in mangrove plants. To investigate the uptake and tolerance of Cd in mangrove plants, two distinct mangrove species, Avicennia marina and Rhizophora stylosa, are characterized by unique apoplast barriers. The results showed that both mangrove plants exhibited the highest concentration of Cd2+ in roots, followed by stems and leaves. The Cd2+ concentrations in all organs of R. stylosa consistently exhibited lower levels than those of A. marina. In addition, R. stylosa displayed a reduced concentration of apparent PTS and a smaller percentage of bypass flow when compared to A. marina. The root anatomical characteristics indicated that Cd treatment significantly enhanced endodermal suberization in both A. marina and R. stylosa roots, and R. stylosa exhibited a higher degree of suberization. The transcriptomic analysis of R. stylosa and A. marina roots under Cd stress revealed 23 candidate genes involved in suberin biosynthesis and 8 candidate genes associated with suberin regulation. This study has confirmed that suberized apoplastic barriers play a crucial role in preventing Cd from entering mangrove roots. [ABSTRACT FROM AUTHOR]

Premise: Leaf subepidermal secretory cavities are a notable trait in Myrtaceae, but their formation is still controversial because of the lack of consensus on their ontogeny among authors. Knowledge about the compounds present in these cavities has grown over the last few years, demonstrating that terpenoid‐rich oils are not their unique content. These two points are the focus of this study on the ontogeny, structure, and contents of secretory cavities in neotropical Myrtaceae. Methods: We used histochemical tests and Raman analysis to verify the basic chemical composition of the cavity contents of nine species. We studied the ontogeny of glands in one species, comparing aldehyde‐fixed tissues and fresh sections mounted in an inert medium. Results: We observed schizogenous development and appearance of the secretory cavities and found that sample processing may induce cell breakdown, which can be misinterpreted as lysigeny. The content of these cavities contains putative terpenes, resins, carbonyl groups, and flavonoids. Conclusions: Our findings support the hypothesis that the lysigenous appearance of the oil glands is a technical artifact. These tissue distortions must be considered when interpreting the development of this type of secretory structure. Moreover, the basic analyses of chemical constituents show for the first time that the glands of neotropical Myrtaceae are potential reservoirs of some compounds such as flavonoids previously reported as novelties for a few other myrtaceous species. Because some of them are non‐lipid compounds, the idea that the glands are just oil repositories is no longer applicable. [ABSTRACT FROM AUTHOR]

Fire hazard is a constant risk in everyday life with the use of combustibles such as polymeric materials, wood, and fabrics, to name a few. Halogenated compounds have been widely used as efficient flame-retardants, often being applied as coatings or impregnations. With growing environmental concerns and regional bans on the use of halogenated flame-retardant compounds, bio-based alternatives are garnering significant research interest. Naturally occurring materials such as eggshells, DNA, and certain proteins have developed a self-defense mechanism against fire over millions of years of evolution. Cork, a naturally occurring biological tissue in outer bark, is of interest as it is often used as a heat shield and moisture repellent, specifically in spacecraft. A deeper look into the chemical structure of cork indicates the presence of suberin, a bio-polyester group that makes up as much as 40% of its chemical composition. These bio-polyester groups play a key role as a protective barrier between the plant and the surrounding external environment. Thus, the role of suberin in plants could be mimicked for the design of biobased flame-retardant materials.

Histological stains are useful tools for characterizing cell shape, arrangement and the material they are made from. Stains can be used individually or simultaneously to mark different cell structures or polymers within the same cells, and to visualize them in different colors. Histological stains can be combined with genetically-encoded fluorescent proteins, which are useful for understanding of plant development. To visualize suberin lamellae by fluorescent microscopy, we improved a histological staining procedure with the dyes Fluorol Yellow 088 and aniline blue. In the complex plant organs such as roots, suberin lamellae are deposited deep within the root on the endodermal cell wall. Our procedure yields reliable and detailed images that can be used to determine the suberin pattern in root cells. The main advantage of this protocol is its efficiency, the detailed visualization of suberin localization it generates in the root, and the possibility of returning to the confocal images to analyze and re-evaluate data if necessary.

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. [ABSTRACT FROM AUTHOR]

Exocarp color is a commercially essential quality for pear which can be divided into two types: green and russet. The occurrence of russet color is associated with deficiencies and defects in the cuticular and epidermal layers, which affect the structure of the cell wall and the deposition of suberin. Until now, the genetic basics triggering this trait have not been well understood, and limited genes have been identified for the trait. To figure out the gene controlling the trait of exocarp color, we perform a comprehensive genome-wide association study, and we describe the candidate genes. One gene encoding the ABCG protein has been verified to be associated with the trait, using an integrative analysis of the metabolomic and transcriptomic data. This review covers a variety of omics resources, which provide a valuable resource for identifying gene-controlled traits of interest. The findings in this study help to elucidate the genetic components responsible for the trait of exocarp color in pear, and the implications of these findings for future pear breeding are evaluated. [ABSTRACT FROM AUTHOR]

Taurus flowering ash (Fraxinus ornus subsp. cilicica) is an endemic tree species in Turkey. The bark of the species was characterized for summative chemical composition, the monomeric composition of polysaccharides, phenolic content, in vitro and ex vivo antioxidant properties of hydrophilic extracts, the composition of lipophilic extractives and suberin, and thermal degradation. The bark has an elevated ash content (17%), primarily composed of calcium, and a noteworthy extractive content (38.9%), predominantly of hydrophilic compounds. The antioxidant activity of the bark extracts is moderate, with an IC50 value of 40 μg/mL and an EC50 value of 230 μg/mL by DPPH and TBARS methods. The lipophilic extractives principally contain fatty acids and diterpenoids. The suberin content is low (1%) and composed primarily of ω-hydroxy acids with 9,10,18 trihydroxyoctadecanoic acid as the major suberin monomer. The lignin content is low (9.8%), and polysaccharides represent 33%. The ignition temperature of the bark is 190 °C, the burnout temperature is 653 °C, and the activation energy in combustion is 29 kJ mol−1. A biorefinery concept was developed considering the bark's chemical and thermal characteristics to convert approximately 90% of the bark mass into valuable chemicals, extracts, functional materials, and additives. [ABSTRACT FROM AUTHOR]

The phytoremediation of soil contaminated with petroleum depends on the association of plants with rhizosphere bacteria capable of promoting plant growth and destroying petroleum hydrocarbonates. In the present work, we studied the effects of "Lenoil" biological product containing bacteria Pseudomonas turukhanskensis IB 1.1, capable of destroying petroleum hydrocarbons on Secale cereale L plants, which previously proved to be resistant to the weak oil pollution of gray forest soil and to the composition of microorganisms in their rhizosphere. The composition of microorganisms in the rhizosphere of rye roots was studied, morphometric parameters of shoots and roots of rye plants were estimated, and hormone concentration was immunoassayed under conditions of 4% petroleum pollution of the soil. Addition of petroleum to the soil increased the content of oligonitrophilic (by 24%) and hydrocarbon-oxidizing (by 33%) microorganisms; however, the content of cellulolytic (by 12.5 times) microorganisms in the rhizosphere decreased. The use of Lenoil led to a further increase in the number of cellulolytic (by 5.6 times) and hydrocarbon-oxidizing (by 3.8 times) microorganisms and a decrease in the number of oligonitrophilic (by 22.7%) microorganisms in the rhizosphere. Under petroleum pollution, the content of auxins (IAA), zeatin riboside, zeatin nucleotide, and zeatin decreased, while the content of abscisic acid (ABA) increased in the shoots of rye plants. Lenoil treatment led to an eight-fold increase in the IAA content in the roots and a decrease in the ABA content in the aerial part and in the roots. It was shown for the first time that the treatment of petroleum-contaminated soil with "Lenoil" increased root mass due to the development of lateral roots, concurrent with high root IAA content. Petroleum pollution increased the deposition of lignin and suberin in the roots, which strengthened the apoplastic barrier and, thus, reduced the infiltration of toxic components. The deposition of suberin and lignin decreased under "Lenoil" treatment, indicating a decrease in the concentration of toxic petroleum components in the soil degraded by the bacteria. Thus, the biological preparation reduced the growth-inhibiting effect of petroleum on rye plants by increasing the content of growth-stimulating phytohormones (IAA and cytokinins) and reducing the content of ABA, justifying the importance of further study of relevant hormones for the improvement of phytoremediation. [ABSTRACT FROM AUTHOR]

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.

Developing polyols from biomass sources contributes to a more circular economy by replacing petroleum-based polyols in the vast production of polyurethanes (PUR). One such potential biomass source could be leftover birch bark from which suberinic acids (SA) can be obtained. The purpose of this study was to identify the best synthesis routes for novel SA-based polyols, obtain rigid PUR foams, and evaluate their competitiveness and potential suitability as thermal insulation material. Novel polyols were synthesized from depolymerized SA by esterification with various functionality and molecular weight alcohols in several molar ratios. The moisture content, hydroxyl and acid values, and apparent viscosity were tested. Free-rise rigid PUR foams from the most suitable SA-based polyol and tall oil-based polyol were successfully prepared, reaching ~20 wt.% total renewable material content in the foam. The obtained rigid PUR foams' morphological, mechanical, and thermal properties were investigated and compared to present foam materials, including commercial foams. The apparent density (~33 kg/m3), as well as the closed cell content (~94%), compression strength (0.25 MPa, parallel to the foaming direction), and thermal conductivity (~0.019 W/(m·K)), approved the competitiveness and potential ability of SA-based rigid PUR foam production as thermal insulation material. [ABSTRACT FROM AUTHOR]

Growth- and health-promoting bacteria can boost crop productivity in a sustainable way. Pseudomonas simiae WCS417 is such a bacterium that efficiently colonizes roots, modifies the architecture of the root system to increase its size, and induces systemic resistance to make plants more resistant to pests and pathogens. Our previous work suggested that WCS417-induced phenotypes are controlled by root cell-type-specific mechanisms. However, it remains unclear how WCS417 affects these mechanisms. In this study, we transcriptionally profiled five Arabidopsis thaliana root cell types following WCS417 colonization. We found that the cortex and endodermis have the most differentially expressed genes, even though they are not in direct contact with this epiphytic bacterium. Many of these genes are associated with reduced cell wall biogenesis, and mutant analysis suggests that this downregulation facilitates WCS417-driven root architectural changes. Furthermore, we observed elevated expression of suberin biosynthesis genes and increased deposition of suberin in the endodermis of WCS417-colonized roots. Using an endodermal barrier mutant, we showed the importance of endodermal barrier integrity for optimal plant-beneficial bacterium association. Comparison of the transcriptome profiles in the two epidermal cell types that are in direct contact with WCS417—trichoblasts that form root hairs and atrichoblasts that do not—implies a difference in potential for defense gene activation. While both cell types respond to WCS417, trichoblasts displayed both higher basal and WCS417-dependent activation of defense-related genes compared with atrichoblasts. This suggests that root hairs may activate root immunity, a hypothesis that is supported by differential immune responses in root hair mutants. Taken together, these results highlight the strength of cell-type-specific transcriptional profiling to uncover "masked" biological mechanisms underlying beneficial plant-microbe associations. This study investigates how the epiphytic plant-beneficial bacterium Pseudomonas simiae WCS417 affects different root cell types of Arabidopsis thaliana using cell-type-specific transcriptomics. Bioinformatic analysis and further experimental validation demonstrate that WCS417 alters cell wall biogenesis and suberin deposition inside the root, potentially facilitating root development and bacterial colonization. Furthermore, this study reveals a role of root hairs in immune activation. [ABSTRACT FROM AUTHOR]

Fire hazard is a constant risk in everyday life with the use of combustibles such as polymeric materials, wood, and fabrics, to name a few. Halogenated compounds have been widely used as efficient flameretardants, often being applied as coatings or impregnations. With growing environmental concerns and regional bans on the use of halogenated flame-retardant compounds, bio-based alternatives are garnering significant research interest. Naturally occurring materials such as eggshells, DNA, and certain proteins have developed a self-defense mechanism against fire over millions of years of evolution. Cork, a naturally occurring biological tissue in outer bark, is of interest as it is often used as a heat shield and moisture repellent, specifically in spacecraft. A deeper look into the chemical structure of cork indicates the presence of suberin, a bio-polyester group that makes up as much as 40% of its chemical composition. These bio-polyester groups play a key role as a protective barrier between the plant and the surrounding external environment. Thus, the role of suberin in plants could be mimicked for the design of biobased flame-retardant materials. [ABSTRACT FROM AUTHOR]

Summary: The root barrier to radial O2 loss (ROL) is a key root trait preventing O2 loss from roots to anoxic soils, thereby enabling root growth into anoxic, flooded soils.We hypothesized that the ROL barrier can also prevent intrusion of hydrogen sulphide (H2S), a potent phytotoxin in flooded soils. Using H2S‐ and O2‐sensitive microsensors, we measured the apparent permeance to H2S of rice roots, tested whether restricted H2S intrusion reduced its adverse effects on root respiration, and whether H2S could induce the formation of a ROL barrier.The ROL barrier reduced apparent permeance to H2S by almost 99%, greatly restricting H2S intrusion. The ROL barrier acted as a shield towards H2S; O2 consumption in roots with a ROL barrier remained unaffected at high H2S concentration (500 μM), compared to a 67% decline in roots without a barrier. Importantly, low H2S concentrations induced the formation of a ROL barrier.In conclusion, the ROL barrier plays a key role in protecting against H2S intrusion, and H2S can act as an environmental signalling molecule for the induction of the barrier. This study demonstrates the multiple functions of the suberized/lignified outer part of the rice root beyond that of restricting ROL. [ABSTRACT FROM AUTHOR]

Summary: Seeds slowly accumulate damage during storage, which ultimately results in germination failure. The seed coat protects the embryo from the external environment, and its composition is critical for seed longevity. Flavonols accumulate in the outer integument. The link between flavonol composition and outer integument development has not been explored.Genetic, molecular and ultrastructural assays on loss‐of‐function mutants of the flavonoid biosynthesis pathway were used to study the effect of altered flavonoid composition on seed coat development and seed longevity.Controlled deterioration assays indicate that loss of function of the flavonoid 3′ hydroxylase gene TT7 dramatically affects seed longevity and seed coat development. Outer integument differentiation is compromised from 9 d after pollination in tt7 developing seeds, resulting in a defective suberin layer and incomplete degradation of seed coat starch. These distinctive phenotypes are not shared by other mutants showing abnormal flavonoid composition. Genetic analysis indicates that overaccumulation of kaempferol‐3‐rhamnoside is mainly responsible for the observed phenotypes. Expression profiling suggests that multiple cellular processes are altered in the tt7 mutant.Overaccumulation of kaempferol‐3‐rhamnoside in the seed coat compromises normal seed coat development. This observation positions TRANSPARENT TESTA 7 and the UGT78D1 glycosyltransferase, catalysing flavonol 3‐O‐rhamnosylation, as essential players in the modulation of seed longevity. [ABSTRACT FROM AUTHOR]

In this study, an analysis of the influence of extractives, suberin and lignocellulosic components on the pyrolysis decomposition and fire reaction mechanisms of a cork oak powder from Quercus suber L. is presented. The summative chemical composition of cork powder was determined. Suberin was the main component at 40% of the total weight, followed by 24% of lignin, 19% of polysaccharides and 14% of extractives. The absorbance peaks of cork and its individual components were further analyzed by means of ATR-FTIR spectrometry. Thermogravimetric analysis (TGA) showed that the removal of extractives from cork slightly increased the thermal stability between 200 °C and 300 °C and led to the formation of a more thermally stable residue at the end of the cork decomposition. Moreover, by removing suberin, a shift of the onset decomposition temperature to a lower temperature was noticed, indicating that suberin plays a major role in enhancing the thermal stability of cork. Furthermore, non-polar extractives showed the highest flammability with a peak of heat release rate (pHRR) of 365 W/g analyzed by means of micro-scale combustion calorimetry (MCC). Above 300 °C, the heat release rate (HRR) of suberin was lower than that of polysaccharides or lignin. However, below that temperature it released more flammable gases with a pHRR of 180 W/g, without significant charring ability, contrary to the mentioned components that showed lower HRR due to their prominent condensed mode of action that slowed down the mass and heat transfer processes during the combustion process. [ABSTRACT FROM AUTHOR]

As a wall polymer, suberin has a multifaceted role in plant development and stress responses. It is deposited between the plasma membrane and the primary cell wall in specialized tissues such as root exodermis, endodermis, phellem, and seed coats. It is formed de novo in response to stresses such as wounding, salt injury, drought, and pathogen attack and is a complex polyester mainly consisting of fatty acids, glycerol, and minor amounts of ferulic acid that are associated to a lignin-like polymer predominantly composed of ferulates. Metabolomic and transcriptomic studies have revealed that cell wall lignification precedes suberin deposition. The ferulic acid esterified to ω-hydroxy fatty acids, synthetized by the feruloyl transferase FHT (or ASFT), presumably plays a role in coupling both polymers, although the precise mechanism is not understood. Here, we use the promoter of tomato suberin feruloyl transferase (FHT/ASFT) fused to GUS (β-glucuronidase) to demonstrate that ferulate deposition agrees with the site of promoter FHT activation by using a combination of histochemical staining and UV microscopy. Hence, FHT promoter activation and alkali UV microscopy can be used to identify the precise localization of early suberizing cells rich in ferulic acid and can additionally be used as an efficient marker of early suberization events during plant development and stress responses. This line can be used in the future as a tool to identify emerging suberization sites via ferulate deposition in tomato plants, which may contribute to germplasm screening in varietal improvement programs. [ABSTRACT FROM AUTHOR]

The intensive use of groundwater in agriculture under the current climate conditions leads to acceleration of soil salinization. Given that almond is a salt-sensitive crop, selection of salt-tolerant rootstocks can help maintain productivity under salinity stress. Selection for tolerant rootstocks at an early growth stage can reduce the investment of time and resources. However, salinity-sensitive markers and salinity tolerance mechanisms of almond species to assist this selection process are largely unknown. We established a microscopy-based approach to investigate mechanisms of stress tolerance in and identified cellular, root anatomical, and molecular traits associated with rootstocks exhibiting salt tolerance. We characterized three almond rootstocks: Empyrean-1 (E1), Controller-5 (C5), and Krymsk-86 (K86). Based on cellular and molecular evidence, our results show that E1 has a higher capacity for salt exclusion by a combination of upregulating ion transporter expression and enhanced deposition of suberin and lignin in the root apoplastic barriers, exodermis, and endodermis, in response to salt stress. Expression analyses revealed differential regulation of cation transporters, stress signaling, and biopolymer synthesis genes in the different rootstocks. This foundational study reveals the mechanisms of salinity tolerance in almond rootstocks from cellular and structural perspectives across a root developmental gradient and provides insights for future screens targeting stress response.

Summary: Wood is the most important repository of assimilated carbon in the biosphere, in the form of large polymers (cellulose, hemicelluloses including glucuronoxylan, and lignin) that interactively form a composite, together with soluble extractives including phenolic and aliphatic compounds. Molecular interactions among these compounds are not fully understood.We have targeted the expression of a fungal α‐glucuronidase to the wood cell wall of aspen (Populus tremula L. × tremuloides Michx.) and Arabidopsis (Arabidopsis thaliana (L.) Heynh), to decrease contents of the 4‐O‐methyl glucuronopyranose acid (mGlcA) substituent of xylan, to elucidate mGlcA's functions.The enzyme affected the content of aliphatic insoluble cell wall components having composition similar to suberin, which required mGlcA for binding to cell walls. Such suberin‐like compounds have been previously identified in decayed wood, but here, we show their presence in healthy wood of both hardwood and softwood species. By contrast, γ‐ester bonds between mGlcA and lignin were insensitive to cell wall‐localized α‐glucuronidase, supporting the intracellular formation of these bonds.These findings challenge the current view of the wood cell wall composition and reveal a novel function of mGlcA substituent of xylan in fastening of suberin‐like compounds to cell wall. They also suggest an intracellular initiation of lignin–carbohydrate complex assembly. See also the Commentary on this article by Oliveira, 238: 8–10. [ABSTRACT FROM AUTHOR]

Introduction: Apple russeting is mainly due to the accumulation of suberin in the cell wall in response to defects and damages in the cuticle layer. Over the last decades, massive efforts have been done to better understand the complex interplay between pathways involved in the suberization process in model plants. However, the regulation mechanisms which orchestrate this complex process are still under investigation. Our previous studies highlighted a number of transcription factor candidates from the Myeloblastosis (MYB) transcription factor family which might regulate suberization in russeted or suberized apple fruit skin. Among these, we identified MdMYB68, which was co-expressed with number of well-known key suberin biosynthesis genes. Method: To validate the MdMYB68 function, we conducted an heterologous transient expression in Nicotiana benthamiana combined with whole gene expression profiling analysis (RNA-Seq), quantification of lipids and cell wall monosaccharides, and microscopy. Results: MdMYB68 overexpression is able to trigger the expression of the whole suberin biosynthesis pathway. The lipid content analysis confirmed that MdMYB68 regulates the deposition of suberin in cell walls. Furthermore, we also investigated the alteration of the non-lipid cell wall components and showed that MdMYB68 triggers a massive modification of hemicelluloses and pectins. These results were finally supported by the microscopy. Discussion: Once again, we demonstrated that the heterologous transient expression in N. benthamiana coupled with RNA-seq is a powerful and efficient tool to investigate the function of suberin related transcription factors. Here, we suggest MdMYB68 as a new regulator of the aliphatic and aromatic suberin deposition in apple fruit, and further describe, for the first time, rearrangements occurring in the carbohydrate cell wall matrix, preparing this suberin deposition. [ABSTRACT FROM AUTHOR]

Suberin, a complex polyester deposited in the seed coat outer integument, acts as a hydrophobic barrier to control the movement of water, ions, and gas. However, relatively little is known about the signal transduction involved in suberin layer formation during seed coat development. In this study, the effect of the plant hormone abscisic acid (ABA) on suberin layer formation in seed coats was investigated by characterizing mutations in Arabidopsis related to ABA biosynthesis and signaling. Seed coat permeability to tetrazolium salt was noticeably elevated in aba1-1 and abi1-1 mutants, but not significantly altered in snrk2.2/3/6, abi3-8, abi5-7, and pyr1pyl1pyl2pyl4 quadruple mutants compared with that in the wild-type (WT). ABA1 encodes a zeaxanthin epoxidase that functions in the first step of ABA biosynthesis. aba1-1 and aba1-8 mutant seed coats showed reduced autofluorescence under UV light and increased tetrazolium salt permeability relative to WT levels. ABA1 disruption resulted in decreased total seed coat polyester levels by approximately 3%, with a remarkable reduction in levels of C24:0 w-hydroxy fatty acids and C24:0 dicarboxylic acids, which are the most abundant aliphatic compounds in seed coat suberin. Consistent with suberin polyester chemical analysis, RT-qPCR analysis showed a significant reduction in transcript levels of KCS17, FAR1, FAR4, FAR5, CYP86A1, CYP86B1, ASFT, GPAT5, LTPG1, LTPG15, ABCG2, ABCG6, ABCG20, ABCG23, MYB9, and MYB107, which are involved in suberin accumulation and regulation in developing aba1-1 and aba1-8 siliques, as compared with WT levels. Together, seed coat suberization is mediated by ABA and partially processed through canonical ABA signaling. [ABSTRACT FROM AUTHOR]

Suberin is a complex polyester biopolymer, and it is practically impossible to estimate the real content of suberin in suberised plant tissues. This indicates the importance of the development of instrumental analytical methods for the comprehensive characterisation of suberin derived from plant biomass for the successful integration of suberinic products into biorefinery production chains. In this study, we optimised two GC-MS methods—one with direct sylilation, and the second with additional depolymerisation, using GPC methods with RI detector and polystyrene calibration and with a three-angle light scattering detector and an eighteen-angle light scattering detector. We also performed MALDI-Tof analysis for non-degraded suberin structure determination. We characterised suberinic acid (SA) samples obtained from birch outer bark after alkaline depolymerisation. The samples were particularly rich in diols, fatty acids and their esters, hydroxyacids and their corresponding esters, diacids and their corresponding esters, as well as extracts (mainly betulin and lupeol) and carbohydrates. To remove phenolic-type admixtures, treatment with ferric chloride (FeCl3) was used. The SA treatment with FeCl3 allows the possibility to obtain a sample that has a lower content of phenolic-type compounds and a lower molecular weight than an untreated sample. It was possible to identify the main free monomeric units of SA samples by GC-MS system using direct silylation. By performing an additional depolymerisation step before silylation, it was possible to characterise the complete potential monomeric unit composition in the suberin sample. For the molar mass distribution determination, it is important to perform GPC analysis. Even though chromatographic results can be obtained using a three- laser MALS detector, they are not fully correct because of the fluorescence of the SA samples. Therefore an 18-angle MALS detector with filters was more suitable for SA analysis. MALDI-Tof analysis is a great tool for the polymeric compound structural identification, which cannot be done using GC-MS. Using the MALDI data, we discovered that the main monomeric units that makes up the SA macromolecular structure are octadecanedioic acid and 2-(1,3-dihydroxyprop-2-oxy)decanedioic acid. This corresponds with GC-MS results, showing that after depolymerisation hydroxyacids and diacids were the dominant type of compounds found in the sample. [ABSTRACT FROM AUTHOR]

Mangrove plants demonstrate an impressive ability to tolerate environmental pollutants, but excessive levels of cadmium (Cd) can impede their growth. Few studies have focused on the effects of apoplast barriers on heavy metal tolerance in mangrove plants. To investigate the uptake and tolerance of Cd in mangrove plants, two distinct mangrove species, Avicennia marina and Rhizophora stylosa, are characterized by unique apoplast barriers. The results showed that both mangrove plants exhibited the highest concentration of Cd2+ in roots, followed by stems and leaves. The Cd2+ concentrations in all organs of R. stylosa consistently exhibited lower levels than those of A. marina. In addition, R. stylosa displayed a reduced concentration of apparent PTS and a smaller percentage of bypass flow when compared to A. marina. The root anatomical characteristics indicated that Cd treatment significantly enhanced endodermal suberization in both A. marina and R. stylosa roots, and R. stylosa exhibited a higher degree of suberization. The transcriptomic analysis of R. stylosa and A. marina roots under Cd stress revealed 23 candidate genes involved in suberin biosynthesis and 8 candidate genes associated with suberin regulation. This study has confirmed that suberized apoplastic barriers play a crucial role in preventing Cd from entering mangrove roots.

The apple is an economically important fruit, and fruit russeting is not conducive to its appearance. Although studies have examined fruit russeting, its mechanism remains unclear. Two apple strains of the F1 hybrid population derived from 'Fuji' and 'Golden Delicious' were used in this study. We found that the skin of russet apples was rough and fissured, while that of non-russet apples was smooth and waxy. Chemical staining, LC- and GC-MS showed that both lignin and suberin were increased in russet apple skin. Meanwhile, genes involved in lignin and suberin synthetic pathways were upregulated in russet apple skin. Additionally, we found many differentially expressed genes (DEGs1) involved in hormone biosynthesis and signaling and stress responses in the two apple strains. We found that WRKY13 may influence russeting by regulating lignin synthesis. Our study identified several candidate metabolites and genes, which will provide a good foundation for further research. [ABSTRACT FROM AUTHOR]

The cork layer present in all dicotyledonous plant species with radial growth is the result of the phellogen activity, a secondary meristem that produces phellem (cork) to the outside and phelloderm inwards. These three different tissues form the periderm, an efficient protective tissue working as a barrier against external factors such as environmental aggressions and pathogen attacks. The protective function offered by cork cells is mainly due to the abundance of suberin in their cell walls. Chemically, suberin is a complex aliphatic network of long chain fatty acids and alcohols with glycerol together with aromatic units. In most woody species growing in temperate climates, the first periderm is replaced by a new functional periderm upon a few years after being formed. One exception to this bark development can be found in cork oak (Quercus suber) which display a single periderm that grows continuously. Quercus suber stands by its thick cork layer development with continuous seasonal growth. Cork raw material has been exploited by man for centuries, especially in Portugal and Spain. Nowadays, its applications have widened vastly, from the most known product, stoppers, to purses or insulating materials used in so many industries, such as construction and car production. Research on how cork develops, and the effect environmental factors on cork oak trees is extremely important to maintain production of good-quality cork, and, by maintaining cork oak stands wealthy, we are preserving a very important ecosystem both by its biodiversity and its vital social and economic role in areas already showing a population declination. [ABSTRACT FROM AUTHOR]

Our previous studies, comparing russeted vs. waxy apple skin, highlighted a MYeloBlastosys (Myb) transcription factor (MdMYB52), which displayed a correlation with genes associated to the suberization process. The present article aims to assess its role and function in the suberization process. Phylogenetic analyses and research against Arabidopsis thaliana MYBs database were first performed and the tissue specific expression of MdMYB52 was investigated using RT-qPCR. The function of MdMYB52 was further investigated using Agrobacterium-mediated transient overexpression in Nicotiana benthamiana leaves. An RNA-Seq analysis was performed to highlight differentially regulated genes in response MdMYB52. Transcriptomic data were supported by analytical chemistry and microscopy. A massive decreased expression of photosynthetic and primary metabolism pathways was observed with a concomitant increased expression of genes associated with phenylpropanoid and lignin biosynthesis, cell wall modification and senescence. Interestingly key genes involved in the synthesis of suberin phenolic components were observed. The analytical chemistry displayed a strong increase in the lignin content in the cell walls during MdMYB52 expression. More specifically, an enrichment in G-Unit lignin residues was observed, supporting transcriptomic data as well as previous work describing the suberin phenolic domain as a G-unit enriched lignin-like polymer. The time-course qPCR analysis revealed that the observed stress response, might be explain by this lignin biosynthesis and by a possible programmed senescence triggered by MdMYB52. The present work supports a crucial regulatory role for MdMYB52 in the biosynthesis of the suberin phenolic domain and possibly in the fate of suberized cells in russeted apple skins. [ABSTRACT FROM AUTHOR]

Wound healing is a postharvest characteristic of potato tubers through accumulating suberin and lignin, which could reduce decay and water loss during storage. This study aimed to explore the impact and mechanisms of sodium silicate on wound healing of potatoes. After being wounded, "Atlantic" potato tubers were treated with water or 50mM sodium silicate. The results showed that sodium silicate treatment accelerated the formation of wound healing structures and significantly reduced the weight loss and disease index of tubers. Furthermore, sodium silicate induced the genes expression and enzyme activity of phenylalanine ammonia lyase (PAL), 4-coumarate: coenzyme A ligase (4CL), and cinnamyl alcohol dehydrogenase (CAD) involved in the phenylpropane metabolism, enhancing the synthesis of the main precursors of suberin polyphenolic (SPP) and lignin, such as coniferyl alcohol, sinapyl alcohol, and cinnamyl alcohol. Meanwhile, the gene expression of StPOD and StNOX was activated, and the production of O2- and H2O2 was promoted, which could be used for injury signal transmission and oxidative crosslinking of SPP monomers and lignin precursors. Besides, antimicrobial compounds, total phenolics, and flavonoids were also induced. We suggest that sodium silicate could promote wound healing by inducing the deposition of SPP, lignin, and antimicrobial compounds in potato tubers. [ABSTRACT FROM AUTHOR]

Drought is the most serious adversity faced by agriculture and animal husbandry industries. One strategy that plants use to adapt to water deficits is modifying the root growth and architecture. Root endodermis has cell walls reinforced with apoplastic barriers formed by the Casparian strip (CS) and suberin lamellae (SL) deposits, regulates radial nutrient transport and protects the vascular cylinder from abiotic threats. Elymus sibiricus is an economically important meso-xerophytic forage grass, characterized by high nutritional quality and strong environmental adaptability. The purpose of this study was to evaluate the drought tolerance of E. sibiricus genotypes and investigate the root structural adaptation mechanism of drought-tolerant genotypes' responding to drought. Specifically, a drought tolerant (DT) and drought sensitive (DS) genotype were screened out from 52 E. sibiricus genotypes. DT showed less apoplastic bypass flow of water and solutes than DS under control conditions, as determined with a hydraulic conductivity measurement system and an apoplastic fluorescent tracer, specifically PTS trisodium-8-hydroxy-1,3,6-pyrenetrisulphonic acid (PTS). In addition, DT accumulated less Na, Mg, Mn, and Zn and more Ni, Cu, and Al than DS, regardless of osmotic stress. Further study showed more suberin deposition in DT than in DS, which could be induced by osmotic stress in both. Accordingly, the CS and SL were deposited closer to the root tip in DT than in DS. However, osmotic stress induced their deposition closer to the root tips in DS, while likely increasing the thickness of the CS and SL in DT. The stronger and earlier formation of endodermal barriers may determine the radial transport pathways of water and solutes, and contribute to balance growth and drought response in E. sibiricus. These results could help us better understand how altered endodermal apoplastic barriers in roots regulate water and mineral nutrient transport in plants that have adapted to drought environments. Moreover, the current findings will aid in improving future breeding programs to develop drought-tolerant grass or crop cultivars. [ABSTRACT FROM AUTHOR]

Summary: Outer protective barriers of animals use a variety of bio‐polymers, based on either proteins (e.g. collagens), or modified sugars (e.g. chitin). Plants, however, have come up with a particular solution, based on the polymerisation of lipid‐like precursors, giving rise to cutin and suberin. Suberin is a structural lipophilic polyester of fatty acids, glycerol and some aromatics found in cell walls of phellem, endodermis, exodermis, wound tissues, abscission zones, bundle sheath and other tissues. It deposits as a hydrophobic layer between the (ligno)cellulosic primary cell wall and plasma membrane. Suberin is highly protective against biotic and abiotic stresses, shows great developmental plasticity and its chemically recalcitrant nature might assist the sequestration of atmospheric carbon by plants. The aim of this review is to integrate the rapidly accelerating genetic and cell biological discoveries of recent years with the important chemical and structural contributions obtained from very diverse organisms and tissue layers. We critically discuss the order and localisation of the enzymatic machinery synthesising the presumed substrates for export and apoplastic polymerisation. We attempt to explain observed suberin linkages by diverse enzyme activities and discuss the spatiotemporal relationship of suberin with lignin and ferulates, necessary to produce a functional suberised cell wall. [ABSTRACT FROM AUTHOR]

The simultaneous occurrence of heat stress and drought is becoming more regular as a consequence of climate change, causing extensive agricultural losses. The application of either heat or osmotic stress increase cell-wall suberization in different tissues, which may play a role in improving plant resilience. In this work, we studied how the suberization process is affected by the combination of drought and heat stress by following the expression of suberin biosynthesis genes, cell-wall suberization and the chemical composition in Arabidopsis roots. The Arabidopsis plants used in this study were at the onset of secondary root development. At this point, one can observe a developmental gradient in the main root, with primary development closer to the root tip and secondary development, confirmed by the suberized phellem, closer to the shoot. Remarkably, we found a differential response depending on the root zone. The combination of drought and heat stress increased cell wall suberization in main root segments undergoing secondary development and in lateral roots (LRs), while the main root zone, at primary development stage, was not particularly affected. We also found differences in the overall chemical composition of the cell walls in both root zones in response to combined stress. The data gathered showed that, under combined drought and heat stress, Arabidopsis roots undergo differential cell wall remodeling depending on developmental stage, with modifications in the biosynthesis and/or assembly of major cell wall components. [ABSTRACT FROM AUTHOR]

This paper presents a wood modification method using renewable and non-toxic suberin monomers (SMs) under heat treatment to improve dimensional stability of wood from fast-growing species. Specimens of poplar (Populus cathayana) wood were impregnated with SMs and then subjected to heat treatment at 180°C for two hours. The untreated wood (Control), suberin monomers impregnated wood (Sub) and suberin monomers impregnated wood with heat treatment (Sub 180°C) were analyzed by scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and Fourier transform infrared spectroscopy (FTIR). The hygroscopicity and dimensional stability of modified wood were evaluated. The results showed that SMs in the treated wood were located in the cell lumen of fibers and vessels, as well as in the cell wall which was bulked. The dimensional stability of SMs modified wood was improved, and this enhancement became more pronounced by a combination with heat treatment. The anti-swelling efficiency of Sub and Sub 180°C treatments were 30.0% and 49.6%, respectively. The presented results showed potential of SMs treatment to develop an effective modification approach and improve dimensional stability of wood of fast-growing species, as well as to promote the reuse of suberin from the bark.

As a wall polymer, suberin has a multifaceted role in plant development and stress responses. It is deposited between the plasma membrane and the primary cell wall in specialized tissues such as root exodermis, endodermis, phellem, and seed coats. It is formed de novo in response to stresses such as wounding, salt injury, drought, and pathogen attack and is a complex polyester mainly consisting of fatty acids, glycerol, and minor amounts of ferulic acid that are associated to a lignin-like polymer predominantly composed of ferulates. Metabolomic and transcriptomic studies have revealed that cell wall lignification precedes suberin deposition. The ferulic acid esterified to ω-hydroxy fatty acids, synthetized by the feruloyl transferase FHT (or ASFT), presumably plays a role in coupling both polymers, although the precise mechanism is not understood. Here, we use the promoter of tomato suberin feruloyl transferase (FHT/ASFT) fused to GUS (β-glucuronidase) to demonstrate that ferulate deposition agrees with the site of promoter FHT activation by using a combination of histochemical staining and UV microscopy. Hence, FHT promoter activation and alkali UV microscopy can be used to identify the precise localization of early suberizing cells rich in ferulic acid and can additionally be used as an efficient marker of early suberization events during plant development and stress responses. This line can be used in the future as a tool to identify emerging suberization sites via ferulate deposition in tomato plants, which may contribute to germplasm screening in varietal improvement programs.

Global environmental change is altering the quality and quantity of plant inputs into soil. However, it is unclear how these long-term changes may fundamentally shift the biogeochemistry of soil carbon in forests. To better understand how varied detrital inputs alter soil organic matter (OM) biogeochemistry and composition at the molecular-level, soil samples were collected from a 20 year detrital manipulation experiment in an old-growth coniferous rainforest in Western Oregon. The experiment includes ambient (control) plots and six treatments: Double Litter, Double Wood, No Roots, No Litter, No Inputs and OA-less (replacement of O and A horizons with B horizon). Total soil carbon and nitrogen, molecular-level OM composition using solid-state 13C nuclear magnetic resonance, and targeted compound extractions were measured. Although soil carbon did not increase with Double Litter and Double Wood, microbial biomass and the decomposition of specific forms of soil OM (i.e., cellulose) increased, likely due to sustained soil priming over 20 years. Mineral (0–10 cm) soil carbon was similar across litter exclusion treatments (No Litter, No Roots, No Inputs), however, soil OM decomposition increased relative to the control. Microbial-derived lipids increased under Double Litter but decreased when above-ground litter was excluded. Notably, needle-derived lipids decreased with above-ground litter exclusion and root-derived compounds did not change under below-ground root exclusion. These results suggested that above-ground litter alters soil carbon biogeochemistry in surface soils to a greater extent than below-ground inputs. This study also demonstrated that long-term soil carbon biogeochemical trajectories were mostly governed by litter quality, quality and microbial processing of above-ground inputs. [ABSTRACT FROM AUTHOR]

Summary: Tomato varieties resistant to the bacterial wilt pathogen Ralstonia solanacearum have the ability to restrict bacterial movement in the plant. Inducible vascular cell wall reinforcements seem to play a key role in confining R. solanacearum into the xylem vasculature of resistant tomato. However, the type of compounds involved in such vascular physico‐chemical barriers remain understudied, while being a key component of resistance.Here we use a combination of histological and live‐imaging techniques, together with spectroscopy and gene expression analysis to understand the nature of R. solanacearum‐induced formation of vascular coatings in resistant tomato.We describe that resistant tomato specifically responds to infection by assembling a vascular structural barrier formed by a ligno‐suberin coating and tyramine‐derived hydroxycinnamic acid amides. Further, we show that overexpressing genes of the ligno‐suberin pathway in a commercial susceptible variety of tomato restricts R. solanacearum movement inside the plant and slows disease progression, enhancing resistance to the pathogen.We propose that the induced barrier in resistant plants does not only restrict the movement of the pathogen, but may also prevent cell wall degradation by the pathogen and confer anti‐microbial properties, effectively contributing to resistance. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]