Green Synthesis of Metallic Nanoparticles from Quercus Bark Extracts: Characterization and Functional Properties.

Quercus species are utilized for their durable wood, providing sustenance for wildlife, conserving biodiversity, and contributing ecological, medicinal, and esthetic benefits to ecosystems and landscapes. In this study, we aimed to use the bark of three Quercus species (Q. dalechampi, Q. fraineto, and Q. petraea) for the synthesis of silver and gold nanoparticles (AgNPs and AuNPs). The aqueous extracts from the bark of Quercus sp. acted both as reducing and stabilizing agent, facilitating the rapid synthesis of AuNPs (AuQD, AuQF, and AuQP) and AgNPs (AgQD, AgQF, and AgQP). The obtained nanoparticles were characterized using UV-vis spectroscopy, TEM, DLS, and FTIR. Characterizations revealed that the nanoparticles exhibited a variety of shapes, such as polygonal, triangular, and spherical forms, with sizes ranging between 14 and 24 nm for AuNPs and 45-70 nm for AgNPs. The total phenolic content was assessed through spectroscopic methods, while several individual phenolic compounds were identified and quantified using UPLC-PDA. Furthermore, we assessed the antioxidant, antibacterial, and antifungal capacities of AuNPs, AgNPs, and raw extracts. The highest antioxidant activity was observed for raw extracts, followed by AgNPs and AuNPs, while the most potent antibacterial and antifungal activity was observed in AgQP. Moreover, cytotoxicity was examined in a human keratinocyte cell line (HaCaT). The results indicated no cytotoxic effects for AuNPs, while AgNPs and the raw extracts exhibited cytotoxic effects after 48 h of incubation. This research underscores the multifaceted utility of Quercus bark extracts in the green synthesis of metallic nanoparticles and their subsequent bioactivity assessment, suggesting promising perspectives for their application in various fields while urging cautious consideration of their cytotoxic implications.

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

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

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

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

Habitat and tree species identity shape aboveground and belowground fungal communities in central European forests.

Introduction: Trees interact with fungi in mutualistic, saprotrophic, and pathogenic relationships. With their extensive aboveground and belowground structures, trees provide diverse habitats for fungi. Thus, tree species identity is an important driver of fungal community composition in forests. Methods: Here we investigate how forest habitat (bark surface vs. soil) and tree species identity (deciduous vs. coniferous) affect fungal communities in two Central European forests. We assess differences and interactions between fungal communities associated with bark surfaces and soil, in forest plots dominated either by Fagus sylvatica, Picea abies, or Pinus sylvestris in two study regions in southwestern and northeastern Germany. Results: ITS metabarcoding yielded 3,357 fungal amplicon sequence variants (ASVs) in the northern and 6,088 in the southern region. Overall, soil communities were 4.7 times more diverse than bark communities. Habitat type explained 48-69% of the variation in alpha diversity, while tree species identity explained >1-3%. NMDS ordinations showed that habitat type and host tree species structured the fungal communities. Overall, few fungal taxa were shared between habitats, or between tree species, but the shared taxa were highly abundant. Network analyses, based on co-occurrence patterns, indicate that aboveground and belowground communities form distinct subnetworks. Discussion: Our study suggests that habitat (bark versus soil) and tree species identity are important factors structuring fungal communities in temperate European forests. The aboveground (bark-associated) fungal community is currently poorly known, including a high proportion of reads assigned to "unknown Ascomycota" or "unknown Dothideomycetes." The role of bark as a habitat and reservoir of unique fungal diversity in forests has been underestimated.

Potential Use of Quercus dalechampii Ten. and Q. frainetto Ten. Barks Extracts as Antimicrobial, Enzyme Inhibitory, Antioxidant and Cytotoxic Agents.

The bark resulted as a by-product after different Quercus sp. processing is a valuable alternative source of phenolic compounds (mainly tannins). Hence, the focus of the present work was to obtain eight extracts from the rhytidome of the less-studied Q. dalechampii and Q. frainetto and characterize them in terms of chemical and bioactive profiles. Ultrasound and microwave-assisted extraction methods were used for the preparation of the extracts. Total phenolic and tannin contents were assessed through classic spectrophotometric methods, while several individual phenolic compounds were identified and quantified using UPLC-PDA. Antioxidant, enzyme-inhibitory, antibacterial, and cytotoxic activities were tested using in vitro assays; additionally being evaluated was the ability of the extracts to inhibit the adherence of MRSA to suture wires. The UPLC analysis confirmed the presence of gallic acid, catechin, taxifolin, vanillic acid, epicatechin, and caffeic acid. The results showed that tested extracts were able to exert cytotoxic effects, at 6% and 3% concentrations, on confluent cells. The tested solutions inhibit α-glucosidase activity and the antibacterial potential suggested a mild to moderate effect against the Gram-positive strains. Overall, the obtained results revealed rich phenolic and tannin contents for the extracts obtained from both species through microwave-assisted extraction, probably responsible for their mild antibacterial and cytotoxic effects.

Qualitative and Quantitative Anatomical Analysis of the Constitutive Bark of Q. ilex x Q. suber Hybrids.

Hybridization and introgression between cork oak (Quercus suber) and holm oak (Q. ilex) have traditionally been reckoned as undesirable processes, since hybrid individuals lack the profitable bark characteristics of cork oak. Nevertheless, a systematic and quantitative description of the bark of these hybrids at the microscopic level, based on a significant number of individuals, is not available to date. In this work we provide such a qualitative and quantitative description, identifying the most relevant variables for their classification. Hybrids show certain features intermediate between those of the parent species (such as phellem percentage in the outer bark, which was approximately 40% as a mean value for hybrids, 20% in holm oak and almost 99% in cork oak), as well as other unique features, such as the general suberization of inactive phloem (up to 25% in certain individuals), reported here for the first time. These results suggest a relevant hybridization-induced modification of the genetic expression patterns. Therefore, hybrid individuals provide a valuable material to disentangle the molecular mechanisms underpinning bark development in angiosperms.

Programmed cell death during the formation of rhytidome and interxylary cork in roots of Astragalus membranaceus (Leguminosae).

Programmed cell death (PCD) plays a critical role throughout the lives of plants, it is regarded as a highly regulated and active process of plant cell death during the times of biotic or abiotic stress. This study aims to provide developmental anatomical characteristics of the interxylary cork formation in the roots of Astragalus. membranaceus var. mongholicus, and to subsequently show cytomorphological evidence that PCD is involved in the development of rhytidome and interxylary cork. The developmental anatomy of rhytidome and interxylary cork of the perennial fresh main root of A. membranaceus var. mongholicus was studied using light microscopy, whereas the PCD in the development of rhytidome and interxylary cork was studied using fluorescence microscopy and transmission electron microscopy. Histologically, it was observed that the parenchyma cells of secondary phloem and xylem in roots recovered their meristematic ability, and later developed into rhytidome and interxylary cork. Cytologically, ultrastructural characteristics such as nucleus malformation, vacuole disappearance, mitochondrial degeneration, and vesicle filling were observed. In roots, the nucleus of the phloem parenchyma cells were terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive from the pre-rhytidome stage to the formation of rhytidome stage and 4',6-diamidino-2-phenylindole dihydrochloride (DAPI)-negative during the mature rhytidome stage. The TUNEL assay of the xylem parenchyma cells showed positive characteristics from the early stage of interxylary cork formation to the interxylary cork formation stage, whereas DAPI-negative characteristics were observed in the mature interxylary cork. Gel electrophoresis showed that DNA cleavage was random. Our results indicated that the formation of the rhytidome and interxylary cork involved the PCD process.

A Critical Review of Phenolic Compounds Extracted from the Bark of Woody Vascular Plants and Their Potential Biological Activity.

Polyphenols are one of the largest and most widespread groups of secondary metabolites in the plants world. These compounds are of particular interest due to their occurrence and the properties they possess. The main sources of phenolic compounds are fruits and vegetables, but lately, more and more studies refer to woody vascular plants, especially to bark, as an important source of phenolic compounds with a potential biological effect. This study aims to bring together information on the phenolic compounds present in the bark of woody vascular plants by discussing extraction methods, the chemical composition of the extracts and potential biological effects. The literature data used in this paper were collected via PubMed (2004⁻2019). Search terms were: bark, rhytidome, woody vascular plant, polyphenols, phenolic compounds, biologic activity, antioxidant, immunostimulatory, antimutagenic, antibacterial, anti-inflammatory, and antitumoral. This paper intends to highlight the fact that the polyphenolic extracts obtained from the bark of woody vascular plants represent sources of bioactive compounds with antioxidant, immunostimulatory, antimutagenic, antibacterial properties, etc. Future research directions should be directed towards identification and isolation of bioactive compounds. Consequently, biologically active compounds obtained from the bark of woody plants could be exploited on an industrial scale.

Developmental anatomy of anomalous structure and classification of commercial specifications and grades of the Astragalus membranaceus var. mongholicus.

The Daodi herb, Huangqi from Shanxi, is derived from the root of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao. Huangqi was divided into different commercial specifications and grades in producing area according to macroscopic features. Developmental anatomy of anomalous structure of the perennial fresh main root of A. membranaceus var. mongholicus was investigated using conventional paraffin section to explore the relationship between anomalous structure and classification of commercial specifications and grades of Huangqi. The results of developmental anatomy showed that parenchyma cells in secondary phloem and secondary xylem of the perennial fresh main root regained meristematic ability, and resulted in anomalous structure. Several layers of additional periderm could sometimes be found in the secondary phloem near the initial periderm, which was called rhytidome. Additional periderm could sometimes be observed in the center of secondary xylem, which was described as "rotten heart." The area of rhytidome and "rotten heart" increased gradually with increasing age. The results of different commercial specifications and grades of Huangqi revealed rhytidome and "rotten heart" mainly existed in Ge-Da-Tou, Er-Dao-Tou, Hong-Lan-Qi, Special Class, First Class, and Second Class. The additional periderm in secondary phloem of Ge-Da-Tou, Er-Dao-Tou, Hong-Lan-Qi, and Special Class could reach five layers; however, that of First Class and Second Class was three layers at most. In summary, this study demonstrated the rules of the development of rhytidome and "rotten heart" in A. membranaceus var. mongholicus. The relationship between anomalous structure and different commercial specifications and grades of Daodi herb Huangqi, was clarified. RESEARCH HIGHLIGHTS: The rhytidome and the development of "rotten heart" have not been reported in Astragalus membranaceus var. mongholicus. At present article, we demonstrated the developmental anatomy of the rhytidome and "rotten heart" existed in the secondary phloem and secondary xylem of the root of A. membranaceus var. mongholicus respectively. The rhytidome and "rotten heart" is different among six commercial specifications and grades of Huangqi.

A comparative transcriptomic approach to understanding the formation of cork.

KEY MESSAGE: The transcriptome comparison of two oak species reveals possible candidates accounting for the exceptionally thick and pure cork oak phellem, such as those involved in secondary metabolism and phellogen activity. Cork oak, Quercus suber, differs from other Mediterranean oaks such as holm oak (Quercus ilex) by the thickness and organization of the external bark. While holm oak outer bark contains sequential periderms interspersed with dead secondary phloem (rhytidome), the cork oak outer bark only contains thick layers of phellem (cork rings) that accumulate until reaching a thickness that allows industrial uses. Here we compare the cork oak outer bark transcriptome with that of holm oak. Both transcriptomes present similitudes in their complexity, but whereas cork oak external bark is enriched with upregulated genes related to suberin, which is the main polymer responsible for the protective function of periderm, the upregulated categories of holm oak are enriched in abiotic stress and chromatin assembly. Concomitantly with the upregulation of suberin-related genes, there is also induction of regulatory and meristematic genes, whose predicted activities agree with the increased number of phellem layers found in the cork oak sample. Further transcript profiling among different cork oak tissues and conditions suggests that cork and wood share many regulatory mechanisms, probably reflecting similar ontogeny. Moreover, the analysis of transcripts accumulation during the cork growth season showed that most regulatory genes are upregulated early in the season when the cork cambium becomes active. Altogether our work provides the first transcriptome comparison between cork oak and holm oak outer bark, which unveils new regulatory candidate genes of phellem development.