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Background: Paper mulberry is one of the most common skin-lightening agents in the beauty industry due to its strong anti-tyrosinase activity. This narrative review aims to summarize the chemical composition, biological activities, and applications of paper mulberry in cosmetics. Method: The literature for this article was acquired from the PubMed, Web of Science, and Google Scholar databases before September 2022. The keywords for searching included “paper mulberry”, “Broussonetia papyrifera”, “skin-lightening”, “skin-whitening”, “depigmentation”, “pharmacological activity”, and “biological activity”. Results: Paper mulberry consists of various components, including flavonoids, tannins, alkaloids, phenols, saponins, coumarins, glycosides, and polysaccharides, which possess a wide range of pharmacological properties. Apart from its anti-tyrosinase activity, paper mulberry and its compounds exhibited anti-inflammatory, antioxidant, antimicrobial, antiviral, anticancer, antidiabetic, anticholinesterase, antigout, antinociceptive, and hepatoprotective effects. Phenols and flavonoids were demonstrated to be the main contributors to the biological activities of paper mulberry. Paper mulberry is widely applied in cosmetics for skin lightening and skin moisturizing purposes and shows potential for application in hair care products due to the hair nourishing effects. The safety of paper mulberry for topical application was proven in clinical studies. Conclusion: The current review provides a better understanding of paper mulberry’s properties and allows us to extend the application of this plant and its bioactive components in cosmetics.

This study aims to investigate the anaerobic digestion (AD) performance of Broussonetia papyrifera, a wide distributed nitrogen-rich and lignocellulosic-like plant, for methane production. The fermentation was conducted with the volatile solid (VS) concentration at 0.5%, 1.0%,1.5%,2.0% and 4.0% under 37°C and 55°C for 30 days, respectively. Results suggested that the feedstock concentration at 2.0% and fermentation temperature at 55°C was optimal for the AD treatment of Broussonetia papyrifera. A maximal specific methane yield was achieved at 277.72 ± 15 mL/g VS, which was 94.71% higher than that for the maximum specific methane under the thermophile condition, likely because that the C/N was more suitable compared with other lignocellulosic biomass. A kinetic study suggested that the modified Gompertz equation was reliable (determination coefficients R2 greater than 0.97) to describe the kinetic behavior of anaerobic digestion of Broussonetia papyrifera. In general, Broussonetia papyrifera was proved to be a suitabe feedstock for anaerobic digestion for methane production. [ABSTRACT FROM AUTHOR]

Thirty-six leaf samples of Sophora davidii, Indigofera amblyantha, and Broussonetia papyrifera were collected during different growth periods. The dynamic changes in nutritional components and secondary metabolites were analysed, and the biological activities of the secondary metabolites were evaluated. The optimal utilization stage of each plant was comprehensively determined. The results showed that S. davidii (the crude protein (CP) content 24.82%, relative feed value (RFV) 186.52)andI.amblyantha (CP 24.28%, RFV 177.54) were premium forage materials and that B. papyrifera (CP 21.65%, RFV 133.84) was a first-level forage. The best period for using the leaves of these three plants as animal feed was the flowering stage. As the growth progressed, the contents of CP, crude fibre (CF), dry matter (DM), and nitrogen-free extract (NFE) decreased, while the CF, neutral detergent fibre (NDF), acid detergent fibre (ADF), and crude ash (Ash) increased. The S. davidii,I. amblyantha, and B. papyrifera contains total flavonoid (0.36-0.70, 1.07-2.87, and 0.59-0.78 mg/g, respectively), tannin (5526.21-7717.77, 4284.03-5818.73, and 6390.66-7640.60 nmol/g, respectively), total phenol (205.62-298.69, 163.22-214.22, and 143.87-214.34 mg/g, respectively), total saponin (0.11-0.22, 0.20-0.39, and 0.10-0.16 ng/g, respectively) and total alkaloid (0.20-0.32, 0.23-0.38, and 0.20-0.36 ng/g, respectively). Moreover, the tannins, total phenols, saponins, flavonoids, and alkaloids in these woody forages scavenged hydroxyl radicals and superoxide anions to varying degrees, and their ability to reduce Fe3+ gradually increased with concentration. These plants inhibited hyaluronidase and albumin denaturation. The alkaloid and flavonoid extracts of S. davidii significantly (P≤0.05) inhibited Staphylococcus aureus with antibacterial diameters of 14.48 and 14.44 mm, respectively. The flavonoid (11.29 mm) and tannin (11.17 mm) extracts of S. davidii significantly (P≤0.05) inhibited Escherichia coli. The alkaloid (12.29 mm) and tannin (13.15 mm) extracts of B. papyrifera, and the flavonoid extract of S. davidii (13.44 mm) and I. amblyantha (10.32 mm) inhibited Bacillus subtilis. The secondary metabolites of these plants possess antioxidant, anti-inflammatory and antibacterial activities. In conclusion, these findings provide new perspectives on the nutritional value and biochemical characteristics of leaves of S. davidii, I. amblyantha, and B. papyrifera, emphasizing their use as valuable feed source for ruminants. [ABSTRACT FROM AUTHOR]

Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent chronic liver diseases. The leaves of Broussonetia papyrifera contain a large number of flavonoids, which have a variety of biological functions. In vitro experiments, free fatty acids were used to stimulate HepG2 cells. NAFLD model was established in vivo in mice fed with high fat diet (HFD) or intraperitoneally injected with Tyloxapol (Ty). At the same time, Total flavonoids of Broussonetia papyrifera (TFBP) was used to interfere with HepG2 cells or mice. The results showed that TFBP significantly decreased the lipid accumulation induced by oil acid (OA) with palmitic acid (PA) in HepG2 cells. TFBP decreased the total cholesterol (TC), the triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and increased high-density lipoprotein cholesterol (HDL C) in serum. TFBP could also effectively inhibit the generation of reactive oxygen species (ROS) and restrained the level of myeloperoxidase (MPO), and enhance the activity of superoxide dismutase (SOD) to alleviate the injury from oxidative stress in the liver. Additionally, TFBP activated nuclear factor erythroid-2-related factor 2 (Nrf2) pathway to increasing the phosphorylation of AMP-activated protein kinase (AMPK). Meanwhile, protein levels of mTORC signaling pathway were evidently restrained with the treatment of TFBP. Our experiments proved that TFBP has the therapeutic effect in NAFLD, and the activation of Nrf2 and AMPK signaling pathways should make sense. • Our experiments proved that the total flavonoids of Broussonetia papyrifera has therapeutic effect in NAFLD, and the activation of Nrf2 and AMPK should make sense. • A promising approach to extract certain compounds from plants for the prevention of NAFLD [ABSTRACT FROM AUTHOR]

More attention has been paid in recent times to the application of Broussonetia papyrifera (BP) silage in ruminants, owing to its nutritional value. This study aimed to characterize the functionality of fermented BP and preliminarily explore its dietary effects on the gut health of laying hens. In this study, we characterized the antioxidant and antibacterial activities, bioactive compound profile, and bacterial community in Lactobacillus plantarum-fermented BP (FBP), as well as its dietary effects on intestinal morphology, microbiota and gene expression of laying hens. Improved contents of protein, total polyphenol and flavonoids as well as antioxidant and antibacterial activities were found after fermentation of BP. Untargeted metabolomics displayed more abundant apigenin, luteolin, diosmetin, and quercetin within the FBP, which may contribute to its functionality. Microbiome demonstrated increased abundance of Firmicutes at the expense of Cyanobacteria phylum, accompanied with raised levels of Lactobacillus genus. The results of a feeding trial showed dietary FBP supplementation increased the serum superoxide dismutase, but down-regulated gene expression of aryl hydrocarbon receptor (AhR), mucin2, and ZO-2, without obviously affecting the intestinal morphology and colonic microbiota. These findings suggest that FBP warrants further investigation as it may serve as a functional dietary supplement in laying hen feed.

The quick adjustments of leaf traits are critical for keeping the survival of plants under dehydration. In this study, we examined the hypotheses that plants would adapt to dehydration by shrinking its mesophyll cells to offset the water loss, or triggering water regulation mechanism caused by enzymes. Leaf structure, elastic modulus (Em), leaf tensity (LT) and leaf density (LD) were determined with detached leaves of Broussonetia papyrifera (L.) Vent. and Morus alba L. at each water loss moment (0, 1, 2, 3, 4 and 5 h). The coupling model between gripping force and LT was established using the Gibbs free energy equation, and the initial LT was determined. The intracellular water availability of M. alba decreased at 4 h, which was earlier than that of B. papyrifera. The intracellular water availability of M. alba was more sensitive than B. papyrifera. Broussonetia papyrifera adapted to dehydration by shrinking its mesophyll cells to offset the water loss, or triggering water regulation mechanism caused by enzymes, i.e., carbonic anhydrase. The sponge parenchyma of B. papyrifera at 3 h decreased by 25.73% of that at control. Morus alba maintained intracellular water availability just by changing the leaf structure. The offset effects through shrinking cells differed between B. papyrifera and M. alba, because the elastic-plastic behavior of their leaves and cells were different. The Em of M. alba was over five-fold higher than that of B. papyrifera. The investigations of water status were more accurate in terms of leaf physical traits instead of water content. [ABSTRACT FROM AUTHOR]

Broussonetia papyrifera (L.) L’Hér. ex Vent. has considerable economic and ecological value and a long history of use in China. In this paper, root cuttings were used as the material to establish an efficient vegetative propagation of B. papyrifera. The results revealed that root segments with a diameter of 1.5~2.0 cm and a length of 20~30 cm were most suitable for shoot regeneration, as these segments had the highest adventitious shoot induction rates (93.3%), strongest adventitious shoots, and highest multiplication coefficients (7.07). With regard to the methods used for root burial, a horizontal burial at a depth of 1~3 cm yielded the best results, in this case, the adventitious shoot induction rate can reach 86.7%. The best substrate combination was perlite: peat: coconut chaff = 1:1:1 (v/v/v), wherein the adventitious shoot induction rate can reach 75.6%. The best sterilization method was mixing soil with carbendazim and soaking the root sections in carbendazim for 30 min, wherein the adventitious shoot induction rate can reach 77.8%. Adding 0.2 mg/L naphthaleneacetic acid (NAA) to 1/4 Hoagland nutrient solution significantly improved the rooting rate of adventitious shoots to 82.2%, and the survival rate of the acclimatized plants was more than 90.0%.

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Abstract A new flavane, bropapyriferol (1), and eleven known ones were isolated from the EtOAc part of Broussonetia papyrifera under the guidance of bioassay. The structure of compound 1 was determined by extensive 1D and 2D NMR, [α]D spectroscopic data and quantum computation. Daphnegiravan F (2) and 5,7,3′,4′-tetrahydroxy-3-methoxy-8,5′-diprenylflavone (3) showed significantly anti-oral microbial activity against five Gram-positive strains and three Gram-negative strains in vitro. Especially, compound 3 was more potent in suppressing Actinomyces naeslundii and Porphyromonas gingivalis (MIC = 1.95 ppm) than the positive control, triclosan. Graphical Abstract

The experiment was to explore the effect of accessories, such as wheat bran, corn flour, molasses, on fermentation quality of Broussonetia papyriferadry powder feed. The control group contained Broussonetia papyrifera powder 100%, the test groupⅠcontained Broussonetia papyrifera powder 90% + bran 10%, the test groupⅡcontained Broussonetia papyrifera powder 85% + bran 10% + corn powder 5%, and the test group Ⅲ contained Broussonetia papyrifera powder 80% + bran 10% + corn powder 5% + molasses 5%. Under the condition of solid-liquid ratio 1 g/mL, fermentation bacteria ( Lactobillis Plantum ∶ Candidum production = 1∶1) 5% and compound enzyme preparation 1‰, room temperature fermentation was carried out to vacuum state. The results showed that the content of lactic acid in test group Ⅲ was significantly higher than that in control group and test group Ⅰ and group Ⅱ ( P <0.05), and the pH value and NH3-N/TN were significantly lower than those in control group and test group Ⅰ and group Ⅱ ( P <0.05), the content of acid soluble protein in test group Ⅲ was significantly higher than control group ( P <0.05), and the content of neutral and acid detergent fiber were significantly lower than control group ( P <0.05). The experiment indicates that adding wheat bran, corn flour, molasses can significantly improve the fermentation quality of Broussonetia papyrifera powder feed. [ABSTRACT FROM AUTHOR]

The experiment was to improve the silage quality of Broussonetia papyrifera, to investigate the effect of formic acid on silage quality and nutrient composition. The experiment took Broussonetia papyrifera as the material, the control group (CK group) was silage directly without additives, and experimental groups were added 0.5, 1.0, 2.0 and 4.0 mL/100 g formic acid, respectively. The content of nutrients composition, pH value and the content of organic acids in Broussonetia papyrifera silage were determined after silaged for 30 d. The results showed that the direct silage treatment of Broussonetia papyrifera made the fermentation quality poor. After the addition of formic acid, the pH value of silage, the content of NDF and ADF decreased significantly (P<0.05), the content of lactic acid, OM and RFV increased significantly (P<0.05). The experiment indicates that adding the minimum concentration of 0.5 mL/100 g formate silage can achieve better quality and nutritional value. [ABSTRACT FROM AUTHOR]

Heavy concentrations of bicarbonates present throughout karst ecosystems necessitate identification of the role of in plant growth, especially when under drought stress. We aim to elucidate effect of different concentrations of on paper mulberry (Broussonetia papyrifera L.) seedlings growth under drought stress. Effects of four bicarbonate concentrations (0, 3, 6, and 9 mM) with simulated mild drought stress on growth, glucose metabolism, photosynthetic traits, self-protective abilities and extent of cell damage were examined. Compared with other treatments, plants under 3 mM bicarbonate with simulated mild drought stress showed the best osmotic adjustment and reactive oxygen species scavenging ability, the highest total activity of glucose catabolism enzymes and glucose-6-phosphate dehydrogenate activity, and the highest growth rate. Specific concentration of reduce negative influence in Broussonetia papyrifera L. under drought stress. Due to both producing more metabolic substrate and activating pentose phosphate pathway, those plants showed stronger resistance, therefore might suffer less harm and retain a better growth status under mild drought stress. This suggests that when specific plants in a karst environment are suffering drought stress, specific concentration of benefit to their growth. [ABSTRACT FROM AUTHOR]

This study was carried out in an attempt to evaluate the impact of Broussonetia papyrifera as a roughage substitute at different levels on carcase traits, growth performance, meat quality and immune performance in Hu ram lambs. Sixty Hu rams (5 months of age, 26.70 ± 2.14 kg body weight) were randomly divided into four groups. The treatments comprised B. papyrifera supplementation at levels of 0% (G0), 30% (G30), 60% (G60) and 100% (G100) of roughage feed. The results suggested that diet supplemented with B. papyrifera (G100 group) caused a higher average weight gain (AWG) and average daily gain (ADG) than those of G0 group. The highest carcase weight was observed in the G60 group. The chemical and physical properties of the longissimus dorsi muscle of Hu rams showed no significant differences (p>.05). For fatty acid, the G60 group had significantly lower content of saturated fatty acids (SFAs), higher contents of monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) and higher PUFA/SFA ratio than other groups (p<.05). In terms of the immune response, G60 and G100 groups significantly showed a significant rise in the immunoglobulins (IgG, IgA and IgM) (p<.05). This study indicated multiple beneficial effects of the inclusion of B. papyrifera (60% of the roughage feed) on the growth performance, carcase traits, meat quality and immune response in Hu rams. It could be used as a high-quality unconventional feedstuff for rams. The addition of Broussonetia papyrifera to the diet resulted in a significant improvement in the average weight gain (AWG) and average daily gain (ADG) of Hu rams. Adding appropriate proportion of B. papyrifera in diets positively modulates fatty acid percentage in the longissimus dorsi muscle, and also increases immunoglobulins (IgG, IgA and IgM) in blood. Broussonetia papyrifera could be used as a high-quality unconventional feedstuff for rams and has a positive effect on their growth performance, meat quality and immune performance. [ABSTRACT FROM AUTHOR]

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Information about plant materials of construction in artefacts advances knowledge about human history, agriculture, trade, migration and adaptation to new environments. Typically, materials identification in artefacts made from plants is problematic, since processing, age, dirt and surface treatments can mask identifying features, while ethical considerations relating to sampling limit the use of some analytical techniques. The study tests the usefulness of polarized light microscopy for identifying the New Zealand and Pacific plant species used to make tapa, indicating birefringent and morphological characteristics that can be used to differentiate fibres at the level Moraceae (Pacific; from genera Artocarpus, Broussonetia and Ficus) and Malvaceae (New Zealand; from genera Hoheria and Plangianthus). [ABSTRACT FROM AUTHOR]

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Copyright of Acta Prataculturae Sinica is the property of Acta Prataculturae Sinica Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In northern Laos, shortened fallows in the slash-and-burn upland rice system are not sustainable either from a production or economic standpoint. This paper evaluates the potential for using the indigenous fast-growing tree, Broussonetia papyrifera L. Vent. (paper mulberry) as an economic fallow crop. The objectives of this study are to assess the inner bark yield of paper mulberry, and to evaluate rice productivity following paper mulberry cultivation. In addition, allometric models for estimating the inner bark yield of paper mulberry are developed. Inner bark yield of paper mulberry stands ranging in age from 9 to 48 months ranged from 10 to 208 g m−2, and increased with age. The models using diameter at breast height and stem height, or diameter at breast height only as independent variables explained well the variation in inner bark yield. When paper mulberry was completely removed during the rice growing season, rice yields in these areas were similar to those following weedy bush fallow. When paper mulberry was allowed to grow, rice yields were negatively correlated to paper mulberry stem density. When paper mulberry lateral branches were pruned back, rice yields could be maintained if the paper mulberry stem density was

Broussonetia papyrifera leaves were successfully converted into new magnetic adsorbents, including pretreatment using ionic liquid and magnetization via a coprecipitation technology. The resultant magnetic adsorbent was characterized with infrared spectroscopy (IR), transmission electron microscopy, scanning electron microscopy, and X-ray diffraction. Then, it was selected to adsorb methylene blue from aqueous solutions. The adsorption behaviors could be described well by pseudo-second-order kinetic and Langmuir isotherm models. The thermodynamic analysis for adsorption displayed that the process was exothermic and feasible spontaneous. Moreover, it exhibited a well reusability in the regeneration test. This adsorbent might be an alternative candidate with the advantageous properties of simplicity, repeatability, and convenient separation for pollutant removal. [ABSTRACT FROM AUTHOR]

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Effective phytoremediation of Mn contaminated soil requires the selection of a species with good manganese tolerance. Broussonetia papyrifera is an important economic plant and pioneer species, it could be well adapted to drought and saline-alkali environment. In order to understand the effect of Mn stress on B. papyrifera , the effects of different concentrations of Mn (0–50 mmol/L) stress on the growth, morphology, Mn tolerance and physiological indexes of the plant were explored. The results showed that the biomass, surface area, length, root volume, tips, forks, and crossings of B. papyrifera reached the maximum at the Mn concentration of 1 mmol/L. Mn content in the tissue and TF in plants increased with the increase of concentration, while the BCF increased first and then decreased, and the maximum BCF was 0.154 at 10 mmol/L. The accumulation of Mn lead to cell membrane lipid peroxidation, which increased toxic substances in plants, resulting in the increase of MDA and PRO, and affected the synthesis of chlorophyll. However, B. papyrifera could effectively alleviate oxidative stress by increasing the activities of antioxidant enzymes (SOD, POD, CAT), protein and soluble sugar. The results suggested that B. papyrifera had a good oxidative stress mechanism to Mn stress and could be used as candidates for remediation of pollution in mining areas. • B. papyrifera can grow well under high Mn concentration (50 mmol/L). • The accumulation ability of Mn in different tissues was leaf > root > stem. • The accumulation of Mn resulting in the increase of MDA and PRO, and affected the synthesis of chlorophyll. • B. papyrifera alleviated oxidative stress by increasing the activities of antioxidant enzymes, soluble protein and sugar. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate whether δ13C values can be used to identify pollen specie in the atmosphere. A Burkard 7-day recording volumetric spore trap was used to collected pollens in the atmosphere in Tainan City, Taiwan, from January 2 to December 28, 2006, and a light microscope was used to identify the pollen species and concentrations. A Burkard cyclone sampler was used to collect particulate matter and an elemental analyzer with an isotope ratio mass spectrometer was used to analyze the δ13C values. Our data showed that the predominate pollen specie in the atmosphere was Broussonetia papyrifera pollen and that the annual average concentration was 27 grains/m3 (pollen season, 36; nonpollen season, 9 grains/m3). The average δ13C value was − 26.19‰ for particulate matter in the atmosphere (pollen season, − 26.00‰; nonpollen season, − 26.28‰). No significant association was observed between δ13C values and Broussonetia papyrifera pollen concentrations. However, the δ13C value in the atmosphere was associated with the levels of Broussonetia papyrifera pollen among the samples with a diameter of particulate matter smaller than 10 μm at a level lower than 40 μg/m3. In addition, the relative contribution of Broussonetia papyrifera pollen to the carbon in the atmosphere using a two end-member mixing models was found to be associated with the Broussonetia papyrifera pollen concentration. In summary, our study suggested that δ13C values can be applied in the assessment of Broussonetia papyrifera pollen specie under specific conditions in the atmosphere. [ABSTRACT FROM AUTHOR]

[ Objectives ] The purpose of ihis experiment was to study ihe effects of Broussonetia papyrifera ( BP) as a crude feed source on the rumen fermentation characteristics, digestion and metabolism, production performance and meat quality of Huanghuai white goat. [Methods]Forty goats(5 months of age)were randomly assigned to four treatments:25% BP(B25) .50% BP(BP50), 75% BP(BP75) and BP0( without adding BP) .respectively, with 5 pens per treatment. The experimental period lasted for 60 days. [Results]The results indicated that adding B. papyrifera in ratio had no significant difference between pi! value( P>0.05)and had very significant difference between NH3-N concentration( P<0.05) .compared with BP0(the control). The NH3-N concentration of adding B. papyrifera groups (BP25,BP50,BP75) increased about 21.30%,25.18% and 38.92% .respectively, compared with that of BP0 group. Microbial protein (MCP)yield of BP75 group increased about 22.82%,compared with that of BP0 group, and there was significant difference between MCP yield of BP75 and BP25 group( P<0.05). No significantly difference was observed for apparent digestibility of acid detergent fiber (ADF)among each group( P>0.05). Apparent digestibility of nutrition of BP25 group was higher than BP50 group and BP75 group, and apparent digestibility of dry matter( DM)of BP25 had significant difference with that of BP75 group( P<0.05) .and apparent digestibility of neutral detergent fiber(NDF)of BP50 group had significant difference with that of BP25 group( P<0.05). Apparent digestibility of nutrition had no significant difference between BPM group and BP0 group(P>0.05). Nitrogen intake, digestible nitrogen and deposition of nitrogen of groups with B. papyri/era in ratio weni significantly higher than those of BP0 group( P<0.05) .nitrogen utilization of BP25 group was supreme, and had significant difference with other groups. Average daily gain( ADG) .carcass weight, and slaughter rate of BP25 group and BP50 group were significantly higher than those of BP0 group(P<0.05). Muscle tenderness and water holding capacity of meat increased with the addition li. papyri/era, cooked meat rate of BP50 group was highest and had significant difference with that of other groups (P<0.05), a' value and I. value of meat color from hind leg and longissimus dorsi of BPW group were higher than those of other groups( P<0.05). [ Conclusions ] B. papyri/era supplementation from 25% to 50%,and matched with com silage could improve rumen fermentation and nitrogen utilization of goats .and thereby it could increase performances and make meat quality better. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate the effects of Broussonetia papyrifera (paper mulberry) invasion and land use on the floristic composition of a dry semi-deciduous forest in Ghana. Forty-five plots (25 m × 25 m each), distributed among three land uses—selectively logged (SL); abandoned farmlands (AF); and an undisturbed reference (RF)—were surveyed. Results showed lower tree species richness (S), diversity (H′), evenness (S) and basal area (BA) in the SL (46, 0.78, 0.32 and 269.12 m2 ha−1, respectively) and AF (40, 0.53, 0.45, and 131.16 m2 ha−1) sites compared to the RF site (79, 2.66, 0.87, 963.72 m2 ha−1). Similar patterns were found at the shrub layer, but no differences were observed at the herb layer. Non-metric multidimensional scaling ordination revealed distinct species composition among the land uses. The two disturbed habitats, SL and AF, were associated with increased B. papyrifera invasion particularly in the overstory, with importance value index and mean relative density of 45 and 65.03%, and 42 and 53.29%, correspondingly. However, the species was only sparsely represented in the RF site. Tree density of B. papyrifera correlated negatively with H′, S, E, BA, and native tree density and richness. These findings highlight the strong link between human land use (i.e., logging and slash-and-burn farming), invasion, and vegetation characteristics, and suggest the need to limit these disturbances to conserve biodiversity within tropical forest ecosystems. [ABSTRACT FROM AUTHOR]

Few studies have investigated the effect of environment on the root-associated microbiome, especially for woody plants in their native environment. The roots and rhizosphere soils of a native woody species (Broussonetia papyrifera) sampled across four different climate types in China were used to elucidate the influence of environment on the root-associated microbiome. Our results showed that the B. papyrifera root-associated microbiome contained abundant Proteobacteria and Basidiomycota, especially Pseudomonas and Rhizobium. The root-associated microbiomes were found to be significantly different under different climate types except for the bacterial community in the rhizosphere, and the proportion of bacterial operational taxonomic units (OTUs) shared among different climate types was lower than that of fungi. More than 50% of the total variance between microbiomes could be explained by 15 environmental factors, six of which, especially soil concentration phosphate and nitrate, had a significant effect. This study provided a comprehensive understanding of the root-associated microbiome of B. papyrifera and further confirmed the effect of environment on the root-associated microbiome of B. papyrifera under different climate types, with some exceptions in the rhizobacterial community and fungal OTUs. Our findings advanced knowledge of the effect of environment through an exploration of environmental factors and found that the nitrogen and phosphorus content represented the key factors. [ABSTRACT FROM AUTHOR]

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Abstract Background Heavy metal contamination is becoming a limitation to the utilization of soil and the distribution of vegetation. In particular, cadmium (Cd) pollution has had a serious impact on the food chain. Broussonetia papyrifera is a widely distributed pioneer tree species of heavy metal contaminated areas with important economic value. However, little is known about the genomic background of the Cd-tolerance mechanism in B. papyrifera. Results The CdCl 2 responsive physiology was evaluated and proved to be involved in antioxidase activity and active oxygen species (ROS) accumulation. The leaf and root transcriptomes derived from B. papyrifera grown under normal and CdCl 2 stress conditions were systematically investigated using the Illumina HiSeq method. A total of 180,678,660 bp (27.1 GB) clean reads were assembled into 589,487 high-quality unigenes, of which 256,025 (43.43% of the total) and 250,251 (42.45% of the total) were aligned in Gene Ontology (GO) and Protein family (Pfam), respectively. A total of 24,414 differentially expressed genes (DEGs) were GO-annotated into 53, 23, 55, and 60 terms from the transcriptomes of root and leaf tissues under Cd stress and control conditions. A total of 117,547 Kyoto Encyclopedia of Genes and Genomes (KEGG) Orthology (KO)-annotated DEGs were enriched in at least 47 KEGG pathway terms among the four comparisons. Many genes encoding important transcription factors (e.g., auxin/indole-3-acetic acid (AUX/IAA), basic helix-loop-helix (bHLH), DNA-binding one zinc finger (Dof), and MYB) and proteins involved in plant-pathogen interactions, phenylpropanoid biosynthesis, plant hormone signal transduction, oxidative phosphorylation, carbon fixation, peroxisomes, flavonoid biosynthesis, and glutathione metabolism, among others, were substantially upregulated under CdCl 2 stress. Conclusions These genes represent important candidates for studying Cd-response mechanisms and molecular biology of B. papyrifera and related species. Our findings provide a genomic sequence resource for functional genetic assignments in B. papyrifera , which will help elucidate the molecular mechanisms of its Cd-stress responses and facilitate the bioremediation of heavy metal contaminated areas via breeding of new stress-tolerant cultivars. Graphical abstract fx1 Highlights • Broussonetia papyrifera respond to Cd stress involving in reactive oxygen species and antioxidant enzyme metabolic pathway. • Abundant important Cd response related genes and regulation pathways that involving in ABA and auxin signal were identified. • The gene resource of B. papyrifera will help elucidate the molecular mechanisms of Cd-stress responses. [ABSTRACT FROM AUTHOR]

Abstract: A new flavane, bropapyriferol (1), and eleven known ones were isolated from the EtOAc part of Broussonetia papyrifera under the guidance of bioassay. The structure of compound 1 was determined by extensive 1D and 2D NMR, [α]D spectroscopic data and quantum computation. Daphnegiravan F (2) and 5,7,3′,4′-tetrahydroxy-3-methoxy-8,5′-diprenylflavone (3) showed significantly anti-oral microbial activity against five Gram-positive strains and three Gram-negative strains in vitro. Especially, compound 3 was more potent in suppressing Actinomyces naeslundii and Porphyromonas gingivalis (MIC = 1.95 ppm) than the positive control, triclosan.Graphical Abstract: [ABSTRACT FROM AUTHOR]

The formation of a common mycorrhizal network (CMN) between roots of different plant species enables nutrient transfers from one plant to another and their coexistence. However, almost all studies on nutrient transfers between CMN-connected plants have separately, but not simultaneously, been demonstrated under the same experimentation. Both conspecific and heterospecific seedlings of Cinnamomum camphora, Bidens pilosa, and Broussonetia papyrifera native to a karst habitat in southwest China were concurrently grown in a growth microcosm that had seven hollowed compartments (six around one in the center) being covered by 35.0-μm and/or 0.45-μm nylon mesh. The Ci. camphora in the central compartment was supplied with or without Glomus etunicatum and 15N to track N transfers between CMN-connected conspecific and heterospecific seedlings. The results showed as follows: significant greater nitrogen accumulations, biomass productions, 15N content, % Ntransfer, and the Ntransfer amount between receiver plant species ranked as Br. papyrifera≈Bi. pilosa > Ci. camphora under both M+ and M−, and as under M+ than under M− for Ci. camphora but not for both Bi. Pilosa and Br. papyrifera; the CMN transferred more nitrogen (15N content, % Ntransfer, and Ntransfer amount) from the donor Ci. camphora to the heterospecific Br. papyrifera and Bi. pilosa, with a lower percentage of nitrogen derived from transfer (%NDFT). These findings suggest that the CMN may potentially regulate the nitrogen transfer from a donor plant to individual heterospecific receiver plants, where the ratio of nitrogen derived from transfer depends on the biomass strength of the individual plants. [ABSTRACT FROM AUTHOR]

Abstract Pollen has long been recognized as a major allergen, having diverse patterns of allergenicity caused by differences in climate, geography, and vegetation. Our research aimed to explore the role of a regionally dominant pollen in Taiwan, Broussonetia papyrifera, on clinical sensitization and daily 5collected and extracted for a skin prick test on 30 volunteers recruited from a medical college. Daily atmospheric pollen levels were measured using a Burkard 7-day volumetric trap. The association between daily atmospheric pollen levels and clinic visits for allergic illness was examined using a generalized additive model with a normal assumption. After excluding four participants with a positive response to a negative control, 10 participants (38.4%) were determined to be sensitive to B. papyrifera pollen extract. The three-day lagged concentration of B. papyrifera pollen exhibited the highest risk of daily asthma visits (relative risk [RR] = 1.166, 95% confidence interval [CI]: 1.014–1.341) and allergic rhinitis visits (RR = 1.119, 95% CI: 0.916–1.367) when the pollen increased equally in magnitude to its mean. Our study is the first to provide evidence indicating that the most dominant airborne pollen in Taiwan, B. papyrifera, plays a major role in sensitization and clinic visits for asthma and allergic rhinitis, thus highlighting the need to integrate aeroallergen monitoring with clinical diagnosis. Graphical abstract Unlabelled Image Highlights • The role of pollen allergy has long been underestimated in Taiwan. • Daily pollen counts and skin prick test of fresh pollen extracts were used. • Broussonetia papyrifera pollen is an important allergen in Taiwan. • The observed association between dominant pollens and clinical admissions for asthma and allergy show that adequate aeroallergens monitoring is needed. [ABSTRACT FROM AUTHOR]

Both manganese-slag and sewage sludge are typical solid wastes, but their utilization is limited. Based on the soil properties, the abovementioned pollutants were combined with Broussonetia papyrifera to treat soil cadmium (Cd) pollution. Three materials (sewage sludge-derived biochar (SSB), Mn-SSB, and Mn-slag (Slag)) were prepared using oxygen-limited pyrolysis technology with Slag and sewage sludge, and the effects of the three materials on the phytoremediation of Cd-polluted soil were investigated. All three materials had distinct morphological characteristics, good functional group structure, specific surface area, and porosity. The adsorption and leaching experiments in the solution indicated that the three materials could not only directly absorb Cd2+ but also release nutrients, such as nitrogen and phosphorus. The soil pH increased significantly (p < 0.05) with the addition of the above environmental remediation materials. Furthermore, the contents of soil organic carbon, available nitrogen, and available phosphorus in soil increased significantly, whereas the electrical conductivity of the soil decreased significantly (p < 0.05). During remediation of Cd-polluted soil by integrating the above materials with B. papyrifera , Slag significantly increased the B. papyrifera biomass, but the effects of SSB and Mn-SSB were not significant. SSB, Mn-SSB, and Slag significantly increased the protein content of B. papyrifera leaves, with Mn-SSB having the most significant effect (p < 0.05). The applications of SSB, Mn-SSB, and Slag reduced the malondialdehyde content and increased the activities of superoxide dismutase and peroxidase, reducing the damage to B. papyrifera. Mn-SSB significantly reduced the Cd content in the roots, stems, and leaves of B. papyrifera , and SSB and Slag promoted Cd enrichment in B. papyrifera. This study realized the comprehensive utilization of Mn-slag and sewage sludge and established a recycling system from solid waste to the treatment of waste soil. [Display omitted] • Environmental remediation materials based on manganese-slag and sewage sludge can not only improve soil properties, but also directly absorb Cd. • The environmental remediation materials can promote the growth of Broussonetia papyrifera , especially the root system. • The prepared environmental remediation materials also can enhance the resistance of Broussonetia papyrifera to Cd stress. [ABSTRACT FROM AUTHOR]

HD-Zip transcription factors are unique to higher plants and contributes to abiotic stress tolerance. In this study, PsHB7 and PsHB12 from the phloem of Pyrus sinkiangensis were cloned and transferred into hybrid Broussonetia papyrifera to evaluate their functions in cold acclimation. PsHB7 and PsHB12 expression in B. papyrifera decreased the rate of water loss, thereby affecting the overall physiology of the plants. After ABA treatment, increases in the water content and soluble protein levels in transgenic lines conferred enhanced freeze resistance. Comprehensive analyses of anti-freezing performance using logistic regression and membership functions yielded consistent results with the physiological experimental data. Furthermore, after natural overwintering, the transgenic lines exhibited less frost damage and the germination time was 20 days earlier than that of the wild-type plants. These results suggest that PsHB7 and PsHB12 play a key role in the negative feedback regulation of ABA signaling, which can balance cellular water efflux and the accumulation of dehydrated proteins. • Hybrid B. papyrifera , a perennial woody plant, provides high-quality forage but is often vulnerable to winter frost damage. • After natural overwintering, the PsHB7 / 12 transgenic lines showed lower LT50. • PsHB7 / 12 transgenic lines exhibited an earlier bud break by 20 d compared to the wild type after natural overwintering. • PsHB7 / 12 play a role in balancing cell water efflux and dehydrin protein accumulation in the ABA signaling pathway. • PsHB7 / 12 may regulate the ABA signal through negative feedback to prevent excessive dehydration of cells under ABA response. [ABSTRACT FROM AUTHOR]

The current study was designed to assess the inhibitory activity of Broussonetia papyrifera-derived polyphenols against 3-chymotrypsin-like and papain-like coronavirus cysteine proteases. The isolated compounds were broussochalcone B (1), broussochalcone A (2), 4-hydroxyisolonchocarpin (3), papyriflavonol A (4), 3′-(3-methylbut-2-enyl)-3′,4,7-trihydroxyflavane (5), kazinol A (6), kazinol B (7), broussoflavan A (8), kazinol F (9), and kazinol J (10). All polyphenols were more potent against papain-like protease (PLpro) than against 3-chymotripsin-like protease (3CLpro); therefore, we investigated their structural features that were responsible for this selectivity. Compound 4 was the most potent inhibitor of PLpro with an IC50 value of 3.7 μM. The active compounds displayed kinetic behaviors, and the binding constants of their interaction with PLpro were determined from surface plasmon resonance analysis. Our results suggest B. papyrifera constituents as promising candidates for development into potential anti-coronaviral agents.

Broussonetia papyrifera is a widely distributed economic tree species, and it is also a pioneer species in adverse environments. In order to investigate the growth and adaptation mechanism of B. papyrifera under cadmium (Cd) contaminated soil, potted experiments were used with six-month treatments to study Cd enrichment and the transportation, morphological and physiological characteristics of B. papyrifera tissues. The results showed that Cd mainly accumulated in the root when the Cd concentration was high (14.71 mg/kg), and the root biomass was significantly reduced by Cd stress although Cd promoted the growth of seedlings. The bioconcentration factors (BCF) increased with the increase in Cd concentration, and reached the maximum value of 0.21 at 14.71 mg/kg. On the contrary, translocation factor (TF) decreased significantly at 8.28–14.71 mg/kg Cd concentration. Cd not only led to the loose arrangement of the xylem vessels of leaves, but also changed the chlorophyll content. However, B. papyrifera could synthesize organic solutes such as soluble protein, soluble sugar and proline to reduce the intracellular osmotic potential. Our study proved that B. papyrifera has good tolerance to Cd stress and is a pioneer tree species for soil and ecological environment restoration.

Broussonetia papyrifera (B.papyrifera), belonging to the Moraceae family, is known to elicit anti-inflammatory, antioxidant, anti-tyrosinase, anticancer, antinociceptive, and antimicrobial effects. The present study has been designed to examine the effects of B. papyrifera extract on hair growth through in vitro and clinical samples. Real-time cell growth assay, T-cell factor/lymphoid enhancer-binding factor (TCF/LEF), activation of signal transducer and activator of transcription-6(STAT6) and STAT3 reporter gene function, and Western blotting was performed to examine whether B. papyrifera regulates the expression of target proteins implicated in the proliferation of human hair follicle dermal papilla (hHFDP) cells. In this human trial, using a phototrichogram, the effect of B. papyrifera on hair growth was examined by reconstitution analysis after shaving the hair of the clinical subject’s dorsal skin. B. papyrifera promoted growth equally in hHFDP cells, which is comparable to that of minoxidil and tofacitinib. Treatment with B. papyrifera extract enhanced the TCF/LEF-luciferase activity and increased the level of β-catenin protein. Moreover, B. papyrifera extract significantly suppressed interleukin-4 (IL4)-induced STAT6 phosphorylation. In clinical trial, using a phototrichogram, we assessed the hair density and total hair counts at 0, 6, and 12 weeks after the use of hair tonic containing B. papyrifera extract. After using the hair tonic for 12 weeks, the total hair count was significantly increased as compared with the subjects at the start date (n = 11). B. papyrifera promotes dermal papilla cells proliferation in vitro and clinically among human volunteers through the regulation of WNT-β-catenin and STAT6 pathways.

The alleviation of cadmium (Cd) toxicity in Broussonetia papyrifera by arbuscular mycorrhizal (AM) fungi are still not completely elucidated. This study investigated the effects of Rhizophagus irregularis on physiological and biochemical characteristics, and molecular regulation in B. papyrifera under different levels of Cd (0, 30, 90 and 270 mg kg-1 Cd) stress. Results showed that (1) AM symbiosis improved the growth and photosynthesis, enhanced ROS levels as stress signaling and maintained ROS balance under low and medium Cd stress. (2) AM symbiosis regulated AsA-GSH cycle to mitigate ROS overproduction under high Cd stress. (3) AM fungus can chelate more Cd under high Cd stress, increasing soil pH and GRSP content. (4) AM plants can fix or chelate more Cd by P in leaves and reserve more P in stems under high Cd stress. (5) AM symbioses increased root net Cd2+ influx and uptake under medium Cd stress but inhibited under high Cd stress, with upregulation of genes related heavy metals (HMs) transport under medium Cd stress and inhibited the transcription of genes related HMs transport under high Cd stress. Therefore, the alleviation mechanisms of Cd toxicity in B. papyrifera by R. irregularis symbiosis depends on the levels of Cd stress. [Display omitted] • AMF promote AsA-GSH cycle to alleviate high Cd stress. • AM plants fixed more Cd by P under high Cd stress. • AMF increased Cd2+ influx under medium Cd stress, but inhibited under high Cd stress. • The detoxification effect of AMF depends on the levels of Cd stress. [ABSTRACT FROM AUTHOR]

Broussonetia papyrifera , known as paper mulberry, is a potential feed stock for bioethanol production because of its cellulose-rich composition. Lignin in dilute acid pretreated Broussonetia papyrifera was fractionated to three different fractions, and their physiochemical properties were determined by FT-IR, GPC and NMR analyses. Different structural characteristics were observed from each lignin fraction. Cellobiohydrolases I (CBH) adsorption to each lignin was understood by the lignin properties. The results showed that aliphatic hydroxyl groups in lignin showed positive correlations with the maximum binding ability of CBH onto lignin samples. Also, the contents of phenolic compounds such as p -hydroxyphenyl benzoate (PB), syringyl (S) and guaiacyl (G) units in the lignin influenced their CBH binding. [ABSTRACT FROM AUTHOR]

* Background and Aims Paper mulberry or Broussonetia papyrifera (L.) L'Hér. ex Vent. (Moraceae) is a dioecious species native to continental South-east Asia and East Asia, including Taiwan, that was introduced to the Pacific by pre-historic voyagers and transported intentionally and propagated asexually across the full range of Austronesian expansion from Taiwan to East Polynesia. The aim of this study was to gain insight into the dispersal of paper mulberry into Oceania through the genetic analysis of herbaria samples which represent a more complete coverage of the historical geographical range of the species in the Pacific before later introductions and local extinctions occurred. * Methods DNA from 47 herbarium specimens of B. papyrifera collected from 1882 to 2006 from different islands of the Pacific was obtained under ancient DNA protocols. Genetic characterization was based on the ribosomal internal transcribed spacer ITS-1 sequence, a sex marker, the chloroplast ndhF-rpl32 intergenic spacer and a set of ten microsatellites developed for B. papyrifera. * Key Results Microsatellites allowed detection of 15 genotypes in Near and Remote Oceanian samples, in spite of the vegetative propagation of B. papyrifera in the Pacific. These genotypes are structured in two groups separating West and East Polynesia, and place Pitcairn in a pivotal position. We also detected the presence of male plants that carry the Polynesian chloroplast DNA (cpDNA) haplotype, in contrast to findings in contemporary B. papyrifera populations where only female plants bear the Polynesian cpDNA haplotype. * Conclusions For the first time, genetic diversity was detected among paper mulberry accessions from Remote Oceania. A clear separation between West and East Polynesia was found that may be indicative of pulses during its dispersal history. The pattern linking the genotypes within Remote Oceania reflects the importance of central Polynesia as a dispersal hub, in agreement with archaeological evidence. [ABSTRACT FROM AUTHOR]

Islamabad is rich in diversified flora. This study was conducted to assess the vegetation communities of Islamabad vicinity. In present study seven different sites were selected in the vicinity i.e. CDA Nursery, F9 Park, Faisal Mosque, Jasmine Garden, near Prime Minister House, near Quaid-e-Azam University and Rawal Lake. A total of 62 major plant species were found in the study area. The dominance of plant species was estimated on basis of importance value. Most dominating species was Brossonetia papyrifera with highest importance value (71.4+35.82). These selected sites were categories on the basis of specie richness indices and Shanen's diversity indices. Jinah Garden was rich in plant species on basis of specie richness.

The current study was designed to assess the inhibitory activity ofBroussonetia papyrifera-derived polyphenols against 3-chymotrypsin-like and papain-like coronavirus cysteine proteases. The isolated compounds were broussochalcone B (1), broussochalcone A (2), 4-hydroxyisolonchocarpin (3), papyriflavonol A (4), 3′-(3-methylbut-2-enyl)-3′,4,7-trihydroxyflavane (5), kazinol A (6), kazinol B (7), broussoflavan A (8), kazinol F (9), and kazinol J (10). All polyphenols were more potent against papain-like protease (PLpro) than against 3-chymotripsin-like protease (3CLpro); therefore, we investigated their structural features that were responsible for this selectivity. Compound4was the most potent inhibitor of PLprowith an IC50value of 3.7 μM. The active compounds displayed kinetic behaviors, and the binding constants of their interaction with PLprowere determined from surface plasmon resonance analysis. Our results suggestB. papyriferaconstituents as promising candidates for development into potential anti-coronaviral agents. [ABSTRACT FROM PUBLISHER]

Background: Broussonetia papyrifera (B. papyrifera), also known as paper mulberry, has been used as a traditional medicine for the treatment of several diseases, including ophthalmic disorders and impotency. However, the biological activity of kazinol A (1) among flavonols isolated from B. papyrifera has not been identified.Purpose: We identified a candidate metabolite for anti-human bladder cancer treatment from B. papyrifera and investigated the possible molecular mechanisms underlying its cytotoxic effects in T24 and cisplatin-resistant T24R2 human bladder cancer cells.Methods: T24 and T24R2 cells were treated with five flavonols from B. papyrifera and their cytotoxic effects were determined using MTT assay, cell cycle analysis, mitochondrial membrane potential, and propidium iodide staining. Autophagy rate was calculated by counting LC3-GFP dots in the cells. All related protein expressions were analyzed by immunoblotting.Results: Compound 1 showed relatively higher cytotoxicity in the human bladder cancer cells, T24 and T24R2, rather than other tissues-originated cancer cells. Compound 1 significantly attenuated cell growth through G0/1 arrest mediated by a decrease in cyclin D1 and an increase of p21. Apoptosis and autophagy induced by compound 1 treatment was accompanied by a modulation of the AKT-BAD pathway and AMPK-mTOR pathway, respectively.Conclusions: Our results suggest that compound 1 induces cytotoxic effects in human bladder cancer cells, including the cisplatin-resistant T24R2. Compound 1 may be a candidate for the development of effective anti-cancer drug on human urinary bladder cancer. [ABSTRACT FROM AUTHOR]

From the ethyl acetate extract of a culture of the endophytic fungus Alternaria species G7 in Broussonetia papyrifera , a new compound altertoxin IV ( 1 ) together with nine known compounds were isolated and identified by means of bioassay-guided fractionation. The structures of these compounds were established on the basis of spectroscopic methods, among which the absolute configuration of compound 1 , a new tetrahydroperylenone derivative, was determined by means of X-Ray Crystallographic analysis. The isolated compounds were subjected to cytotoxic activity against three human cancer cell lines (A549, MG-63, and SMMC-7721). Compound 2 showed significant cytotoxic activities against tested cell lines, with IC 50 values of 1.47, 2.11 and 7.34 μg/mL, respectively. Additionally, compound 4 also exhibited significant cytotoxic activities against cell lines MG-63 and SMMC-7721, with IC 50 values of 0.53 and 2.92 μg/mL. Endophytic fungi Alternaria from B. papyrifera might be promising sources of natural bioactive and novel metabolites. [ABSTRACT FROM AUTHOR]

The peopling of Remote Oceanic islands by Austronesian speakers is a fascinating and yet contentious part of human prehistory. Linguistic, archaeological, and genetic studies have shown the complex nature of the process in which different components that helped to shape Lapita culture in Near Oceania each have their own unique history. Important evidence points to Taiwan as an Austronesian ancestral homeland with a more distant origin in South China, whereas alternative models favor South China to North Vietnam or a Southeast Asian origin. We test these propositions by studying phylogeography of paper mulberry, a common East Asian tree species introduced and clonally propagated since prehistoric times across the Pacific for making barkcloth, a practical and symbolic component of Austronesian cultures. Using the hypervariable chloroplast sequences of 604 samples collected from East ndhF-rpl32 Asia, Southeast Asia, and Oceanic islands (including 19 historical herbarium specimens from Near and Remote Oceania), 48 haplotypes are detected and haplotype cp-17 is predominant in both Near and Remote Oceania. Because cp-17 has an unambiguous Taiwanese origin and cp-17-carrying Oceanic paper mulberries are clonally propagated, our data concur with expectations of Taiwan as the Austronesian homeland, providing circumstantial support for the "out of Taiwan" hypothesis. Our data also provide insights into the dispersal of paper mulberry from South China "into North Taiwan," the "out of South China-Indochina" expansion to New Guinea, and the geographic origins of post-European introductions of paper mulberry into Oceania. [ABSTRACT FROM AUTHOR]