Detection of Hybrids in Willows (Salix, Salicaceae) Using Genome-Wide DArTseq Markers.

The genus Salix, comprising some 400-500 species, is important in various alluvial or wet habitats of the northern hemisphere. It is a promising crop for applications such as biomass production, biofuels, or environmental projects. Clear species delimitation is crucial in ecology, biotechnology, and horticulture. DArTseq markers, a genome-wide technique, were tested for species and hybrid identification. A total of 179 willow samples were analysed, including six species of Salix subgen. Salix and four species of Salix subgen. Vetrix, including those used in biomass crop production, representing important European taxa. Identification of species-specific markers, clustering analyses (principal coordinate analysis, neighbor-joining) and Bayesian methods (Structure) unambiguously identified putative hybrids. In addition to demonstrating the high efficiency of DArT-seq markers in identifying willow hybrids, we also opened-up new questions about hybridisation processes and systematics. We detected unidirectional hybridisation between S. alba and S. fragilis, forming backcross hybrids, and we rejected the hypothesis that S. fragilis does not occur naturally in Europe. Further, the isolated position of Salix triandra within the genus was confirmed.

Curtobacterium salicis sp. nov., isolated from willow tree stems in Washington state.

Curtobacterium sp. strain WW7 is a Gram-positive, non-motile, orange rod-shaped bacterium isolated from branches of wild willow (Salix sitchensis) trees. The WW7T strain has optimum growth in the temperature range between 25 and 30 °C, a pH range of 6-7.7, and tolerates up to 5.5% (w/v) of NaCl. The genome sequencing of strain WW7T revealed a genome size of approximately 3.8 Mbp and a G + C content of 71.3 mol%. The phylogenomic analyses support the WW7T affiliation to a novel Curtobacterium lineage, with Curtobacterium herbarum being the closest type-strain. Chemotaxonomic analysis indicates that the carbon sources assimilation profile of strain WW7T was similar to the type strains, i.e. Curtobacterium luteum, Curtobacterium albidum, and Curtobacterium flaccumfaciens, while no assimilation of the organic acids succinate, alpha-ketobutyrate, mono methyl-succinate, and lactate was observed. Finally, fatty acid methyl ester analysis identifies anteiso-C15:0 and anteiso-C17:0 as major cellular fatty acids which is a common feature for members of the Curtobacterium genus. Based on the results of phylogenomic and chemotaxonomic analyses, strain WW7T represents a novel Curtobacterium lineage, for which the name Curtobacterium salicis sp. nov. is proposed. The type strain is WW7T (DSM 34805T-NRRL B-68078T).

Flooding regimes alleviate lead toxicity and enhance phytostabilization of salix: Evidence from physiological responses and iron-plaque formation.

Aggravated metal pollution in wetland and riparian zones has become a global environmental issue, necessitating the identification of sustainable remediation approaches. Salix exhibits great potential as a viable candidate for metal(loid) remediation. However, the underlying mechanisms for its effectiveness in different flooding regimes with Pb pollution have not been extensively studied. In this study, fast-growing Salix×jiangsuensis 'J172' was selected and planted in different Pb polluted soils (control, 400 and 800 mg ∙ kg-1) under non-flooded and flooded (CF: continuous flooding and IF: intermittent flooding) conditions for 60 days. This study aimed to explore the effects of flooding on Salix growth performance, physiological traits, and the relationship between Pb uptake/translocation and root Fe plaques. Salix×jiangsuensis 'J172' exhibited excellent tolerance and adaptation to Pb pollution with a tolerance index (TI) exceeding 0.6, even at the highest Pb levels. Moreover, the TIs under flooded conditions were higher than that under non-flooded conditions, suggesting that flooding could alleviate Pb toxicity under co-exposure to Pb and flooding. Leaf malondialdehyde (MDA) exhibited a dose-dependent response to Pb exposure; however, CF or IF mitigated the oxidative damage induced by Pb toxicity with decreased MDA content (2.2-11.9%). The superoxide dismutase and peroxidase activities were generally enhanced by flooding, but combined stress (flooding and Pb) significantly decreased catalase activity. Pb was predominantly accumulated in Salix roots, and flooding markedly increased root Pb accumulation by 19.2-173.0% compared to non-flooded condition. Additionally, a significant positive correlation was observed between the iron (Fe) content of the root plaque and root Pb accumulation, indicating that the formation of Fe plaque on the root surface could enhance the phytostabilization of Pb in Salix. The current findings highlight that fast-growing woody plants are suitable for phyto-management of metal-polluted wetlands and can potentially minimize the risk of metal mobility in soils.

Phenolic Compound, Antioxidant, Antibacterial, and In Silico Studies of Extracts from the Aerial Parts of Lactuca saligna L.

Medicinal plants are considered a major source for discovering novel effective drugs. To our knowledge, no studies have reported the chemical composition and biological activities of Moroccan Lactuca saligna extracts. In this context, this study aims to characterize the polyphenolic compounds distributed in hydro-methanolic extracts of L. saligna and evaluate their antioxidant and antibacterial activities; in addition, in silico analysis based on molecular docking and ADMET was performed to predict the antibacterial activity of the identified phenolic compounds. Our results showed the identification of 29 among 30 detected phenolic compounds with an abundance of dicaffeoyltartaric acid, luteolin 7-glucoronide, 3,5-di-O-caffeoylquinic acid, and 5-caffeoylquinic acid with 472.77, 224.30, 196.79, and 171.74 mg/kg of dried extract, respectively. Additionally, antioxidant activity assessed by DPPH scavenging activity, ferric reducing antioxidant power (FRAP) assay, and ferrous ion-chelating (FIC) assay showed interesting antioxidant activity. Moreover, the results showed remarkable antibacterial activity against Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, and Listeria monocytogenes with minimum inhibitory concentrations between 1.30 ± 0.31 and 10.41 ± 0.23 mg/mL. Furthermore, in silico analysis identified three compounds, including Apigenin 7-O-glucuronide, Quercetin-3-O-glucuronide, and 3-p-Coumaroylquinic acid as potent candidates for developing new antibacterial agents with acceptable pharmacokinetic properties. Hence, L. saligna can be considered a source of phytochemical compounds with remarkable activities, while further in vitro and in vivo studies are required to explore the main biological activities of this plant.

Quantitative distribution of flavan-3-ols, procyanidins, flavonols, flavanone and salicylic acid in five varieties of organic winter dormant Salix spp. by LC-MS/MS.

Willow trees (Salix spp.) exhibit remarkable genetic and phenotypic diversity, yielding a broad spectrum of bioactive compounds, notably valuable phenolic compounds such as condensed tannins (phenolic polymers), flavonoids, salicylic glucosides, and phenolic compounds. These enhance the economic value of willow crops and make them suitable for circular bioeconomy. Phenolic compounds known for their diverse applications as antioxidants, antimicrobial agents, pharmaceuticals, nutraceuticals and antiseptics and more, find a natural source in willow. This study aimed to elucidate the composition of 12 flavonoids and salicylic acid in different segments of five organic winter dormant willow species (S. daphnoides, S. fragilis, S. dasyclados, S. viminalis, and S. dasyclados x viminalis) using quantitative analysis and providing valuable insights into their high-value phenolic compounds. Separation into buds, wood and bark segments allowed for a precise characterization of the location of certain phenolic compounds and quantification using LC-MS/MS techniques. LC-MS/MS is an analytical technique known for its increased sensitivity and chromatographic precision. Among the findings, catechin emerged as the predominant flavan-3-ol in all Salix species, with the highest concentration in the buds of Salix viminalis (7.26 mg/g DM). Naringenin exhibited species-specific variations, with S. dasyclados and S. viminalis recording the highest levels. Salicylic acid concentrations peaked in S. dasyclados (5.38 mg/g DM) and S. daphnoides (4.43 mg/g DM), particularly within the bark. When evaluating other individual flavonoids and total polyphenol content (TPC), disparities between buds, bark, and wood became evident, with wood consistently displaying the lowest content. Notably, the higher concentration of polyphenolic compounds in willow bark can be attributed to its susceptibility to external threats and its role as a robust defense mechanism against pathogens and herbivores. This study underscores the significance of diverse willow species as a source of high-value phenolic compounds, distributed differentially across plant parts and species. This knowledge holds promise for their potential applications in the circular bioeconomy.

The effect of peat replacement in horticulture media by willow (Salix viminalis L.) biomass compost for cucumber transplant production.

This research evaluated the usefulness of horticultural substrates prepared on the basis of compost from chipped willow without additives and with the addition of nitrogen and decomposing mycelium of the cellulose-lignin fraction of wood in the cultivation of cucumber seedlings. The produced composts were mixed in different proportions: mixture 1 (W1) - the proportion of compost without additives and compost prepared with the addition of nitrogen and mycelium was in the ratio of 50:50, mixture 2 (W2) - the proportion of compost without additives and compost prepared with the addition of nitrogen and mycelium was in the ratio of 75:25. The starting mixtures were used to prepare horticultural substrates with different components (peat - P, vermicompost - V) and additives: basaltmeal - B, biochar from deciduous wood - C. The components were added in varying proportions. A total of 29 different substrates were subsequently tested in the study. Plant showed that the traits assessed varied to a greater extent under the effect of the test factors than at earlier growth stages. It was demonstrated that cucumber grown on substrates with 75% or 50% willow compost had a unit weight at the same statistical level as when grown on peat substrate (P). The plants with the highest unit weight (8.5- 10.4 g), belonged to the same homogeneous group and derived from sites W1P1B2, W2P1, W1P1B1, W2P2, W1P1C1, P, W1P1, W2B1, W2P2B2. High-quality cucumber transplant should characterise well develop, optimal height-to-stem thickness ratio, short hypocotyl, thick green leaves and cotyledons.

Comparative performance of Populus spp. and Salix spp. for growth, nutrition, and heavy metal uptake in a wastewater hydroponic system.

This research aimed to ascertain the growth, biomass, and phytoremediation capacity of poplars and willow cultivated using wastewater in a hydroponic system. The cuttings were exposed to two water treatments for eight weeks: (1) tap water supply with 1/4 strength Hoagland's solution (TW) as a control and (2) urban raw wastewater with 1/4 strength Hoagland's solution (WW) in a completely randomized experimental design and growth parameters and nutrient and heavy metal content distribution in various plant tissues were assessed. Our results suggest that compared with the TW, seven studied species/clones revealed an increase in growth and biomass parameters (stem height by 16.4%, and root, shoot, and total biomass by 37.3%, 26.9%, and 29.3%, respectively) caused by WW treatment, except the root length and stem diameter that remained the same under two water treatments. Poplars and willow showed a remarkable variability in growth and biomass parameters, with the highest stem diameter, stem height, and root, shoot, and total biomass in Populus nigra L. 62.154. Expression of growth responses to WW treatment with tolerance indices (Tis) indicated the tested poplars and willow as highly tolerant (Ti >100%) with no significant differences among them based on this index. Plant analysis showed that WW treatment increased the concentrations of nutrients and heavy metals in the shoots and roots. Overall, under both water treatments, the capability of the tested species/clones to uptake and accumulate micro-nutrients (except copper (Cu) in shoots) and heavy metals (except chromium (Cr) in shoots) in the plant tissues varied noticeably. However, for all tested plants, the roots had higher concentrations of micro-nutrients (iron (Fe), zinc (Zn), and Cu concentrations in the roots were about 36.8%, 107.6%, and 30.1% of that in the shoots, respectively) and heavy metals (nickel (Ni), Cr, and lead (Pb) concentrations in the roots were about 115.3%, 344.2%, and 198.9% of that in the shoots, respectively), suggesting their capability for micro-nutrients and heavy metals rhizofiltration. Concerning obtained results under hydroponic culture, it can be concluded that these poplars and willow might be promising candidates for wastewater applications. However, data obtained by a hydroponic system need to be confirmed in pot and field experiments.

There are limited studies on the reactions of Salicaceae plants in a wastewater hydroponic system. This research can be considerable for preliminary evaluation of Salicaceae plants to estimate their effectiveness in contaminated soils. Furthermore, the growth, biomass, and phytoremediation responses of Salicaceae plants differ to wastewater application, so this work is important to evaluate the responses of seven different poplar and willow species/clones to municipal wastewater application.

Impacts of willow (Salix babylonica L.) leaf extract on growth, cecal microbial population, and blood biochemical parameters of broilers.

The investigation examined the use of willow leaf extract (WLE) on broiler chickens, examining carcass characteristics, cecal microbiota, antioxidants, and blood parameters. In 4 groups of 300 chicks, a basal diet was given for 5 wk, and the first treatment was basal diet (C). The diets for the remaining 3 treatments (WLE150, WLE300, and WLE450) contained 150, 300, and 450 mg of willow leaf extract /kg, respectively. The study found that birds fed willow leaf extract supplements had significantly greater body weight (BW), body weight gain (BWG), and enhanced feed conversion ratio (FCR) vs. the control group. Birds fed at 450 mg/kg food showed the greatest growth features, carcass weight, liver weight, lower abdominal fat, better low-density lipoprotein (LDL), and high-density lipoprotein (HDL) concentrations, and highest hematological characteristics. Chickens fed diets supplemented with varied doses of willow leaf extract showed significantly increased antioxidant enzyme activity, with higher amounts of glutathione peroxidase (GPx) activity, superoxide dismutase (SOD), total antioxidant capacity (TAC), and lower malondialdehyde (MDA). However, in the study, birds fed a diet supplemented with 450 mg of willow leaf extract per kg meal showed a significant drop of 13.02%, which found no significant variations in hazardous bacteria (Escherichia coli) across 2 treatments (WLE150 and WLE300). In addition, the study discovered that birds fed with varied doses of willow leaf extract had fewer cecum infections (Staphylococci aureus). We conclude that using willow at a level of 450 mg/kg diet can significantly enhance the BWG, FCR, antioxidant levels and beneficial bacteria activity besides the condition of broiler chicken's general health.

Psyllids in Natural Habitats as Alternative Resources for Key Natural Enemies of the Pear Psyllids (Hemiptera: Psylloidea).

The pear psyllids (Cacopsylla spp.; Psylloidea) comprise ~24 species of sap-feeding insects distributed in Europe, temperate Asia, and (as introductions) in the Americas. These pear-specialized insects are among the most damaging and difficult to control pests in orchards. Biological control increasingly is being used to replace or partially replace insecticidal management of pear psyllids. Many key natural enemies of pear psyllids regularly occur in non-orchard habitats on native plants. The presence of beneficial species both in orchard and non-orchard habitats (here referred to as "spillover") has prompted suggestions that native plants and their associated psyllids should be conserved as alternative resources for natural enemies of pear psyllids. The expectation is that the natural enemies will move from those habitats into psyllid-infested orchards. This review shows that psyllids in native habitats are important resources for several key predators and parasitoids of pear psyllids. These resources are critical enough that some beneficials exhibit an almost nomadic existence as they move between plant species, tracking the seasonal appearance and disappearance of psyllid species. In contrast, other natural enemies show minimal or no spillover between orchard and non-orchard habitats, which likely is evidence that they exhibit limited movement at best between orchard and non-orchard habitats. To show conclusively that spillover also indicates that a beneficial species disperses between native habitats and orchards requires difficult research on insect movement. This review concludes with a brief discussion of these difficulties and possible solutions.

Contrasting effect of pristine, ball-milled and Fe-Mn modified bone biochars on dendroremediation potential of Salix jiangsuensis "172" for cadmium- and zinc-contaminated soil.

Bone biochar (BC) has a high capacity for the immobilization of potentially toxic elements (PTEs); however, its effect on dendroremediation efficiency remains unclear. Therefore, this study aimed to determine the effects of various concentrations (0, 0.5, 1, and 2 wt%) of BC, ball-milled BC (MBC), and Fe-Mn oxide-modified BC (FMBC) on soil properties, plant growth, and metal accumulation in Salix jiangsuensis "172" (SJ-172) grown in cadmium (Cd)- and zinc (Zn)-contaminated soil. BC and MBC promoted the photosynthetic rate, mineral element absorption, and plant growth of SJ-172, whereas FMBC inhibited the growth of SJ-172. Different biochars greatly influenced the concentrations of Cd and Zn in tissues of SJ-172. BC and MBC elevated the Cd levels, whereas FMBC decreased the Cd content in the leaves, stems, and cuttings of SJ-172. Unlikely, BC, MBC and FMBC show no evident change to the Zn concentration in the aboveground tissues of SJ-172, while decreased root Cd and Zn content compared with the control. MBC, at a 2.0% application rate, significantly increased the translocation factors of Cd (55.0%) and Zn (40.87%), whereas BC and FMBC demonstrated no significant effects compared with the control (P > 0.05). Moreover, 2.0% BC and MBC increased Cd and Zn accumulation in SJ-172 by 28.40 and 41.14, and 25.89 and 36.16%, respectively, whereas 2.0% FMBC reduced Cd and Zn accumulation by 53.20% and 13.18 %, respectively, compared with the control. The phytoremediation potential of SJ-172 for Cd- and Zn-contaminated soils was enhanced by MBC and BC, whereas it was lowered by FMBC compared to the control. These results provide novel insights for the application of fast-growing trees assisted by biochar amendments in the dendroremediation of severely PTEs-contaminated soil.

Expression of Sailx suchowensis SsIRT9 enhances cadmium accumulation and alters metal homeostasis in tobacco.

Cadmium (Cd) contamination in soils is of great concern for plant growth and human health. Willow (Salix spp.) is a promising phytoextractor because of its high biomass production. However, as a non-hyperaccumulator, willow has a low competitive ability in extraction of Cd. Thus, improving Cd concentrations in developing tissues is one of the primary tasks. Here, our study uncovers a novel SsIRT9 gene from Sailx suchowensis which manipulates plant Cd accumulation. SsIRT9 was more highly expressed in willow roots than other SsIRT genes. As a plasma membrane-localized protein, when expressed in yeast, SsIRT9 retarded cell growth more severely than other SsIRT proteins in the presence of Cd. Furthermore, SsIRT9 was cloned and expressed in tobacco and SsIRT9 did not affect plant growth. In hydroponic experiments, SsIRT9 lines displayed higher Cd in the shoots than the wild type. When grown in Cd-contaminated soils, Cd levels in transgenic tobacco increased by 152-364% in roots and by 135-444% in shoots, demonstrating significant superiority in Cd accumulation over other functional IRT/ZIP transporters. Moreover, expressing SsIRT9 in tobacco altered metal homeostasis, especially manganese and zinc. Taken together, we envision that SsIRT9 expression in plants is a promising strategy for upgrading extraction of Cd from soils.

Genome-wide identification and expression analysis of xyloglucan endotransglucosylase/hydrolase genes family in Salicaceae during grafting.

BACKGROUND: Poplar (Populus cathayana)and willow (Salix rehderiana) are important fast-growing trees in China. Grafting plays an important role in improving plant stress resistance and construction of ornamental plants. It is found that willow scions grafted onto poplar rootstocks can form ornamental plants. However, this grafted combination has a low survival rate. Many studies have reported that the xyloglucan endotransglucosylase/hydrolase (XTH) family plays an important role in the healing process of grafts. RESULTS: A total of 38 PtrXTHs and 32 SpuXTHs were identified in poplar and willow respectively, and were classified into three subfamilies. Tandem duplication was the main reason for the expansion of the PtrXTHs. Grafting treatment and Quantitative real time PCR (RT-qPCR) analysis revealed that five XTH genes differentially expressed between self-grafted and reciprocal grafted combinations. Specifically, the high expression levels of SrXTH16, SrXTH17, SrXTH25, PcXTH22 and PcXTH17 may contribute to the high survival rate of the grafted combination with willow scion and poplar rootstock. Subcellular localization identified that the SrXTH16, SrXTH17, SrXTH25, PcXTH17 and PcXTH22 proteins were located on the cell walls. Transcription factors (NAC, MYB and DOF) may regulate the five XTH genes. CONCLUSIONS: This study provides a new understanding of the roles of PcXTH and SrXTH genes and their roles in grafting. Our results will give some hints to explore the molecular mechanisms of PcXTH and SrXTH genes involved in grafting in the future.

In Vitro Anti-Tumor and Hypoglycemic Effects of Total Flavonoids from Willow Buds.

Salix babylonica L. is a species of willow tree that is widely cultivated worldwide as an ornamental plant, but its medicinal resources have not yet been reasonably developed or utilized. Herein, we extracted and purified the total flavonoids from willow buds (PTFW) for component analysis in order to evaluate their in vitro anti-tumor and hypoglycemic activities. Through Q-Orbitrap LC-MS/MS analysis, a total of 10 flavonoid compounds were identified (including flavones, flavan-3-ols, and flavonols). The inhibitory effects of PTFW on the proliferation of cervical cancer HeLa cells, colon cancer HT-29 cells, and breast cancer MCF7 cells were evaluated using an MTT assay. Moreover, the hypoglycemic activity of PTFW was determined by investigating the inhibitory effects of PTFW on α-amylase and α-glucosidase. The results indicated that PTFW significantly suppressed the proliferation of HeLa cells, HT-29 cells, and MCF7 cells, with IC50 values of 1.432, 0.3476, and 2.297 mg/mL, respectively. PTFW, at different concentrations, had certain inhibitory effects on α-amylase and α-glucosidase, with IC50 values of 2.94 mg/mL and 1.87 mg/mL, respectively. In conclusion, PTFW at different doses exhibits anti-proliferation effects on all three types of cancer cells, particularly on HT-29 cells, and also shows significant hypoglycemic effects. Willow buds have the potential to be used in functional food and pharmaceutical industries.

Functional analysis of Salix purpurea genes support roles for ARR17 and GATA15 as master regulators of sex determination.

The Salicaceae family is of growing interest in the study of dioecy in plants because the sex determination region (SDR) has been shown to be highly dynamic, with differing locations and heterogametic systems between species. Without the ability to transform and regenerate Salix in tissue culture, previous studies investigating the mechanisms regulating sex in the genus Salix have been limited to genome resequencing and differential gene expression, which are mostly descriptive in nature, and functional validation of candidate sex determination genes has not yet been conducted. Here, we used Arabidopsis to functionally characterize a suite of previously identified candidate genes involved in sex determination and sex dimorphism in the bioenergy shrub willow Salix purpurea. Six candidate master regulator genes for sex determination were heterologously expressed in Arabidopsis, followed by floral proteome analysis. In addition, 11 transcription factors with predicted roles in mediating sex dimorphism downstream of the SDR were tested using DAP-Seq in both male and female S. purpurea DNA. The results of this study provide further evidence to support models for the roles of ARR17 and GATA15 as master regulator genes of sex determination in S. purpurea, contributing to a regulatory system that is notably different from that of its sister genus Populus. Evidence was also obtained for the roles of two transcription factors, an AP2/ERF family gene and a homeodomain-like transcription factor, in downstream regulation of sex dimorphism.

Energy performance of compressed biomethane gas production from co-digestion of Salix and dairy manure: factoring differences between Salix varieties.

Biogas from anaerobic digestion is a versatile energy carrier that can be upgraded to compressed biomethane gas (CBG) as a renewable and sustainable alternative to natural gas. Organic residues and energy crops are predicted to be major sources of bioenergy production in the future. Pre-treatment can reduce the recalcitrance of lignocellulosic energy crops such as Salix to anaerobic digestion, making it a potential biogas feedstock. This lignocellulosic material can be co-digested with animal manure, which has the complementary effect of increasing volumetric biogas yield. Salix varieties exhibit variations in yield, composition and biomethane potential values, which can have a significant effect on the overall biogas production system. This study assessed the impact of Salix varietal differences on the overall mass and energy balance of a co-digestion system using steam pre-treated Salix biomass and dairy manure (DaM) to produce CBG as the final product. Six commercial Salix varieties cultivated under unfertilised and fertilised conditions were compared. Energy and mass flows along this total process chain, comprising Salix cultivation, steam pre-treatment, biogas production and biogas upgrading to CBG, were evaluated. Two scenarios were considered: a base scenario without heat recovery and a scenario with heat recovery. The results showed that Salix variety had a significant effect on energy output-input ratio (R), with R values in the base scenario of 1.57-1.88 and in the heat recovery scenario of 2.36-2.94. In both scenarios, unfertilised var. Tordis was the best energy performer, while the fertilised var. Jorr was the worst. Based on this energy performance, Salix could be a feasible feedstock for co-digestion with DaM, although its R value was at the lower end of the range reported previously for energy crops.

A Comparative Analysis of Phytochemicals, Metal Ions, Volatile Metabolites in Heart Wood, Stem Bark and Leaves of Salix alba L. along with in Vitro Antioxidant, Antacid, Antimicrobial Activities for Sake of Environment Conservation by Substitution of Stem Bark With Leaf.

The genus of Salix is used in food, medicine and nutraceuticals, and standardized by using the single marker compound Salicin only. Stem bark is the official part used for the preparation of various drugs, nutraceuticals and food products, which may lead to overexploitation and damage of tree. There is need to search substitution of the stem bark with leaf of Salix alba L. (SA), which is yet not reported. Comparative phytochemicals viz. Salicin, Procyanidin B1 and Catechin were quantified in the various parts of SA viz. heart wood (SA-HW), stem bark (SA-SB) and leaves (SA-L) of Salix alba L.by using newly developed HPLC method. It was observed that SA-HW and SA-L contained far better amount of Salicin, Procyanidin B and Catechin as compared to SA-SB (SA-HW~SA-L≫SA-SB). Essential and toxic metal ions of all three parts were analysed using newly developed ICP-OES method, where SA-L were founded as a rich source of micronutrients and essential metal ions as compared to SA-SB and SA-HW. GC-MS analysis has shown the presence of fatty acids and volatile compounds. The observed TPC and TFC values for all three parts were ranged from 2.69 to 32.30 mg GAE/g of wt. and 37.57 to 220.76 mg QCE/g of wt. respectively. In DPPH assay the IC50 values of SA-SB, SA-HW, and SA-L were 1.09 (±0.02), 5.42 (±0.08), and 8.82 (±0.10) mg/mL, respectively. The order of antibacterial activities against E. coli, S. aureus, P. aeruginosa, and B. subtilis strains was SA-L>SA-HW>SA-SB with strong antibacterial activities against S. aureus, and B. subtilis strains. The antacid activities order was SA-L>SA-SB>SA-HW. The leaves of SA have shown significant source of nutrients, phytochemicals and medicinal properties than SA-HW and SA-SB. The leaves of SA may be considered as substitute of stem bark to save the environment or to avoid over exploitation, but after the complete pharmacological and toxicological studies.

Willow Bark (Salix spp.) Used for Pain Relief in Arthritis: A Meta-Analysis of Randomized Controlled Trials.

This study intends to assess the analgesic effects, physical facilitation, and safety of willow bark use in patients with arthritis. Our study was conducted based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. PubMed, Scopus, EMBASE, Web of Science, Cochrane, and ClinicalTrials.gov were searched for relative randomized controlled trials (RCTs) describing the efficacy or adverse events of willow bark in patients with arthritis until 12 April 2023. We used Cochrane ROB 2.0 and the Grading of Recommendations, Assessment, Development, and Evaluations system to evaluate the quality of studies and evidence. The meta-analysis was carried out by the fix-effects model. This study included five studies with six RCTs consisting of 329 patients with arthritis. The results showed significant differences in pain relief and improvement in physical status for patients with arthritis between willow bark treatment and placebo groups, and no significant differences in the risk of all adverse events in patients with arthritis between willow bark treatment and placebo. Owing to the potential bias, the certainty and evidence of our findings are still inadequate. Therefore, further RCTs are needed to confirm our results.

Utilization of Torrefied and Non-Torrefied Short Rotation Willow in Wood-Plastic Composites.

The torrefaction process is widely used in the energy field, but the characteristics of the torrefied wood also have positive effects on the production of wood plastic composites. In this study, short-rotation shrub willow was torrefied at 225 and 300 °C and incorporated into polypropylene composites filled with changing levels of weight percent (wt%) of non-torrefied and torrefied (5, 15, 25, and 40 wt%) wood. Nine different formulations were extruded for mechanical, thermal, and water absorption properties. The tensile properties of composites were not affected by any level of torrefaction, while higher flexure properties were in favor of lower wt% of torrefied wood. The slowest rate of thermal degradation was confirmed for the highest wt% of torrefied wood with a torrefaction temperature of 300 °C. In contrast, the presence of torrefied wood in composites did not show a difference in crystallization or melting temperatures. The most noticeable contribution of torrefaction temperature and wt% was found for water-absorbing properties, where the higher torrefaction temperature and largest wt% of torrefied wood in the composite resulted in decreased water uptake.

The oldest known clones of Salix herbacea growing in the Northern Apennines, Italy are at least 2000 years old.

PREMISE: Dominant in many ecosystems around the world, clonal plants can reach considerable ages and sizes. Due to their modular growth patterns, individual clonal plants (genets) can consist of many subunits (ramets). Since single ramets do not reflect the actual age of genets, the ratio between genet size (radius) and longitudinal annual growth rate (LAGR) of living ramets is often used to approximate the age of clonal plants. However, information on how the LAGR changes along ramets and how LAGR variability may affect age estimates of genets is still limited. METHODS: We assessed the variability of LAGR based on wood-section position along the ramets and on the duration of the growing season on three genetically distinct genets of Salix herbacea growing in the Northern Apennines (Italy). We compared genet ages estimated by dividing genet radius by the LAGRs of its ramets. RESULTS: LAGR increased significantly from the stem apex to the root collar; indicating that ramet growth rate decreased with time. Furthermore, a difference of ca. 2 weeks in the onset of the growing period did not impact LAGR. Considering the high LAGR variability, we estimated that the three genets started to grow between ~2100 and ~7000 years ago, which makes them the oldest known clones of S. herbacea even considering the most conservative age estimate. CONCLUSIONS: Our findings indicate that analyzing ramets at the root collar provides an integrative measurement of their overall LAGR, which is crucial for estimating the age of genets.

Large effect of phosphate-solubilizing bacteria on the growth and gene expression of Salix spp. at low phosphorus levels.

Phosphorus is one of the most important nutrients required for plant growth and development. However, owing to its low availability in the soil, phosphorus is also one of the most difficult elements for plants to acquire. Phosphorus released into the soil from bedrock quickly becomes unavailable to plants, forming poorly soluble complexes. Phosphate-solubilizing bacteria (PSB) can solubilize unavailable phosphorus-containing compounds into forms in which phosphorus is readily available, thus promoting plant growth. In this study, two willow species, Salix dasyclados cv. Loden and Salix schwerinii × Salix viminalis cv. Tora, were inoculated with two selected bacterial strains, Pantoea agglomerans and Paenibacillus spp., to evaluate the plant growth parameters and changes in gene expression in the presence of different concentrations of tricalcium phosphate: 0 mM (NP), 1 mM (LP), and 2 mM (HP). Inoculation with PSB increased root, shoot and leaf biomass, and for the HP treatment, significant changes in growth patterns were observed. However, the growth responses to plant treatments tested depended on the willow species. Analysis of the leaf transcriptomes of the phosphate-solubilizing bacterium-inoculated plants showed a large variation in gene expression between the two willow species. For the Tora willow species, upregulation of genes was observed, particularly for those involved in pathways related to photosynthesis, and this effect was strongly influenced by bacterial phosphate solubilization. The Loden willow species was characterized by a general downregulation of genes involved in pathway activity that included ion transport, transcription regulation and chromosomes. The results obtained in this study provide an improved understanding of the dynamics of Salix growth and gene expression under the influence of PSB, contributing to an increase in yield and phosphorus-use efficiency.