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River management practices have altered the hydrological regimes of many rivers and also altered the availability of regeneration niches for riparian species. We investigated the impact of changed hydrological regimes on the sex ratios and the Spatial Segregation of the Sexes (SSS) in the dioecious species Salix myrsinifolia Salisb.—phylicifolia L. and S. lapponum L. by studying the free-flowing Vindel River and the regulated Ume River in northern Sweden. We surveyed sex ratios of these species in 12 river reaches on the Vindel River and in 17 reaches on the Ume River. In addition, we surveyed the sex and location above mean river stage of 1,002 individuals across both river systems to investigate the SSS of both species. Cuttings were collected from male and female individuals of S. myrsinifolia—phylicifolia from both rivers and subjected to four different water table regimes in a greenhouse experiment to investigate growth response between the sexes. We found an M/F sex ratio in both river systems similar to the regional norm of 0.62 for S. myrsinifolia—phylicifolia and of 0.42 for S. lapponum. We found no evidence of SSS in either the free-flowing Vindel River or the regulated Ume River. In the greenhouse experiment, hydrological regime had a significant effect on shoot and root dry weight and on root length. Significantly higher shoot dry weights were found in females than in males and significantly different shoot and root dry weights were found between cuttings taken from the two rivers. We concluded that changed hydrological regimes are likely to alter dimensions of the regeneration niche and therefore to influence sex ratios and SSS at an early successional stage, making it difficult to find clear spatial patterns once these species reach maturity and can be sexed.

Fluctuating asymmetry (FA) is often proposed as an early warning indicator of subtle changes in plant functioning. Here, we tested whether leaf FA responds consistently to the alleviation of cold stress in three boreal willow species—Salix caprea, S. myrsinifolia and S. phylicifolia. We enclosed 10 naturally growing individuals of each species in open‐top chambers at budburst and compared their leaf traits to those of unenclosed control plants after leaf development had ceased. All measurements were conducted blind to treatment. Willows in open‐top chambers showed a 9% increase in specific leaf area, indicating that the 1°C–2°C warming within chambers affected leaf development. However, neither leaf length nor FA responded significantly to the warming treatment. FA also did not differ among species or individual plants, suggesting that it may reflect statistical noise rather than a reliable biological signal in this context. These findings add to growing concerns that many reported FA responses to environmental change may result from confirmation bias—an issue that can be mitigated by adopting blind measurement protocols. We tested whether leaf fluctuating asymmetry (FA) responds to experimental warming in three boreal willow species using open‐top chambers and blind measurements. While warming increased specific leaf area, neither leaf length nor FA showed significant responses, and FA did not differ among species or individuals. These results suggest that FA may represent statistical noise rather than a reliable indicator of environmental stress.

Earlier studies have shown that the bark of Salix L. species (Salicaceae family) is rich in extractives, such as diverse bioactive phenolic compounds. However, we lack knowledge on the bioactive properties of the bark of willow species and clones adapted to the harsh climate conditions of the cool temperate zone. Therefore, the present study aimed to obtain information on the functional profiles of northern willow clones for the use of value-added bioactive solutions. Of the 16 willow clones studied here, 12 were examples of widely distributed native Finnish willow species, including dark-leaved willow ( S. myrsinifolia Salisb.) and tea-leaved willow ( S. phylicifolia L.) (3 + 4 clones, respectively) and their natural and artificial hybrids (3 + 2 clones, respectively). The four remaining clones were commercial willow varieties from the Swedish willow breeding program. Hot water extraction of bark under mild conditions was carried out. Bioactivity assays were used to screen antiviral, antibacterial, antifungal, yeasticidal, and antioxidant activities, as well as the total phenolic content of the extracts. Additionally, we introduce a fast and less labor-intensive steam-debarking method for Salix spp. feedstocks. Clonal variation was observed in the antioxidant properties of the bark extracts of the 16 Salix spp. clones. High antiviral activity against a non-enveloped enterovirus, coxsackievirus A9, was found, with no marked differences in efficacy between the native clones. All the clones also showed antibacterial activity against Staphylococcus aureus and Escherichia coli , whereas no antifungal ( Aspergillus brasiliensis ) or yeasticidal ( Candida albicans ) efficacy was detected. When grouping the clone extract results into Salix myrsinifolia , Salix phylicifolia , native hybrid, artificial hybrid, and commercial clones, there was a significant difference in the activities between S. phylicifolia clone extracts and commercial clone extracts in the favor of S. phylicifolia in the antibacterial and antioxidant tests. In some antioxidant tests, S. phylicifolia clone extracts were also significantly more active than artificial clone extracts. Additionally, S. myrsinifolia clone extracts showed significantly higher activities in some antioxidant tests than commercial clone extracts and artificial clone extracts. Nevertheless, the bark extracts of native Finnish willow clones showed high bioactivity. The obtained knowledge paves the way towards developing high value-added biochemicals and other functional solutions based on willow biorefinery approaches.

In the northern boreal zone, revegetation and landscaping of closed mine tailings are challenging due to the high concentrations of potentially toxic elements; the use of nutrient-poor, glacigenic cover material (till); cool temperatures; and short growing period. Recycled waste materials such as biochar (BC) and composted sewage sludge (CSS) have been suggested to improve soil forming process and revegetation success as well as decrease metal bioavailability in closed mine tailing areas. We conducted two field experiments in old iron mine tailings at Rautuvaara, northern Finland, where the native mine soil or transported cover till soil had not supported plant growth since the mining ended in 1989. The impacts of CSS and spruce ( Picea abies )–derived BC application to till soil on the survival and growth of selected plant species ( Pinus sylvestris , Salix myrsinifolia , and grass mixture containing Festuca rubra , Lolium perenne , and Trifolium repens ) were investigated during two growing seasons. In addition, the potential of BC to reduce bioaccumulation of metals in plants was studied. We found that (1) organic amendment like CSS markedly enhanced the plant growth and is therefore needed for vegetation establishment in tailing sites that contained only transported till cover, and (2) BC application to till soil-CSS mixture further facilitated the success of grass mixtures resulting in 71–250% higher plant biomass. On the other hand, (3) no effects on P. sylvestris or S. myrsinifolia were recorded during the first growing seasons, and (4) accumulation of metals in cover plants was negligible and BC application to till further decreased the accumulation of Al, Cr, and Fe in the plant tissues. Graphical abstract

In boreal woody plants, concentrations of defensive phenolic compounds are expected to be at a high level during the juvenile phase and decrease in maturity, although there is variation between plant species. Females of dioecious species, like most of the Salicaceae, are expected to invest their resources in defense and reproduction, while males are expected to be more growth-oriented. We studied age- and sex-dependent changes in leaf and stem phenolics, and in height and diameter growth in a dioecious Salix myrsinifolia plants over a seven-year time period. In addition, we registered flowering as well as rust damage in the leaves. From the first year and throughout ontogenetic development from juvenile to adult phases, there was no significant change in the concentrations of any of the studied compounds in the leaves of S. myrsinifolia. In the stems, the concentrations of six out of 43 identified compounds decreased slightly with age, which may be partly explained by dilution caused by the increment in stem diameter with age. The fairly steady chemistry level over seven years, accompanied by moderate genotypic phenolic variation, indicates important roles of chemical defenses against herbivory for this early-successional species.

Salicinoids are well-known defense compounds in salicaceous trees and careful screening at the population level is warranted to fully understand their diversity and function. European aspen, Populus tremula, is a foundation species in Eurasia and highly polymorphic in Sweden. We exhaustively surveyed 102 replicated genotypes from the Swedish Aspen collection (SwAsp) for foliar salicinoids using UHPLC-ESI-TOF/MS and identified nine novel compounds, bringing the total to 19 for this species. Salicinoid structure followed a modular architecture of a salicin skeleton with added side groups, alone or in combination. Two main moieties, 2'-cinnamoyl and 2'-acetyl, grouped the SwAsp population into four distinct chemotypes, and the relative allocation of salicinoids was remarkably constant between different environments, implying a highly channeled biosynthesis of these compounds. Slightly more than half of the SwAsp genotypes belonged to the cinnamoyl chemotype. A fraction synthesized the acetyl moiety alone (∼7%) or in combination with cinnamoyl (∼2%), and close to forty percent lacked either of the two characteristic moieties, and thus resemble P. tremuloides in their salicinoid profile. The two most abundant chemotypes were evenly distributed throughout Sweden, unlike geographical patterns reported for SwAsp phenology traits, plant defense genes, and herbivore community associations. Here we present the salicinoid characterization of the SwAsp collection as a resource for future studies of aspen chemical ecology, salicinoid biosynthesis, and genetics.

We studied the induction of salicylates in mature leaves of Salix myrsinifolia Salisb. (Salicaceae) following severe wounding by a specialist leaf-beetle Phratora vitellinae L. (Chrysomelidae). Levels of individual salicylates and aromatic amino acids and their total levels were determined in leaves at different developmental stages. Induction of salicylates depended on: (1) the individual compound; (2) the developmental stage of the plant organ; and (3) the genotype of the plant. Induction of salicylates was systemic: levels of salicylates rose in unwounded young immature and mature leaves, but no local response was detected in wounded leaves. In addition, there were clear clonal variations in both the constant and induced levels of salicylates: clones with the highest levels of salicylates were also most capable of increasing this level in response to herbivore attack i.e. no trade-off between constant and induced levels was detected. Furthermore, the levels of three aromatic amino acids, Phe, Tyr and Trp, increased in immature leaves of herbivore-affected plants, which may indicate induction of enzymes of the shikimate pathway by wounding. The increase in salicylates was suggested to be a consequence of an increased rate of synthesis rather than that of translocation. The induced levels of salicylates did not affect the subsequent feeding of highly specialized P. vitellinae. However, the ability to increase levels of salicylates may reduce grazing by generalist herbivores which are not able to tolerate high levels of salicylates.

We explored the inter- and intrapopulation variability in the larval defensive chemistry of the leaf beetle Chrysomela lapponica with respect to the salicylic glycoside (SG) content of its host species. Secretions of larvae from three populations associated in nature with SG-poor willows contained nearly twice as many components and 40-fold higher concentrations of autogenously produced isobutyrates and 2-methylbutyrates than secretions of larvae from three populations associated with SG-rich willows, which in turn had 200-fold higher concentrations of host-derived salicylaldehyde. Reciprocal transfer experiments showed that the larvae from populations associated with SG-rich willows did not produce appreciable amounts of butyrates on either SG-rich or SG-poor willows, while populations feeding on several SG-poor willow species retained the ability for efficient sequestration of SGs, along with their ability to produce high amounts of isobutyrates and 2-methylbutyrates. Only the populations associated with SG-poor willows demonstrated among-family variation in the composition of defensive secretion and differential responses of individual families to willows with alternative SG levels, which can be seen as the prerequisites for shifting to novel hosts. These non-specialized populations show a dual defensive strategy, which corresponds to the ancestral state of this species, while populations that fully depend on host-derived toxins (feeding on SG-rich willows) or have lost the ability to produce salicylaldehyde (feeding on birches) are most deviant from the ancestral state. The results indicate that defensive strategies may differ between populations within a species, and suggest that this variation reduces extinction risks and maintains the high ecological diversity and wide distribution of C. lapponica .

Coffee and willow are known as valuable sources of biologically active phytochemicals such as chlorogenic acid, caffeine, and salicin. The aim of the study was to determine the interactions between the active compounds contained in water extracts from coffee and bark of willow (Salix purpurea and Salix myrsinifolia). Raw materials and their mixtures were characterized by multidirectional antioxidant activities; however, bioactive constituents interacted with each other. Synergism was observed for ability of inhibition of lipid peroxidation and reducing power, whereas compounds able to scavenge ABTS radical cation acted antagonistically. Additionally, phytochemicals from willow bark possessed hydrophilic character and thermostability which justifies their potential use as an ingredient in coffee beverages. Proposed mixtures may be used in the prophylaxis or treatment of some civilization diseases linked with oxidative stress. Most importantly, strong synergism observed for phytochemicals able to prevent lipids against oxidation may suggest protective effect for cell membrane phospholipids. Obtained results indicate that extracts from bark tested Salix genotypes as an ingredient in coffee beverages can provide health promoting benefits to the consumers; however, this issue requires further study.

• Elevated carbon dioxide (CO₂), temperature (T) and ultraviolet-B (UV-B) radiation may affect plant growth and secondary chemistry in different directions, but the effect of the combination of the three factors has seldom been tested. • Here, we grew four dark-leaved willow (Salix myrsinifolia) clones under combinations of ambient or elevated CO₂, T and UV-B radiation in top-closed chambers for 7 wk. • Elevated UV-B had no effects on growth or phenolic compounds, and there were no significant interactions between UV-B, CO₂ and T. CO₂ alone increased most growth parameters, but the magnitude of the effect varied among the clones. Total phenolics increased at elevated CO₂, whereas they decreased at elevated T. The responses varied between the clones. • The results imply that dark-leaved willow are fairly resistant to the applied three-factor climate change, probably because of high constitutive defense. However, the interactions between clone and climate change factors implies that some clones are more susceptible than the species as a whole.