Vegetation influence on ectomycorrhizal inoculum Available to sub-arctic willow (Salix lapponum L.) planted in an upland Site

Summary

Restoration of scrub and woodland in deforested upland sites is an important conservation activity. However, little is known about the mycorrhizal colonisation potential of upland soils or the factors that influence the distribution of mycorrhizal inoculum. We investigated the effect of existing vegetation on mycorrhizal colonisation potential for a sub-arctic willow (Salix lapponum) by planting uninoculated cuttings into plotsrepresenting two upland habitats with either grassand herbs (‘grass’) or Vaccinium myrtillus (‘vaccinium’) and assessing mycorrhizal colonisation after 14 months using morphological and molecular techniques. From 40 willow cuttings (20 in each habitat), DNA sequences of rive ectomycorrhizal (EcM) fungal taxa were recovered: Laccaria proxima, Thelephora terrestris, Hebeloma sp., ‘Thelephoraceae sp.’ and ‘Pezizales sp.’. Cuttings in the ‘grass’ habitat were dominated by Laccaria proxima and ‘Pezizales sp.’ and in the ‘vaccinium’ habitat by Thelephora terrestris which was absent from the ‘grass’ habitat. There were no significant differences between habitats in frequency of EcM inoculum (overall percentage of cuttings colonised = 70%) or colonisation potential (overall mean percentage of root tips colonised per cutting = 20 %). These data suggest that the mycorrhizal colonisation potential and diversity of fungi available to willow in these upland soils are low and planted willow may benefit from inoculum enhancement.     

Growth Response of Coyote Willow (Salix exigua) Cuttings in Relation to Alluvial Soil Texture and Water Availability

A common approach to re‐establishing cottonwood–willow habitat along regulated rivers is through installing dormant, rootless cuttings, yet there is little published information exploring floodplain characteristics that optimize growth of southwestern riparian willows planted in this manner. The goal of this project was to evaluate relationships between growth attributes of Salix exigua and soil texture and soil water availability. Monitoring plots were established in five willow swales planted with dormant S. exigua cuttings along the banks of the Middle Rio Grande in central New Mexico. Data analysis revealed significantly higher aerial cover, height, and stem density for S. exigua plants installed in plots with intermediate levels (15–25%) of fine textured soils distributed through the soil profile. Similar relationships were found in relation to soil water availability. Regression analysis of percent fines and available water at different depth increments provided limited explanation of variability in willow growth attributes at different plots. Findings indicate that S. exigua plants established from cuttings can achieve heights and aerial cover values similar to naturally established willow bars if the floodplain soil profile contains intermediate levels of fine textured soils and the maximum depth to groundwater is within 1.5 m of the ground surface. Where sites are dominated by coarse sand, S. exigua growth may be improved if maximum depth to groundwater is within 1 m of the ground surface.     

Characterization of eight polymorphic microsatellites in endangered willow Salix hukaoana

Salix hukaoana is an endangered riparian tree that is found and known only in several isolated sites in eastern Japan, and that has been suffering from the loss and fragmentation of its habitat. To investigate the genetic variation and structures of this willow in these sites, eight polymorphic microsatellites with three to eight alleles were characterized. Cross‐species amplification was successful for many of the loci using the congeners Salix sachalinensis and Chosenia arbutifolia. This set of microsatellites will be useful for the study of the spatial genetic structures of S. hukaoana and other coexisting willows suffering from habitat fragmentation in riparian forests.     

Salix kaptarae sp. nov. (Salicaceae) from Crete

A new species of willow from Crete, named Salix kaptarae Cambria, C. Brullo & Brullo, is described and illustrated. Morphologically, it shows close relationships with species belonging to the S. apennina group, mainly with respect to the leaf shape, and the hairy ovary, capsule and base of stamina filaments. Considerations on its ecology, leaf micro‐morphology and conservation status are also provided.     

Population genetic structure of native versus naturalized sympatric shrub willows (Salix; Salicaceae)

Vegetative propagation of an introduced species can contribute significantly to its ability to spread and become naturalized, potentially in competition with native species. This study focused on the naturalization of a willow shrub, Salix purpurea, which was introduced to the United States from Europe and is commonly sympatric with the native shrub willow, S. eriocephala. Both species are capable of vegetative and sexual reproduction, but little is known about their relative frequency, nor the impact of clonal propagation on population-level genetic diversity. We analyzed genotypes at several microsatellite loci in 993 individuals belonging to 30 subpopulations of S. eriocephala and 28 subpopulations of S. purpurea in areas of sympatry across three watersheds to compare their genetic diversity and genetic structure. Our results revealed six subpopulations of S. purpurea containing plants with identical multilocus genotypes, while clonal individuals were rare among S. eriocephala populations. These species are dioecious with relatively high levels of heterozygosity, but S. eriocephala had much higher allelic diversity and genotypic diversity than did S. purpurea. These results strongly suggest that vegetative propagation has contributed to the naturalization of S. purpurea and has resulted in higher levels of genetic differentiation among S. purpurea populations than among native S. eriocephala populations.     

Inoculation of willows (Salix spp.) with ectomycorrhizal fungi on mined boreal peatland

Inoculation with ectomycorrhizal fungi was explored as a means to improve productivity of experimental short-rotation plantations of the willowsSalix viminalis andSalix dasyclados for biomass production on surface-mined peatlands in northern Finland. Both willow species formed ectomycorrhizas withAmanita spp.,Cortinarius purpurascens, Entoloma nidorosum, otherEntoloma spp.,Hebeloma crustuliniforme, H. pusillum, Laccaria bicolor, andPaxillus involutus in greenhouse experiments.Field trials on a mined peatland site revealed (after one growing season) statistically significant growth stimulation after inoculation due to mycorrhiza formation in both willow species: plants inoculated withEntoloma were sometimes twice as large as control plants. However, such effects were observed only in plots receiving normal phosphate fertilization as opposed to low phosphate application, and were not consistent from season to season. With the inoculum of other species (Cortinarius, Hebeloma andPaxillus) some evidence of growth enhancement was found in the field, but these results were sometimes attributable to non-symbiotic effects of inoculation.     

Conservation of montane willow scrub in Scotland

Summary

The community known as montane willow scrub has been almost totally destroyed in Scotland by human activities. The species of montane willows are not regarded as threatened by the conventional criteria, but the continued effects of grazing, combined with their sparse distribution in suboptimal habitat and dioecious habit, suggest that the long term threat to some species may have been underestimated. Conservation measures are needed urgently if we are to restore areas of montane willow scrub. It will be necessary to use fencing to provide protection from grazing, and also it may well be necessary to do some planting to provide enough individuals for effective pollination and regeneration to occur.     

The impacts of pollination mode,plant characteristics and local density on the reproductive success of a scarce plant species, <Emphasis Type="Italic">Salix arbuscula</Emphasis>

Pollination failure represents one stage at which sexual reproduction of plants may be limited. It is therefore important to understand the pollination mechanism of the plant and how this may be affected by plant and population characteristics. This study examined the reproductive biology of a scarce montane willow species, Salix arbuscula, as part of a programme to determine ecological strategies for the conservation of montane willow scrub, an endangered habitat in the UK. The relative importance of insect and wind pollination, and the role of plant characteristics including plant size, catkin length, number of flowers and local density in determining reproductive success (number of fruit set) were investigated in three populations. Insect exclosures were used to determine the pollination mechanism. Our results suggest that S. arbuscula is predominantly insect-pollinated. In naturally pollinated plants, catkins with a higher proportion of ripe fruit were longer. Plants with more than two males in the local area and unbrowsed plants had a higher proportion of ripe fruit per catkin. The extent of wind pollination was very low, but differed between populations. The success of wind-only pollination was dependent on the number of males nearby, indicating that local density is more important in this type of pollination. Pollination mechanism and fruit set varied between the three populations observed, demonstrating the importance of multi-site comparisons. The number of males in the surrounding area had a positive effect on fruit set in both types of pollination, suggesting that Allee effects are likely to be operating in this species.     

Genetic characterization of Salix alba L. and Salix fragilis L. by means of different PCR-derived marker systems

Abstract

Salix alba L. and Salix fragilis L. are two closely related willow species whose phenotypic features, showing a large and continuous variation, have a low diagnostic value for identifying pure species and interspecific hybrids. In this paper, the effectiveness of different multilocus PCR-based molecular markers, such as I-SSRs, RAPDs and AFLPs in detecting genetic polymorphisms able to discriminate the two willow species was evaluated by analysing a set of 12 reference samples. Three genetic similarity indexes, Dice, Jaccard and Simple Matching coefficient, were used for all possible pairwise comparisons of individuals, revealing the same trend of variation within and between species when different marker systems were used. Cluster analysis, based on Dice genetic similarity coefficient, clustered the individuals of S. alba and S. fragilis into two distinct subgroups, indicating that the gene pools are well differentiated. Moreover, a number of private alleles for each marker system allowed the discrimination of the two species because always present only in one of the two. The utility of different marker systems in discriminating willow species was evaluated by the Polymorphism Information Content (PIC) and the Marker Index (MI) parameters. The variation of Dice's indexes obtained from a different number of experiments in relation to the marker systems is discussed.     

Leaf beetle grazing does not induce willow trichome defence in the coppicing willow Salix viminalis

Abstract 1 Willows are frequently attacked and defoliated by adult leaf beetles (Phratora vulgatissima L.) early in the season and the plants are then attacked again when new larvae emerge. The native willow Salix cinerea has previously been shown to respond to adult grazing by producing new leaves with an increased trichome density. Subsequent larval feeding was reduced on new leaves. This type of induced plant response may reduce insect damage and could potentially be utilized for plant protection in agricultural systems. 2 Here, we investigated if the willow species most commonly used for biomass production in short rotation coppice, Salix viminalis, also responds to adult beetle grazing by increasing trichome density. Larval performance and feeding behaviour on plants previously exposed to adult beetles was compared with that on undefoliated control plants in a greenhouse. 3 We found an overall decrease in trichome density within all the plants (i.e. trichome density was lower on new leaves compared to that for older basal leaves on S. viminalis). However, leaves of beetle defoliated plants had a higher trichome density compared to control plants. Larval growth and feeding was not affected by this difference between treatments. Larvae appeared to remove trichomes when feeding on S. viminalis, a behaviour that might explain the lack of difference between treatments.     

Interactions between the willow beetle Phratora vulgatissima and different genotypes of Salix viminalis

Abstract 1 Planting of species mixtures is a strategy for the non‐chemical management of willow beetles in short‐rotation coppice willows. However, the relatively susceptible Salix viminalis genotypes and their hybrids dominate current high‐yielding willows. Interactions between Phratora vulgatissima and different genotypes of S. viminalis were examined under laboratory conditions to determine if these genotypes exhibit genetic differences in susceptibility to willow beetle damage. 2 Seven S. viminalis genotypes and four hybrids were tested for the feeding preference of adult P. vulgatissima, larval performance and within‐season plant response to manual defoliation (50% and 75%). 3 The feeding preference of adult beetles, the growth rate of larvae, and the weight at 33 days of larvae and pupae differed significantly between genotypes. Genotypes also differed significantly in their height and weight responses to mechanical defoliation. Two genotypes were taller, with longer internodes, after defoliation than were undamaged plants. Two hybrids and their S. viminalis parent showed no significant reduction in final dry weight between 0% and 50% defoliation treatments. 4 Susceptibility of genotypes to adult feeding was not correlated with their tolerance to defoliation in terms of weight or height responses; however, larval growth rate on genotypes was negatively correlated both with final height and number of leaves after 75% defoliation and with the susceptibility of genotype to adult feeding. 5 Salix viminalis showed genetic differences for all parameters tested. This suggests that the planting of a mixture of these genotypes would contain genetic differences with respect to host susceptibility to P. vulgatissima. Some parameters showed similarities between a hybrid and its parent, whereas others showed differences between siblings. This offers potential for effective breeding of desirable traits.     

Microsatellite markers for diverse Salix species

Forty‐six microsatellites were isolated from an enriched library of Salix burjatica and tested on 20 individuals (of nine species/hybrids) from the National Willows Collection (IACR‐Long Ashton Research Station, UK). Twenty‐nine were monomorphic, gave multilocus or unscorable patterns, or were duplicates. The remaining 17 microsatellites gave 2–22 alleles/locus. Three microsatellites successfully cross‐amplified in 31 additional Salix species. A further six were tested on panels comprising 6–25 individuals from the 31 species. Cross‐amplification was successful in all cases. These results suggest that the microsatellites isolated here should prove useful for population studies in a wide range of Salix species.     

Approaches on vegetative propagation of difficult-to-root <Emphasis Type="Italic">Salix caprea</Emphasis>

In the framework of a willow rust research project, it was necessary to include vegetatively propagated plant material of selected sallow trees (Salix caprea L.) into biotests for identification of pathotypes. Since it was not possible to root sufficient clonal plants by conventional cutting propagation, the applicability of tissue culture methods was tested. From 10 selected donor trees of Salix caprea newly sprouted shoots were harvested and transferred to nutrient media after surface disinfection. The cultures were grown at 20--22 °C, illuminated with warm-white fluorescent tubes. The majority of shoot tips and nodal segments died during the first month, but only with nursery-grown plants this was caused by bacteria contamination. Two clones could be established easily on hormone-free medium. Five clones could be initiated only after repeated subcultures on various media variants. Three clones failed completely. Different basic media compositions were tested and Woody Plant Medium, supplemented with 0.1% activated charcoal, proved to be best for most of the sallow clones. Well developed rooted plantlets were used in vitro for microcutting propagation. The resulting plants were transferred to soil and could be included in the rust screening program after acclimatising. The applicability of micropropagation for selected Salix caprea donor trees was strongly depending on the genotype. But the comparison of results from microcuttings with conventional cutting propagation showed that these methods were successful for different clones each.     

The influence of phenolic compounds on the suitability of three Salix species as hosts for the willow beetle Phratora vulgatissima

The phenolic glycosides salicin and salicortin were found to influence larval growth and development rates and adult feeding preference of Phratora vulgatissima in laboratory feeding studies. Salicortin was more toxic to larvae than salicin, and none of the third instar larvae fed on Salix viminalis leaves amended with 1.52% (fresh mass) salicortin pupated. Condensed tannins (proanthocyanadins) did not affect larval performance. It was concluded that Salix burjatica resistance to willow beetle is due to the high levels of salicortin which occur in leaves of this species.     

Seasonal dynamics of dry weight, growth rate and root/shoot ratio in different aged seedlings of Salix caprea

Willows (e.g. Salix caprea L.) are deciduous and richly branched shrubs or small trees. Salix caprea shows a high adaptability to different habitat conditions. One way of evaluating this adaptability is to measure willow biomass and production. Young plants of S. caprea were sampled from the bottom of an artificial lagoon in which sediments removed from the local Vajgar fishpond were deposited. The bottom of the lagoon was overgrown by vegetation dominated by seedlings of the willows S. caprea and S. aurita. Willows grew in the lagoon at average density of 58 plants per m². The biomass production and growth of S. caprea were determined for 15 samples (collected from 315 individuals) during the growing season. Annual net dry matter production in the whole community was estimated for 2.7 kg m⁻². Willows are generally considered to be fast-growing plants. The highest RGR of willows recorded by us was 0.03 to 0.04 g g⁻¹ d⁻¹ both in the stems and roots. This value was often recorded from July to August.     

Selection ofAzolla forms resistant to the water lily aphid,Rhopalosiphum nymphaeae Life history of Rhopalosiphum nymphaeae

Genotypic and environmentally determined differences inSalix viminalis L. suitability forDasineura marginemtorquens (Bremi) (Diptera: Cecidomyiidae) growth and survival were investigated in laboratory and field experiments. Earlier studies have documented high neonate larval mortality on certainS. viminalis genotypes. Here we present results from laboratory experiments which show no difference in larval establishment behaviour on resistant and susceptible willow genotypes. These data indicate that larvae are not able to detect the plant characteristic responsible for resistance. In a field study insect performance was measured on willow shoots of dissmilar size. Adult body size was positively correlated to willow shoot length. In turn, body size was positively correlated to potential fecundity. These results are discussed in relation to observed high densities ofD. marginemtorquens in vigorously growing willows cultivated for biomass production.     

The Proportion of Three Foundation Plant Species and Their Genotypes Influence an Arthropod Community: Restoration Implications for the Endangered Southwestern Willow Flycatcher

As part of a restoration project, multiple genotypes of two tree species, Fremont cottonwood (Populus fremontii) and Goodding's willow (Salix gooddingii), and one shrub species, Coyote willow (S. exigua), were experimentally planted in different proportions at the Palo Verde Ecological Reserve near Blythe, California, U.S.A. These common woody plant species are important to the endangered southwestern willow flycatcher, providing perch, nesting, and foraging habitat. We conducted this study to evaluate plant species proportion and plant genotype effects on the arthropod community, the prey base for the endangered southwestern willow flycatcher. Three patterns emerged. First, plant species proportions were important; the arthropod community had the greatest richness and diversity (H′) when Goodding's willow proportion was high and Fremont cottonwood proportion was lower; that is, fewer Fremont cottonwoods are required to positively affect overall arthropod diversity. Second, we found significant genotypic effects, for all three plant species, on arthropod species accumulation. Third, while both planting proportion and genotype effects were significant, we found that the effect of planting proportion on arthropod richness was about twice as large as the effect of plant genotype. This shows that both plant species proportions and genotype should be utilized in restoration projects to maximize habitat heterogeneity and arthropod richness. Similar studies can determine which planting proportion and specific genotypes may result in a more favorable arthropod prey base for the southwestern willow flycatcher and other species of concern. Greater attention to planting design and genotype can result in significant gains in diversity at little or no additional project cost.     

Variation in the uptake of Arsenic, Cadmium, Lead, and Zinc by different species of willows Salix spp. grown in contaminated soils

The experiment assessed the variability of in seven clones of willow plants of high biomass production (Salix smithiana S-218, Salix smithiana S-150, Salix viminalis S-519, Salix alba S-464, Salix ’Pyramidalis’ S-141, Salix dasyclados S-406, Salix rubens S-391). They were planted in a pots for three vegetation periods in three soils differing in the total content of risk elements. Comparing the calculated relative decrease of total metal contents in soils, the phytoextraction potential of willows was obtained for cadmium (Cd) and zinc (Zn), moderately contaminated Cambisol and uncontaminated Chernozem, where aboveground biomass removed about 30% Cd and 5% Zn of the total element content, respectively. The clones showed variability in removing Cd and Zn, depending on soil type and contamination level: S. smithiana (S-150) and S. rubens (S-391) demonstrated the highest phytoextraction effect for Cd and Zn. For lead (Pb) and arsenic (As), the ability to accumulate the aboveground biomass of willows was found to be negligible in both soils. The results confirmed that willow plants show promising results for several elements, mainly for mobile ones like cadmium and zinc in moderate levels of contamination. The differences in accumulation among the clones seemed to be affected more by the properties of clones, not by the soil element concentrations or soil properties. However, confirmation and verification of the results in field conditions as well as more detailed investigation of the mechanisms of cadmium uptake in rhizosphere of willow plants will be determined by further research.     

Using prey availability to evaluate Lower Colorado River riparian restoration

The Lower Colorado River Multi‐species Conservation Program (MSCP) is charged with restoring habitat for 26 species such as the southwestern willow flycatcher (Empidonax traillii extimus) impacted by water development projects on the river. As of 2015, the MSCP had spent $200 million to create 1,200 ha of habitat at nine sites, but the benefits to these insectivorous birds and other target species have not been quantified. Many MSCP projects emphasized riparian plantings of willow (Salix exigua, Salix gooddingii) and cottonwood (Populus fremontii) on high terraces disconnected from the river. We documented prey availability for insectivores in constructed habitats as an indicator of restoration effectiveness. Using sticky traps as a proxy to estimate aerial insect flux, we found the number of aquatic insects, proportion of aquatic insects, total number of insects, and number of insect orders were all significantly lower in MSCP plantation sites than at the river's edge. Riparian restoration sites over 100 m from the river had only 4% of the aquatic insects, 20% of the total insects, and only half as many insect orders as sites adjacent to the river. Thus, food availability and overall habitat quality for insectivores are likely low in restoration sites that are distant from the river.     

Plant phenology-mediated indirect effects: The gall midge opens the phenological window wider for a leaf beetle

We examined whether larvae of the gall midge Rabdophaga rigidae (Diptera: Cecidomyiidae) can modify the seasonal dynamics of the density of a leaf beetle, Plagiodera versicolora (Coleoptera: Chrysomelidae), by modifying the leaf flushing phenology of its host willow species, Salix serissaefolia and Salix eriocarpa (Salicaceae). To test this, we conducted field observations and a laboratory experiment. The field observations demonstrated that the leaf flushing phenology of the willows and the seasonal dynamics of the beetle density differed between shoots with stem galls and shoots without them. On galled shoots of both willow species, secondary shoot growth and secondary leaf production were promoted; consequently, leaf production showed a bimodal pattern and leaf production periods were 1 to 2 months longer than on non‐galled shoots. The adult beetle density on galled shoots was thus enhanced late in the season, and was found to change seasonally, synchronizing with the production of new leaves on the host willow species. From the results of our laboratory experiment, we attributed this synchrony between adult beetle density and willow leaf flush to beetles’ preference to eat new leaves rather than old. Indeed, beetles consumed five times more of the young leaves when they were fed both young and old leaves. These results indicate that stem galls indirectly enhance the adult beetle density by enhancing food quality and quantity late in the beetle‐feeding season. We therefore conclude that midge galls widen the phenological window for leaf beetles by extending the willows’ leaf flush periods.