Clonal variation in survival and growth of hybrid poplar and willow in an in situ trial on soils heavily contaminated with petroleum hydrocarbons

Species and hybrids between species belonging to the genera Populus (poplar) and Salix (willow) have been used successfully for phytoremediation of contaminated soils. Our objectives were to: 1) evaluate the potential for establishing genotypes of poplar and willow on soils heavily contaminated with petroleum hydrocarbons and 2) identify promising genotypes for potential use in future systems. We evaluated height, diameter, and volume after first year budset by testing 20 poplar clones and two willow clones. Unrooted cuttings, 20 cm long, were planted in randomized complete blocks at 0.91- x 0.91-m spacing at Gary, IN, USA (41.5 degrees N, 87.3 degrees W). Four commercial poplar clones (NM6, DN5, DN34, and DN182) were planted as 20- and 60-cm cuttings. Sixty-cm cuttings exhibited greater height and diameter than 20-cm cuttings; however, we recommend continued use and testing of different combinations of genotype and cutting length. We identified promising genotypes for potential use in future systems and we recommend allocating the majority of resources into commercial poplar clones, given their generalist growth performance. However, further utilization and selection of experimental clones is needed. Specific clones rather than genomic groups should be selected based on the geographic location and soil conditions of the site.     

Field note: successful establishment of a phytoremediation system at a petroleum hydrocarbon contaminated shallow aquifer: trends, trials, and tribulations

We report the establishment of a mixed hybrid poplar (Populus spp.) and willow (Salix spp.) phytoremediation system at a fuel-contaminated site. Several approaches were used to balance competing goals of cost-effectiveness yet successful tree establishment without artificial irrigation or trenching. Bare root and unrooted cuttings were installed using either: (1) 1.2 m deep holes excavated with an 8 cm diameter auger using a direct-push rig and backfilled with the excavated, in situ soil; (2) 1.2 m deep holes created with a 23 cm diameter auger attached to a Bobcat rig and backfilled with clean topsoil from offsite; and (3) shallow holes between 15-30 cm deep that were created with a 1.3 cm diameter rod and no backfill. Tree mortality from initial plantings indicated contaminated zones not quantified in prior site investigations and remedial actions. Aquifer heterogeneity, underground utilities, and prior remediation infrastructure hampered the ability of the site to support a traditional experimental design. Total stem length and mortality were measured for all planted trees and were incorporated into a geographic information system. Planting early in the growing season, augering a larger diameter hole, and backfilling with clean, uncontaminated topsoil was cost effective and allowed for greater tree cutting growth and survival.     

Taming the Beast: Managing Hydrology to Control Carolina Willow (Salix caroliniana) Seedlings and Cuttings

Historically, wetlands along the St. Johns River, Florida, were dominated by herbaceous marshes. However, in the last 50 years many areas transformed to shrub‐dominated wetlands, at the same time a system of levees and canals was constructed to control flooding. We tested the role of water management in controlling Carolina willow (Salix caroliniana), a native shrub that accounts for most of this shift. We assessed survival and growth of seedlings and cuttings on four artificial islands. We planted willow seedlings and cuttings at the spring waterline and at three higher levels (+17.5, +35, and +50 cm) and evaluated their responses to natural hydrologic fluctuations. Overall, seedlings had lower survival than cuttings. Highest mortality occurred during summer floods and willows greater than 50 cm above marsh surface had the highest survivorship. Surviving seedlings attained similar height and biomass among elevations, but the cuttings had greater stem diameter, stem height, and biomass at higher elevations. In the second experiment, we planted seedlings and short (25 cm) and tall (50 cm) cuttings at the waterline and at three higher levels (+25, +35, and +50 cm) in artificial ponds with controlled water levels. Before flooding, seedlings at the highest elevation suffered some mortality due to desiccation, but after flooding, they had the highest survival. Elevation did not affect cutting survival, but those at the lowest elevation had the greatest height and biomass. Hydrologic manipulation can be a powerful tool to control willow establishment. However, its success depends on timely and prolonged inundation or water drawdown .     

Developmental quantitative genetics of growth in Populus

 Nursery growth and dry weight were analyzed for F₂ genotypes derived from Populus trichocarpa×P. deltoides that have been field tested with clonal replicates in three different environments. The correlations between nursery and plantation performance differed among the environments, with higher values at Boardman and Clatskanie (both planted with rooted cuttings) than Puyallup (planted with unrooted cuttings). At Puyallup, nursery height was more strongly associated with plantation growth than were nursery diameter and dry weight. Yet, this finding was not supported by QTL mapping. A single overdominant QTL on linkage group G affected the stem height of both seedlings and resprouts in the nursery but showed nonsignificant LOD scores for plantation height from ages 1 to 5 at Puyallup. A total of four QTLs were identified for nursery diameter, one of which on linkage group O also controlled plantation basal area at all ages. Two important nursery QTLs on linkage groups B and G were used to estimate the relative efficiency of marker-assisted selection for plantation productivity. Despite the fact that they were not detected in the plantation stage, these two QTLs could significantly increase the proportion of the phenotypic variance explained by plantation QTLs. Received: 5 February 1998 / Accepted: 1 April 1998     

Factors governing survival of black willow (Salix nigra) cuttings in a streambank restoration project

A field study was conducted at Little Topashaw Creek in northern Mississippi, aimed at expanding the limited database on the survivorship of Salix nigra (black willow) cuttings planted on riparian restoration sites. We tested the hypothesis that sediment moisture availability (deficit, excess) as mediated by sediment texture and depth to the prevailing water table is a major factor governing black willow survival during the initial stage of establishment following transplanting. Replicated plots were established across elevational gradients and a range of soil texture. Each plot contained 16 planted cuttings (2.5 cm diameter × 2.5 m length). Plot depth to water table, soil texture, and soil redox potential were measured. Plant gas exchange, leaf chlorophyll content, growth, and survival were monitored periodically over two growing seasons. Survival was best at low elevation compared to cuttings planted at mid- and high elevations. Poor survival and growth were noted for cuttings that encountered sediment moisture deficits in plots with coarse texture while the best cutting survival was recorded for intermediate sand content plots. Results indicated that plot location on the bank and soil texture are two important factors that influence riverbank restoration success. Therefore, any riparian restoration plan should include careful assessment of these factors prior to undertaking such efforts.     

Effects of Propagule Phenology (Non-Dormant Versus Dormant) and Planting System (Vertical Versus Horizontal) on Growth Performance of Willow Clones Grown Under Different Weeding Regimes

To assess the effects of propagule phenology and planting system on growth performance of three willow clones grown under different weeding regimes, a field experiment was performed in central Sweden 2014–2016. Freshly harvested (non-dormant) and cold-stored (dormant) cuttings (planted vertically) and billets (planted horizontally) from willow clones Tordis, Tora, and Jorr were planted in weeded and in unweeded plots. Sprouting was significantly higher for willows grown from non-dormant (74%) than dormant (58%) propagules and for cuttings (84%) compared with billets (42%). Survival was higher for willows from non-dormant propagules in weeded (71%) compared with unweeded (63%) plots, willows from dormant propagules in weeded (72%) compared with unweeded (60%) plots, and for willows from cuttings (93%) compared with billets (39%). During 2014–2016, aboveground biomass production was significantly higher for willows from cuttings (11.71 t DW ha^?1) than from billets (6.13 t DW ha^?1), grown in weeded (15.29 t DW ha^?1) than in unweeded (2.55 t DW ha^?1) plots, and differed significantly among willow clones (11.48, 9.27, and 6.01 t DW ha^?1 for Tordis, Tora, and Jorr, respectively). In this study, (i) planting with cold-stored and freshly harvested willow propagules was equally successful and therefore cold storage could be potentially avoided and replaced with planting of freshly harvested propagules in early spring; however, (ii) in terms of measured growth performance parameters, willows grown from cuttings performed better than grown from billets; and (iii) weed competition significantly reduced survival and aboveground biomass production, confirming that weed control during establishment of willow is crucial.     

Responses of Nondormant Black Willow (Salix nigra) Cuttings to Preplanting Soaking and Soil Moisture

The use of willow cuttings for streambank stabilization is a common practice in riparian ecosystems throughout the United States. Many environmental factors govern the outcome of such planting. However, other factors such as preplanting treatments, planting methods, and physiological status of cuttings (dormant vs. actively growing) may also be crucial in determining the survival of willow cuttings. Actively growing (nondormant) Black willow (Salix nigra) cuttings, 30 cm in length and 1 cm in diameter at the base, were subjected to three soaking treatments (0, 7, and 15 days) prior to planting. Following the initial treatment, cuttings were grown in a greenhouse in pots under three soil moisture regimes (well‐watered but not flooded, permanently flooded, and intermittently flooded). Plant gas exchange, growth, biomass, and survival were measured. Results demonstrated that soaking for 7 days was beneficial to early development of cuttings in the well‐watered (control) soil moisture regime, enhancing percent bud flush and survival significantly. However, 15 days of soaking proved to be detrimental to survival of cuttings irrespective of soil moisture regimes. Results also demonstrated that the beneficial effects of 7‐day soaking were limited to the well‐watered soil moisture regime but not to the flooded or intermittently flooded regimes. Soaking nondormant cuttings may be worthwhile if the planting site is likely to present ample soil moisture but nonflooded conditions to the transplanted cuttings.     

Cadmium accumulation and tolerance in Populus nigra and Salix alba

Rooted cuttings of Populus nigra L. clone Poli and Salix alba L. clone SS5 were treated for three weeks with 50 μM CdSO₄ in nutrient solution. The willow showed a far higher Cd tolerance, expressed as tolerance index (Ti), than the poplar in both roots and leaves. The root Cd content was higher in poplar than in willow, whereas in leaves the opposite was found. As a consequence, the translocation factor (Tf) revealed a greater ability of Cd transport in willow than in poplar. Cd treatment enhanced cysteine, γ-glutamylcysteine and reduced glutathione contents in roots of both species, whereas in leaves they were only enhanced in poplar. Furthermore, only poplar leaves showed an enhanced content of phytochelatins, whereas malic and citric acids rose in response to Cd only in the willow leaves. Cd treatment increased putrescine, spermidine and spermine contents in both roots and leaves of the willow, whereas in poplar only the putrescine content was enhanced in roots.     

Effects of Cutting Phenology (Non-dormant Versus Dormant) on Early Growth Performance of Three Willow Clones Grown Under Different Weed Treatments and Planting Dates

To assess the effects of cutting phenology on early growth performance of three willow clones grown under different weed treatments and planting dates, freshly harvested (non-dormant) and cold-stored (dormant) cuttings from willow clone Tora, Jorr, and Olof were planted in bucket experiment outdoors in central Sweden on five planting dates (May–June 2013) with or without a model weed (spring barley). Non-dormant cuttings sprouted faster than dormant cuttings when planted early in the season. For cuttings planted later in the season, bud sprouting was affected only by willow clone. Aboveground biomass production was affected by cutting phenology, planting date, clone, and weed treatment. When planted on May 3 and May 10, biomass produced from non-dormant and dormant cuttings did not differ, while willows grown from dormant cuttings produced 59% more aboveground biomass than willows grown from non-dormant cuttings when planted on May 24–June 16. Tora produced on average 12% more biomass than Jorr and Olof, and weed competition reduced aboveground biomass production on average with 36%. The ability of willow to suppress weeds (WSA) was 26 (non-dormant cuttings) and 12% (dormant cuttings) higher for willows planted on May 3 compared with WSA of willows grown from cuttings planted later in the season. The ability to tolerate competition from weeds (WT) was 51 and 52% lower for willows grown from non-dormant and dormant cuttings planted late in the season compared with WT of willows planted earlier in the season. We conclude that planting with long-term cold storage of willow cuttings can be replaced with planting freshly harvested cuttings when planting is performed in early season, and that weed competition strongly reduces biomass production. Weed control during the establishment phase is crucial in order to maximize willow biomass production.     

Predicting within-family variability in juvenile height growth of <Emphasis Type="Italic">Salix</Emphasis> based upon similarity among parental AFLP fingerprints

Willow is being developed as a crop for biomass plantations in the Northeast and North-central United States, but has only recently been the subject of controlled breeding to generate improved genotypes. Maximizing variability among progeny within full-sib families produced by controlled pollination may increase the probability of producing willow clones exhibiting desirable extreme phenotypes. Yet, predicting combinations of parents yielding highly variable progeny is not currently possible. Controlled pollinations were completed among 15 Salix eriocephala clones and the resulting progeny were vegetatively propagated and planted in a greenhouse progeny test. Heights of rooted cuttings were measured after 4 months of growth. Genetic similarity among parents was estimated based on 77 polymorphic AFLP bands. Strong negative correlation (r = -0.88) was detected between mean female-parent similarity indices and the standard deviation of height among half-sib progeny from those females. Parent combinations that had relatively low similarity indices tended to produce progeny that had greater variability in height. This negative relationship suggests that AFLP fingerprints of S. eriocephala parents may be useful for predicting parent combinations that will yield families with large variability.     

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.     

Effects of Pre-Planting Soaking on Growth and Survival of Black Willow Cuttings

Black willow (Salix nigra) uses periodic flood events for dispersal of vegetative propagules, subjecting them to periods of soaking before their deposition along the streambank. It was hypothesized that this life history trait results in optimal conditions for willow growth and survival. To test this hypothesis, a greenhouse experiment was conducted using 1.2‐m‐long black willow cuttings (posts) with a basal diameter of 5 cm. Cuttings were subjected to three soaking treatments (0, 3, and 10 days) and then grown under four soil moisture regimes (control, drought, permanently flooded, and intermittently flooded). Growth, biomass, and survival were recorded. Results showed that soaking posts for 10 days was most beneficial in the control soil moisture regime, enhancing root, shoot, leaf, and total biomass production. Shoot abundance and growth were also enhanced by 10 days of soaking in the control and permanently flooded moisture regimes. Finally, survival increased significantly in control and drought moisture regimes in response to the 10‐day soaking treatment. Results clearly demonstrated that soaking had significant effects on willow post success when evaluated across all soil moisture regimes. Posts subjected to the 10‐day soaking treatment consistently had greatest growth and biomass and displayed a doubling of the survival rate. Therefore, soaking willow cuttings before planting is a simple, inexpensive technique that may be used to bolster streambank restoration success.     

Phytoremediation of Metal Contaminated Soil Using Willow: Exploiting Plant-Associated Bacteria to Improve Biomass Production and Metal Uptake

Short rotation coppice (SRC) of willow and poplar is proposed for economic valorization and concurrently as remediation strategy for metal contaminated land in northeast-Belgium. However, metal phytoextraction appears insufficient to effectuate rapid reduction of soil metal contents. To increase both biomass production and metal accumulation of SRC, two strategies are proposed: (i) in situ selection of the best performing clones and (ii) bioaugmentation of these clones with beneficial plant-associated bacteria. Based on field data, two experimental willow clones, a Salix viminalis and a Salix alba x alba clone, were selected. Compared to the best performing commercial clones, considerable increases in stem metal extraction were achieved (up to 74% for Cd and 91% for Zn). From the selected clones, plant-associated bacteria were isolated and identified. All strains were subsequently screened for their plant growth-promoting and metal uptake enhancing traits. Five strains were selected for a greenhouse inoculation experiment with the selected clones planted in Cd-Zn-Pb contaminated soil. Extraction potential tended to increase after inoculation of S. viminalis plants with a Rahnella sp. strain due to a significantly increased twig biomass. However, although bacterial strains showing beneficial traits in vitro were used for inoculation, increments in extraction potential were not always observed.     

Bark morphological and chemical features are differentially correlated with disease resistance and yield in hybrid poplar taxa

In the southeastern United States, the establishment of short-rotation intensively cultured plantations of hybrid poplar has been hindered by its susceptibility to stem cankers. We evaluated the tradeoffs between biomass yield and disease tolerance in hybrid poplar genotypes belonging to P. deltoides × P. maximowiczii (DM), P. deltoides × P. nigra (DN), P. trichocarpa × P. maximowiczii (TM), and P. deltoides × P. deltoides (DD) taxa. We hypothesized that canker resistant genotypes will have thicker bark but bark thickness and biomass yield will be negatively correlated. After two growing seasons, the DD genotypes developed thicker bark compared to the genotypes of other taxa and bark thickness was not correlated with biomass yield in the DD genotypes (R² = 0.002). However, in the TM, DM, and DN genotypes, bark thickness was negatively correlated with biomass yield (R² = 0.33–0.77). Disease incidence studies revealed that the DM genotypes were most susceptible to canker whereas no disease was detected in DD genotypes. Furthermore, bark analysis conducted by Fourier transform infrared spectroscopy coupled with multivariate analysis showed that that DD genotypes to be chemically separate from the three hybrid genotypes and that bark chemistry was correlated with canker disease incidence. Taken together, these results reveal that it is possible to generate hybrid poplar genotypes with thicker bark, disease resistance, and higher biomass yields. This insight should guide further efforts to develop genetically improved hybrid poplar genotypes, both in terms of biomass yield and disease tolerance, for cultivation in the southeastern United States. Hybrid poplar cultivation in southeastern United States is hindered by its susceptibility to stem cankers. We evaluated tradeoffs between yield and canker disease resistance in various hybrid poplar genotypes. After two growing seasons, the DD genotypes showed disease resistance and developed thicker bark that was chemically distinct from the other genotypes. Bark thickness was not correlated with yield in the DD genotypes but was negatively correlated with yield in the other genotypes. These results will guide the development of hybrid poplar genotypes that are both disease resistant and high yielding for cultivation in the southeastern United States.     

Resprouting Ability of Dry Forest Tree Species after Disturbance Does Not Relate to Propagation Possibility by Stem and Root Cuttings

Tropical dry forest tree species are recognized for their high resprouting ability after disturbance. We tested whether species that commonly produce root and stem suckers can be propagated by large stem and root cuttings, a useful method for landscape restoration programs. We performed four experiments: (1) In a greenhouse, we tested the propagation of six species using large stem cuttings collected from early successional sites. We used the following treatments: (i) dry season collection and planting; (ii) dry season collection, storage in humid soil, and wet season planting; (iii) wet season collection and planting; and (iv) wet season collection and planting after treatment with commercial NAA auxin. (2) Stem cuttings of Myracrodruon urundeuva were planted in a pasture during the rainy season after either NAA, IBA, or no auxin treatment. (3) As a control experiment, we also planted cuttings of Spondias mombin, a species known for successfully regenerating from cuttings. (4) Root cuttings of six species were collected in recently plowed pastures and planted in the greenhouse with and without treatment with NAA auxin. No root cuttings rooted. Only M. urundeuva and Astronium fraxinifolium stem cuttings rooted. Maximum success was obtained for stem cuttings collected and planted in the dry season (23%). Only 13% of M. urundeuva had sprouted by the 15th month of the field experiment. As a result, large cuttings are not recommended for propagation of the studied species. Future studies should include development of suitable methods of root harvesting and prospection of traditional knowledge for species selection.     

Development of salix and populus mycorrhizae in metallic mine tailings

Summary Roots of two year old willow and poplar planted as cuttings in copper and iron mine tailings were stained intact and examined microscopically for mycorrhizal development. Incidence of mycorrhizae was related to tree vigor and to chemical and physical properties of the tailings. Results showed that no mycorrhizal development occurred on roots of these tree species in the copper tailings; inoculations with natural forest soil extract failed to initiate this symbiosis. Tops and roots of the willow and poplar were stunted and survival was poor. Roots in the iron mine tailings developed extensive ectomycorrhizae and trees showed good growth. Presence of mycorrhizae was confirmed and external morphology examined with the scanning electron microscope. Aseptic jack pine and naturally mycorrhizal white pine roots were used in microscopic comparisons with the hardwoods. re]19760220     

Interactive effects of biochar and an organic dust suppressant for revegetation and erosion control with herbaceous seed mixtures and willow cuttings

Land reclamation efforts on “non‐soil” substrates frequently involve additions of organic material coupled with soil‐stabilizing dust suppressants to control erosion, but formal experiments, particularly of interactive effects, are lacking. We examined the effects of Entac?, a tall oil pitch emulsion, and pyrolyzed wood waste‐derived biochar on early plant establishment, growth, and survivorship, in 2 factorial greenhouse experiments: 1 using a seed mix widely used for roadside erosion control (the grasses Festuca rubra, Poa pratensis, Lolium perenne, and white clover [Trifolium repens]) and 1 using rooted cuttings of sand bar willow (Salix exigua). Biochar additions of 5, 10, and 20 t/ha enhanced biomass of clover by 14–250%, but had no significant effects on grasses. Entac additions inhibited early establishment and growth of both clover and grasses at all biochar treatment levels. Entac plus biochar treatments significantly enhanced survivorship of willows following a drought treatment in a greenhouse trial. In a field trial, Entac treatments had positive effects on willow diameter growth. Collectively, these results suggest that better soil moisture retention is the principal mechanism responsible for enhanced survivorship and growth of planted willows treated with Entac and biochar on sand substrates. Given Entac's inhibitive effects on early establishment of herbaceous species, and the abilities of Entac and biochar in combination to mitigate drought stress, these treatments may be particularly well suited for ecological restoration efforts using planted tree stock on coarse‐textured substrates and where exposure and ingress of non‐native herbaceous species are of concern.     

Field performance of <Emphasis Type="Italic">Theobroma cacao</Emphasis> L. plants propagated via somatic embryogenesis

Somatic embryogenesis is an in vitro clonal propagation method with potential to contribute to the improvement of cacao varieties. Before using this technology for commercial production, it is essential that somatic embryogenesis-derived plants be tested in field conditions. Therefore, we established a field test at Union Vale Estate, Saint Lucia. Thirty- to 50-yr-old trees were selected for clonal propagation as potentially high yielding based on local farmers observations. Clonal plants were propagated in vitro from immature flowers by embryogenesis and micropropagation. Multiple plants from nine genotypes were acclimated to greenhouse conditions then returned to Saint Lucia and planted in a field. Orthotropic rooted cuttings and locally propagated open pollinated seedlings were also planted for a total of 214 trees. Growth data were collected every 4–6 mo. including: stem diameter, stem height, length of the longest jorquette branch, number of jorquette branches, and dates of first flowering and fruiting. At 4.5 yr after planting in the field there were no major differences in all growth parameters among the propagation methods evaluated with exception of the orthotropic rooted cuttings. Trees grown from seeds were slightly taller then trees propagated by the other methods. Trees propagated as orthotropic rooted cuttings exhibited smaller average stem diameters, shorter stem heights to the jorquette, and shorter jorquette branches. We concluded that somatic embryo-derived plants demonstrated normal phenotypes in field conditions and have growth parameters similar to plants propagated by traditional methods.     

Establishment of Moose Browse on Four Growth Media on a Proposed Mine Site in Southcentral Alaska

The objective of this study was to evaluate seven woody plant species on four growth media for their potential contribution to moose habitat and establishment of viable plant communities on a proposed mine site in southcentral Alaska. Populus balsamifera (balsam poplar), Salix alaxensis (feltleaf willow), S. barclayi (Barclay willow), S. bebbiana (Bebb willow), Alnus tenuifolia (thinleaf alder), Betula papyrifera (paper birch), and Picea glauca (white spruce) were selected for their functions in moose habitat, ease of propagation, and presence in the existing native vegetation. Three native soils were selected for biological characteristics such as different potential to form mycorrhizae and to regenerate local plant species, both of which are governed partly by existing vegetation. The fourth growth medium, glacial till or overburden, was expected to have little or no biological activity. A mining disturbance was simulated on three sites by removing existing vegetation from the plots, stripping the native soils, and then spreading these soil materials over the respective study plots. Rooted cuttings of the Salicaceae and nursery seedlings of the other species were planted in each of the four growth media. Height and survival of all plant species were greater on the three soil media than on the glacial till during the second and third years. Percentage of ectomycorrhizal infection on transplants was similar among growth media, although lower ectomycorrhizal infection occurred on volunteers in grassland soils than in the other growth media. Browsable plants were produced within three years on the disturbed native soils but not on the glacial till.     

Efficient in vitro regeneration systems for Vaccinium species

Efficient protocols for shoot regeneration from leaf explants suitable for micropropagation as well as for the development of transgenic plants were developed for blueberry (Vaccinium corymbosum) and lingonberry (Vaccinium vitis-idaea) cultivars. Nodal segments were used to initiate in vitro shoot cultures of lingonberry cultivar ‘Red Pearl’ and southern highbush blueberry cultivar ‘Ozarkblue’. In order to develop an optimized regeneration procedure, different types and concentrations of plant growth regulators were tested to induce adventitious shoot regeneration on excised leaves from micropropagated shoots of both cultivars. The effect on percentage regeneration and number of shoots per explant was investigated. Results indicated that zeatin was superior to TDZ and meta-topolin in promoting adventitious shoot formation. A concentration of 20 μM zeatin was most effective in promoting shoot regeneration in both cultivars, in case of ‘Red Pearl’ along with 1 μM NAA. Shoots were either allowed to root in vitro on medium containing IBA or NAA or ex vitro in a fog tunnel. IBA was superior to NAA for induction of root development in vitro in both Vaccinium cultivars. Ex vitro rooting under high humidity was tested with cuttings from mature field-grown plants, from acclimatized tissue culture derived plants and with unrooted in vitro proliferated shoots planted directly. It was found that in vitro shoots rooted better under fog than cuttings from the other plant sources and rooting was equivalent to that achieved in vitro.