In vitro interactions between bacteria isolated from Sitka spruce stumps and Heterobasidion annosum

Culture medium composition affected antagonism by bacterial isolates from Sitka spruce (Picea sitchensis) stumps against Heterobasidion annosum. Fifty percent of bacterial isolates inhibited H. annosum growth on sporulation agar or yeast–dextrose–peptone agar; only 10% of isolates caused inhibition on both media. Proportions of isolates inhibiting H. annosum varied with stump age; fewer isolates from 4‐ or 6‐year‐old stumps exhibited antagonism than isolates from older or younger stumps. Fifteen isolates showing antagonism on sporulation agar were tested against H. annosum in spruce wood cubes. None of the bacterial isolates alone caused a significant weight reduction in inoculated cubes. Relative inoculation times of bacterial isolates and H. annosum had an effect on weight loss in interactions; simultaneous inoculation with isolates and H. annosum inhibited weight loss caused by H. annosum compared with bacteria‐free controls. Inoculation with bacterial isolates 10 days before H. annosum had no effect on the decay rate. In contrast, inoculation with H. annosum 10 days before bacteria increased weight loss of cubes by 200% relative to cultures lacking bacteria. The effect of a mixed bacterial inoculum on weight change in 0.2‐mm spruce wood slips co‐inoculated with H. annosum, Resinicium bicolor, Hypholoma fasciculare, Stereum sanguinolentum or Melanotus proteus differed between different fungi.     

Persistence of a biocontrol strain of Phlebiopsis gigantea in conifer stumps and its effects on within‐species genetic diversity

Fungal isolations and genetic fingerprinting were used to determine whether Phlebiopsis gigantea stump treatment against Heterobasidion annosum sl. using a single genotype (Rotstop) would affect the genetic diversity of P. gigantea populations. The survival time of P. gigantea was longer in Norway spruce (Picea abies) stumps compared to Scots pine (Pinus sylvestris) as no isolates were obtained from pine stumps 6 years after treatment, whereas in about half of the spruce stumps the fungus was still present. The usage of Rotstop did not seem to increase the occurrence of the fungus 5 years after the treatment in fresh (1‐year‐old) untreated stumps within the same forest stands. All the isolates from the 6‐year‐old treated spruce stumps were identical in genotype with the Rotstop‐strain, whereas all isolates from the fresh untreated spruce and pine stumps differed from it. Within the treated pine stand, the biocontrol usage seemed to have caused a slight reduction in genetic markers not related to Rotstop, but there were no statistically significant differences between the marker frequencies and the local natural population. Thus, Rotstop is not likely to cause any immediate threat to the genetic diversity of P. gigantea.     

Effects of Mounds and Stumps on the Root Architecture of Sitka Spruce on a Peaty Gley Restocking Site

Root systems from Sitka spruce trees planted at stump, awayfrom stumps and on mounds were extracted from a restocking site14 years after planting. Compass direction and diameter of allmain roots were measured and the position of the centre of diameter,a concept analogous to the centre of mass, was calculated. Themagnitude and direction of the centre of diameter from the stemcentre were used in the statistical analysis of root systemsymmetry. The presence of a stump close to the planting positionprevented the development of a uniform root system and causedconsiderable clustering of roots away from the stump. Root systemsof trees planted away from old stumps or on mounds were moreuniform, although there was enhanced root growth in the directionof the mound centre for those trees planted on the side of themound. The implications for tree stability are discussed andalternatives to planting close to stumps are suggested. Received 3 September 1990.     

Growth of Phlebiopsis gigantea in wood of seven conifer species

Heterobasidion parviporum and Heterobasidion annosum are widely distributed root‐rot fungi that infect conifers throughout Europe. Infection of conifer stumps by spores of these pathogens can be controlled by treating fresh stumps with a competing non‐pathogenic fungus, Phlebiopsis gigantea. In this study, growth of three Latvian strains of P. gigantea and the biological control agent ‘Rotstop’ strain was evaluated in stem pieces of Norway spruce, Scots pine, lodgepole pine, Douglas‐fir, Weymouth pine, Siberian larch and Sitka spruce. The growth rates of one H. parviporum and one H. annosum isolate were also measured in the same stem pieces. The growth rate of P. gigantea varied greatly in wood of different conifer species. It was higher in the three pine species, lower in Norway spruce and lowest in Sitka spruce and Siberian larch, and in Douglas‐fir, this fungus did not grow. The largest area of wood occupied by P. gigantea was in lodgepole pine. Growth of Latvian isolates of P. gigantea in the wood of Pinus and Picea species was comparable to that of the Rotstop isolate. Consequently, stump treatment with local P. gigantea isolates should be recommended. However, our results suggest that Douglas‐fir stump treatment against Heterobasidion by P. gigantea may be ineffective and other stump treatment methods should be considered.     

Effect of planting Scots pine around Norway spruce stumps on the spread of Heterobasidion coll.

Management of a Norway spruce stand planted on a site infected by Heterobasidion coll. is problematic because the fungus spreads vegetatively from the colonized stumps of the previous generation to the new seedlings. Growing of mixed stand with more resistant tree species has been suggested to decrease the economic losses caused by butt rot in Norway spruce trees. The mechanistic simulation model Rotstand describing the spread of Heterobasidion coll. in coniferous stands of southern Finland was used to study the effect of planting Scots pines around colonized clear‐felling stumps of Norway spruce of the previous generation. Planting of Scots pines in clusters around colonized stumps markedly decreased the butt rot of Norway spruce trees at the age of 20 years and at clear felling. If the same number of Scots pines were planted randomly, the effect was weak. When the average diameter of colonized clear‐felling stumps was 30 cm, a Scots pine circle with a radius of 3 m resulted in the highest soil expectation value (SEV) at 2% discounting rate, whereas with 40‐cm stump diameter, a 4‐m radius produced the highest SEV. When the proportion of Heterobasidion parviporum in the old colonized stumps was 50% instead of 95%, planting pines around colonized stumps still clearly decreased the butt rot at the age of 20 years and in final felling.     

Fungi occurring in the roots and basal parts of one‐ and two‐year‐old spruce and pine stumps

The occurrence of fungi in the underground parts of 1‐ and 2‐year‐old conifer stumps (Pimis sylveslris and Picea abies) was studied in two clear‐felled areas of eastern Uppland in Sweden, and most fungal isolates were identified to species.

Decay fungi (Basidiomycetes) were more frequent in spruce stumps than in pine stumps, and their incidence was highest in 2‐year‐old stumps. The occurrence of blue‐stain fungi and hyaline mycelia did not differ significantly between stumps of different species or age. Within stumps, decay fungi were most frequent in large roots close to the cut surface, whereas blue‐stain fungi and hyaline mycelia were most abundant in small roots. The occurrence of yeasts, bacteria and fast‐growing moulds was also recorded.     

Effects of benomyl and captan on growth and mycorrhizal colonization of Sitka‐spruce (Picea sitchensis) and ash (Fraxinus excelsior) in Irish nursery soil

The effects of two fungicides (benomyl and captan ‐ at recommended doses and up to three soil drenches) on root development and mycorrhizal colonization of Sitka‐spruce (Picea sitchensis) and ash (Fraxinus excelsior) were examined after 20 weeks growth, under environmentally controlled conditions, in soil from an Irish tree nursery. Although four mycorrhizal types have been found on Sitka‐spruce at the nursery, only one ectomycorrhizal type (Piceirhiza horti‐inflata) was identified on the short roots in this study. An inoculant (Vaminoc: MicroBio Ltd, Hemel Hempstead, UK) was used to inoculate ash and 20‐week‐old seedlings had a higher arbuscular mycorrhizal (AM) colonization in comparison with uninoculated controls. Multiple applications (2–3) of benomyl reduced the length of root and shoot and shoot dry mass of Sitka‐spruce, whereas in ash, it only depressed root length. Benomyl decreased the numbers of ectomycorrhizas of Sitka‐spruce and arbuscular mycorrhizal colonization of Vaminoc‐inoculated ash. A single application of captan stimulated root length and ectomycorrhizal colonization of Sitka‐spruce and root dry mass in ash compared with uninoculated controls. Applications of captan reduced arbuscular mycorrhizal colonization of Vaminoc‐inoculated ash to levels near to those of uninoculated controls. Of the two fungicides used, benomyl had the most deleterious effect on root length and mycorrhizal colonization of Sitka‐spruce and ash.     

Effect of inoculum density and borate concentration in a stump treatment trial against Heterobasidion annosum

The effectiveness of disodium octaborate tetrahydrate (DOT) as a stump treatment chemical for Sitka spruce in Britain was tested on six occasions by inoculating treated and untreated stumps with basidiospores of Heterobasidion annosum at three concentrations ranging from an average 49 viable spores/ml to 4.9 × 10⁵/ml of water. The extent of colonization of heartwood by H. annosum was measured and, along with the incidence of infected stumps, provided a measure of the combined effects of spore concentration and of the two DOT treatments (15 and 30 g/m²) on the trial results. On untreated stumps, both the incidence of infection and the cross‐sectional area of stump heartwood colonized by the fungus increased with inoculum density. The same effect was evident in treated stumps, but it was reduced by increasing DOT application. Infection was at its lowest in stumps treated with DOT at 30 g/m², being entirely absent from those 60 stumps that were inoculated with the fewest spores. The implications of these findings for the design of trials of control agents that rely on artificial inoculation with H. annosum and for the selection of dose rates to use in harvesting operations are discussed.     

Bacteria inhabiting artificially inoculated xylem of Picea abies

Bacteria inhabiting the xylem of Norway spruce (Picea abies (L.) H. Karsten) were investigated. The trees had been wounded and artificially inoculated with fungi and bacteria obtained from wounds of naturally infected spruce. One and five growing seasons after inoculation the Gram‐negative bacterial population present in the stem of inoculated trees were analysed.

The Gram‐negative bacteria isolated from the trees were identified on the basis of morphological, biochemical and physiological tests and whole‐cell fatty acid composition. The predominant strains were Enterobacteriaceae fermenter strains (E. agglomerans or E. sakazakii), fluorescent and yellow pigmented Pseudomonas, Acinetobacter and Moraxella spp. All Gram‐positive bacteria were Bacillus species.

The Gram‐negative bacteria of Norway spruce differed from the Gram‐positive species in possessing stronger lipolytic activity and in their ability to utilize pine resins for growth. Gram‐positive bacteria were generally able to utilise cellulose and hemicellulose, whereas among the Gram‐negative bacteria only one xylanolytic (yellow Pseudomonas) strain was found.     

Spatial distribution and persistence of Heterobasidion parviporum genets on a Norway spruce site

Spatial distribution of Heterobasidion genets over a period of ca 50 years in two successive generations of Norway spruce (Picea abies) was unravelled. The genets were first identified in 1993 in a naturally regenerated 43‐year‐old spruce stand that had been thinned the previous winter. Heterobasidion parviporum was found in 17.5% of the old stumps of the previous spruce generation. Nine genets were identified on the study plot; seven of them were present in old stumps of the previous tree generation and two only in the new spruce generation. Eighteen spruce trees of the new generation were infected, 15 of them by vegetative growth of genets originating from the old stumps. The study plot was investigated again in 2005. No new genets had been established after thinning, and three old genets had died out. The remaining genets had infected five new trees, most likely from the thinning stumps of diseased trees. At the age of 56 years, 16.1% of the residual spruces were infected by Heterobasidion. The results of this study suggest that if spore infection to stumps of spruce can be prevented, the decay frequency caused by H. parviporum will not necessarily increase in successive generations.     

Rhizomorph production and stump colonization by co‐occurring Armillaria cepistipes and Armillaria ostoyae: an experimental study

In managed spruce forests, Armillaria cepistipes and A. ostoyae are efficient stump colonizers and may compete for these resources when they co‐occur at the same site. The aim of this experiment was to quantify the mutual competitive ability of the two Armillaria species in producing rhizomorphs and in colonizing Norway spruce (Picea abies) stumps. Five isolates of A. cepistipes and two isolates of A. ostoyae were simultaneously inoculated pair‐wise into pots containing a 4‐year‐old spruce seedling. For comparison, each isolate was also inoculated alone. One year after inoculation, stumps were created by cutting down the seedlings. Six months after creation of the stumps, rhizomorph production and stump colonization were assessed. Armillaria spp. were identified from 347 rhizomorphs and 48 colonized stumps. Armillaria cepistipes dominated both as rhizomorphs in the soil and on the stumps. Nevertheless, A. ostoyae was relatively more frequent on the stumps than in the soil and A. cepistipes was relatively more frequent in the soil than on the stumps. In both species, the ability to colonize the stumps in simultaneous inoculations was significantly reduced compared with single inoculations. In respect to rhizomorph production, simultaneous co‐inoculations had a slightly stimulatory effect on A. cepistipes and no significant effect on A. ostoyae. Our study suggests a rather neutralistic co‐existence of A. cepistipes and A. ostoyae as rhizomorphs in the soil. Concerning the ability to colonize stumps, the two species experience a mutual negative effect from the interaction, probably because of interspecific competition.     

Incidence of Heterobasidion annosum in precommercial thinning stumps in coastal British Columbia

Coniferous stumps in 83 stands in coastal British Columbia were sampled 3-5 years after precommercial thinning. The percentage of stumps and surface area colonized by Heterobasidion annosum were determined for 25 stumps of each species in each 5-cm diameter class present in each stand. There were significant differences among species in the percentages of stumps and surface area colonized, with Douglas-fir (Pseudotsuga menziesii) having the lowest values, amabilis fir (Abies amabilis) and Sitka spruce (Picea sitchensis) the highest and western hemlock (Tsuga heterophylla) being intermediate. For stumps of each species 5–20 cm in diameter, both the percentage of stumps and surface area colonized increased with increasing diameter. In stumps that were grafted to an adjacent tree, there was decreased incidence of H. annosum for Douglas-fir and Sitka spruce and increased incidence for western hemlock and amabilis fir. There were trends in the percentage of stumps and area colonized for season of thinning and biogeoclimatic subzones with the values for most species decreasing as the amount of precipitation increased. Colonization of precommercial thinning stumps by H. annosum occurs throughout the coastal region of British Columbia, and this will increase the amount of inoculum and will likely increase the incidence of butt rot. The results of this study suggest that the increase in inoculum can be minimized by thinning before age 15, by cutting only trees less than 10 cm in diameter and by thinning during low risk seasons.     

Effectiveness of treatment of Norway spruce stumps with Phlebiopsis gigantea at different rates of coverage for the control of Heterobasidion

The natural establishment of the root and butt rot causing fungus Heterobasidion annosum s.l. on Norway spruce (Picea abies) thinning stumps treated with Phlebiopsis gigantea was investigated on seven sites in southern Sweden. The trees were cut during summertime and the stumps were treated with different patterns simulating the effect of mechanical stump treatment with a single‐grip harvester. Sampling was conducted 3 and 12 months after treatment. At both samplings, the best control was obtained when 100% of the stump surface was covered by P. gigantea: in contrast, untreated control stumps showed the highest incidences of H. annosum s.l. infection at both sampling times. However, 30 and 26% of the fully covered stumps at the first and second samplings, respectively, were diseased, and question the efficacy of treating Norway spruce stumps with this biological control agent in Sweden.     

Susceptibility of Sitka spruce and grand fir trees to decay by Heterobasidion annosum

The susceptibility of Sitka spruce (Picea sitchensis) and grand fir (Abies grandis) to decay by Heterobasidion annosum was compared on a site with a previous history of serious disease. The incidence of decay 15–16 years after planting was 1.5% in grand fir, compared with 34.8% in Sitka spruce. These results are broadly similar to those reported from other trials. More information is required for older trees in Britain but the evidence so far available suggests that grand fir is more resistant to H. annosum than Sitka spruce. Planting grand fir on severely infested sites may therefore significantly reduce the serious losses that could be expected with Sitka spruce. In 14 out of 271 decayed Sitka spruce H. annosum was absent from the middle or base of the decay column but present at the top. Approximately 3% of infected spruce showed features suggesting some degree of resistance. Resin bleeding was observed at the base of 23.2% of the infected spruce, and the mean height of the decay column in these trees was significantly greater (0.96 m) than in those without this feature (0.66 m). The inoculum for infection was provided by stumps of the previous crop, and the genets of H. annosum that were present in some infected trees were also identified in stumps. The majority of infected trees contained only a single genet but three trees contained two genets. In this young crop individual genets were smaller than those found elsewhere; the largest extended to include six trees.     

Evaluation of the biological control agent Rotstop in controlling the infection of spruce and pine stumps by Heterobasidion in Latvia

The biological control agent Rotstop^® composed of a suspension of spores of Phlebiopsis gigantea (Fr.) Jül. is widely used for protecting conifer stumps from aerial infection by Heterobasidion species. The efficacy of Rotstop application on Norway spruce (Picea abies) and Scots pine (Pinus sylvestris) stumps was determined in several locations and at different seasons in Latvia. Mean efficacy in controlling natural infection by Heterobasidion spp. in spruce stumps was 64%, calculated on the basis of number of infected stumps, and 89%, calculated on the basis of area of infected wood on sample discs cut from the stumps. Corresponding proportions for pine were 82% and 95%. The results show that Rotstop can be successfully used for stump treatment in Latvia, although improved efficacy is desirable, particularly in spruce. A Latvian isolate of P. gigantea, selected from numerous isolates in preliminary tests, was included in one experiment and was shown to be as effective as the Rotstop isolate. In untreated spruce stumps Heterobasidion spp. and P. gigantea were present in the same stump three times more frequently than in untreated pine stumps. Heterobasidion spp. infection in untreated spruce stumps was low when P. gigantea covered more than 10% of stump dissection.     

Stump infection by Armillaria in first-rotation conifers

Information about the entry of Armillaria into first-rotation pine and spruce stands was obtained by searching for infected stumps, rhizomorph systems or trees that had been killed. In pines Armillaria foci were very rare. In pure Norway spruce Armillaria lutea and A. mellea were detected in stumps but rhizomorphs did not extend into the soil; in Norway spruce mixed with oak, by contrast, A. lutea sometimes produced extensive rhizomorph systems. In Sitka spruce small groups of trees had been killed by A. ostoyae. All foci investigated in conifers contained different genotypes of Armillaria and probably originated from spore infection of stumps created by thinning. Some implications of these findings are discussed.     

White spruce regeneration following a major spruce beetle outbreak in forests on the Kenai Peninsula, Alaska

Between 1987 and 2000, a spruce beetle (Dendroctonus rufipennis) outbreak infested 1.19 million ha of spruce (Picea spp.) forests in Alaska, killing most of the large diameter trees. We evaluated whether these forests would recover to their pre-outbreak density, and determined the site conditions on which spruce germinated and survived following the spruce beetle outbreak in forests of the Anchor River watershed, Kenai Peninsula, Alaska. White spruce (Picea glauca) and Lutz's spruce (Picea × lutzii), a hybrid between white and Sitka spruce (Picea sitchensis), dominate the study area. We measured the pre- and post-outbreak density of spruce in 108 3 m × 80 m plots across the study area by recording all live trees and all dead trees >1.5 m tall in each plot. To determine the fine scale site conditions on which spruce germinated and survived, we measured ground surface and substrate characteristics within 20 cm circular plots around a subset of post-outbreak spruce seedlings. The density of post-outbreak spruce (855/ha) was adequate to restock the stands to their pre-outbreak densities (643/ha) for trees >1.5 m tall. We could not accurately estimate recovery for pre-outbreak spruce seedlings because dead seedlings may have decayed in the 5–18 years since the beetle outbreak occurred. At the fine scale, spruce that germinated post-outbreak grew on a wide variety of substrates including downed log, stump, mesic organic mat, peat, hummocks and mineral soil. They exhibited a strong preference for downed logs (53%) and stumps (4%), and most (91%) of the downed logs and stumps that spruce rooted on were heavily decayed. This preference for heavily decayed logs and stumps was especially evident given that their combined mean cover was only 2% in the 3 m × 80 m plots. Within the 3 m × 80 m plots, spruce seedling survival was negatively correlated with bluejoint (Calamagrostis canadensis) litter cover.     

Controlling and predicting the spread of heterobasidion annosum from infected stumps and trees of picea abies

Two Norway spruce stands with heavy infections of Heterobasidion annosum were clear‐cut in 1957 and 1959 in Sweden. The stumps were extracted, the soil sifted to remove most of the roots, and young Norway spruce were planted. After 25 and 28 years, H. annosum had infected 1 % and 2% of trees on plots where stumps had been removed and 17 % and 12% of the trees on control plots, respectively. Several of the H. annosum clones fruiting on old‐growth stumps were also detected in decayed, standing trees. The same fungal clone was found to be infecting adjacent trees from several old‐growth stumps. In addition to old stumps, stumps from recent thinnings and diseased living trees were traced as infection sources. Their relative importance in spreading disease was estimated. Disease risk predictions based on the distance of a tree from various infection sources correlated well with observed frequencies of rot.     

Stocks and decay dynamics of above- and belowground coarse woody debris in managed Sitka spruce forests in Ireland

Coarse woody debris (CWD) has become recognised as an important component of the carbon (C) pool in forest ecosystems. In Ireland, managed Sitka spruce (Picea sitchensis (Bong) Carr.) forests account for 52.3% of the total forest estate. To determine the stock and decay dynamics of above and belowground CWD, field surveys using fixed area sample plots, were conducted in six even-aged Sitka spruce stands, representing the young, intermediate and mature stages of a typical commercial rotation. The volume, mass, density loss and C:N ratio of all CWD types (logs, stumps, and coarse roots) were determined using a five-decay class (DC) system. The decay rates and half life of CWD was also determined. To estimate CWD coarse root mass; roots associated with stumps classified in different decay classes were excavated. The coarse roots were categorised into small (2-10 mm), medium (10-50 mm) and large (>50 mm) diameter classes.CWD C-mass ranged from 6.98 to 18.62 Mg ha⁻¹ and was highest in an intermediate forest (D35), while the aboveground volume varied from 6.31 to 42.27 m³ ha⁻¹. Coarse roots accounted for 21% to 85% of the total CWD C-pool in the surveyed stands. The total CWD C-mass was poorly correlated with the number of thinning events (R² = 0.29), when data from D35 was excluded. The density loss was significant in logs (45%), stumps (58%), and small- (38%), medium- (50%) and large roots (38%) as decay progress from DC 0 to 4. There was a 46%, 41%, 51%, 72% and 57% decline in C:N ratio of logs, stumps, small-, medium- and large roots, respectively, as decay progressed from DC 0 to 4. The density decay rates were 0.059, 0.048 and 0.036 kg m⁻³ year⁻¹ for logs, stumps and coarse roots, respectively. The size classification of roots did not significantly affect their decay rate. The half life (50% decomposition) of CWD was estimated has 12-, 14- and 19 years for logs, stumps and roots of Sitka spruce. Regression curves showed a strong correlation between the density and C:N ratio (R² = 0.69, 0.74 and 0.93 for logs, stumps and coarse roots, respectively). The long term storage of C and its slow rate of decomposition make CWD a vital structural and functional component of the CWD C-pool and a major controller of forest ecosystem C-retention.     

The spreading of the S type of Heterobasidion annosum from Norway spruce stumps to the subsequent tree stand

The occurrence of Heterobasidion annosum in stumps and growing trees was investigated on 15 forest sites in southern Finland where the previous tree stand had been Norway spruce (Picea abies) infected by H. annosum, and the present stand was either Scots pine (Pinus sylvestris), lodgepole pine (Pinus contorta), Siberian larch (Larix siberica), silver birch (Betula pendula) or Norway spruce 8–53 years old. Out of 712 spruce stumps investigated of the previous tree stand, 26.3% were infected by the S group and 0.3% by the P group of H. annosum. The fungus was alive and the fruit bodies were active even in stumps cut 46 years ago. In the subsequent stand, the proportion of trees with root rot increased in spruce stands and decreased in stands of other tree species. On average, one S type genet spreading from an old spruce stump had infected 3.0 trees in the following spruce stand, 0.5 trees in lodgepole pine, 0.3 trees in Siberian larch, 0.05 trees in Scots pine and 0.03 trees in silver birch stand. Although silver birch generally was highly resistant to the S type of H. annosum, infected trees were found on one site that was planted with birch of a very northern provenance.