The efficacy of selected biological and chemical control agents against Heterobasidion abietinum on Abies cilicica

The efficacy of 30% aqueous urea solution, borax powder and spore suspensions of Phlebiopsis gigantea and Trichoderma harzianum against establishment of artificially inoculated Heterobasidion abietinum on Abies cilicica was tested both in the field and in a simulated stump treatment experiment carried out in a growth chamber. In the field, in each of the four selected stands 20 fresh stumps per control agent were treated and 20 stumps were left as untreated controls. In two of the stands, the treatments were applied in June and in the two others in November. Stumps were sampled twice, at 6‐ and 12‐months after treatment. In the growth chamber, 10 log pieces per treatment and control were used, and sampling performed after 6‐week incubation. Results of the urea and borax treatments were consistent between the experiments; the mean efficacies were 98.8 and 99.4% in the growth chamber, and 80.2–91.3 and 89.4–90.1% in the two samplings of the field experiment, respectively. Despite the high efficacies of the P. gigantea (85.9%) and T. harzianum (97.5%) treatments in the growth chamber, efficacies of these biological control agents in the field were 47.1–49.2 and 61.3–65.5%, respectively.     

Heterobasidion annosum infection of Picea abies following manual or mechanized stump treatment

Heterobasidion annosum (Fr.) Bref. s. l. colonization following thinning was studied in 1246 stumps of Picea abies (L.) Karst. trees from 14 previously unthinned stands in Sweden. Treatments included mechanized and manual application of (1) 35% urea solution, (2) Phlebiopsis gigantea (Fr.) Jül. spores, and (3) 5% solution of disodium octaborate tetrahydrate (DOT), and untreated stumps, cut in the summer and winter. Compared with untreated stumps cut in the summer, all stump treatments and winter cuttings significantly reduced the colonized stump area 6–7 weeks after thinning by 88–99%. Mechanized stump treatment provided as good protection as manual treatment against H. annosum infections. The probability of spore infection (p _ij ) was reduced by 53–83% in mechanized treatment and 79–98% in manual treatment compared with untreated summer thinning. In terms of p _ij , urea had significantly higher control efficacy than P. gigantea and manual treatment performed better than mechanized treatment.     

Effects of Stump-treatment Substances for Root-rot Control on Ground Vegetation and Soil Properties in a Picea abies forest in Sweden

There is increasing commercial interest in treating stumps to restrict the spread of root rot [Heterobasidion annosum (Fr.) Bref.] when thinning Swedish coniferous forests during the summer. Both chemical and biological substances are used for this purpose. During the treatment, however, a large proportion of the substance applied is spilled beside the stumps. A field study was conducted on the effects of stump-treatment substances on various ground-vegetation species in a Picea abies (L.) Karst. forest in Sweden. Three different substances commonly used in forestry were studied: urea solution (0.23 kg N m^-2), borate solution (10 g B m^-2) and a fungal preparation of Phlebiopsis gigantea (Fr.) Jül. spores (1 g spores m^-2). The principal objectives were to assess whether any of the substances were harmful to plants and whether plant species differed in their sensitivity. Both borate and urea solution caused severe damage to most ground-vegetation species tested. Bryophytes were affected more strongly than vascular plants and urea was slightly more toxic than borate. Treatments with P. gigantea caused no obvious damage. The size and persistence of chemical changes in the soil induced by the treatments were also analysed. Transient changes were apparent in topsoil where borate or urea had been added. Very high concentrations of B were initially observed where borate had been applied, and even after 1 yr they were slightly higher than the threshold concentration at which plant injuries are expected. Urea treatment initially resulted in a pH increase of 2 units and a substantial increase in soil ammonium content. After 1 yr these effects had largely disappeared, although some increase in ammonium was still detectable.     

The effect of Phlebiopsis gigantea and urea stump treatment against spore infection of Heterobasidion spp. on hybrid larch (Larix × eurolepis) in southern Sweden

The efficacy of stump treatment with 40% urea solution and spore suspension of Phlebiopsis gigantea against primary infection by Heterobasidion spp. on hybrid larch (Larix × eurolepis) was tested in two field studies in southern Sweden. In the first study, stumps treated with urea or P. gigantea were sampled by cutting a cross‐section disc after 3 months following exposure to natural conditions, and in the second study, stumps treated with only P. gigantea were sampled after 2 months. Spore traps made from hybrid larch, Norway spruce (Picea abies) and Scots pine (Pinus sylvestris) were used to estimate the abundance of ambient spores of Heterobasidion spp. in the second study. Urea significantly reduced infection frequency of Heterobasidion spp. compared to the control. Phlebiopsis gigantea was less effective at reducing infection frequency in the first study than in the second study. Infections were caused by both H. parviporum and H. annosum sensu stricto. The amount of H. annosum infection in proportion to Heterobasidion spp. was significantly lower on the stumps treated with P. gigantea compared to the control; however, the proportion of H. parviporum on the treated stumps was not higher than the control in the first study. Spore traps made of Scots pine had a significantly higher frequency of infection than Norway spruce and hybrid larch. Only the spore traps made of hybrid larch showed significant correlation with the control stumps in terms of relative infected area. Conclusively, it seems prudent to protect hybrid larch stumps from primary infection by Heterobasidion spp., and both urea and P. gigantea can be recommended as stump treatment agents on hybrid larch, even if urea seemed to present more stable results.     

Relationship between stump treatment coverage using the biological control product PG Suspension,and control of Heterobasidion annosum on Corsican pine,Pinus nigra ssp. laricio

The relationship between the proportion of the stump surface covered by the biological stump treatment agent PG Suspension, containing Phlebiopsis gigantea and its efficacy against the pathogen Heterobasidion annosum sensu stricto was studied during a first thinning of Corsican pine (Pinus nigra ssp. laricio) in Thetford Forest, UK. PG Suspension was manually applied to 100%, 75%, 50% or 0% of the surface of 150 stumps. Spores of H. annosum were inoculated onto 75 of the stumps, and the remaining stumps exposed to natural airborne spore deposition. The relationship between coverage and efficacy was found to be quantitative. Covering all the stump surface with PG Suspension completely excluded the pathogen, whereas stumps not treated with PG Suspension (the 0% treatment) became infected with H. annosum. Partial (75%) PG Suspension coverage resulted in the pathogen colonizing 40% of stumps following artificial inoculation with H. annosum, and just 7% of stumps exposed to ambient H. annosum spore infection. Decreasing levels of coverage allowed increasing areas of the stump surface to be colonized by H. annosum. Some small gaps in coverage were closed by lateral growth of P. gigantea, but it is recommended that operators aim for full stump coverage to give complete protection against H. annosum.     

A trial of disodium octaborate tetrahydrate for the control of Heterobasidion annosum

The extent of heartwood infection of Sitka spruce Picea sitchensis stumps following inoculation with basidiospores of Heterobasidion annosum was significantly reduced by application of 2% and 4% solutions of disodium octaborate tetrahydrate (DOT) immediately after felling. Infection of sapwood was so rare in all treatments, including controls, that the effectiveness of DOT in this tissue could not be judged. The results of this trial suggest that DOT may be toxic to H. annosum at lower levels of borate loading than previously thought possible. Over the 10 months of the experiment the borate loading in the stumps appeared to reach an equilibrium that was independent of the initial concentration applied. Stump-surface fruiting of Melanotus proteus, a potential fungal competitor of H. annosum, was not inhibited by these treatments.     

Presence of Heterobasidion infections in Norway spruce stumps 6 years after treatment with Phlebiopsis gigantea

Natural colonization by the root and butt rot causing fungi Heterobasidion spp. on Norway spruce (Picea abies) stumps following thinning and treatment with the biological control agent Phlebiopsis gigantea was investigated on three sites in southern Sweden 6 years after treatment. The fully treated stumps and control stumps were excavated and sampled to compare the survival of Heterobasidion spp. in the long term. Six years post‐treatment, 47 and 11% of untreated and treated stumps, respectively, had Heterobasidion infection. There was no difference in the relative infected area in discs collected from the butt and the roots for the different treatments. Control efficacy was 83% for treated stumps. After 6 years, there were no apparent differences between the remaining infections in treated compared with those in untreated stumps regarding the number of colonies, their size or relative infection area. Although infections, 3 months after treatment with P. gigantea, were significantly fewer and smaller than in untreated stumps, Heterobasidion inoculum can survive for at least 6 years in the stump and, when it does, constitute a risk for neighbouring 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.     

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.     

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.     

Armillaria ostoyae in managed coniferous forests in Kastamonu in Turkey

Although several Armillaria species have been reported in Turkey, there is little information about their ecology in Turkish forests. In this study, we investigated five forest stands, approximately 5–74 ha in size, in Kastamonu province in the Black Sea Region of Turkey for the presence of Armillaria species in stumps and logs. The stands were mixed Abies nordmanniana ssp. bornmülleriana and Pinus sylvestris forests managed using a selective cuttings system; the proportion of fir in the total number of stems and stumps ranged from 36 to 98%. Based on sequence analysis of the internal transcribed spacer and intergenic spacer regions of the rDNA, all rhizomorphs sampled from the stumps and logs were of Armillaria ostoyae. The size of the genets was estimated with random amplified microsatellites analysis of the isolates and ranged from single stumps to approximately 450 m². One to seven genets were found in each stand. These results indicate that the genets had arisen from spores and vegetative spread was limited on most sites.     

Efficacy of different concentrations of Rotstop® and Rotstop®S and imperfect coverage of Rotstop®S against Heterobasidion spp. spore infections on Norway spruce stumps

Two Rotstop^® preparations, one containing a Phlebiopsis gigantea strain from Finland (Rotstop F) and the other one from Sweden (Rotstop S), were used in different concentrations to treat thinning stumps of Picea abies against spore infections by Heterobasidion spp. in southern Sweden. Trees were cut on three sites during the summer of 2004, and 285 stumps were treated manually ensuring 100% coverage. Spore concentrations in the treatment suspensions were ca 5 × 10⁶ and 10 × 10⁶ spores/l, and approximately 10 ml of suspension was applied per 100 cm² of stump surface. An additional 31 stumps on one of the sites were treated mechanically with Rotstop S; in this treatment the spore concentration was high, about 20 × 10⁶ spores/l, but the coverage was incomplete. Three months later there was a significant reduction in frequency and relative areas of Heterobasidion spp. infections on stumps with manual treatment compared with the untreated stumps. However, there was no significant difference between the preparations or the concentration of active ingredient in terms of their reduction of Heterobasidion infection. Mechanical treatment with incomplete coverage failed to control infection. Therefore, in conditions of moderate Heterobasidion spore load in the environment there seems to be no reason to use higher concentrations of P. gigantea in the treatment of spruce stumps.     

A comparison of the cost-effectiveness of different eucalypt cut-stump control management options to reduce competition from coppice regrowth during stand establishment in Mpumalanga,South Africa

Selected cut-stump (Cut_surface) and basal frill (Basal_frill) treatments were tested on Eucalyptus macarthurii cut stumps that had previously been coppiced multiple times. Treatments included the application of herbicide (triclopyr as an amine salt, 360 g L^?1), either to the cut-surface or to a basal frill (cuts made through the bark and into the cambium at the base of the tree). These treatments were integrated with non-chemical control methods, where seedlings were planted either between each stump row or within the row, to determine whether such treatments could further reduce the level of coppicing. Stump mortality for both the Basal_frill and Cut_surface treatments were equally effective, with 87% of the stumps killed when assessed at 14 weeks following initial treatment. No significant treatment differences were detected for the various growth variates of the stand at rotation end (stocking, basal area and volume). This can be attributed to a lack of competition from any coppice regrowth due to good control on each occasion. As there were no significant differences between the treatments, the Cut_surface treatment would be preferable due to the lower overall costs. However, for this method to be most effective, it is important that the herbicide be applied to the cut-surface of the felled tree within one day of felling. This may pose a safety issue, especially with mechanised felling of a stand. Further research on extending the period for application to the cut-surface could be beneficial. However, if reduced herbicide use is a prerequisite, coppice regrowth can be manually removed.     

Community of Aphyllophorales and Root Rot in Stumps of Picea abies on Clear-felled Forest Sites in Lithuania

The community of Aphyllophorales fungi in stumps of Picea abies (L.) Karst. and the occurrence of root rot caused by Heterobasidion annosum were investigated at 38 clear-felled sites in Lithuania. Fruit-bodies were recorded on 36.0% of the 3924 examined stumps. The most common were Bjerkandera adusta, Phlebiopsis gigantea, Fomitopsis pinicola, Gloeophyllum sepiarium, Stereum sanguinolentum, Trametes zonata and H. annosum. All species occurred in stumps both with and without established root rot, but B. adusta, F. pinicola, G. sepiarium, T. zonata and Trichaptum abietinum occurred less often in stumps containing root rot, while H. annosum was encountered more often. Greater numbers of fruit-bodies were found on stumps cut in summer than on those cut in winter. The fruiting became most abundant 3-4 yrs after the trees were felled. The abundance of nearly all Aphyllophorales correlated positively with the stump diameter, and on larger stumps, fruit-bodies of several different fungi were more common. The number of stumps containing H. annosum root rot in different sites varied from 9.8% to 68.8%, and was 27.6% on average. The incidence of root rot correlated neither with the age nor with the density of the felled stand, but it correlated negatively with the proportion of deciduous trees within a stand.     

Development of suckers by two‐year‐old birch (Betula pendula Roth) at different temperatures and light intensities

Two‐year‐old seedlings of Betula pendula Roth of three provenances were cut to stump heights of 0 and 10 cm. The birch stumps were exposed to different light levels (25–400 μEm^?2s^?1) and temperatures (6–24°C) for 100 days in climate chambers. Birches were also cut down to seven stump heights (0, 2, 5, 10, 15, 20 and 25 cm) and exposed to light intensity 25 and 200 μEm^?2s^?1 at 12/6°C for 100 days in climate chambers. In the third experiment 10 cm stumps of birch were exposed to different light intensities (10–400 μEm^?2s^?1) for 30 days in a greenhouse at 20°C. Starch content in root systems was analyzed before and after treatment. Ten stumps per treatment were used in the experiments except the third experiment where 15 stumps were used. The number of sprouting stumps was correlated with light intensity and temperature. None of the stumps exposed to 6 or 9°C produced suckers. At stump height 0 cm fewer sprouting stumps were produced than at 10 cm. The mean height of suckers was higher the higher the temperature both on 0 and 10 cm stumps. There were differences between provenances in height growth. The number of suckers per sprouting stump was not related to temperature or light intensity. Starch content in root systems of 10 cm high stumps was 4.0% compared with 14.7% in root systems of non‐stumped birch plants after 30 days in the greenhouse. Starch content decreased from 4.0 to 3.0% with decreasing light intensity (400–10 μEm^?2S^?1). The number of suckers and their mean height were correlated with starch content depending on light intensity.     

Modelling the incidence of Heterobasidion annosum butt rots and related economic losses in alpine mixed naturally regenerated forests of northern Italy

A total of 2257 recently felled stumps were sampled from 22 mixed naturally regenerated forest stands in the Aosta Valley, western Italian Alps, and examined for Heterobasidion annosum butt rots. Disease incidence ranged from 6% to 71% depending on sites, and H. annosum accounted for 92% of the total number of diseased stumps. H. annosum incidence was significantly higher (Chi‐squared multiple comparison tests, p < 0.05) on Norway spruce (44%) than on the other tree species (silver fir = 18%, larch = 12% and Scots pine = 6%). Based on the information on the airborne inoculum composition of the fungus, all the three European species of H. annosum were present, with a variable frequency, depending on site. A partial least squares regression analysis showed that the relative abundance of Norway spruce and four variables describing the size of trees were the best predictors for the incidence of H. annosum butt rots. A model having these variables as predictors was developed (r² = 0.75; p < 0.001) and successfully validated on five additional forest stands. An estimate of the losses in yield and value of timber associated with the disease, i.e. direct losses, was also carried out in the forests included in the survey. The percentage of direct financial losses was either comparable or lower than the average level of disease incidence.     

Modern aspects of biological control of Fomes and Armillaria1

Biological methods can help to control root-infecting fungi such as Fomes annosus and Armillaria mellea. For example, inoculation of pine stumps with the fungus Peniophora gigantea is widely used against F.annosus in Britain. Experiments arc in progress to determine whether similar methods can be used for stumps of other conifers. The main sources of infection by Armillaria are stumps of broad-leaved trees, which are often treated chemically to prevent regrowth. Some of these treatments promote colonization by saprophytic fungi that compete well with Armillaria. In experiments, direct inoculation of stumps combined with chemical treatment has given encouraging results.     

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.     

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.     

Urea treatment reduced Heterobasidion annosum s.l. root rot in Picea abies after 15 years

Stump protection using chemical or biological agents is the main control measure against root and butt rot caused by Heterobasidion annosum s.l. in northern and temperate conifer forests. Long-term effects of urea treatment of stumps are poorly known and here we describe a 15-year study of urea treatment on the rot incidence on Norway spruce (Picea abies). We also tested the effect of urea protection on tree growth and on the resistance of stands against strong winds. Four treatments were made in two replications in two first-rotation P. abies stands in southern Sweden; after first-thinning stumps were (i) treated with urea 35% (w/v), (ii) artificially infected with H. annosum conidia, (iii) half urea treated and half artificially conidia infected, (iv) untreated, therefore naturally infected. After 15 years, the trees were sampled at 20 cm above ground using an increment borer and observed for presence of rot and, following incubation, presence of H. annosum conidia. Tree growth was calculated by measuring the diameter before and after the treatment. Urea treated plots showed the lowest incidence of rot (3%) as well as of H. annosum incidence (0%). Conidia treatment showed the highest incidence of rot (68%), its incidence was higher than that observed in natural infection treatment (43%), but did not differ from that of the 50% conidia treatment (47%). On about 30% of the rotted trees we observed conspicuous H. annosum fruiting. We did not observe growth reduction associated with tree rot. H. annosum was the only fungus observed associated with rotted trees which suggest that it was responsible for most of the rot observed in the investigated plots. Urea treated plots showed the lowest incidence of windthrown trees, and 59% of the windthrown tree incidence among the plots was explained by the incidence of H. annosum. Urea can be regarded as a reliable long-term protection method against root and butt rot of Norway spruce.