Infection of Picea sitchensis and Pinus contorta stumps by basidiospores of Heterobasidion annosum

Infection of Pinus contorta and Picea sitchensis stumps by basidiospores of Heterobasidion annosum is extremely variable, both within and between sites, but in general P. sitchensis stumps are less sus-ceptible than those of P. contorta. Measurement of the cross-sectional area occupied by H. annosum on each stump provides a more sensitive test of species susceptibility than assessment of the proportion of stumps infected. P. sitchensis stumps become infected on a variety of soils but there is evidence to suggest that infection may be reduced by high rainfall. In some infected stumps, H. annosum is confined to the lower stump tissues. Its absence from the upper portion of the stump may be due to replacement by other micro-organisms or, alternatively, physical conditions in the upper stump tissues may prevent its continued survival after infection has taken place. In both species, but more commonly in P. sitchensis, some stumps remain alive for at least two years after felling, particularly on peat soils, due to the presence of root grafts with neighbouring trees. Results for P. sitchensis suggest that infection occurs more readily in living stumps than in those which die rapidly after felling. The viability of H. annosum basidiospore suspensions can be determined more accurately and more rapidly on a selective agar medium than on conifer stem sections.     

Intraspecific variation in Heterobasidion annosum for growth in sapwood of Picea abies and Pinus sylvestris

Two greenhouse experiments were conducted to study intraspecific variation in growth of the root rot fungus Heterobasidion annosum in living host sapwood. In experiment 1, Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) seedlings were inoculated with H. annosum isolates, 14 each of the S-and P-intersterility groups, collected from various parts of Sweden. In pine, the P-group isolates were more virulent than the S-group isolates both in terms of infection frequency, induced mortality rate (p < 0.05), and fungal growth in sapwood (p < 0.05). In spruce, the P-group isolates were also more virulent on average, but the difference was not statistically significant. Both S and P isolates had a higher infection frequency and a significantly longer sapwood growth on spruce than on pine. The P-group caused higher mortality on pine than on spruce. The length of the lesion in the inner bark was strongly correlated with fungal growth in spruce, but not in pine where the lesions were short or absent. In experiment 2, ten Norway spruce clones were inoculated with 18 S-isolates, originating from nine live-decayed trees and from nine spore-infected stumps in a single Norway spruce stand. The objective was to test whether any selection for growth rate in sapwood was detectable among individuals of H. annosum originating either from stumps or trees. The results gave no support for such selection since no difference in sapwood growth between the two groups of isolates was found.     

Biological control of Heterobasidion annosum on Pinus pinea by Phlebiopsis gigantea

The efficacy of a native isolate of Phlebiopsis gigantea in protecting the stumps of Pinus pinea against spore infection by Heterobasidion annosum s.str. was investigated. In preliminary tests carried out in stem pieces of P. pinea in confined environment, the efficacy of the isolate was compared with Rotstop^®, the commercial formulation prepared from a north European P. gigantea strain. Both showed a fully protective effect against artificial inoculation of H. annosum. Moreover, in stump treatment experiments carried out in a coastal P. pinea forest close to Rome, the native P. gigantea was effective against heavy airborne inoculum of H. annosum. Some natural infection by P. gigantea occurred in the stumps, but it was unable to control the pathogen. Random amplified microsatellite analysis allowed to recognize the presence of different P. gigantea strains in the forest.     

Liberation of Heterobasidion annosum conidia by airflow

The effect of speed and humidity of the airflow and the míst sprayed with airflow on the liberation of conidia of Heterobasidion annosum from pure cultures was studied in a small windtunnel. The conidia were liberated more easily in humid than in dry airflow. Liberation was most abundant when mist was added to the airflow. Threshold speed for the liberation of conidia was calculated to vary between 1.8 m/s and 3.9 m/s. According to the results, conidia of H. annosum may be liberated into the air by wind gusts associated with high humidity or mist and thus the conidia may have a role in aerial dispersal, although this is probably small compared with basidiospores.     

Compartmentalization of decay associated with Heterobasidion annosum in roots of Pinus resinosa

Decay associated with Heterobasidion annosum was compartmentalized in roots of Pinus resinosa.     

Infection of roots of Norway spruce (Picea abies) by Heterobasidion annosum

Wounding of excised roots of Norway spruce [Picea abies (L.) Karst.] caused a rapid accumulation of phenolic compounds, decrease of starch content and of certain enzyme activities in the ray cells. Infection of the wounds by Heterobasidion annosum (Fr.) Bref. intensified these processes, caused oxidation of the phenols, and induced activity of acid phosphatase and β-glucosidase in the ray cells of the infected area. The reactions were oxygen-dependent. Intra- and extracellular enzymes were detected in the wood-infecting hyphae. Other wood-growing fungi caused similar reactions in the wood as H. annosum.     

Infection studies with Heterobasidion annosum on young trees of Picea abies

Infection studies with Heterobasidion annosum were carried out on 15-year-old, cloned spruces between 1971 and 1978. The inoculum was introduced into the sapwood of the test plants at the base of the stem. All inoculations, conducted at different times of the year, were uniformly successful. The pathogen spread through the xylem tissue rapidly during the vegetation period, but from early autumn onwards gradually disappeared over the following two years. After this period, the fungus could no longer be detected.     

S and P intersterility groups in Heterobasidion annosum; Infection frequencies through bark of Picea abies and Pinus sylvestris seedlings and in vitro growth rates at different oxygen levels

Seedlings of Norway Spruce and Scots Pine were inoculated with S and P strains of the root rot fungus Heterobasidion annosum. Infection frequencies through the living bark layers were not related to subgroup affiliation in neither spruces nor pines. Growth rates in vitro were not related to growth rates in vivo. Oxygen concentration had a stronger effect on the growth rates of P than on S strains.     

Growth rates of Heterobasidion annosum s.s. and H. parviporum in functional sapwood of Pinus sylvestris and Picea abies

Growth rates of H. annosum s.s. and H. parviporum were investigated in the functional sapwood of young Pinus sylvestris and Picea abies plants as an indicator of the relative susceptibilities of the hosts to these pathogens. The stems of 520 five‐year‐old P. abies and 321 four‐year‐old P. sylvestris plants were inoculated and the extent of infection determined 16 weeks later. H. annosum s.s. grew further than H. parviporum in P. sylvestris sapwood, while in P. abies, no differences between the two Heterobasidion spp. were found. Both H. annosum s.s. and H. parviporum spread faster in the sapwood of P. abies than in P. sylvestris. There was high within‐host species variation in growth rates for both P. sylvestris and P. abies suggesting it may be possible to identify tree genotypes with lower susceptibility.     

North American P type of Heterobasidion annosum shows pathogenicity towards Pinus halepensis in Italy

During a survey on the distribution of Heterobasidion root rot, the North American P group of H. annosum was recorded causing root rot on Aleppo pine, Pinus halepensis, in the gardens of a historical villa in Rome. This is the first report of Heterobasidion root rot on P. halepensis in its natural distribution area, the Mediterranean Basin.     

Analysis of microtubule and microfilament distribution in Pinus sylvestris roots following infection by Heterobasidion species

Cytoskeletal dynamics play a crucial role in pathogen recognition and cell defence during the initial interactions between an invader and plant host. The aim of the work reported here was to characterize how Heterobasidion annosum s.s., Heterobasidion parviporum, and Heterobasidion abietinum affect the microtubules and microfilaments of Pinus sylvestris root cells 12‐, 24‐, 48‐, and 96‐h post‐inoculation. Inoculation of P. sylvestris with H. parviporum or H. abietinum, which have a lower specificity for P. sylvestris than H. annosum s.s, resulted in greater reorganization of host microtubules during the early stages of interaction than inoculation with the more specific H. annosum s.s. In some infected cells, spots of actin aggregates were observed. Disruption of cytoskeletal components by the application of specific cytoskeletal inhibitors facilitated the entry of the H. parviporum and H. abietinum into roots. These results suggest that the P. sylvestris cytoskeleton plays a role in the host response in the initial stages of the host–pathogen interaction.     

In vitro inhibition of Heterobasidion annosum by Phaeotheca dimorphospora

The antagonistic fungus Phaeotheca dimorphospora has been tested in vitro on agar plates and wood discs to evaluate its potential use against P (Eastern Canada) and S (Finland) intersterility groups strains of Heterobasidion annosum using two methods. Production of antifungal metabolites by P. dimorphospora was demonstrated on agar plates as well as on Pinus resinosa and Picea abies discs using the bi‐layer technique. Diffusible metabolites inhibited the growth of the pathogen on both substrates. Direct confrontation on wood discs showed that a 7‐day period of advanced growth by P. dimorphospora completely prevented the colonization of the tissue by H. annosum. Control was partial when the antagonist had only one day of advanced growth. Growth inhibition was superior against S group strains with less than 1% of the samples colonized by H. annosum compared with 22% for P group strains. Moreover, P. dimorphospora was isolated in more than 83% of the samples in the absence of the pathogen in both wood species. In vitro tests demonstrated that P. dimorphospora is able to colonize red pine as well as Norway spruce tissue and to use wood components for the production of antifungal metabolites.     

Pathogenicity of Spanish isolates of Heterobasidion annosum s. s. in Pinus pinaster seedlings

The aim of this study was to test the pathogenicity of two Spanish isolates of Heterobasidion annosum sensu stricto in 2‐year‐old Pinus pinaster seedlings. Two types of inocula (woodchips and sawdust) were used to infect the seedlings by two different routes (stem inoculation and soil infestation). The mortality rates of the stem‐inoculated seedlings differed significantly from controls, but those of the seedlings infected via soil infestation did not differ. For both types of inoculation, the lesions were longer, and wilting symptoms were more severe in the seedlings inoculated with H. annosum than in control seedlings. For stem inoculation, biomass allocation did not differ significantly between the infected and control seedlings. However, the percentage of fine roots was lower in seedlings infected via soil infestation than in the control seedlings. To our knowledge, this is the first pathogenicity test with H. annosum isolates and P. pinaster.     

Two studies to assess the risk to Pinus sylvestris from Heterobasidion spp. in southern Sweden

Abstract

Pine stumps are not being treated against Heterobasidion spp. in Sweden. To determine whether they should be, the frequency of stump infections and the species of Heterobasidion involved were investigated in nine newly thinned pure Scots pine (Pinus sylvestris L.) stands in southern Sweden. The incidence of Heterobasidion was measured in roots of standing Scots pine in another 15 stands. Infections by both H. annosum (Fr.) Bref. s.s. and H. parviporum Niemelä & Korhonen were numerous in stumps six months after a summer thinning. The pathogen, mostly H. annosum, was found in 44 of 60 sampled root systems, from 14 of the 15 stands. Twenty of the infected pines were assessed as healthy on the basis of crown symptoms while 24 trees had defoliated crowns. Infected root systems were most frequent among trees with thin crowns in stands on former agricultural land, where previous thinnings had been carried out during the growing season when airborne spores are plentiful. The study suggests that stump treatment may be a profitable way to reduce disease development in Scots pine monocultures on sandy soils as well as in mixtures with Norway spruce [Picea abies (L.) Karst.] on any soil.     

An intersterility group of Heterobasidion annosum specialized to Abies alba

A third intersterility group of H. annosum in Europe has been identified from Italy. It is specialized to Abies alba., and hence is called the F (fir) group. It is closely related to the European S group, but is sexually incompatible with the geographically adjacent S population occurring in the Alps.     

Variation in Heterobasidion annosum detected by Random Amplified Polymorphic DNAs

The production of Random Amplified Polymorphic DNAs (RAPDs) by Polymerase Chain Reaction (PCR) was used to detect variation in the isolates of Heterobasidion annosum with various geographical origins. Specific RAPD products were detected for each of the intersterility groups: the European S, F, P and the North American S group. There was considerably more polymorphism found among European S than P isolates. The analysis of RAPDs was shown to be a simple and a fast way to generate DNA markers specific to the previously established intersterility groups and thus useful for diagnostic purposes.     

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.