Comparison of the virulence of Armillaria cepistipes and Armillaria ostoyae on four Norway spruce provenances

The basidiomycetes Armillaria cepistipes and Armillaria ostoyae frequently occur in the same forest stand. In this study, we determined the virulence of 20 isolates of A. cepistipes and 16 isolates of A. ostoyae on four different provenances of 2‐year‐old Norway spruce (Picea abies). Within 30 months after inoculation, 1.1 and 19.1% of the seedlings inoculated with A. cepistipes and A. ostoyae, respectively, had died or were dying. The incidence of dead and dying seedlings varied between 3 and 49% among the A. ostoyae isolates. The virulence of an isolate was positively correlated to its ability to produce rhizomorphs. One Norway spruce provenance showed significantly lower susceptibility to A. ostoyae than the other three. Rhizomorphs of both Armillaria species were attached to the root surface. The attached rhizomorphs of A. ostoyae, however, were associated with significantly more lesions. The virulence of the isolates was not correlated with their wood‐degrading capability for either of the Armillaria species.     

Epidemiology of Armillaria root disease in Douglas‐fir plantations in the cedar‐hemlock zone of the southern interior of British Columbia

Results are presented from several studies on the epidemiology of Armillaria ostoyae in Douglas‐fir plantations in the interior cedar‐hemlock (ICH) biogeoclimatic zone of British Columbia. Two plantations were monitored for mortality by A. ostoyae and other agents for 35 years after establishment. In these and other plantations ranging in age from 7 to 32 years, one or more of the following factors were determined: source of inoculum, mode of spread and characteristics of lesions on roots of excavated trees; symptom expression in relation to tree age and damage to the root system and years from initial infection to death on trees killed by the fungus. Mortality from A. ostoyae began in both plantations about 5 years after planting, reaching 30% in one and 11% in the other after 35 years. The spatial pattern of mortality was similar to that reported from New Zealand, France and South Africa; however, the temporal pattern differed, beginning later and, instead of declining, continuing at a nearly constant rate to the present. To age 10, nearly all infections were initiated by rhizomorphs; as plantations aged, the proportion of infections occurring at root–root contacts increased. In seven plantations, in moist and wet subzones of the ICH, from 23 to 52% of Douglas‐firs had root lesions, with the higher incidences occurring on moist sites. The occurrence of aboveground symptoms, reduced leader growth and basal resinosis, was related to the percentage of root length colonized by A. ostoyae on trees with more than 30% of root length killed. Average time from infection to death increased from 1 to 2 years at age 6 to 22 years at age 33. The outlook for timber yield from Douglas‐fir plantations in the cedar‐hemlock zone is discussed. Management alternatives for reducing damage from A. ostoyae when regenerating sites are reviewed.     

Traumatic resin canals as markers of infection events in Douglas‐fir roots infected with Armillaria root disease

In response to an infection, traumatic resin canals (TRCs) are formed in the roots of many conifers, which may be used to determine the timing and sequence of infection events essential for epidemiological studies of root diseases. Juvenile Douglas‐fir (Pseudotsuga menziesii) tree roots at coastal and interior sites in British Columbia were wounded at various times of the year or were inoculated with an isolate of Armillaria ostoyae, and root sections were taken to determine the timing and extent of TRC formation. Naturally infected Douglas‐fir were also examined to determine the extent of the TRCs in infected and uninfected roots on infected trees and in the lower stem. Wounds made in March and October had poor or no TRC formation while the summer wounds responded strongly and were associated with resin soaking. Roots wounded in October did not respond until the following year in all trees except one. Trees produced TRCs and resin soaked tracheids at all times in response to the fungal inoculations. The most striking difference between wounding or fungal inoculation was the multiple bands of TRCs produced in response to the fungus. TRCs at natural A. ostoyae infections were found 92% of time in roots at the stem junction and 74% of the time in the stem at soil line. TRCs were produced in uninfected roots on infected trees but disappeared with increasing distance from the initiating lesion. TRCs can be used to time yearly and seasonal root infections when they can be traced from an identified lesion.     

Rhizomorph growth habit,saprophytic ability and virulence of 15 Armillaria species

The rhizomorph branching habit in soil, competitive saprophytic ability and virulence were determined for 15 species of Armillaria from Europe, North America and Australia and New Zealand. In soil, rhizomorphs of northern hemisphere species branched either monopodially or dichotomously, whereas all five species from Australia and New Zealand branched dichotomously. The dry weight of rhizomorphs produced in soil by isolates of a species and by species was very variable. Species with monopodially branched rhizomorphs had significantly higher saprophytic colonization scores than dichotomously branched species and scores were significantly higher in Garry oak than Douglas‐fir segments and in fresh than autoclaved segments. The damage to Douglas‐fir seedlings caused by isolates of most dichotomously branched species was significantly greater than that caused by monopodially branched species. Species producing dichotomously branched rhizomorphs were more aggressive than monopodially branched species, killing 80% (vs. 17%) of seedlings that died during the first year of the 2‐year experiment.     

Virulence and double‐stranded RNA in Sphaeropsis sapinea

Forty wildtype isolates of Sphaeropsis sapinea were grouped into the morphotypes A and B based on previously defined differences in cultural and morphological criteria as well as restriction sites for Dde I and Bst UI endonucleases in nuclear ribosomal DNA amplicons. Thirteen of 20 type A isolates and nine of 20 type B isolates contained detectable dsRNA (55%) of different molecular weight and size. dsRNA was transmitted into conidia at a frequency of 71–100%. By selecting single conidia, dsRNA‐free subcultures were obtained from six of 22 isolates containing dsRNA. Pathogenicity tests on expanding buds of landscape trees of three species of Pinus showed highly significant statistical interactions between isolate virulence, Pinus species, and year. Pine species‐year had a profound impact on virulence. The pattern in the interactions was revealed by principal component analysis of the interaction sums of squares of the anova (Additive Main Effects and Multiplicative Interaction; AMMI). Pinus sylvestris was highly interactive in its susceptibility to S. sapinea with seasonal effects. P. nigra and P. resinosa were more stable. The interactivity analysis was used to apportion interaction to specific isolates to improve the accuracy of the estimates of virulence. Estimates of the relative virulence of isolates were predicted over five different Pinus species‐years. Isolates were ranked in virulence and interactivity using the AMMI model. This model permitted mean separation tests of the relative virulence among isolates over the combined Pinus species‐years. One isolate was identified as potentially having dsRNA‐mediated hypovirulence based on the significantly greater virulence of its isogenic, dsRNA‐free subculture, as expressed over the three Pinus species and 2 years. Type A isolates containing dsRNA ranged from stable to highly interactive and from low to high in virulence. Type B isolates containing dsRNA were similar in interactivity but virulence ranged from avirulent to moderate, seldom exceeding the mean for S. sapinea. dsRNA‐free isogenic subcultures tended not to express higher virulence than their dsRNA‐containing parent strains but often changed in interactivity. Therefore, in one year a dsRNA‐free subculture might be more virulent than its dsRNA‐containing parent. In another year the dsRNA‐free subculture might be less virulent.     

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.     

Infection of Norway spruce container seedlings by Gremmeniella abietina

First‐ and second‐year containerized Norway spruce seedlings were inoculated with conidia of type A (large tree type) and type B (small tree type) of Gremmeniella abietina var. abietina at different times during the summer. The appearance of symptoms after artificial inoculation and natural infection on spruce seedlings were recorded the following spring and compared with the disease symptoms on Scots pine seedlings. The proportion of diseased seedlings after inoculation reached as high as 80%. The susceptible period during the summer began later on the first‐year seedlings than on the second‐year seedlings, and was similar for the pine seedlings. Susceptibility of first‐year seedlings was highest in August and on second‐year seedlings in July. The accumulated temperature sum, relative humidity and height growth for first‐ and second‐year seedlings was assessed. Natural infection in 2002 caused more disease on pine than on spruce seedlings. Experimental thinning of seedlings had no effect on disease incidence. In a preliminary comparison between the ability of A and B types to cause disease in Norway spruce seedlings, type B caused more damage than type A after inoculation. However, type A caused a high disease frequency in other experiments in this study. Symptoms on Norway spruce seedlings often first occurred in the mid‐section of the shoot, and were similar to those observed on pine seedlings: needles turned brown, starting at the needle base, in the spring following inoculation. On first‐year spruce, diseased needles were shed rapidly, in contrast to a slower rate of shedding on first‐year pine seedlings. Pycnidia developed about 2 years after inoculation (on pine 1 year after inoculation). On Norway spruce seedlings the lower part of the shoot, including the lateral shoots, often remained alive. The experiments show that G. abietina can cause disease on containerized Norway spruce seedlings under nursery conditions in Finland. The coincidence of spore dispersal, seedling susceptibility and predisposing factors are important in disease development.     

Canadian conifers as hosts of the pinewood nematode (Bursaphelenchus xylophilus): Results of seedling inoculations

Seedlings of 22 species of conifers from across Canada were inoculated with m and r form isolates of the pinewood nematode (Bursaphelenchus xylophilus). In an experiment made under ambient (summer‐fall) temperatures in a shadehouse at Victoria, British Columbia, Canada, 8 of the 22 conifer species were killed by the nematodes, but mortality was low, i.e. 4–30%. Pines (eastern white, Jack and red) were more susceptible than other conifers. Yellow cypress, eastern white cedar, western red cedar and western hemlock were not killed by the nematodes. In a second experiment made at elevated temperatures (30°C‐16h long days, 25°C‐8 h long nights) in a greenhouse, 18 of the 22 conifer species died following nematode inoculation. Again, pines (lodgepole, eastern white, western white and red) were among the most susceptible tree species and the four conifers that were unaffected in the first experiment were not killed. Tamarack and western larch, both immune at ambient temperatures, were the two most susceptible conifers at elevated temperatures. Compared to ambient temperatures, seedlings at elevated temperatures died quicker and contained more nematodes. M and r form nematodes were equally pathogenic in both experiments.     

Root growth and water use efficiency of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and lodgepole pine (Pinus contorta Dougl.) seedlings

One-year-old Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and lodgepole pine (Pinus contorta Dougl.) seedlings were grown for 17 weeks in 100-cm deep, 7.8-liter containers. Two Douglas-fir provenances, one from a wet and one from a dry site in coastal British Columbia, and two lodgepole pine provenances, one from a wet and one from a dry site in interior British Columbia, were grown in wet (522% water content) or dry (318% water content) peat/vermiculite soil in a factorial design. Each container was sealed so that water loss occurred only through the seedling. Five harvests were made at three to five week intervals and water use, dry matter increment, root length and root weight were determined at each harvest. Stomatal conductance and shoot water potentials were measured during the last 12 weeks of the experiment. Lodgepole pine seedlings had greater dry matter production, water use, stomatal conductance and new root length than Douglas-fir seedlings. New root weight of lodgepole pine seedlings exceeded that of Douglas-fir seedlings during the last five weeks of the experiment, and specific root length (root length per unit root weight) of new roots was higher for lodgepole pine seedlings throughout the experiment. Douglas-fir seedlings showed higher water use efficiency (WUE) than lodgepole pine seedlings, and both species showed higher WUE in the dry soil treatment. Douglas-fir seedlings had lower water potentials and higher water uptake rates per unit of new root length than lodgepole pine seedlings, although water uptake rates per unit of root dry weight showed little difference between species. Soil water treatment influenced specific root length of new roots, water uptake per unit of new root length, and WUE in Douglas-fir seedlings more than in lodgepole pine seedlings.     

Virulence of Bursaphelenchus mucronatus to pine seedlings and trees under field cownditions

Bursaphelenchus mucronatus is a parasitic nematode of pine that is widely distributed in the natural pine forests of Asia and Europe. It has a very similar morphology and biology to that of Bursaphelenchus xylophilus, the causal agent of pine wilt disease, but has generally been considered to be non‐pathogenic to pine. However, in some provinces of China, B. mucronatus has been isolated from dead pine trees rather than B. xylophilus. Previous studies have shown that B. mucronatus can induce the death of pine seedlings under glasshouse conditions. To investigate the virulence of B. mucronatus, 2‐year‐old seedlings of Pinus massoniana and Pinus elliottii were inoculated with one of six isolates of B. mucronatus under field conditions in April 2014 and their condition was monitored over a year. The virulence of the six B. mucronatus isolates differed on the three host species: P. elliottii seedling mortality ranged from zero to six of the 18 inoculated seedlings, whereas P. massoniana seedling mortality ranged from four to 12 of the 18 inoculated seedlings. Three B. mucronatus isolates that appeared to cause different levels of mortality among the seedlings were used to inoculate 12‐year‐old Pinus thunbergii trees in August 2014. The trees were monitored for a year, during which time between 4 and 12 of the 18 inoculated trees in each treatment wilted and died. The average monthly temperature during the test period appeared to be similar to that of the historical average in the test areas; however, both study sites experienced above‐average rainfall. This study demonstrated that B. mucronatus has potential virulence on pine trees and provided experimental evidence that high temperatures or drought stress is not essential for the virulence of B. mucronatus.     

The interaction between competition in interior Douglas-fir plantations and disease caused by Armillaria ostoyae in British Columbia

Interior Douglas-fir trees in plantations were assessed for size differences related to the level of diseased neighbours infected with Armillaria ostoyae. The four Douglas-fir stands studied ranged from 25- to 34-year-old, and represented the oldest accessible planted stands in the Interior Cedar Hemlock (ICH) ecosystem in British Columbia. Twenty-three to 25, 10-m radius plots were established in each stand. The spatial coordinates, total height, and diameter at breast height of all live and dead trees in the plots were recorded. Subject trees whose competitors were contained in the 10-m radius plots were also identified. Trees were pulled out of the soil using a mechanical excavator and the root systems were surveyed for evidence of infection by A. ostoyae. Stem disks were taken from each tree at 1.3 m above the ground for a determination of basal area. Increasing proportion of diseased trees in the plots resulted in less total plot basal area, but did not affect the mean basal area or height. Individual subject tree basal area was negatively related to the level of disease in surrounding competitors, opposite to expectations; however, diseased subject trees had reduced height and basal area compared to disease-free subject trees. Increasing competition reduced both the height and basal area of the trees, while regular distribution of all trees increased both total and mean plot basal area but not height. Disease incidence at the plot level and in individual subject trees was mainly affected by the neighbourhood conditions in which it grew, and was also related to disease intensity in the tree root systems. Although disease may alter resource partitioning among trees, the utilization of these resources is mostly limited by the increasing disease incidence as the stands age, the higher probability of larger trees being diseased with time, the occurrence of dead trees in clumps, and the high probability that dead trees will eventually infect live neighbours. The widespread belowground incidence of A. ostoyae in the ICH, its rapid colonization of stumps, and its wide host range can reduce site potential in managed stands.     

Blight of conifer seedlings caused by Colletotrichum gloeosporioides

In a growth chamber experiment, 70-day-old seedlings of 10 conifer species were inoculated with Colletotrichum gloeosporioides conidia to determine the host range of the fungus. Based on the percentage of seedlings affected and the disease severity on individual seedlings, the order of most to least susceptibility was: western hemlock (WH), mountain hemlock, western larch, Sitka spruce, Engelmann spruce, Douglas-fir (coastal form, then interior form), white spruce and ponderosa pine; lodgepole pine and western red cedar remained unaffected. Inoculation of WH needles showed that within 24 h C. gloeosporioides conidia germinate and appressoria (penetration structures) form. A growth chamber study demonstrated that the pathogen can infect WH at needle wetness periods as short as 15 min; number of needles affected was higher at 0.5 h, but did not increase further even when wetness was extended up to 8 h. The results are discussed in relation to blight management of greenhouse-grown conifer seedlings.Portion of a Bachelor of Science (Honors) thesis, Department of Biology, University of Victoria, Victoria, British Columbia.     

Studies on anastomosis groups of Rhizoctonia solani isolates causing disease in two forest nurseries in Poland

Thirty‐eight isolates of Rhizoctonia spp. were isolated from Scots pine (Pinus sylvestris) seedlings with damping‐off symptoms, originating from two forest nurseries in central‐west Poland (Wronczyn and Jarocin) and from diseased seedlings grown in soil from Wronczyn nursery. Majority of these isolates (79%) had multinucleate cells and were identified as Rhizoctonia solani. The remaining isolates were recognized as binucleate Rhizoctonia spp. R. solani isolates were characterized using hyphal anastomosis and were divided into five anastomosis groups (AG). The most prevalent was AG5 (37% of isolates), followed by AG2‐1 (30%) and 27% of the isolates were identified as AG4. Groups AG1‐IB and AG2‐2 were only represented by single isolates. The virulence recorded as mortality (in percentage) was comparatively high for binucleate and multinucleate isolates of Rhizoctonia spp. Sequence analysis of the polymerase chain reaction (PCR)‐amplified internal transcribed spacer (ITS) rDNA region was used for phylogenetic analysis. The dendrogram showed that isolates were distinctly separated based on their AG types and there was no relationship between pathogenicity on Scots pine seedlings and the AG to which the isolates belong to. The results are discussed with respect to pathogenic potential of the various AG groups.     

Reaction of Armillaria ostoyae to forest soil microfungi

Fungi isolated from the oak (Quercus robur) rhizosphere were tested for their effects on rhizomorph formation and growth of 16 isolates of Armillaria ostoyae sampled in three localities in western Poland. The number of rhizomorphs, number of rhizomorph apices, and rhizomorph length and weight increased most in the presence of Penicillium lanosum, Penicillium notatum, Cylindrocarpon destructans, Penicillium spinulosum and Mycelium radicis atrovirens α and, to a lesser extent, in the presence of Nectria grammicospora. Inhibition of rhizomorph formation was caused by Trichoderma hamatum and Trichoderma viride in two A. ostoyae isolates and by M. radicis atrovirens α and P. spinulosum in one A. ostoyae isolate. It is suggested that variation in sensitivity to microbial stimulation within A. ostoyae is associated with the environmental and nutritional conditions of its original habitat. Isolates from nutrition‐rich localities, with 20% of the land area covered by deciduous trees, were particularly susceptible to stimulation by rhizosphere fungi.     

Growth rate and shoot: Root allometry in Pseudotsuga menziesii (Mirb.) Franco and Pinus contorta Dougl. Seedlings raised under two photoperiodic regimes

Six provenances of both Pseudotsuga menziesii (Mirb.) Franco and Pinus contorta Dougl. from a range of locations in the interior of British Columbia between latitudes 49° and 58° were grown for 80 days in controlled environment chambers programmed to provide daylengths comparable with those at either latitude 48°, or latitude 58°, during the period from mid‐June to early September. Neither shoot : root ratio nor whole plant dry weight of P. menziesii were affected by photoperiod. However, P. contorta seedlings, which unlike P. menziesii set bud during the experiment, had a greater dry weight and a higher shoot:root ratio (p<0.05) under longer daylengths. The results appear relevant to the interpretation of effects of cross‐latitudinal provenance transfer on plantation productivity.     

Relationship between nuclear DNA markers and physiological parameters in Sitka x interior spruce populations

Eight populations of Sitka spruce (Picea sitchensis (Bong.) Carr.) and interior spruce (Picea glauca (Moench) Voss x Picea engelmannii Parry ex. Engelm.) seedlings were sampled from a zone of Sitka-interior spruce introgression in British Columbia, Canada. Restriction fragment length polymorphisms of the nuclear ribosomal RNA genes (rDNA) were used to define species-specific hybridization patterns for the Sitka spruce and interior spruce populations. Hybridization was estimated from an index based on the relative abundance of polymorphic rDNA combining bands for each population. Sitka x interior hybrid seedlings had an index value for the relative abundance of interior spruce rDNA (Si-rDNA) ranging from 0.07 (Lower Nass; the most westerly collected source) to 0.95 (Bulkley Valley low-elevation seed orchard). During shoot elongation, osmotic potential at saturation (Psi(sat)) and turgor loss point (Psi(tlp)) increased, whereas total turgor (Psi(PTotal)) decreased. After bud set in the summer and throughout the fall, Psi(sat) and Psi(tlp) decreased, whereas Psi(PTotal) increased. At all times of year, populations with a higher Si-rDNA index had lower Psi(tlp) and Psi(sat) and higher Psi(PTotal) than populations with a lower Si-rDNA index. During the fall, Sitka x interior hybrid seedlings exhibited a seasonal decline in the temperature causing 50% needle electrolyte leakage (LT(50)) and in the critical temperature indicating the initial point of freezing injury. Seedlings with a higher Si-rDNA index had lower LT(50) and critical temperature values indicating greater freezing tolerance in the fall. Throughout most of the year, seedling population Si-rDNA index was related to the degree of drought and freezing tolerance.     

Susceptibility of six pine species to Gremmeniella abietina isolates from Spain

Inoculation experiments were performed in order to evaluate the virulence of Gremmeniella abietina isolates from Spain on the main pine species planted in the Iberian Peninsula, as well as the influence of seedling age on this virulence. Two different experiments were carried out with four isolates of G. abietina from Spain. The greenhouse experiments consisted of seedling inoculations. One‐ and 2‐year‐old seedlings of the following five pine species were used: Pinus halepensis, Pinus pinea, Pinus pinaster, Pinus sylvestris and Pinus uncinata; also, 1‐year‐old seedlings of P. nigra were inoculated. The relative necrosis length (RNL) caused by the pathogen after 130 days was used as a response variable. The laboratory experiments were performed on 2‐ to 6‐year‐old internodes of the above pine species excluding P. uncinata. The necrosis length after 6 weeks of incubation was measured. The results have shown that all G. abietina isolates were pathogens on seedlings of these six pine species and seedlings of P. halepensis were consistently the most susceptible ones, although it is important to take into account that all the isolates used in the present work were isolated from P. halepensis, the only pine species in Spain where G. abietina has been recovered up to now. The susceptibility of the other pine species depended on the age of the seedlings.     

The influence of carbon nutrition on Armillaria ostoyae growth and phenolic degradation

Concentrations of 1, 5 and 25 g C L^?1 as glucose, fructose and sucrose were added to basal media containing 0 or 1.0 mM catechol orwa-hydroxybenzoic acid. Media were poured in petri dishes and inoculated with one of three isolates of Armillaria ostoyae(Romagn.) Herink. Armillaria ostoyae isolates usually had greater colony size and biomass yield when glucose was the carbon source as compared to fructose or sucrose. When compared to A. ostoyae growth on basal media alone, 1.0 mM catechol and /wvi-hydroxybenzoic acia added to basal media inhibited A. ostoyae growth. In a second experiment, catechol andpara-hydroxybenzoic acid degradation by three A. ostoyae isolates growing on 1, 5 and 25 g C L^?1 glucose, fructose or sucrose were measured radiometrically. Carbon source and concentration had no effect on the degradation of pzra-hydroxybenzoic acid or catechol by Armillaria ostoyae. Results of these experiments suggests that A. ostoyae can grow faster when at high carbon concentration but cannot more effectively degrade catechol or para-hydroxybenzoic acid.     

Survival,early growth and impact of damage by late-spring frost and winter desiccation on Douglas-fir seedlings in southern Sweden

Introduction of non-native species, such as Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), can be a means of mitigating the effects of climate change by meeting the growing demand for biomass and high quality wood. The aim of this study was to investigate early growth, survival and damage from late-spring frost and winter desiccation. A provenance trial with four coastal and three interior provenances of Douglas-fir originating from British Columbia, Canada, was established in Southwest Sweden (56°43′N, 13°08′E). Seedling height, length of the leading shoot, and occurrence of frost damage, were measured after one, three, and six growing seasons. Timing of bud break in spring was also observed. The interior Douglas-fir were more frequently damaged by late-spring frost compared to the coastal Douglas-fir. The interior Douglas-fir still had a higher survival after six growing seasons compared to the coastal variety. All provenances were damaged by winter desiccation, but the provenances originating from the coastal area were more severely damaged. Choice of variety may reduce the risk for either late-spring frost or winter desiccation.     

Virulence of Fusarium oxysporum and F. commune to Douglas‐fir (Pseudotsuga menziesii) seedlings

Fusarium species can cause damping‐off and root rot of young conifer seedlings, resulting in severe crop and economic losses in forest nurseries. Disease control within tree nurseries is difficult because of the inability to characterize and quantify Fusarium spp. populations with regard to disease potential because of high variability in isolate virulence. Fusarium isolates were collected from healthy and diseased seedlings of Douglas‐fir (Pseudotsuga menziesii) and western white pine (Pinus monticola) from a nursery in Idaho, USA. Molecular markers such as DNA sequences (mitochondrial small subunit and nuclear translation elongation factor 1‐alpha) and amplified fragment length polymorphism were used to identify isolates as either F. oxysporum or F. commune. In addition, diagnostic primers were developed to detect and distinguish F. commune from F. oxysporum. In vitro and greenhouse virulence tests were completed on Douglas‐fir germinants and seedlings. For Douglas‐fir germinants and seedlings, F. oxysporum isolates generally caused less severe symptoms, whereas most F. commune isolates caused mortality through damping‐off. This is the first report of direct evidence that F. commune can cause damping‐off disease on Douglas‐fir seedlings under greenhouse conditions.