The aggressiveness of Sphaeropsis sapinea on Austrian pine varies with isolate group and site of infection

The aggressiveness of the A and B isolate groups (morphotypes) of Sphaeropsis sapinea on shoot tips and lower stems of Austrian pine (Pinus nigra) was compared. The distinct differences in aggressiveness between the two S. sapinea groups, previously reported for other conifers, are confirmed for Austrian pine. However, the relative aggressiveness of the B group isolate varied by site of infection. Although the B group isolate of S. sapinea was not aggressive on shoot tips of Austrian pine, it was aggressive on stems of this host.     

Risk from Sirococcus conigenus to understory red pine seedlings in northern Wisconsin

Interest in development of multicohort stands of red pine (Pinus resinosa) in the Great Lakes region of North America prompted an investigation of the potential impact of the shoot blight pathogen Sirococcus conigenus (syn. S. strobilinus) on understory red pine seedlings. In May 2002 and 2003 healthy, 1‐year‐old red pine seedlings were planted in the understory of a maturing red pine plantation in northern Wisconsin in an area with a history of presence of this pathogen. Occurrence of shoot blight symptoms was recorded periodically during the summer, and in each year seedlings were harvested in fall and examined for signs of shoot blight pathogens. By fall 2002 and 2003, respectively, shoot blight incidence was 89% and 98% and most seedlings were dying. Pycnidia with conidia of S. conigenus were present on almost all of the symptomatic seedlings. The conifer shoot blight and canker pathogen Diplodia pinea (syn. Sphaeropsis sapinea) was also detected, though less frequently. Pycnidia of S. conigenus tended to be found more frequently on symptomatic current year's shoots than symptomatic previous year's shoots; the opposite was true for pycnidia of D. pinea. Risk from S. conigenus to understory red pine seedlings should be considered in any plans for development of multicohort red pine stands in areas where the pathogen is present.     

Association of Sphaeropsis sapinea with insect‐damaged red pine shoots and cones

The association of the shoot blight and canker pathogen Sphaeropsis sapinea with red pine (Pinus resinosa) shoots and cones damaged by insects (especially Dioryctria sp.) was investigated. Samples from a single plantation approximately 35 years old, in Sauk Co., Wisconsin and also from three plantations, between approximately 40 and 50 years old, located in an area of pine shoot moth activity in the preceding year in Adams Co., Wisconsin were visually examined. Samples were arbitrarily collected from trees felled in the first plantation in May. Pycnidia of S. sapinea and insect damage were observed on 56 of 91 (62%) of closed cones and 17 of 165 (7%) of previous year's shoots. In the absence of insect damage, pycnidia of the pathogen were identified only on eight of 91 (9%) closed cones and never on previous year's shoots. In each of the other three plantations, 10 trees were located at intervals along transects in mid‐June; one branch from the lower half of the crown per tree was pruned off, and both current and previous year's shoots were examined. Insect damage and S. sapinea pycnidia were too rare on current year's shoots to draw any conclusions. Insect damage occurred on 20–40% of over 2000 previous year's shoots that were examined, but pycnidia of the pathogen were identified on only about 5%. Although infrequent, S. sapinea was identified in association with insect‐damaged previous year's shoots from these three plantations three times more frequently than those without insect damage. Random amplified polymorphic DNA (RAPD) markers from eight randomly selected isolates were consistent with the A group of S. sapinea, which can be aggressive on red pine. This ability to exploit insect‐damaged shoots may facilitate long‐term persistence of S. sapinea at low disease incidence and severity. The potential role of insect wounds as infection courts and insects as vectors of this important pathogen of pines deserves further study.     

An effective medium for isolating Sphaeropsis sapinea from asymptomatic pines

The pathogen Sphaeropsis sapinea can persist in stems and branches of asymptomatic pines and can later induce disease when triggered by host stress. Several experiments were conducted to test if:(i) medium amended with tannic acid (TA) can increase the frequency of cultural detection of this shoot blight and canker pathogen from asymptomatic red pine (Pinus resinosa) stems, and (ii) S. sapinea can persist in asymptomatic red pine in the field following artificial inoculation. TA (0.5% w/v) in 2% (w/v) water agar proved to be the best medium for isolation of S. sapinea among a larger number of tested media. The addition of TA had little or no effect on the growth of two group A and two group B isolates of S. sapinea. However, when TA was added, 11 other fast‐growing fungal isolates from stems/branches of red or jack pines (P. banksiana) were inhibited and grew more slowly (p < 0.05) than both S. sapinea groups. The TA‐amended medium improved cultural detection of S. sapinea from 2‐year‐old, asymptomatic red pine nursery seedlings compared with two other methods used for the cultural detection of S. sapinea (32%vs. 8.5% and 18% recovery; p < 0.001 and p = 0.031, respectively). A field test using the TA‐amended media established that S. sapinea can persist asymptomatically in red pine trees for at least 1 year. This medium significantly reduces the frequency of false‐negatives from asymptomatic field material.     

Sphaeropsis shoot blight and altered nutrition in red pine plantations treated with paper mill waste sludge

During 1993 progressive, severe shoot blight and canker disease in crowns of mature, merchantable red pines (Pinus resinosa) in central Wisconsin was noted in plantations in which paper mill waste sludge previously had been applied. For eight treated plantations and six non‐treated plantations, incidence of shoot blight attributed to the pathogenic fungus Sphaeropsis sapinea (syn. Diplodia pinea) was quantified during 1993 or 1994. Foliage and soil samples also were collected for analyses. Sphaeropsis shoot blight was more frequent in treated plantations than in the non‐treated stands (means of 81% of trees and 10.2% of shoots compared with 7% of trees and 0.1% of shoots, respectively). Consistent with other reports of damage caused by some diseases of conifers in situations of altered host nutrition, mean foliar N concentrations were higher in treated plantations (1.61%) than non‐treated plantations (1.31%) (p < 0.001). Mean foliar Zn, Mn, Cu, and Al concentrations were lower in treated plantations than non‐treated plantations, and mean soil P, Ca, and Mg concentrations were higher in treated plantations than non‐treated plantations.     

Diplodia shoot blight and asymptomatic persistence of Diplodia pinea on or in stems of jack pine nursery seedlings

Diplodia pinea (syn. Sphaeropsis sapinea) is known as a major cause of damage to red pine (Pinus resinosa) seedlings in nurseries. The fungus can also be a latent pathogen of red pine seedlings, persisting in the absence of gross symptoms and later proliferating under conditions that induce host stress. In the fall of 2004, three nurseries in Wisconsin were surveyed to determine the potential for the occurrence of Diplodia shoot blight on jack pine (Pinus banksiana) seedlings and the persistence of D. pinea on or in asymptomatic seedlings of this species. Incidence of shoot blight was quantified in five 1 m long segments of an interior row in each of two survey areas in each nursery. The pathogen was identified on symptomatic seedlings collected in these areas on the basis of presence of characteristic pycnidia and conidia. Five groups of 20 asymptomatic seedlings were also collected in each of the two survey areas in each nursery. A segment of the lower stem/root collar from each of these asymptomatic seedlings was surface‐disinfested and culturally assayed using tannic acid agar. The mean incidence of shoot blight (as high as 9%) and mean frequency of cultural detection from asymptomatic seedlings (as high as 20%) were greatest in proximity to red pine windbreaks which are a source of inoculum. Only D. pinea was confirmed from subsets of symptomatic and asymptomatic seedlings which were tested using mt SSU rDNA polymerase chain reaction (PCR) primers that allow differentiation of D. pinea from D. scrobiculata and other fungi in the genus Botryosphaeria and related anamorphic fungi. Jack pine seedlings inoculated with D. pinea isolates obtained from asymptomatic nursery seedlings developed shoot blight symptoms in greenhouse trials. Thus, the ability of D. pinea to damage jack pine seedlings in nurseries has been documented and the potential for virulent strains of this latent pathogen to be distributed on asymptomatic jack pine seedlings from nurseries has been confirmed.     

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.     

Surveys for asymptomatic persistence of Sphaeropsis sapinea on or in stems of red pine seedlings from seven Great Lakes region nurseries

The fungus Sphaeropsis sapinea persists on or in stems of asymptomatic red pine (Pinus resinosa) nursery seedlings, and proliferates to cause collar rot and mortality after planting. In the spring of 2002, seven nurseries were surveyed to determine the potential range in frequency of asymptomatic persistence: three operated by Wisconsin Department of Natural Resources (DNR), two by Minnesota DNR, one by Michigan DNR, and one by USDA Forest Service (in Michigan). At each nursery five groups of 20 asymptomatic red pine seedlings were collected near an inoculum source (red pine windbreak), if present, and five groups of 20 asymptomatic seedlings were collected away from such a source (1400 seedlings total). A segment of the lower stem/root collar from each seedling was surface disinfested and incubated on tannic acid agar. Transfers were made from resulting colonies and the pathogen identified from pycnidia and conidia produced in culture. The pathogen was identified from asymptomatic seedlings collected in all Wisconsin and Minnesota nurseries, but was never detected from seedlings from the Michigan DNR or USDA Forest Service nurseries. Frequencies of detection were greater (as high as 88%) from asymptomatic seedlings near red pine windbreaks including diseased trees than from seedlings distant from such windbreaks. A subset of isolates from asymptomatic seedlings was characterized using inter‐simple sequence repeat–polymerase chain reaction analysis. Most isolates were the A group of S. sapinea, but B group isolates (recently named Diplodia scrobiculata) were also obtained from one nursery. One Minnesota nursery was more extensively sampled in 2003, with 17–44 groups of five asymptomatic red pine seedlings collected in four separate fields (525 seedlings total). The mean frequency of detection of the pathogen in these four fields ranged from 40 to 71%. Persistence of S. sapinea on or in asymptomatic seedlings continues to be problematic, not only because of the potential for subsequent seedling mortality, but also as a means by which a pathogen may be widely distributed.     

Frost hardening in first-year eastern larch (Larix laricina) container seedlings

Eastern larch (Larix laricina [du Roi] K. Koch) container seedlings were tested to determine shoot frost hardiness development under short or long days and warm (15 to 25 °C) or cool (10/5 °C, day/night) temperatures, to aid in the development of greenhouse hardening strategies. Seedlings were sampled sequentially over time (25 seedlings per week) from a population of 1000 trees. Frost hardiness increased significantly after one week of fluctuated over the next 6 weeks, and increased thereafter through week 14. Seven weeks of warm, intermittent short days, followed by 6 weeks of cool, continuous short days, resulted in greater frost hardiness than 13 weeks of warm, intermittent short days. In contrast, seedlings exposed to 7 weeks of warm, intermittent short days, followed by six weeks of warm, long days were significantly less frost hardy. Stems with needles attached had lower Index of Injury than stems without needles.     

Variation among New Zealand isolates of Sphaeropsis sapinea

Sphaeropsis sapinea is a pathogen of many coniferous species and causes significant losses to the plantation forestry industries of many countries. New Zealand isolates of S. sapinea were examined for colony and morphological characteristics, and pathogenicity to Pinus radiata seedlings. Considerable variation was observed in colony growth form and colour, and in the growth rates on four agar media. The conidial dimensions, carbon and nitrogen utilization, and pathogenicity also varied among the isolates. The variation observed was such that the isolates could not be placed within previously described distinct morphotypes and the present results concur with previous studies cautioning the use of morphological criteria alone to partition the species. These results contrast markedly with previous reports and seemingly indicate a change in the New Zealand S. sapinea population. This difference may reflect the re‐introduction or spread of additional isolates or a shift in the structure of the resident population.     

Resistance of Pinus contorta to the European race of Gremmeniella abietina

Pinus contorta seedlings, together with Pinus resinosa and Pinus banksiana seedlings, were planted adjacent to 25‐year‐old red pine trees infected by the European (EU) race of Gremmeniella abietina. Resistance to this race was assessed over 5 years. All P. resinosa seedlings were dead after that period while 65% of P. contorta and 86% of P. banksiana seedlings appeared resistant to the disease. The tip blight that occurred on P. contorta was slightly longer than that observed on P. banksiana. In microscopy, one, two, or even more suberized boundaries were seen to be initiated near the surface of the shoot at the base of healthy needles where they extended downward in the direction of the vascular cambium. Suberized boundaries occasionally crossed the xylem and joined together in the pith region to form continuous barriers around necrotic tissues. However, in most cases, these suberized barriers were not continuous across the shoot and compartmentalization was then completed by other barriers mainly constituted of parenchyma cells and xylem tracheids that accumulated phenolic compounds. Meristematic‐like cells were observed adjacent to the necrophylactic periderm. Tissue regeneration, restoration of cambial activities and formation of traumatic resin canals also seemed to be associated with the defence system of P. contorta against the EU race of G. abietina.     

Virulence of two Finnish Gremmeniella abietina types (A and B)

Thirteen Finnish Gremmeniella abietina isolates of types A and B were compared for differences in their virulence. Three kinds of inoculations were made: one with conidia on young Scots pine (Pinus sylvestris) seedlings and two with mycelia on the stems and shoots of young Scots pine trees. Inoculations with conidia were carried out in August 1992 and inoculations with mycelia were carried out six times between 11 August and 20 October, 1992. The experiments were evaluated in the late spring and early summer of 1993. The results showed that there was a difference in virulence between the two types. In the conidial inoculations type A infected 34.7% and type B infected 11.0% of the inoculated seedlings. For mycelial inoculations with type A the mean canker (stems) and necrosis (shoots) lengths were 19.3 mm and 8.6 mm longer, respectively, than with type B inoculations. In shoot inoculations there also was a clear difference between the two types in the number of such inoculations where no symptoms were observed. For type B shoot inoculations there was no fungal growth in 21.5% whereas for type A inoculations the figure was only 3.7%.     

Seedling shoot,needle and bud development in three provenances of Pinus sylvestris and Pinus contorta cultivated in northern Sweden

The patterns of current‐year shoot, needle and terminal bud elongation in seedlings of three Scots pine (Pinus sylvestris L.) and three lodgepole pine (Pinus contorta Dougl. var. latifolia Engelm.) provenances were compared during the third and fourth growing seasons after planting. Lodgepole pine produced longer shoots and buds than did Scots pine, mainly because lodgepole pine formed more stem units and elongated at a faster rate. Stem unit length and the duration of shoot and bud elongation differed relatively little between species and provenances. Lammas or polycyclic growth occurred in some lodgepole pine provenances, but not in any Scots pine provenance, and was associated with enhanced shoot elongation. Needle elongation commenced earlier, proceeded at a faster rate, and was greater in lodgepole pine than in Scots pine, but ceased about the same time in all species and provenances. The heat sum required to attain 50% of final length was lower for shoots and needles in lodgepole pine than in Scots pine, and for shoots in northern provenances than in southern ones. Mitotic activity in the apical meristem of the terminal bud, which occurred less than one week after the seedlings were free from snow, started and ceased about the same time in each species, but was higher in lodgepole pine than in Scots pine early in the shoot elongation period.     

Epiphytology of Scleroderris lagerbergii in a kettle frost pocket

Both the fungus Scleroderris lagerbergii Gremmen and summer frost were implicated in the death of seedlings of Pinus resinosa Ait. and Pinus banksiana Lamb. [=P. divaricata (Ait.) Dumont] in the Goulais Kettle Frost Pocket in central Ontario, Canada. This fungus was responsible for the preponderance of early seedling mortality, with the survival differential between inoculated and uninoculated seedlings decreasing slightly with increasing elevation in the depression. A mechanism whereby inoculum is recirculatcd to the seedlings by wind vortices was identified and described.     

Inducing long-shoot growth for vegetative propagation of western larch

This study evaluated methods to release from suppression both short shoots on lower stems and developing buds on leaders of western larch (Larix occidentalis Nutt.) seedlings, in order to increase the number of long shoots suitable for stem-tip cuttings. Treatments included application of the cytokinin N-6-benzyladenine (BA), horizontal stem bending, and decapitation on container-grown 1.5-year-old seedling ortets. The rate of release of suppressed growth of short shoots on the basal stem segments increased from 15% to 83% after spraying with 200 ppm BA and doubled on unsprayed horizontally-bent stems. Syllepsis on the upper stems increased by 19% following decapitation and by 32% after a combination of decapitation and BA treatments. Although these treatments resulted in large increases in the numbers of long shoots available for stem cuttings, the rootability of BA-treated shoots needs assessing before use in operational programs.     

Susceptibility of cones and seeds to fungal infection in a pine (Pinus spp.) collection

Summary Thirty‐one fungi, members of Ascomycota and Deuteromycota, were isolated from cones/seeds of 28 hosts (Pinus spp.) originating from East Asia, Europe and North America, and growing at the Montreal Botanical Garden, Canada. A total of 21 taxa of these isolated fungi are considered pathogens. The damage severity was most prevalent on Diploxylon pines of European provenance (Pinus nigra, Pinus mugo and Pinus sylvestris), and one indigenous North American species (Pinus albicaulis). Twelve fungi occurred on different hosts, i.e. Sphaeropsis sapinea (12), Herpotrichia juniperi (8), Truncatella hartigii (7), Ceuthospora sp. (5), Fusarium, spp. (5), Pestalotiopsis funerea (3), Phomopsis sp. (3), Valsa spp. (anamorph. Cytospora) (3), Diaporthe sp. (3), Fusicoccum sp. (2) and Sirococcus strobilinus (2). Sphaeropsis sapinea, Herpotrichia juniperi, Phomopsis conorum, Truncatella hartigii, Tubercularia sp. and Valsa spp. were related to high cone and seed damage. For the most frequently observed Sphaeropsis sapinea damage, the subgenus Haploxylon was less susceptible than Diploxylon, and P. sylvestris less than P. nigra and P. mugo. Pinus resinosa showed more tolerance to S. sapinea. Truncatella hartigii was more frequently associated with necrotic lesions on cones/seeds in comparison with Pestalotiopsis funerea. The results are discussed in the context of arboretum, seed orchard, nursery and quarantine management.     

Variation in lateral shoot elongation patterns and hybrid vigor in full-sib families and interspecific hybrids of larch

Time of shoot initiation and cessation, and rate and duration of lateral shoot elongation of full-sib families of Larix decidua Mill., L. laricina (Du Roi) C. Koch, and interspecific hybrids between L. decidua and L. kaempferi (Lamb.) Carrière, L. kaempferi x decidua, L. laricina x decidua, and L. laricina x kaempferi were explored in relation to hybrid vigor determined by height growth after 5 years. Height growth of L. laricina and L. decidua families was significantly less than height growth of the interspecific hybrids. Differences were observed for all lateral shoot elongation traits among L. laricina, L. decidua, and the interspecific hybrids. Lateral shoots of L. laricina families were the first to both initiate and cease elongation, and grew for the shortest period of time. Overall, the length of the shoot elongation period and date of cessation of shoot growth were significantly correlated with total height growth after 5 years, but the time of shoot initiation was not. Rate of elongation of lateral shoots was strongly correlated with growth increment in 1996, but was weakly correlated with total height growth after 5 years. Hybrid vigor in Larix seems to be partly a function of late cessation or increased duration of shoot elongation, or both.     

Conifer species influence Heterobasidion irregulare sporulation on stem discs exposed to natural inoculum and root wood decay in soil‐block tests

The fungal pathogen Heterobasidion irregulare has caused substantial damage to pine species in much of the south‐eastern United States, but the potential for damage from Heterobasidion root disease to forest tree species of the Great Lakes Region has not been thoroughly investigated. In each of three trials in two infested stands, fresh stem discs of six native conifer species were exposed to natural inoculum and examined for the asexual Spiniger stage of H. irregulare after incubation in the laboratory. In the first trial, the fungus was infrequently observed on discs, and differences among species were not found. But in the other two trials, detection of the pathogen was much more frequent on discs of tamarack (Larix laricina), red pine (Pinus resinosa) and white spruce (Picea glauca) than those of northern white cedar (Thuja occidentalis), eastern hemlock (Tsuga canadensis) and balsam fir (Abies balsamea). In a second study, the ability of an isolate of H. irregulare to decay root sapwood of these same species was compared using a soil‐block decay test. After 16 weeks of incubation, weight loss due to decay was greatest for red pine (23.2%) and larch (19.1%) and least for balsam fir (1.3%). Differences in the ability of H. irregulare to infect and saprophytically utilize substrate of these important tree species should prompt additional investigation of the relative risk posed to forests of the Great Lakes Region.     

The effect of late season damage on the apical bud development of Scots pine seedlings

The effect of Lygus rugulipennis Popp. feeding and artificial damage on the apical bud of Scots pine (Pinus sylvestris L.) late in the growing season was studied on two-year-old seedlings. Lygus feeding in late July and August caused inhibition of bud formation in the subsequent year's shoots, which led to loss of apical dominance and formation of interfascicular buds. Mechanical damage caused by piercing the apical bud with a needle in July, August and October produced scars and malformations on subsequent year's shoots and buds, but did not inhibit bud formation. Damage to the apical meristem could not be detected visually on the dormant bud before shoot elongation.     

Grass interference limits resource availability and reduces growth of juvenile red pine in the field

The effects of competing grasses on resource availability, growth and ecophysiological characteristics of 3-0 red pine (Pinus resinosa Ait.) seedlings were examined the first two years following outplanting in Anoka County, Minnesota, USA. Equal numbers of seedlings were planted into suppressed and undisturbed grass communities in a sandy soil. Grass suppression was maintained throughout the first growing season, but partially discontinued thereafter on the site. During the first field season interference from grass reduced pine seedling root collar diameter, needle length, number of new root tips, and lateral root length by over 40%. Mean pre-dawn needle water potential was 0.55 MPa lower in undisturbed grass plots during a brief drought in year one, but otherwise water stress was not significantly (p=0.05) influenced by grass interference. The presence of grass also reduced, up to 50%, the photosynthetically active radiation reaching the seedling canopy. At the end of year one, total biomass N, P, K, and Ca content were significantly (p=0.05) less in seedlings growing in the undisturbed grass community. Nitrogen was deficient in seedlings growing in grass. After two growing seasons, seedling shoot length (p=0.03), root collar diameter (p=0.001), and needle length (p=0.001) were significantly less (40, 54 and 20%, respectively) for seedlings growing in undisturbed grass. Seedling growth reductions induced by grass competition were associated with multiple environmental stressors in the field and not restricted to water stress as was observed in earlier studies with pine species at low and mid-latitude sites.