Environmentally induced changes in the distribution of disease susceptibility phenotypes in Pinus radiata

Pinus radiata (Monterey pine) is highly susceptible to Fusarium circinatum, the cause of pitch canker, but heritable variation in resistance to this disease has been documented in P. radiata. In this study, the distribution of susceptibility phenotypes (=lesion lengths) was assessed by inoculating P. radiata trees with a spore suspension of F. circinatum. The results show the distribution of susceptibility phenotypes in naïve seedlings to be normally distributed, both in a breeding population and in a population established from seed collected in a native forest. This pattern aligns with expectations for a quantitatively inherited trait. However, after 2.5 years of growth in a forest, variation in susceptibility is no longer normally distributed but rather is positively skewed. Thus, exposure to the forest environment results in a distribution that is enriched for relatively resistant individuals. A similarly skewed distribution is observed in standing trees in a native population of P. radiata. The observed change in distribution cannot be attributed to ontological effects on resistance to pitch canker, because tree age accounts for almost none of the variation in susceptibility. We hypothesize that the change in distribution of susceptibility phenotypes is due primarily to infections and infection attempts by the microbiota resident in the forest.     

High genetic diversity of Fusarium circinatum associated with the first outbreak of pitch canker on Pinus patula in South Africa

The disease known as pitch canker results from infection of Pinus species by the fungus Fusarium circinatum. This fungus also causes a serious root disease of Pinus seedlings and cuttings in forestry nurseries. Pinus radiata and P. patula are especially susceptible to the pathogen, but there are no records of pitch canker on P. patula in established plantations. To date, only planting material of this tree species in nurseries or in plantations at the time of establishment have been infected by F. circinatum. Symptoms of pitch canker have recently emerged in an established P. patula plantation in South Africa and this study sought to determine whether the symptoms were caused by F. circinatum. Isolates from cankers were identified as F. circinatum using morphology and DNA-based diagnostic markers. Microsatellite markers were then used to determine the genetic diversity of a collection of 52 isolates. The entire population included 17 genotypes representing 30 alleles, with a greater number of genotypes collected from younger (three- to six-year-old) than older (12- to 19-year-old) trees. Both mating types of F. circinatum were present, but no evidence of sexual recombination was inferred from population genetic analyses. This is the first record globally of pitch canker on P. patula trees in managed plantations. It is of significant concern to South Africa, where P. patula is the most important Pinus species utilised for plantation forestry.     

Systemic induced resistance in Monterey pine

The pathogenic fungus Fusarium circinatum causes pitch canker of pines. This study shows that Monterey pine (Pinus radiata), one of the most economically important pine species in the world and the main host in California, responds to infection by Fusarium circinatum in a manner consistent with systemic induced resistance. Repeated mechanical inoculations of the same trees in the field produced progressively smaller lesions over a period of 2 years, with mean lesion length decreasing significantly from 2.89 ± 0.42 cm to 1.04 ± 0.17 cm. In the greenhouse, predisposing inoculations with the pathogen induced a significant lesion length reduction, from 5.5 ± 0.21 cm in control trees to 4.46 ± 0.36 cm in predisposed trees over a period of 6 weeks. Under constant environmental conditions in a growth chamber, predisposing inoculations also induced a significant reduction in lesion size, from 3.01 ± 0.15 cm to 2.55 ± 0.18 cm over a period of 4 weeks. This is the first unequivocal report of systemic induced resistance in a conifer.     

Variation in pitch canker resistance among provenances of <Emphasis Type="Italic">Pinus patula</Emphasis> and <Emphasis Type="Italic">Pinus tecunumanii</Emphasis> from Mexico and Central America

Pinus patula and high-elevation (HE) sources of P. tecunumanii exhibit intermediate levels of resistance to pitch canker (Fusarium circinatum), compared to extremely resistant species such as P. oocarpa, and extremely susceptible species such as P. radiata. Seedlings from 20 P. patula provenances and 15 HE P. tecunumanii provenances were artificially inoculated with the pitch canker fungus at 21 and 12 weeks of age, respectively, and assessed for resistance 12–20 weeks later. There was important provenance variation in pitch canker resistance for both species. The 20-week LiveStem percentage ranged from 70.3% to 43.6% among the P. patula provenances and 59.6% to 11.7% among HE P. tecunumanii provenances. There was a geographic pattern to the provenance variation, and in both species, low altitude sources demonstrated more resistance than those from high elevation. Provenance variation in pitch canker resistance could be useful when making selection and breeding decisions with these species.     

Resistance levels of Spanish conifers against Fusarium circinatum and Diplodia pinea

Pitch canker, caused by Fusarium circinatum, and Diplodia shoot blight, caused by Diplodia pinea, are both damaging to pines (Pinus spp.) grown in plantations throughout the world, including Spain. To assess the potential for interspecific differences in susceptibility to contribute to the management of pitch canker and Diplodia shoot blight in the Atlantic region of Spain, the present study was undertaken to characterize the susceptibility of six pine species (P. sylvestris, P. nigra, P. pinaster, P. radiata, P. halepensis and P. pinea) and Douglas‐fir (Pseudotsuga menziesii) to F. circinatum and D. pinea. Based on inoculations of 2‐year‐old trees, Ps. menziesii, P. pinea and P. nigra were the most resistant to F. circinatum, with lesion lengths ranging from 3.7 to 21.5 mm, 2.2 to 12.6 mm and 2.8 to 30.9 mm, respectively. At the other extreme, Pinus radiata was the most susceptible, sustaining lesions that ranged from 8.5 to 74.8 mm in length. Pinus sylvestris, P. pinaster and P. halepensis showed an intermediate response to F. circinatum. Broadly similar results were observed in inoculations with D. pinea, with Ps. menziesii being relatively resistant and P. radiata being highly susceptible. Consistent with these results, field surveys revealed no pitch canker in stands of Ps. menziesii and low severity of Diplodia shoot blight, whereas P. radiata was severely affected by both diseases. Our findings suggest that selection of appropriate species can greatly reduce the risk of damage from two important canker diseases affecting pine plantations in the Atlantic region of Spain. Furthermore, intraspecific variation in susceptibility implies that selection may allow for the enhancement of resistance in otherwise susceptible species.     

Assessment of the effect of Nectria flute canker on wood quality within mature <Emphasis Type="Italic">Pinus radiata</Emphasis> using multiple methods

Nectria flute canker is a fungal disease that results in cankers on Pinus radiata stems. The disease, caused by the pathogen Neonectria fuckeliana, is associated with commercial pruning operations that limit knot defect to a central core to promote growth of high-quality clearwood. Nectria flute canker is present in commercial plantation forests in parts of New Zealand and Chile. Symptoms of the disease, bark cracks, stem flattenings or depressions and flute cankers, were identified on 25 mature Pinus radiata trees. Trees were felled, and discs crosscut from within, and adjacent to, the disease symptoms. The discs were photographed, and three different methods were used to determine the extent of Nectria damage beyond the defect core into the valuable clearwood sheath. These methods included a visual assessment of photographs of internal damage and two quantitative methods, based on calculations of defect extent using image processing software. Overall, flute cankers were associated with the greatest degree of encased bark and severe decay pockets, and stem flattenings the least. None of the flattened stems had internal decay. The average damage to the clearwood sheath due to flute canker was 43%, whilst that for bark cracks and flattenings was 19 and 8%, respectively. Similar results were achieved using all three methods, and the preferred method would depend on the context of the study. Using a combination of all three methods provides a robust method for measuring the damage to internal wood quality by stem pathogens and other abiotic agents.     

Assessment of molecular detection of Fusarium circinatum in insects and passive spore traps in Pinus radiata plantations

Fusarium circinatum is the causal agent of pitch canker, a destructive disease that threatens natural and planted pine forests around the world. Although pitch canker has caused problems in Spain and Portugal, concerning Europe as a whole, the fungus is not established across the pine distribution area. Its dispersion by wind and/or insect vectors could nevertheless play a role in the colonization of currently uninfected stands. It is therefore crucial to develop monitoring tools for its detection. To this end, we assessed the molecular detection of the pathogen in environmental samples of bark beetles and passive spore traps, collected in two infected Pinus radiata plantations in Basque country, Spain. The spread pattern of F. circinatum was assessed by an experimental design that included insect and spore traps installed at the centre, at the edge and outside the plots. Our results showed that F. circinatum was detected in both types of samples, at almost all collection dates. In both type of samples, positive detections were mainly found at the centre of the plots, a lower proportion at the edge, and very few outside. This suggests that long‐distance dispersion of Fusarium circinatum does not rely on wind spore dispersal neither on insect flight. Our study also shows that molecular methods are a powerful tool to monitor the pathogen in environmental samples.     

A severe canker disease of Corymbia ficifolia caused by Quambalaria coyrecup in native and urban forests of Western Australia

A serious canker disease is affecting the health of Corymbia ficifolia in urban areas of Perth, Western Australia. Quambalaria coyrecup was frequently associated with diseased trees and its presence confirmed by morphological characters and DNA sequencing. A number of Quambalaria species have previously been identified as causal agents of canker and shoot blight diseases on a range of Eucalyptus and Corymbia species. It was therefore the aim, using glasshouse studies, to determine whether Q. coyrecup was the primary causal agent of the cankers on C. ficifolia and whether other Quambalaria species could also be associated with the disease. All seedlings inoculated with Q. coyrecup produced canker symptoms within 1 month after inoculation. Canker lesions typical of those observed in the field also occurred in the four subsequent months, after which time the trial was terminated. Inoculation with Q. cyanescens and Q. pitereka (isolated from C. ficifolia shoots) did not result in lesion development. This study establishes that Q. coyrecup is a serious pathogen of C. ficifolia and that wounding is required for canker development to occur. The management of Quambalaria canker must therefore include the minimization of artificial wounding of trees in the nursery and field.     

Genetic variability of the ‘bark canker resistance’ character in several natural provenances of Cupressus sempervirens

Several Aegean (Greece) and Anatolian (Turkey) cypress provenances were studied for resistance variability to bark canker, a disease caused by the fungal pathogen Seiridium cardinale. The investigation also examined whether the low disease rate within the natural area of cypress was due to genetic or geographic‐climatic reasons. Results demonstrated strong variability for the ‘bark canker resistance’ character, in particular for trees within families. As trees from the provenances studied were not found to have genetic superiority for bark canker resistance, the above‐mentioned low disease rate could be due to geographic‐climatic barriers that inhibit the development of the fungus or its ability to infect the host. Several half‐sib progenies exhibited high resistance, suggesting that this character is totally inherited through the maternal line. Should this finding be confirmed by further research, it would facilitate the task of genetic improvement for resistance, allowing progenies of resistant trees to be obtained.     

First report of Teratosphaeria gauchensis causing stem canker of Eucalyptus in Kenya

Teratosphaeria stem canker is an important disease of Eucalyptus species in many parts of the world where these trees are intensively propagated in plantations. Symptoms similar to those of Teratosphaeria stem canker were observed on Eucalyptus grandis and a E. grandis × E. camaldulensis hybrid clone in the Central Highlands of Kenya. Symptomatic bark samples were collected from two sites and the associated fungus isolated and identified using DNA sequence analyses of multiple gene regions. The pathogen was identified as Teratosphaeria gauchensis. This represents the first report of the disease and the pathogen in Kenya.     

Differential responses of Central American and Mexican pine species and Pinus radiata to infection by the pitch canker fungus

Seedlings from a wide array of 23 species, varieties,and geographic races were screened for resistance topitch canker using artificial inoculation in agreenhouse. Seed to represent these taxa weregenerally collected in natural stands. In addition,seedlings from 79 families of P. radiata fromcommercial populations from Chile and New Zealand werescreened in a separate experiment. There was littlevariation in resistance among the commercial P.radiata families, with over 98% mortality. Similarresults were obtained with native P. radiatapopulations. All close relatives of P. radiata(sub-section Patula) were very susceptible. However,a number of closed-cone pine species (specifically,from sub-section Oocarpa) were generally veryresistant, with survival approaching 100%. Inaddition, in P. oocarpa, P. jaliscana, and P. tecunumanii from low elevation provenances anextremely high frequency of trees suffered no apparentdamage from the pathogen (92%, 85% and 80%,respectively).     

Testing of selected South African <Emphasis Type="Italic">Pinus</Emphasis> hybrids and families for tolerance to the pitch canker pathogen, <Emphasis Type="Italic">Fusarium circinatum</Emphasis>

Plantations of Pinus spp. constitute approximately 50% of the South African forestry industry. The first aim of this study was to develop a reliable inoculation technique to screen Pinus spp., for tolerance to infection by F. circinatum, which threatens pine forestry in South Africa. Inoculation of branches was compared with stem inoculations and we considered the number of branches or trees required to obtain statistically significant results. Furthermore, variation in the susceptibility of some Pinus families, clones and hybrids was considered. Results showed that branch inoculations were closely correlated with those from stem inoculations, and that it is important to consider branch and stem diameters when assessing susceptibility of trees. Subsequent trials using branch inoculations showed significant differences in F. circinatum tolerance amongst a range of pine species and hybrids of potential interest to forestry in South Africa. Significant differences in susceptibility were also found among clones of two P. radiata families. The most tolerant trees were P. elliottii × caribaea and P. patula × oocarpa hybrids, while the most susceptible species were P. patula, P. greggii and hybrids of these two. This is the first trial considering the susceptibility of Pinus hybrids, Pinus clones and some P. patula provenances, and the results indicate excellent potential for breeding for tolerance to pitch canker in South Africa. Application The accurate selection of disease tolerant planting stock for the South African forestry industry is crucially important for the continued sustainability of this important industry. The work described here provides valuable information on an artificial inoculation technique that will assist the industry in screening trees for tolerance to the pitch canker fungus, F. circinatum. It also provides some indication of the relative susceptibility of a number of Pinus spp., hybrids and families currently being evaluated in the country.     

Variation in susceptibility of six pine species and hybrids to pitch canker caused by Fusarium circinatum

Six pine species or hybrids were tested for susceptibility to pitch canker caused by Fusarium  circinatum. Pinus  densiflora, Pinus  thunbergii, Pinus  x rigitaeda (Pinus  rigida × Pinus  taeda), P. rigida × P. x rigitaeda, Pinus  echinata and Pinus  virginiana were inoculated with three spore loads (50, 500 and 5000 per tree) of F. circinatum. External symptoms, lesion length, and the frequency of reisolation of the fungus were investigated. External symptoms were greatest in P. echinata, followed by P. virginiana, however, P. densiflora was not susceptible to F. circinatum. Based on mean lesion lengths, the six pine species or hybrids differed significantly (p < 0.01) in susceptibility to pitch canker. Pinus  echinata sustained the longest lesions, whereas P. densiflora sustained the shortest lesions. The effect of inoculum density was not significant among three spore treatments within species (p = 0.17), although lesion length was slightly greater at higher spore loads over all pine species. The fungus was reisolated from inoculated stems of all pine species tested, even on trees showing little or no damage from the disease. Additional studies are needed to further explore the basis for resistance to pitch canker.     

Eutypella canker of maple: first report from Germany and situation in Austria

Eutypella canker of maple, caused by Eutypella parasitica (which is native to North America), is reported for the first time from Germany. From 2013 to 2015, this perennial canker disease was recorded on 105 maple trees in Munich. Six maple species were affected: Acer pseudoplatanus, A. campestre, A. platanoides, A. cappadocicum, A. heldreichii ssp. trautvetteri and A. hyrcanum. Occurrence on the latter three species represents new host records for E. parasitica. In Austria, Eutypella canker was newly discovered on two trees at a second locality in 2011, and it is now known to occur on seven A. pseudoplatanus trees at two localities, which are separated nearly 150 km. A. pseudoplatanus was the most frequent host of E. parasitica in Munich and Austria, which is in agreement with previous studies in Europe. The identity of the causative pathogen as E. parasitica was verified by ITS rDNA sequencing of fungal cultures obtained from cankers in Munich and at both Austrian localities. The presence of large and old cankers in both countries suggests that introduction of E. parasitica dates back a long time, probably several decades. The new records of Eutypella canker in Germany and Austria show that the disease is more widely distributed in central Europe than previously recognized.     

Exposure to a pine pathogen enhances growth and disease resistance in Pinus radiata seedlings

Most studies of Fusarium circinatum, the cause of pitch canker in pines, have focused on its activity as a pathogen. However, recent findings indicate that this fungus can colonize roots of Pinus radiata without inducing symptoms. Contrary to expectations, this study revealed that seedlings grown in infested sand grew more rapidly than seedlings not exposed to F. circinatum, based on root and shoot biomass, with modifications to root system architecture, including increased mycorrhizal root development. These effects were dependent on inoculum density and duration following growth in infested rooting medium. Plants exposed to F. circinatum expressed elevated resistance to stem infections, which significantly decreased the incidence of mortality; as above, effects were dependent on inoculum density. Resistance to stem infections was also enhanced in seedlings that emerged through infested litter, as occurs in native stands. Beneficial to neutral interactions of F. circinatum with its host suggest that the life history of this fungus may be more complex than previously recognized, with activities similar to non‐pathogenic endophytes. The potential for non‐lethal infections by F. circinatum to induce resistance in seedlings may influence dynamics of stand establishment. Overall, these results indicate that pathogenic organisms with asymptomatic states may have cryptic ecological functions that extend beyond the impacts of disease.     

A thirteen‐year study on the impact of a severe canker disease of Corymbia calophylla,a keystone tree in Mediterranean‐type forests

Worldwide, forests and woodlands have shown progressive declines in health as a result of global environmental changes in combination with local anthropogenic drivers. This study examined the incidence and progression of a canker disease of marri (Corymbia calophylla) caused by the endemic fungal pathogen Quambalaria coyrecup at three paired forest and anthropogenically disturbed sites in the southwest of Western Australia over 13 years. At the time of plot establishment in 2001, cankers were present on trees at all six sites with 22.7% of the assessed trees cankered. By 2014, cankers had led to the death of 6.7% of the trees, and an additional 10.0% of the trees developed cankers during this time. A further 2.3% of trees died due to causes other than canker, resulting in a final figure of 65.0% of trees remaining alive and free of cankers for the duration of the survey period. Canker incidence was significantly greater on trees present at anthropogenically disturbed sites (along roadsides and in paddocks) than forest trees (35.3% increasing to 50.7%, 10.0% increasing to 14.7%, respectively). Trunk diameter at breast height, tree height and crown ratings were not correlated with canker presence. This long‐term study provides evidence of the increasing severity of this canker pathogen and the impact it is having on the survival of marri.     

Pitch canker ratings of longleaf pine clones correlate with Fusarium circinatum infestation of seeds and seedling mortality in containers

The incidence and severity of pitch canker was rated among 16 clones of mature longleaf pine (Pinus palustris) in a seed orchard and cones were collected from five of those clones across the range of pitch canker disease ratings. Seed were extracted and commercially processed by clone and the percentages contaminated with Fusarium circinatum determined. Seed from each clone were sown either without treatment or with one of three fungicide treatments in a soil‐less mix at a commercial container nursery to evaluate the effects of F. circinatum (syn. F. subglutinans) on seed and seedling survival. The percent of seed with F. circinatum correlated with the pitch canker ratings in the orchard for the year of collection (r = 0.88, p = 0.05) and, when sown without fungicide treatment, with the number of seedlings produced (r = ?0.94, p = 0.01) and with seedling mortality after germination (r = 0.92, p = 0.02). The same orchard clones were more symptomatic of pitch canker through three annual surveys. Fusarium circinatum was isolated from a higher percent of seed from more symptomatic clones and a lower percent of their seed produced plantable seedlings. We propose that removing such clones from seed collections should reduce seedling mortality at the nursery.     

Alternative pine hybrids and species to Pinus patula and P. radiata in South Africa and Swaziland

Through the collaborative efforts of companies affiliated with the International Program for Tree Improvement and Conservation (Camcore), a number of pine hybrids have been produced over the last decade. Many of these have been planted in trials across southern Africa that broadly represent winter and summer rainfall areas, with the latter ranging from warm to cold temperate sites. The five-year survival and growth of the hybrids and other pines in 12 of these trials were compared with Pinus radiata in the winter rainfall, and P. patula in the summer rainfall, regions where these species have been planted extensively. Except for the highest altitude site, where freezing conditions are common, the survival of most hybrids and tropical pines was better than P. patula or P. radiata. This was, in part, attributed to their improved tolerance to the pitch canker fungus, Fusarium circinatum, which was present in the nursery at the time of planting. In the winter rainfall area, the P. elliottii × P. caribaea hybrid, P. maximinoi and, surprisingly, the P. patula hybrids performed well. In the summer rainfall regions, hybrids with tropical parents such as P. caribaea, P. oocarpa and P. tecunumanii were more productive in the subtropical/warm temperate zone and, with increasing elevation, those hybrids crossed with P. patula performed relatively better. The P. patula × P. tecunumanii hybrid, particularly when crossed with low-elevation P. tecunumanii, performed exceptionally across most sites.     

Potential of Phytophthora pinifolia to spread via sawn green lumber: a preliminary investigation

Phytophthora pinifolia causes the needle and shoot disease of Pinus radiata in Chile known as Daño Foliar del Pino. Although P. pinifolia is primarily a needle pathogen, there are concerns that it might be spread to new environments via the export of contaminated timber. In order to determine whether P. pinifolia can enter or persist in green sawn lumber, its presence in lumber produced from trees exposed to the pathogen for at least four years was examined. Green lumber produced from the infected trees, and green wood samples artificially exposed to P. pinifolia inoculum, were analysed by making extensive isolations on Phytophthora selective media. In addition, PCR was conducted using species-specific primers developed for P. pinifolia. Results of the study showed that the green sawn lumber taken from trees infected by P. pinifolia, or green lumber exposed in infected pine plantations, displayed no evidence of the pathogen surviving in this material.