Pedicularis and Castilleja are natural hosts of Cronartium ribicola in North America: a first report

White pine blister rust disease, caused by the introduced pathogen Cronartium ribicola, has severely disrupted five‐needled pine ecosystems in North America. A 100‐year effort to manage this disease was predicated in part on the premise that the pathogen utilizes only species of Ribes (Grossulariaceae) as alternate hosts on this continent. The current study presents the first conclusive demonstration that some species in the family Orobanchaceae (Pedicularis racemosa and Castilleja miniata) are functioning as alternate hosts in a natural ecosystem of North America. This finding has implications for improving our understanding of epidemiology, pathogen adaptation and host–pathogen interactions within white pine blister rust.     

Host removal as a potential control method for Phytophthora cinnamomi on severely impacted black gravel sites in the jarrah forest

Removal of living plants from an area of Eucalyptus marginata (jarrah) forest on black gravel sites infested with Phytophthora cinnamomi significantly reduced subsequent pathogen recovery. Vegetation, including trees and annual and herbaceous perennial plants, was killed on the sites by herbicide application. To determine whether this treatment efficiently eliminated P. cinnamomi, soil samples were seasonally collected and baited to test for the presence of the pathogen. There were no recoveries on treated sites in autumn, 28 months after removal of all vegetation by herbicide application. To test whether this was the result of the complete elimination of the pathogen or whether inoculum remained, regrowth on sites was not controlled after this period leading to the re‐establishment of annual and herbaceous perennial species, some of which are hosts of P. cinnamomi. Recovery of P. cinnamomi after plant regrowth on the formerly treated sites indicated that for complete pathogen removal, sites need to remain free of vegetation for longer than 28 months. Overall, however, this study confirms that the pathogen is a weak saprophyte, and withdrawal of host material for a period of time may make eventual rehabilitation of these sites possible.     

Mechanisms and metabolomics of the host–pathogen interactions between Chestnut (Castanea species) and Chestnut blight (Cryphonectria parasitica)

Chestnut blight is a stem‐girdling disease of Castanea caused by the fungal pathogen Cryphonectria parasitica. Chestnut blight affects all Castanea species to some degree. In Asian species, chestnut blight is a commercially relevant disease which primarily affects nut production. In American and European species, chestnut blight has caused significant declines in wild populations and continues to negatively affect nut production in the European chestnut (C. sativa). Despite the profound effect of this disease in the Castanea genus, very little is known concerning the factors involved in the host–pathogen interaction between C. parasitica and its Castanea hosts. This review summarizes information on known mechanisms and metabolites involved in the host–pathogen interaction and contributes original information on the pathogen in relation to susceptible and putatively resistant genotypes with a view to furthering research that will promote a better understanding of this devastating disease and enable its control.     

Field ready: Development of a rapid LAMP-based colorimetric assay for the causal agent of white pine blister rust,Cronartium ribicola

The invasive fungal pathogen Cronartium ribicola causes white pine blister rust which is considered one of the most destructive diseases of five-needle (white) pines in North America. The disease has a life cycle that requires two hosts: white pines and Ribes spp., although other non-Ribes species, including Castilleja and Pedicularis, have been demonstrated as alternate hosts as well. Detection of this disease can be difficult because of the ephemeral nature of sporulation on pine hosts with ambiguity in other symptoms, and the alternate hosts for C. ribicola can also be an alternate host for other pine rust species. We used the previously published C. ribicola genome and species-specific real-time PCR assay to develop a field-ready loop-mediated isothermal amplification (LAMP) specific colorimetric assay for this pathogen. Specificity results across regionally identified pine rust pathogens showed the assay is highly specific to C. ribicola and can detect as little as 40 pg of pathogen DNA. We also developed a simple DNA extraction method that works with several tissue types (bark/phloem, aeciospores, and urediniospores/telia) to prepare the DNA samples for the LAMP assay. The DNA extraction and LAMP assay take ~70 min to complete and require a relatively small investment in equipment. This tool enables quick and efficient detection of white pine blister rust.     

Detection of Chalara fraxinea from tissue of Fraxinus excelsior using species‐specific ITS primers

Chalara fraxinea (teleomorph: Hymenoscyphus albidus) is known as a serious pathogen of Fraxinus excelsior, causing massive dieback of trees in Europe. The fungus is able to cause latent infections, and has been previously detected as an endophyte in asymptomatic tissues. Chalara fraxinea is a slow grower in culture, and is thus likely to be overgrown by faster growing fungi whenever pure culture isolations are being attempted. This study reports species‐specific ITS primers allowing fast and reliable detection of the pathogen directly from infected tissues of F. excelsior.     

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.     

Identification and distribution of Armillaria species associated with an oak decline event in the Arkansas Ozarks

Forests in the Ozark Mountains of northern Arkansas recently experienced a widespread oak decline event. Armillaria, a root rot fungus, has been associated with other oak decline events and may have been an important contributing factor to tree mortality in this event. Although Armillaria has been identified from the Ozark Mountains in Missouri, it has never been investigated in the Arkansas Ozarks. Molecular diagnostic techniques were used in this study to identify species of Armillaria present on roots removed from dead trees of two common oak species, northern red oak, Quercus rubra L., and white oak, Q. alba L., from three geographic areas and on three topographic positions – ridges, south‐ and west‐facing benches. Armillaria(A. mellea, A. gallica or A. tabescens) was identified from 31% of root samples taken from 102 trees in seven of nine sample plots. Armillaria mellea, occurred most often (20 samples, both oak species on seven plots) followed by A. gallica (10 samples, northern red oak only on four plots), and A. tabescens occurred twice (on northern red oak in a single plot). Thus, all three Armillaria species occurred on northern red oaks while A. mellea was the only species recovered from white oaks. Results varied by topographic position with samples from tree roots on ridges having the fewest positive identifications, one of 29. West‐facing benches had the highest positive samples with 20 of 41 testing positive and trees on south‐facing benches were intermediate with 11 of 32 samples from infected trees. This study documents the occurrence of three species of Armillaria in the Arkansas Ozarks and their association with oak mortality resulting from an oak decline event coupled with a red oak borer, Enaphalodes rufulus, outbreak. Further, it documents some potential variation in host/pathogen combinations and forest site conditions.     

Involvement of Cylindrocarpon destructans in root death of Pinus sylvestris seedlings: Pathogenic behaviour and predisposing factors

The common rhizospheric fungus Cylindrocarpon destructans was investigated in relation to its role in root death of Pinus sylvestris in Nordic nurseries and plantations. Laboratory methods were developed for studying similar root problems as well as the early effects of phytotoxicity and fungal infection. Seedlings grown under standardized optimal conditions were exposed to controlled stress (known to occur in nurseries), with or without C. destructans in the rhizosphere. Low light conditions, anaerobic root environment, and fungicide treatment were each found to predispose pine seedlings to invasion by the pathogen. The pathogen was very sensitive to competition as well as antagonism on the root, and fungicide‐induced inhibition of antagonists such as Trichoderma spp. also increased the severity of attack by the pathogen. To compete successfully, the pathogen would have to invade and dominate weakened roots prior to the arrival of saprophytes. Toxic metabolites produced by the pathogen weakened or killed nearby root tissues, and pathogen metabolites seemed to prevent saprophytes from taking hold in the infected roots. Such heavily infected, dead roots may act as inoculum sources allowing the pathogen to invade adjacent living roots, even healthy ones. Dead roots left in nursery soil after earlier harvests may act as reservoirs of inoculum for long periods and pose a threat to new plants.     

Population structure of Chrysoporthe austroafricana in southern Africa determined using Vegetative Compatibility Groups (VCGs)

Chrysoporthe austroafricana is one of the most damaging pathogens of Eucalyptus trees in southern Africa. It also occurs on non‐native Tibouchina granulosa trees and native Syzygium species. Additional isolates of the pathogen from previously unstudied countries in the region have become available from survey studies. The aim of this study was to use VCGs to consider the diversity in populations of isolates collected in various countries in southern Africa (Malawi, Mozambique, Namibia, South Africa and Zambia) and from different hosts. We also wanted to determine whether there are shared VCGs among these countries and hosts in southern Africa and establish a VCG tester strain database. Results showed a high diversity amongst isolates from different countries and hosts, but suggested little movement of VCGs among countries or hosts based on the available isolates. A total of 108 VCG tester strains were identified for southern Africa.     

Elm yellows: A phytoplasma disease of concern in forest and landscape ecosystems

Elm yellows (EY) is a lethal or decline phytoplasma disease that affects several Ulmus (elm) species and hybrids, which is widespread in North America and Europe. The symptoms vary among the elm species. In those native to North America, main symptoms include epinasty, chlorosis, premature casting of the leaves, yellow to brown discoloration of the phloem in the roots and stem and tree death that usually occurs within 1 or 2 years from the appearance of foliar symptoms. In contrast, affected trees of European and Asian species are primarily characterized by witches’ broom as a specific symptom, do not show phloem discoloration and are less prone to decline. The disease is caused by a relatively genetically homogeneous phytoplasma, the EY agent “Candidatus Phytoplasma ulmi,” a member of the EY phytoplasma group or 16SrV group, subgroup 16SrV‐A. In nature, this pathogen exhibits a high plant host specificity. The elm leafhopper Scaphoideus luteolus is the only confirmed vector of EY phytoplasma in North America, whereas Macropsis mendax has been reported as a natural vector in Northern Italy. However, other insect vectors are likely to be involved in its natural spread. Phytoplasmas of other taxonomic groups or 16SrV subgroups, which are known to infect a wide range of plant hosts, have been identified in naturally infected elm trees. However, the pathological relevance of these “non‐elm” phytoplasmas needs to be confirmed in many cases. Their detection is mainly based on the highly sensitive nested PCR assays, while pathological data are lacking. This study summarizes, within the framework of a single comprehensive review, the current knowledge of EY. Gaps in knowledge of this disease and prospects for future research are also critically discussed.     

Phytophthora ramorum is a generalist plant pathogen with differences in virulence between isolates from infectious and dead‐end hosts

Variation in virulence was examined among isolates of Phytophthora ramorum from epidemiologically important or infectious (non‐oak) and transmissive dead‐end (oak) hosts from North America. Twelve isolates representative of the genetic, geographic and host range of P. ramorum in the western United States were inoculated on leaves of Umbellularia californica (bay laurel or bay) and stems of Quercus agrifolia (coast live oak). In spite of extreme genetic similarity among the isolates employed, and even within the same genotype, significant differences in lesion size were measured, suggesting virulence in this pathogen is also controlled by epigenetic factors. A strong positive correlation between lesion size on bay laurel and coast live oak provides experimental evidence P. ramorum is a generalist pathogen that lacks host specificity. Isolates from non‐transmissive oaks were significantly less pathogenic both on oaks and bays than isolates from infectious hosts. These results are essential to further our understanding of the epidemiology and evolutionary potential of this pathogen. A quantitative differential in virulence of isolates from hosts with different epidemiological roles has been described for many animal diseases, but is a novel report for a plant disease.     

Tree chitinases – stress‐ and developmental‐driven gene regulation

In recent years, a considerable number of studies have harnessed the power of genomics to decipher the role of pathogenesis‐related (PR) proteins in plant defence against various biotic and abiotic stresses. Chitinases are PR antifungal proteins expressed constitutively at low levels in plants and induced during biotic pressures and are demonstrated to be involved in the plant defence responses. Remarkable induction of chitinase enzymes by various abiotic agents (salicylic acid, jasmonic acid, ethylene and ozone) and biotic components (pathogens, insect pest, fungal cell wall components and oligosaccharides) is well demonstrated in plants. Several reviews on plant chitinase expression during host–pathogen interaction are available for annual species, whilst reports of their expression in tree species are limited to a few woody perennials: Populus, Pinus, Picea, Eucalyptus, Castanea and Pseudotsuga. The aim of this paper is to review the induction of chitinase during various stresses and developmental processes in forest tree species.     

Occurrence of Diplodia corticola,including new oak host records,in Wisconsin,USA

In the United States, diseased oaks (Quercus species) exhibit tip blight, branch and stem cankers, and dieback often attributed to Diplodia species or related fungi. Emergence of Diplodia corticola as a pathogen of European oaks, and reports of this fungus in the eastern and western United States, prompted re‐examination of strains from Wisconsin. These had been obtained in the late 1990s and early 2000s and previously identified only as Diplodia species. Nuclear ribosomal ITS sequences were obtained from the Wisconsin strains and analysed with other sequences from GenBank. Wisconsin strains confirmed as D. corticola were from northern red oak (Q. rubra), black oak (Q. velutina), white oak (Q. alba) and bur oak (Q. macrocarpa). Other strains from oaks in Wisconsin were D. mutila and D. seriata. Wound inoculation of northern red, white and bur oak seedlings with D. corticola in a greenhouse resulted in shoot death and stem lesions, from which the pathogen was reisolated. We conclude that D. corticola has been present in the northcentral United States for at least two decades and report two previously unrecognized hosts of this pathogen: white oak and bur oak. The roles of D. corticola, related fungi and influences of other environmental factors in deterioration of oak health in North America merit additional investigation.     

Host specificity of co‐infecting Botryosphaeriaceae on ornamental and forest trees in the Western Balkans

The Botryosphaeriaceae is a diverse family of endophytes and fungal pathogens of mainly woody plants. We considered the host range and distribution of these fungi by sampling diseased ornamental and forest trees and shrubs in Serbia, Montenegro, Bosnia and Herzegovina, spanning a Mediterranean and a Continental climatic region. In total, ten Botryosphaeriaceae species were identified in the Western Balkans and with the exception of Sphaeropsis visci and Phaeobotryon cupressi, which occurred on one host, all the species had a broader host range. Phaeobotryon cupressi was found only in the Mediterranean region and S. visci, Dothiorella sp., Dothiorella sarmentorum and Diplodia seriata were present only in the Continental region. Pathogenicity tests were conducted on a variety of hosts from which the Botryosphaeriaceae species were isolated. These included leaves and/or stems of seedlings of 21 hosts, and cut leaves and/or branches of six hosts. Moreover, stems of seedlings of Chamaecyparis lawsoniana, Cedrus deodara, Picea omorika, Pinus patula and Eucalyptus grandis were inoculated as hosts from which some or all of the Botryosphaeriaceae species used for inoculation were not isolated. Inoculations showed that the majority of these fungi could also co‐infect hosts other than those from which they were isolated. The results suggest that most of the species have broad host ranges and can potentially cause disease on a broad range of tree species under certain conditions.     

New and unexpected host associations for Diplodia sapinea in the Western Balkans

Diplodia sapinea is an important pathogen of pine trees in plantations and urban areas in many parts of the world. This pathogen has recently also been isolated from diseased Cedrus atlantica, C. deodara and Picea omorika planted as ornamentals across the Western Balkans. The aim of this study was to consider the host range of D. sapinea in Serbia and Montenegro. Diplodia sapinea was identified from a broader collection of Botryosphaeriaceae from the Western Balkans region, based on the DNA sequence data for the internal transcribed spacer (ITS) rDNA and the translation elongation factor 1α (TEF 1‐ α). The D. sapinea isolates were obtained from sixteen tree species in the genera Abies, Cedrus, Chamaecyparis, Juniperus, Picea, Pinus, Pseudotsuga and Fagus. Four species represented new hosts in the Balkans, and this is the first report of D. sapinea from F. sylvatica anywhere in the world. Pathogenicity tests were conducted on the tree hosts from which D. sapinea was isolated, as well as on P. abies, Thuja occidentalis, Prunus laurocerasus, Eucalyptus grandis and P. patula. Inoculations were made on seedlings in the field, in the greenhouse or on freshly detached branches. Inoculations on P. pungens, P. omorika, P. abies, P. menziesii, A. concolor, P. nigra and P. sylvestris resulted in death of the seedlings 5–16 weeks after inoculation. Diplodia sapinea produced lesions on J. horizontalis and P. patula seedlings and F. sylvatica cut branches. Reciprocal inoculations showed that D. sapinea is not a pine‐specific pathogen, causing disease on tree species, including those from which it had not been isolated. Not surprisingly, the pathogen was most aggressive on some species of Pinaceae.     

Extent of colonization by Raffaelea quercivora of artificially inoculated living and gamma‐ray‐sterilized seedlings of two Japanese and three American oak species

Mass mortality of Fagacean tree species caused by Raffaelea quercivora has occurred widely in Japan. Because conidia or other propagules of the pathogen have not been found in infected trees, pathogen spread is assumed to occur primarily by hyphae. To clarify the relationship between hyphal growth of the pathogen within trees and their vessel arrangements, we examined two native Japanese oaks, Quercus crispula and Quercus glauca, and three exotic American oaks, Quercus coccinea, Quercus palustris and Quercus rubra. Quercus glauca is a radial‐porous species, whereas the other four species have a ring‐porous wood structure. Hyphal growth within inoculated potted living seedlings and in cut, sterilized stem segments of these species was examined microscopically after fungal inoculation. Water conductance in the seedlings was examined using transverse stem sections. The proportion of non‐conductive sapwood in Q. crispula, Q. coccinea and Q. palustris differed between inoculation and control treatment, being much higher in inoculated seedlings. The proportions were positively correlated with the extent of the hyphal growth. In sterilized stem segments, the extent of fungal colonization varied among the foreign ring‐porous species Q. coccinea, Q. palustris and Q. rubra. It is hypothesized that the extent of colonization by R. quercivora reflects the extent of non‐conductive sapwood irrespective of tree species, but is little affected by vessel arrangements.     

Involvement of Phytophthora species in white oak (Quercus alba) decline in southern Ohio

This study was initiated to investigate the possible role of Phytophthora species in white oak decline (Quercus alba) in southern Ohio at Scioto Trail State Forest. Surveys demonstrated the presence of four species of Phytophthora including one novel species. By far, the most common species was P. cinnamomi; P. citricola and P. cambivora were isolated infrequently. In few instances, P. cinnamomi was isolated from fine roots and necroses on larger roots. No special pattern of incidence was found, but P. cinnamomi was more commonly isolated from greater Integrated Moisture Index values suggesting moist lower bottomlands favour this Phytophthora species. When tree crown condition was examined relative to the presence of Phytophthora, no significant association was found. However, roots of declining P. cinnamomi‐infested trees had 2.5 times less fine roots than non‐infested and healthy trees, which was significantly different. The population densities of P. cinnamomi from declining trees were significantly greater than from healthy trees, suggesting increased pathogen activity that has the potential to cause dieback and decline and possibly the cause of a reduced fine root amount found on declining trees.     

Arbuscular mycorrhizal association of indigenous agroforestry tree species and their infective potential with maize in the rift valley, Ethiopia

Tree species in agroforestry are important source of inoculum for companion agricultural crops. Agroforestry trees can serve as a source of Arbuscular mycorrhiza (AM) inoculants to intercropped annuals. We studied spore abundance, root colonization of Albizia gummifera (J.F. Gmel.) and Croton macrostachyus (Hochst Ex Del.) trees and their effect on colonization of maize. Soil and root samples were collected from field standing trees from under and outside the canopy of trees and maize crops in the main rainy season. The number of spore count was significantly higher under the canopy of A. gummifera (791/100 g of dry soil) and C. macrostachyus (877/100 g of dry soil) trees than outside the canopy (547 and 588/100 g of dry soil, respectively). The level of root colonization of C. macrostachyus (45 %) was higher than A. gummifera (41 %). Root colonization of maize crops grown under the canopy of A. gummifera and C. macrostachyus trees was significantly higher than outside the canopy (P < 0.001). Maize seedlings grown on non-sterilized soils collected under and outside the canopy of A. gummifera and C. macrostachyus trees recorded higher root colonization, plant height, shoot and root dry weight than grown on sterilized soils (P < 0.001). The percentage of AM colonized roots of Zea mays seedlings was significantly positively correlated with the number of spore counts for field soils. The rhizospheres of indigenous agroforestry perennial species are important source of inoculum for annuals. The integration of perennials and annuals in an agroforestry system enhances the maintenance of soil quality in the tropics.     

Chrysoporthe puriensis causing canker and mortality in Pleroma mutabile in the Atlantic Forest,Brazil

Fungi of the Cryphonectriaceae family are globally known to be tree pathogens. In Brazil, several species of Chrysoporthe have been found causing stem and branch cankers in Pleroma (= Tibouchina) spp. Recently, Chrysoporthe puriensis was described as a new species causing stem and branch cankers in Pleroma granulosum, Pleroma candolleanum, and Pleroma heteromallum, all native species of the Melastomataceae family. During an investigation to collect isolates of the Cryphonectriaceae family in an important Brazilian biome, the Atlantic Forest, in Serra do Mar, structures typical of Chrysoporthe species were found in a different host, Pleroma mutabile. Fruiting bodies present in the bark of these trees were collected and isolated. The isolates were submitted on morphological characterization and phylogenetic analyses of the internal transcribed spacer (ITS) and β-tubulin gene regions using Maximum likelihood, Bayesian inference and Maximum parsimony methods. The isolates collected, for this study, were identified as C. puriensis. Pathogenicity tests on seedlings of Pleroma species and Eucalyptus clones revealed C. puriensis can infect and cause canker in these plant species as mortality. The results demonstrate the importance of delimiting the C. puriensis collection range to track its dissemination in other hosts. No host specificity was observed in the inoculation tests, suggesting this is an important finding, the pathogen causes diseases and mortality in several plants of the Atlantic Forest. Additionally, the pathogen can affect others hosts, such as Eucalyptus clones in commercial plantations.