Serpula himantioides,Heterobasidion annosum and Calocera viscosa as butt rot fungi in a Danish Douglas‐fir stand

A scattered wind fall of 50 trees in a 46‐year‐old stand of Douglas‐fir (Pseudotsuga menziesii) revealed Serpula himantioides, Heterobasidion annosum and Calocera viscosa as decay causing fungi. Sixty‐four percent of the stumps had visible rot on the stump surface. Mainly, a central brown rot was seen, caused by either S. himantioides or C. viscosa combined with eccentrically placed spots of incipient H. annosum rot. Subsequently, the appearance of fruitbodies disclosed that in total 76% of the wind thrown trees were associated with one or more of the three decay fungi. S. himantioides was present in a surprisingly large number of trees (54%), and the rot column extended up to 2 m into the stem. This investigation represents the first known incidence of S. himantioides and C. viscosa causing root and butt rot in living conifers in Denmark.     

Butt rot in conifers caused by Serpula himantioides (Fr.) Karst.

Serpula himantioides was found to be a causal agent of butt rot in Douglas fir and Japanese larch in northern Germany. The decay started predominantly from tap roots and main side roots and extended few meters into the stem. In final stages of rot the basal heartwood was completely destroyed and the residual width of healthy sapwood was reduced. Laboratory tests revealed a tolerance by the fungus to extremely acid substrates.     

Tinctoporellus epimiltinus,a causal fungus of butt rot of Japanese cypress

One of the causal fungi of butt rot of Chamaecyparis obtusa in Kyushu Island, Japan, was identified as Tinctoporellus epimiltinus from cultural characters, and by mating and inoculation tests. The cultural characters of the isolates obtained from butt rot of C. obtusa and T. epimiltinus isolated from the basidiocarp tissue and from decayed wood were examined. The morphological characters and chemical reactions of the isolates from butt rot were typical for T. epimiltinus. Di‐mon mating tests were performed between five monospore isolates of T. epimiltinus and five heterokaryotic isolates from the butt rot. All of the monospore isolates examined were heterokaryotized by the isolates from butt rot. Tinctoporellus epimiltinus isolates were inoculated on roots of 24‐year‐old C. obtusa. Butt rot was observed on all inoculated trees 1–2 years after the inoculation, and the inoculated fungus was re‐isolated from the decayed parts of the inoculated trees.     

Occurrence of brown root rot in Ocean Expo Park,Okinawa, Japan: Infection route and preventive control measures

Ocean Expo Park, established in 1976 on subtropical Okinawa Island, Japan, has approximately 8000 planted or naturally grown trees in an area of 77.2 ha. Windfall of these trees occurs mainly due to typhoons; wood chips made from the trunks and roots of windfall trees are commonly used as mulching material throughout the park areas, despite possible infection by pathogenic fungi such as Phellinus noxius, the pathogen causing brown root rot. P. noxius is distributed in tropical and subtropical areas worldwide and has a broad host range. The fungus can infect host plants with mycelia via root-to-root contact or with basidiospores. Here, we report the current status of tree damage caused by brown root rot in the park and infer the infection routes of P. noxius based on microsatellite markers. Among a total of 294 trees surveyed in the park, P. noxius was isolated from 13 trees of five species. P. noxius was observed more frequently on Ficus microcarpa than on other tree species. Ficus superba var. japonica was identified as a new host species, and Bombax ceiba was first recorded as a host plant of P. noxius in Japan. Among the 13 P. noxius isolates from the diseased trees, 2 isolates from adjacent diseased trees were considered to be of the same genet, while the other 11 isolates were different genets, indicating that infection by basidiospores occurred frequently among trees in the entire park, although spread via root systems also occurred between at least two adjacent trees. An identical genet was not detected at distant locations, which suggests that the disease does not spread via chips. However, mulching with chips can lead to wood decay due to decaying fungi other than P. noxius. Therefore, in accordance with the precautionary principle, the park currently prohibits the chipping and mulching of windfall or damaged trees and is considering disposal methods such as burning.     

Armillaria species distribution on symptomatic hosts in northern hardwood and mixed oak forests in western Massachusetts

Armillaria spp. are some of the most important forest pathogens in mixed hardwood forests of southern New England, yet their role as prominent disturbance agents is still not fully appreciated. We investigated the distribution of Armillaria species across eight separate stands of northern hardwood and mixed oak forests in western Massachusetts. We were specifically interested in the Armillaria species distribution from live, symptomatic hosts and not in determining overall incidence in the forest. From 32 plots (16 within each forest type), 320 isolates were collected. Armillaria was routinely encountered causing disease of live trees. In total, 89% (286/320) of all isolations came from live hosts exhibiting symptoms of root and butt rot. Overall, A. gallica was the dominant species in each forest type, making up 88/160 (55%) isolates from northern hardwood and 153/160 (96%) of all isolations from mixed oak stands. However, northern hardwood forests showed much greater species diversity, as A. calvescens, A. gemina, A. ostoyae, and A. sinapina were all found. At one site, a northern hardwood forest surrounding a high elevation spruce-fir forest, A. ostoyae was the most abundant species encountered. All five Armillaria species were found causing disease of live hosts, including A. gemina, a species considered by some as weakly virulent. Armillaria gallica was found on 22/23 tree species’ sampled, and was found most often causing butt rot.     

Root and butt rot of Todo fir (Abies sachalinensis) caused by Heterobasidion annosum s.l. in Hokkaido,Japan

The occurrence and symptoms of root and butt rot were examined in a 35 × 30 m plot of 68‐year‐old Todo fir plantation in Hokkaido, Japan. Forty‐seven percent of the cut stumps were decayed and 52% of the decayed stumps showed similar decay characteristics with yellowish orange to light brown colouration and expanded pockets in the heartwood. Morphological characteristics of the pure cultures isolated from the decay were similar to the cultures isolated from basidiocarps of Heterobasidion annosum sensu lato, found on fallen logs outside of the research site. Also DNA analysis based on the combined data set of three gene loci (glyceraldehyde 3‐phosphate dehydrogenase, heat shock protein 80–1 and elongation factor 1‐alpha genes) showed that the isolates from the decay are included in the same clade with the Japanese H. annosum s.l. isolates. They form a subclade to H. parviporum (the European S group of H. annosum s.l.). This is the first report of molecular determination of H. annosum s.l. isolated from root and butt rot in a plantation in Japan.     

Phylogenetic and pathogenicity evaluation of the marasmioid fungus Marasmius palmivorus causing fruit bunch rot disease of oil palm

Marasmius palmivorus is a marasmioid fungal species that exhibits parasitic behaviour, although most marasmioids are rarely parasitic. The fungus has been reported to cause fruit bunch rot disease of oil palm and coconut, but only a few studies on its pathogenic behaviour are available, particularly on oil palm. Hence, there is a need to assess the ability of the fungus to act as a pathogen and to study its molecular evolution and taxonomy. Nine isolates of M. palmivorus were successfully isolated from basidiocarps and diseased fruitlets of oil palm collected from oil palm plantations and were morphologically characterized on potato dextrose agar (PDA) followed by molecular identification based on nucleotide sequence alignments of internal transcribed spacer (ITS) regions of ribosomal RNA (rRNA) gene clusters with sequences from GenBank. Koch's Postulates confirmed that M. palmivorus could infect oil palm fruitlets with symptoms similar to those observed for bunch rot disease. Molecular phylogenetic studies using nucleotide sequences of ITS and the nuclear ribosomal large subunit (nLSU) showed that isolates sharing the same phenotypic characteristics of Marasmius palmivorus and Marasmiellus palmivorus are monophyletic and share a common ancestor. The fungus has also been shown to be more closely related to the genus Marasmius than Marasmiellus; therefore, we support retention of the taxon name of the pathogen causing bunch rot disease of oil palm as Marasmius palmivorus.     

Phylogenetic and differentiation analysis of the trunk rot pathogen Fomitiporia tsugina in North America

Fomitiporia hartigii s.l. is an important trunk rot pathogen of conifers throughout the Northern Hemisphere. In North America, this pathogen primarily attacks Tsuga, but is also found on Abies, Picea and Pseudotsuga. Previous research showed that isolates of F. hartigii from North America represent a distinct phylogenetic species, known as F. tsugina. However, that conclusion is based on limited data. To better understand the phylogenetic relationships of F. tsugina in North America, a phylogenetic analysis was performed using three loci (internal transcribed spacer, nuclear large subunit and tef1) with 23 isolates originating from the northern United States, Canada and central Europe. North American isolates formed a monophyletic group with significant statistical support, confirming previous reports that F. tsugina represents a unique phylogenetic species, distinct from European F. hartigii s.s. Population subdivision between isolates of F. tsugina from eastern (Massachusetts, Michigan, New Hampshire, New York and Wisconsin) and western (British Columbia, Colorado, Oregon and Washington) North America was investigated. Tests of gene flow and genetic differentiation based on region of origin detected significant variation (F_ST = 0.761; K_ST = 0.625, p < 0.01), suggesting gene flow between the two populations may be limited. Neutrality tests revealed significant, negative departures from the standard neutral model, which could indicate that a purifying or stabilizing selection has maintained low levels of polymorphisms in the population, perhaps favouring an advantageous phenotype. Further studies are required to better understand the occurrence of F. tsugina on Abies and Picea in boreal forests outside the natural range of Tsuga.     

Local epidemiology of the wood decay agent Laetiporus sulphureus in carob stands in Sicily

The basidiomycete Laetiporus sulphureus (Bull) Murrill is a forest pathogen causing brown cubical heart rot in a broad range of host trees. Despite its wide distribution and importance, studies aimed at understanding the epidemiology of the fungus in specific areas or hosts are lacking. In this study, an incidence of L. sulphureus as high as 34% was determined through molecular analysis of wood samples collected from 70 carob (Ceratonia siliqua L.) trees in the South‐west of Sicily, Italy. A phylogenetic analysis of Internal Transcribed Spacer (ITS) sequences indicated that all carob isolates belonged to the cluster E of Laetiporus taxonomy. Ten molecular markers based on Single Sequence Repeats (SSRs) designed on the L. sulphureus genome were developed, and isolates were genotyped through High‐resolution Melting (HRM) analysis. High gene diversity (0.581), no correlation between fungal genotype and host tree species and significant correlation between spatial and genetic distance were observed, suggesting an important role of basidiospores in the epidemiology of the fungus and a risk of transmission from a host tree species to the others. Finally, from a prognostic perspective, significant differences among isolates in terms of mycelial growth suggest that in addition to an accurate identification, a phenotypic characterization of isolates affecting trees may also be important.     

Pathogenicity of Malaysian Phytophthora palmivora on cocoa,durian, rubber and oil palm determines the threat of bud rot disease

The epidemic of bud rot disease affecting oil palm in Colombia is primarily caused by Phytophthora palmivora. The pathogen has a cosmopolitan presence that includes Southeast Asia, but to date, bud rot has not been reported in this region. This study provides an overview of the potential risk of Malaysian P. palmivora isolates cross‐infecting other host species, including cocoa, durian, rubber and Malaysian oil palm planting materials (Dura × Pisifera, D × P). On cocoa pods, the durian isolate PP7 caused dark brown necrotic lesions. Detached leaf bioassays showed that P. palmivora isolates PP3 and PP7 infected different hosts, except rubber foliage without wounding. Inoculation tests on cocoa, durian and rubber seedlings caused brown necrotic lesions when stems were wounded, with 10% mortality in cocoa and durian at 17 days post‐inoculation (dpi). However, no further infection was observed, and lesions closed within 14–28 dpi on the non‐wounded seedlings. Pathogenicity tests of oil palm seedlings inoculated with isolates PP3 and PP7 indicated that Malaysian P. palmivora isolates were not pathogenic to oil palms based on localized infection observed only through wounding. Overall, the work demonstrated that Malaysian P. palmivora isolates were able to cross‐infect multiple hosts but did not show severe infections on oil palms.     

Fungal colonization patterns in necrotic rootstocks and stem bases of dieback‐affected Fraxinus excelsior L.

European ash (Fraxinus excelsior) trees currently face the major threat of ash dieback caused by an invasive fungus, Hymenoscyphus fraxineus. Collar rots in F. excelsior have been increasingly associated with infections by this pathogen. However, the aetiology of the collar rots is still unclear and remains heavily debated. In contrast to most studies of this kind, entire rootstocks of four diseased ash trees were dug out to examine necrotic tissues in these rootstocks and stem bases in detail and to sample necrotic wood for fungal isolation. With the aid of morphological and molecular identification techniques, five to twelve fungal taxa were detected per tree. Members of the Nectriaceae family and Botryosphaeria stevensii, the causal agent of stem and branch cankers on many tree species, were frequently isolated from outer xylem. In contrast, H. fraxineus was the dominating species in interior wood layers. Microsatellite genotyping of 77 H. fraxineus isolates helped to identify up to six different genotypes per tree. The role of H. fraxineus and other isolated fungi in the aetiology of ash collar rots are discussed.     

Urea treatment reduced Heterobasidion annosum s.l. root rot in Picea abies after 15 years

Stump protection using chemical or biological agents is the main control measure against root and butt rot caused by Heterobasidion annosum s.l. in northern and temperate conifer forests. Long-term effects of urea treatment of stumps are poorly known and here we describe a 15-year study of urea treatment on the rot incidence on Norway spruce (Picea abies). We also tested the effect of urea protection on tree growth and on the resistance of stands against strong winds. Four treatments were made in two replications in two first-rotation P. abies stands in southern Sweden; after first-thinning stumps were (i) treated with urea 35% (w/v), (ii) artificially infected with H. annosum conidia, (iii) half urea treated and half artificially conidia infected, (iv) untreated, therefore naturally infected. After 15 years, the trees were sampled at 20 cm above ground using an increment borer and observed for presence of rot and, following incubation, presence of H. annosum conidia. Tree growth was calculated by measuring the diameter before and after the treatment. Urea treated plots showed the lowest incidence of rot (3%) as well as of H. annosum incidence (0%). Conidia treatment showed the highest incidence of rot (68%), its incidence was higher than that observed in natural infection treatment (43%), but did not differ from that of the 50% conidia treatment (47%). On about 30% of the rotted trees we observed conspicuous H. annosum fruiting. We did not observe growth reduction associated with tree rot. H. annosum was the only fungus observed associated with rotted trees which suggest that it was responsible for most of the rot observed in the investigated plots. Urea treated plots showed the lowest incidence of windthrown trees, and 59% of the windthrown tree incidence among the plots was explained by the incidence of H. annosum. Urea can be regarded as a reliable long-term protection method against root and butt rot of Norway spruce.     

Hosts and distribution of the intersterility groups of Heterobasidion annosum in the highlands of Greece

The intersterility groups of 127 pure cultures of H, annosum collected from different host trees in 12 mountain areas in Greece were identified. The F group commonly caused butt rot on the fir species Abies cephalonica and Abies borisii regis. It was the only type of H. annosum found in pure fir forests. The P group caused serious root rot in pinus sylvestris stands in north-eastern Greece. In more southern mountain areas it often colonized stumps of pinus nigra but seldom killed this tree species. The S group was found in natural forests of Picea abies in northern Greece, causing butt rot of spruce. In mixed forests, the intersterility groups of H. annosum were found relatively often in stumps of tree species other than their main hosts, although some host preference seemed to occur also in stump colonization.     

First report of Phyllosticta spinarum causing arborvitae foliar blight in Russia

Thuja occidentalis is a popular and widely planted tree in Russia. However, it is susceptible to arborvitae foliar blight, which can compromise the long-term health of trees in the landscape and in nursery production. Leaves with arborvitae needle blight were collected in 2019 in the foothill zone of the Republic of Adygea, and two isolates of Phyllosticta sp. were obtained. Multilocus analyses were performed with sequences of the internal transcribed spacer and 28 S rDNA of the ribosomal DNA, partial actin gene, partial RNA-polymerase II gene, and partial translation elongation factor 1-alpha. Based on phylogenetic analyses combined with morphological features the isolates were identified as Phyllosticta spinarum. Following inoculation of the original host species both isolates produced symptoms similar to those observed in naturally infected leaves. Koch's postulates were then fulfilled by re-isolating P. spinarum from the inoculated leaves. These findings represent the first confirmed detection of P. spinarum causing arborvitae needle blight in Russia.     

Occurrence of Heterobasidion basidiocarps on cull pieces of Norway spruce left on cutting areas and in mature spruce stands

Fruiting of Heterobasidion on cull pieces and stumps of Norway spruce was investigated in cutting areas and mature spruce stands located in southern Finland. Cull pieces of variable size and showing butt rot were left on three clear‐cut areas and in one thinned stand. Additionally, a part of the cull pieces was transported to mature forest sites with closed canopy. During the succeeding 3–4 years the cull pieces were investigated annually for sporocarps of Heterobasidion, and the area of actively sporulating pore layer of each sporocarp was measured. Root bases of spruce stumps in the logging areas were excavated and sporocarps found on the stumps also measured. At the onset of the experiment, Heterobasidion spp. were isolated from 76% of the cull pieces showing butt rot; 85% of the isolates were identified as H. parviporum and 15% as H. annosum s.s. During the following 3–4 years sporocarps were found on 20% of the 1938 cull pieces where Heterobasidion butt rot was initially detected visually. Sporocarp formation was promoted by advancement of butt rot, increasing cull piece diameter and end‐to‐end ground contact, but restricted by the colonization of the cull piece by Stereum sanguinolentum. Between‐site differences were significant but could not be explained by differences in tree cover. At the end of the investigation period the average sporulating area of Heterobasidion sporocarp per metre of cull piece was higher than the average sporulating area per stump at three of four logging sites. Hence, leaving cull pieces containing Heterobasidion butt rot at logging areas in southern Finland can considerably increase local production of Heterobasidion spores.     

Unravelling the Phellinus pini s.l. complex in North America: a multilocus phylogeny and differentiation analysis of Porodaedalea

Phellinus sensu lato (s.l.) is a complex of segregate genera that act as aggressive pathogens of woody plants. Nearly all of the genera in this complex have unresolved taxonomies, including Porodaedalea, which is one of the most important trunk rot pathogens of coniferous trees throughout the northern hemisphere. In an attempt to elucidate the species within Porodaedalea, a multilocus phylogenetic analysis was performed with partial sequences from four loci (internal transcribed spacer, nuclear large subunit, tef1 and rpb2) using 41 isolates that originated from North America and Europe. For reference, we analysed the neotype isolates of Porodaedalea pini and P. chrysoloma. Our results confirmed that Porodaedalea pini s.s. and P. chrysoloma s.s. are unique phylogenetic species that do not occur in North America. We detected two discrete clades of Porodaedalea originating from the southwestern and southeastern United States. Isolates from these regions grouped with significant statistical support and represent undescribed taxa. With the exception of P. cancriformans, our analyses revealed monophyly among 28 isolates originating from the northern United States, Canada and Fennoscandia, a group we have labelled the ‘Holarctic group’. Holarctic group isolates were collected from Larix, Picea, Pinus, Pseudotsuga and Tsuga and were presumed to represent at least four morphological species (P. gilbertsonii, P. laricis, P. pini s.l. and P. piceina). Tests of gene flow and genetic differentiation detected significant differences among Holarctic group isolates by region of origin, and three subgroups were designated: (i) Atlantic‐Boreal; (ii) Interior; and (iii) Pacific. Neutrality tests using the Holarctic group demonstrated significant departures from the standard neutral model of evolution and could indicate that a diversifying selection has maintained rare phenotypes in the population, which has fostered taxonomic confusion in Porodaedalea.     

Characterization and pathogenicity of six Cytospora strains causing stem canker of wild apple in the Tianshan Forest,China

Cytospora species are capable of causing destructive cankers of stems belonging to a wide range of woody plant species. In severe cases, cankers may lead to dieback of twigs and branches. Little is known about the Cytospora species causing canker disease of wild apple (Malus sieversii) trees in the Wild Fruit Forest Reserve in Tianshan Forest, Xinjiang Uygur Autonomous Region, China. In this study, six Cytospora isolates belonging to two species were isolated from cankerous lesions of wild apple twigs. Based on multi‐locus phylogenetic analysis using three DNA markers (ITS, tef1‐α and tub2) and morphological characterization, these isolates were identified as Cytospora mali and Cytospora parasitica. Temperature trials (15, 20, 25 and 30°C) showed that the optimal growth temperature for six isolates was 25°C. At a variety of temperatures, C. mali isolates tended to grow faster than isolates of C. parasitica, with the C. mali isolate, EGI1 performing better than others with regard to growth rate. Morphological observations showed that these species exhibited a single locule without conceptacles, and the conidia length was 3–5 μm. In vitro inoculation trials of twigs and leaves of M. sieversii seedlings revealed that the C. mali isolates were highly virulent phytopathogenic fungi, whereas the C. parasitica isolates were less virulent. The isolate EGI1 was the most virulent isolate among the six isolates. This paper presents the first report of pathogenic Cytospora spp. of the M. sieversii Tianshan Wild Fruit Forest Reserve of Yili, Xinjiang in China. It will aid in the understanding of how apple tree cankers are induced and provide disease management guidelines for M. sieversii forest conservation.     

An integrative study of Sakia sisanganensis n. sp. (Rhabditida: Tylenchidae) from Sisangan forest,Iran, and new morphological observations for the genus

Further data on the morphology (the lip region characters) and phylogeny of the genus Sakia are presented. The new observations were based on scanning electron microscopy (SEM) and light microscopy (LM). A new species, Sakia sisanganensis n. sp., was recovered from rotten wood of a dead beech tree (Fagus orientalis) in northern Iran, herein described and illustrated based on an integrative approach, that is morphological, morphometric and molecular characters. The new species is characterized by a combination of the following features: fine transverse striae and vestigial single band in the lateral field in SEM. Labial area dorso‐ventrally flattened. Oral region with two concentric hexagonal plates, the inner one apparently elevated. Amphidial openings short, slit‐like. Stylet delicate. Median bulb fusiform to spindle‐shaped with weak valvular apparatus. Spermatheca functional. Tail filiform with faintly pointed tip and males common. The new species was morphologically compared with four known species of the genus, viz., S. alii, S. arboris, S. castori and S. indica, all having indistinct lateral fields. Molecular phylogenetic analyses were performed based on the small subunit ribosomal RNA gene (SSU rDNA). In the Bayesian inference (BI), S. sisanganensis n. sp. with two isolates was strongly supported as a sister taxon of a clade harbouring S. arboris + Lelenchus species. However, in the maximum likelihood (ML) analysis, the new species formed a clade with S. arboris, thus supporting the reciprocal monophyly of the genera Sakia and Lelenchus. Accordingly, the test of monophyly was performed (using Bayes factor) and the results did not reject the monophyly of sakia (i.e., S. sisanganensis n. sp. and S. arboris as sister taxa) based on the currently available data.     

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.     

Molecular phylogeny of Ganoderma lucidum isolates collected from northern India

Ganoderma lucidum has been found to be a pathogen on a wide range of tree species in India. Because of large variations found in both macro‐ and microscopic characters, traditional identification and characterization methods are unreliable for this genus. We analysed nrDNA internal transcribed spacer (ITS) sequences of 52 isolates in the G. lucidum complex by the parsimony method, finding 17 lineages. The maximum parsimony analysis showed that the isolates of G. lucidum from the same host species had high similarity in ITS sequences, compared to isolates from other tree species. The result was also supported by high values of population differentiation parameters such as N_ST and F_ST values. Genetic analysis suggests the occurrence of host‐mediated genetic differentiation in G. lucidum isolates, which requires further investigation.