Histopathology of Pinus taeda needles infected by Ploioderma lethale

The histopathology of the host/parasite relationship of Pinus taeda needles infected by the Ascomycete Ploioderma lethale and the morphology of the fructifications of the pathogen were investigated. Amphigenous elliptical teleo‐ and anamorph stromata developed only on dead portions of affected needles. Insertion of the teleo‐ and anamorph stromata in the host needle tissue was subepidermal, and the stromata were covered by a shiny to dull black clypeus. Teleomorph stromata frequently coalesced. Abnormal host tissue was primarily confined to tissues external to the endodermis in the symptomatic portion of affected needles, where a collapse of the mesophyll cells was exhibited. An obvious narrowing of affected needles was observed at the distinct juncture of dead and green tissue. Inter‐ and intracellular hyphae colonized the collapsed mesophyll cell regions, endodermal, transfusion, vascular tissues, and resin ducts. Intercellular hyphae and an interface were present in the living mesophyll cells of the green needle tissue subtending the dead terminal portion of affected needles.     

Adelges aphids and fungi causing premature defoliation of Larch

Meria laricis invaded needles of Larix X eurolepis at the feeding sites of Adelges spp and caused pre-mature needle fall. The position of the feeding site on the needle determined the time of their fall. Among other fungi isolated from needles attacked by aphids were Cladosporium herbarum, Lophodermitim laricinum, Aureobasidium pullulans, Stereum sp., Thyssanophora penicillioides, Botrytis cinerea and Epicoccum sp. The feeding sites on the needles, the remaining unattacked parts of the needles and the naturally senescent needles were found to be colonized by different fungi at different times.     

Uni- and binucleate Rhizoctonia spp. co-existing on the roots of Norway-spruce seedlings suffering from root dieback

Rhizoctonia fungi were isolated from the roots of 2-year-old nursery-grown Norway-spruce seedlings displaying root-dieback symptoms. The most frequently isolated species, a uninucleate Rhizoctonia sp., was found to co-exist with binucleate Rhizoctonia in the same root system of several seedlings. All the uninucleate isolates anastomosed with each other forming a single anastomosis group with common cultural characteristics. Binucleate Rhizoctonia isolates were divided into several, morphologically dissimilar anastomosis groups (AG-I, R. spp.). In a pathogenicity test under sterile conditions, isolates belonging to the uninucleate Rhizoctonia sp. infected all root regions, particularly the root tips, resulting in a stunted root-system morphology, as was also observed in the isolation material. Binucleate Rhizoctonia spp. colonized only basal root regions, occasionally infecting cortical cells with monilioid hyphae, and had no effect on root growth.     

Fungi associated with Cyclaneusma needle cast in Scots pine in the west of Poland

Needles of Pinus sylvestris with and without symptoms of Cyclaneusma needle cast, from the west of Poland, were examined for abundance and diversity of fungi using Illumina sequencing. Fungal communities were dominated by Ascomycota (93.6%–98.6% of OTUs). Basidiomycota and non‐culturable fungi were less frequent. Needles were colonized by 260 taxa. Ascomycota, Basidiomycota and Glomeromycota were represented by 149, 39 and 1 taxa. Abundance of fungi was least in 1.5‐year‐old needles attached to twigs and greatest in fallen 2‐year‐old needles. Fungal communities had least diversity in 1.5‐year‐old needles and most diversity in 0.5‐year‐old needles of current growth. It was found that (a) the most common fungi were the needle pathogens Cyclaneusma minus, Lophodermium spp. and Sydowia polyspora; (b) less common potential pathogens were Cenangium ferruginosum, Coniothyrium complex, Desmazierella acicola, Neocatenulostroma germanicum and species in the genera Neodidymelliopsis, Pestalotiopsis, Phoma, Pleurophoma and Pyrenochaeta; (c) common primary or secondary saprotrophs included species of Alternaria, Aureobasidium, Beauveria, Cladophialophora, Cladosporium, Epicoccum, Exophiala, Lecanicillium, Penicillium, Cryptococcus and Kwoniella; (d) Lophodermium was represented mostly by Lophodermium pinastri which occurred 4–72 times more frequently than Lophodermium seditiosum; (e) frequencies of C. minus and C. ferruginosum were lower in the 0.5‐year‐old symptomless needles, increased in the symptomatic and symptomless 1.5‐year‐old needles and decreased after needle fall; (f) frequency of L. seditiosum was highest in 0.5‐year‐old needles; (g) frequency of L. pinastri increased with needle age whereas S. polyspora increased after needle fall; (h) lower frequency of L. pinastrii was associated with higher frequency of S. polyspora. It was concluded that Cyclaneusa needle cast in Poland may be caused by C. minus accompanied by C. ferruginosum, L. seditiosum, L. pinastrii and S. polyspora. Participation of Coniothyrium spp., Epicoccum nigrum, Pestalotiopsis spp. and Phoma spp. in the disease progress cannot be excluded.     

Occurrence and pathogenicity of fungi in necrotic and non-symptomatic shoots of declining common ash (<Emphasis Type="Italic">Fraxinus excelsior</Emphasis>) in Sweden

Currently, massive dieback of Fraxinus excelsior is observed in countries of eastern, northern and central Europe, and the reasons for it are unclear. The aims of the present work were (a) to study fungal communities in declining F. excelsior crowns; (b) to clarify role of fungi in the decline. Shoots from symptomatic crowns were collected in four localities in central Sweden, and distributed into the following categories: (a) visually healthy; (b) initial necroses; (c) advanced necroses; (c) dead tops. The most frequently isolated fungi were Gibberella avenacea, Alternaria alternata, Epicoccum nigrum, Botryosphaeria stevensii, Valsa sp., Lewia sp., Aureobasidium pullulans and Phomopsis sp., and these taxa were consistently found in shoots of all four symptomatic categories. Forty-eight taxa of other fungi were isolated, and fungal diversity was not exhausted by the sampling effort. The same taxa of fungi were dominant in F. excelsior shoots of different symptomatic categories, and moderate to high similarity of fungal communities was observed in shoots despite the symptoms. Forty-four isolates from 24 fungal taxa were used for artificial inoculations of 277 1-year-old F. excelsior seedlings in bare root nursery. After 2 years, only four fungi caused symptomatic necroses of bark and cambium: A. alternata, E. nigrum, Chalara fraxinea and Phomopsis sp. The most pathogenic was C. fraxinea, inducing symptoms on 50% of inoculated trees, while three other fungi caused necroses on 3–17% of inoculated trees. Infection biology of C. fraxinea and environmental factors determining susceptibility of F. excelsior to decline deserve future investigations.     

Methods of inoculating Acer spp., Populus tremuloides, and Fagus grandifolia logs for commercial spalting applications

One of the most promising wood value-added processes currently under development is spalting, where pigment is added to wood via fungal colonization. Previous studies have shown laboratory level spalting to be achievable and highly predictable. However, large-scale spalting for potential commercial applications introduces a substantial number of additional variables which impact the spalting process. To test the potential of commercial-scale spalting, Acer saccharum, Fagus grandifolia, and Populus tremuloides logs were inoculated with multiple known spalting fungi utilizing both liquid spray cultures and live dowel pin cultures. Many of the fungi that successfully produce spalting in small, sterile cultures also produced significant amounts in large logs, with many spalting patterns identical to those found in small-scale testing. Pairings of Trametes versicolor/Scytalidium cuboideum and Xylaria polymorpha/Xylaria polymorpha (different isolates) produced significant amounts of zone lines. In addition, the method of inoculation impacted the amount of spalting: more zone lines were produced when fungi were introduced via plugs, while more stain was produced when liquid cultures were sprayed onto the logs. These results indicate that many of the standard spalting fungi are suitable for large-scale applications; however, the inoculation method appears to be a vital component for successful spalting under a restricted time schedule.     

Colonization and lignin decomposition of pine needle litter by Lophodermium pinastri

The colonization and lignin decomposition of pine (Pinus thunbergii and Pinus densiflora) needle litter by an endophytic fungus, Lophodermium pinastri, was examined with field observations, a field experiment and laboratory experiments. In pine needles collected from the field, needle mass per length and lignin content were lower in needle portions bearing Lophodermium fruiting bodies than in the remaining needle portions, whereas total carbohydrate content was not different between them. Total and live hyphal lengths were greater in needle portions bearing Lophodermium fruiting bodies than in the remaining needle portions. Lophodermium fruiting bodies were not formed on sterilized needles after a 6‐month incubation on the forest floor, whereas they formed on 20% of non‐sterilized needles, indicating that this fungus can only colonize live needles on the branch. In pure culture decomposition tests, mass loss of lignin was detected for several isolates of L. pinastri, but was variable among isolates and between the needles of two pine species. A comparison of the rDNA‐internal transcribed spacer sequences between L. pinastri isolates of P. thunbergii and P. densiflora indicated that the two groups were phylogenetically separated. This study is the first to show that L. pinastri has the ability to decompose lignin in dead pine needles.     

The biology of Lophodermella conjuncta Darker on Corsican pine needles

The biology of Lophodermella conjuncta on Corsican pine needles was studied. Results of a spore exclusion experiment and histological examination of needles indicated that infection of young current years needles occurred and that the fungus first colonized only the endodermis. Infection remained latent until symptoms developed, from hyphae penetrating the mesophyll from the endodermis, in July of the year following infection. Ascocarps were produced on second year needles by mid-August. Aseospores were trapped throughout the year.Infected needles were cast prematurely during the third season. The phenomenon of latent infection of pine needles by some pine needle fungi is discussed.     

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.     

Quantification of Phaeocryptopus gaeumannii colonization in Douglas‐fir needles by ergosterol analysis

Current assessments of infection levels of Phaeocryptopus gaeumannii, the incitant pathogen of Swiss needle cast disease on Pseudotsuga menziesii, typically rely on surveys of abundance of fruit bodies on diseased needles. The relationship between this measure and internal fungal colonization is unknown. In this article, a series of experiments to determine whether ergosterol can be used to quantify P. gaeumannii internal colonization within Douglas‐fir needles is reported. It was found that ergosterol content in seven commonly occurring Douglas‐fir foliar fungi is proportionally related to biomass, and in P. gaeumannii this relationship is not affected by age of the culture. Furthermore, at four sites tested, P. gaeumannii was the most common fungus species isolated from Douglas‐fir needles, accounting for approximately 50% of the isolations. Ergosterol content in these needles was best related to P. gaeumannii despite the presence of other fungi. The strong relationship between ergosterol and P. gaeumannii is attributed to its greater contribution to total fungal biomass compared with all other fungi present within Douglas‐fir needles.     

Über Pyrenochaeta parasitica spec. nov., die Nebenfruchtform von Herpotrichia parasitica (Hartig) E. Rostrup (= Trichosphaeria parasitica Hartig

Pyrenochaeta parasitica spec. nov., the conidial stage of Herpotrichia parasitica (Hartig) E. Rostrup (= Trichosphaeria parasitica Hartig). This study contributes to the systematic position of the fungus Herpotrichia parasitica (Hartig) E. Rostrup, found on the needles of silver fir (Abies alba Mill.) and describes its Pyrenochaeta conidial stage.     

Fungi infecting Pinus sylvestris needles of various ages

The studies were carried out in three 8–11 year old stands of Pinus sylvestris in Southwestern Poland. They included the isolation of fungi from infection spots which were sampled after 1, 1, 5 and 2, 5 years on living needles developed in 1978 on lower and upper branches of tree crowns. There were 28 fungal species isolated from such needles. An attempt was made to evaluate the role of the 5 most numerous species in causing needle cast of Pinus sylvestris.     

Modelling of pH effects and CIE L*a*b*colour spaces of beech wood-inhabiting fungi by NIRS

A combination of statistical techniques of analyses were used to evaluate the potential of International Commission on Illumination (CIE) lightness (L*), redness (a*) and yellowness (b*) colour space system and near-infrared spectroscopy (NIRS) to assess surface changes in relation with progressive decay of beech (Fagus grandifolia Ehrh.) by wood-inhabiting lignicolous fungi Inonotus hispidus, Trametes versicolor and Xylaria polymorpha. pH effects based modelling predictions of beech earlywood and latewood tissues were also included. Multivariate analysis techniques included response surface optimization, sample-specific standard error of prediction (SEP) method and projection to latent structures partial least squares (PLS) regression. Strong statistical relationships were derived for pH predictions with R² values ranged: from 0.77 to 0.84 for I. hispidus; from 0.77 to 0.84 for T. versicolor and from 0.83 to 0.91 for X. polymorpha. R² values for CIE-based L*a*b* colour space measurements ranged: from 0.43 to 0.69 (L*), 0.66 to 0.76 (a*), 0.42 to 0.53 (b*) for I. hispidus; from 0.59 to 0.69 (L*), 0.69 to 0.79 (a*), 0.64 to 0.79 (b*) for T. versicolor; and from 0.51 to 0.75 (L*), 0.89 to 0.94 (a*), 0.85 to 0.89 (b*) for X. polymorpha. Multivariate technical analysis (response surface analysis, sample-specific SEP, PLS regression) of CIE L*a*b* system and NIRS results should be able to characterize pH effects and surface changes of wood spalted by lignicolous fungi as a quick and reliable non-destructive method relevant to wood-spalting concerns and the forest products industry.     

Über Sukzession und pathogene Eigenschaften Kiefernnadeln bewohnender Pilze

Succession and pathogenic properties of fungi inhabiting pine needles . Isolates from healthy and dying pine needles of various age permit conclusions about the relationship of some needle fungi to their host. Of the six most prevalent fungi, only Lopbodermium seditiosum has a pathogenic nature.     

Untersuchungen zur Biologie,Morphologie und Verbreitung von Herpotrichia parasitica (Hartig) E. Rostrup (vormals Trichosphaeria parasitica Hartig)

Studies on the biology, morphology and spread of Herpotrichia parasitica (Hartig) E. Rostrup (formerly Trichosphaeria parasitica Hartig). Part II: Method of infection, hosts and pure culture. H. parasitica (Hartig) E. Rostrup (conidial stage Pyrenocbacta parasitica Preyer et van der Aa) overwinters by means of both types of fruit bodies on the needles of hosts but also by means of mycelium which grows along the under sides of shoots during the growing season, reaches the buds and penetrates under the outer bud scales. Spread of the fungus is thus independent of spores. As well as Abies alba, other silver firs are attacked. Contact infection with mycelium on Picea sitchensis was also observed. Studies with pycnospores showed that germination on needles of host species varied with humidity. In culture the fungus grew best on 2 % malt agar, pH 6 at 21°C.     

Seasonal patterns of colonization of Norway spruce needles by Lophodermium piceae

Summary Colonization of living Norway spruce needles by Lophodermium piceae was studied in southern Sweden. One‐month to 3‐year‐old needles were assessed for frequency of infection. Needles appearing in the growing season of 1996 were monitored and sampled at 1 to 4 week intervals over a period of 14 months. Infection frequency of the current‐year needles increased from around 20% in July to approximately 40% in August 1996. This level had then doubled to approximately 80% after another growing season in August 1997. Multiple infections were rare in the needles during the first growing season, but after the second growing season at least 70% of the needles had multiple infections. The average number of infections in 9‐, 21‐ and 33‐month‐old needles were 1.3, 6.3 and 8.4, respectively. The size of the individual mycelia was estimated to be small in all needles, none of the individuals could be isolated from more than two adjacent needle sections, about 1.5 mm long. In conclusion, the number of L. piceae infections per needle increased during ageing of needles but mycelial growth is probably postponed until needle senescence and death.     

Decline‐associated Phaeoacremonium species occurring on forest trees in the north of Iran

Between 2013 and 2015, several surveys were conducted throughout forest areas in the north of Iran with the aim of isolating and identifying fungi present on trees showing decline symptoms. Fungal species, isolated from symptomatic wood tissues of various tree genera, including Zelkova, Acer, Parrotia, Quercus, Diospyros, Fagus, Fraxinus, Populus, Pinus, Pterocarya, Gleditsia, Carpinus, Ulmus, Crataegus, Alnus, Mespilus, Punica, Cydonia, Cupressus and Prunus, were morphologically identified and characterized. A molecular identification based on comparisons of DNA sequences of the β‐tubulin gene allowed us to identify seven species belonging to Phaeoacremonium; namely, Phaeoacremonium parasiticum, Pm. alvesii, Pm. minimum, Pm. iranianum, Pm. scolyti, Pm. fraxinopennsylvanicum and Pm. croatiense were isolated and identified. Phaeoacremonium minimum was the most common species and isolated from Gleditsia caspica, Parrotia persica, Cydonia oblonga and Alnus glutinosa. Pathogenicity on selected plants using a mycelium plug inoculation method revealed that most of these species are pathogenic to all the tested trees, with Pm. parasiticum and Pm. minimum being the most pathogenic species. According to host plant species and geographical distribution, the majority of Phaeoacremonium species found represent new records. This is also the first report of Pm. croatiense in Iran and outside Croatia.     

Pathogenic fungi associated with pre‐ and post‐emergence seedling blight of pine and cypress in Fars Province,Iran

Seed and root rot of pine and cypress seedlings cause heavy annual losses to forest nurseries in Fars Province. Root and crown samples of various species of conifers, such as Tehran pine (Pinus eldarica), brutia pine (Pinus brutia), Arizona cypress (Cupressus arizonica), Shirazian cypress (Cupressus sempervirens var. fastigiata), common cypress (C. sempervirens var. horizontalis) and oriental arborvitae (Thuja orientalis), showing chlorosis, necrosis, stunted growth, defoliation and root and crown rot symptoms were collected from pine and cypress nurseries across Fars province at various time intervals. Infected tissues were washed and cultured on acidified potato dextrose agar (PDA) and corn meal agar (CMA) amended with Delvocide and ampicillin, with and without surface sterilization, respectively, and incubated at 25°C for 3–5 days. Decaying seeds of all plant species were also collected from nursery seed stocks and cultured on PDA. Hyphal tip isolates were used for further studies. Species of Phytophthora, Pythium, Rhizoctonia and Fusarium were isolated from symptomatic seedlings of pine and cypress at different times during the growing season. Pathogenicity of isolates of Phytophthora, Pythium and Rhizoctonia was confirmed on seedlings of all plant species, whereas Fusarium sp. isolated from the seeds of Tehran pine was only pathogenic on seedlings of Tehran pine and Arizona cypress. Virulent isolates identified as Phytophthora nicotiana var. parasitica, Pythium ultimum, Pythium paroecandrum, Rhizoctonia solani and binucleate Rhizoctonia‐like fungus caused root and crown rot of pine and cypress seedlings. Pathogenic isolates of Fusarium identified as Fusarium proliferatum also caused pre‐emergence seed decay of pine and cypress. Isolates of Phytophthora and Pythium were the most virulent of these fungal isolates.     

Morphological and molecular characterization of Fusarium oxysporum and Fusarium avenaceum causing a novel needle blight of Picea asperata in China

Picea asperata is a unique tree species in China. It has important economic and ecological values. In this study, the disease symptoms resembling needle blight, a new disease of P. asperata (10- to 15-year-old), were detected in a forest area in Tianshui City, Gansu Province, China, in August 2020. Affected needles turned yellow, the colour became darker at the junction of infected and healthy areas, the needles wilted and even fell off the tree. Through pathogenicity tests, the two pure fungi strains isolated from diseased needles caused distinct needle blight symptoms on P. asperata, with symptoms similar to those observed in the forest. It was observed that they had sickle-shaped conidia. Subsequently, multilocus phylogenetic analysis of partial sequences of the internal transcribed spacer region (ITS), translation elongation factor 1-α (TEF-1α) and RNA polymerase II second major subunit (RPB2) genes was performed. They were found closely clustered with Fusarium oxysporum and F. avenaceum, respectively. Based on morphological and molecular biological determinations, the two pathogens were identified as F. oxysporum and F. avenaceum. To our knowledge, this is the first report of needle blight caused by F. oxysporum and F. avenaceum on P. asperata in China.     

In vitro screening of biocontrol and biodegradation potential of selected hymenomycetes against Ganoderma boninense and infected oil palm waste

The antagonistic and enzymatic activities of non‐pathogenic hymenomycetes naturally occurring on oil palm (Elaeis guineensis) trunks were assessed to determine their potential as biocontrol agents against G. boninense. Twenty‐five Basidiomycota were isolated on a Ganoderma‐selective medium and identified using internal transcribed spacer (ITS) sequencing. The three fungi with potentially higher antagonistic activity against G. boninense, based on the percentage inhibition of radial mycelial growth (PIRG) in dual culture, were Pycnoporus sanguineus (84%), Trametes lactinea (82%) and Grammothele fuligo (81%). In addition to these three isolates, Lentinus tigrinus and a Rigidoporus sp. were also able to invade sterilized oil palm blocks previously colonized by G. boninense despite having lower PIRG values. Colonization of oil palm blocks by G. fuligo, P. sanguineus, Rigidoporus sp., T. lactinea and L. tigrinus resulted in mass losses of 32.50, 30.78, 27.64, 25.20 and 19.33%, respectively, probably due to an ability to produce one or more lignocellulolytic enzyme. Further work is needed to determine the enzymatic degradation potential of these fungi under natural conditions and their possible use in the prevention of colonization of oil palm by G. boninense.