Pathogenicity of fungi associated with ash dieback symptoms of one‐year‐old Fraxinus excelsior in Montenegro

In addition to Hymenoscyphus fraxineus, two fungi identified as Diaporthe eres aff. and Fusarium sambucinum aff. were also isolated from necrotic bark lesions on declining one‐year‐old Fraxinus excelsior in a forest stand in Montenegro. To examine their involvement in ash decline, a pathogenicity test was performed using under bark inoculations on one‐year‐old Fraxinus excelsior. Hymenoscyphus fraxineus was included as comparison. All three fungal species proved highly pathogenic towards one‐year‐old seedlings although lesion sizes differed significantly between the different species. Hymenoscyphus fraxineus was most aggressive, followed by F. sambucinum aff., while D. eres aff. caused the smallest lesions. This study demonstrates for the first time the ability of isolates in the D. eres and F. sambucinum species complexes to cause decline on one‐year‐old common ash seedlings.     

Characterization of Corynelia uberata Fr., a putative fungal pathogen of Podocarpus falcatus in Ethiopian forests

Corynelia spp. are ascomycetes belonging to the order Coryneliales and are thought to be obligate parasites of trees in the Podocarpaceae. The aims of this study were to determine the disease intensity of Corynelia infection on Podocarpus falcatus in Ethiopian forests and verify the identity of Corynelia spp. from Ethiopia and other countries using morphological and molecular methods. Disease surveys were conducted in P. falcatus forest areas at Adaba‐Dodola, Bushoftu, Menagesha, Shashamane and Wondo Genet in Ethiopia between 2009 and 2011, and samples were collected for morphological and molecular studies. Additional dried specimens morphologically collected as C. uberata, C. portoricensis and C. tropica from Podocarpus species in Kenya, South Africa, Puerto Rico and New Zealand were also characterized. Morphologically, the South African specimen (F‐006479) of C. uberata had significantly larger ascospores when compared with the other specimens. There was a high sequence similarity (99–100%) in the internal transcribed spacer and 5.8S (ITS‐5.8S) region among the studied C. uberata sequences. Cloning and amplification of the insert spanning partial small ribosomal unit (SSU) and ITS‐5.8S regions of ribosomal DNA validated the unidentified ITS‐5.8S region as the sequence of C. uberata by inferring the reference sequence of SSU rDNA of C. uberata in GenBank. Both neighbour‐joining and/or maximum parsimony methods placed ITS‐5.8S and SSU rDNA sequences of Corynelia spp. at the basal position of the clade Eurotiomycetidae. C. uberata was found to be a potential pathogen on leaves, fruits and young stems of P. falcatus in Ethiopia.     

昆虫病原真菌对荔枝蝽的致病性

采用1 0×107conidia·ml-1孢子悬浮液浸染方法,从白僵菌和绿僵菌的10个菌中筛选出1株杀虫活性较高的僵菌Ma01菌株,它对荔枝蝽成虫的感染致死率达73 3%。同时野外杀虫活性试验表明,利用2 0×108conidia·ml-1也取得了明显的杀虫效果。     

Growth and ecophysiology of seedlings of Podocarpus falcatus in plantations of exotic species and in a natural montane forest in Ethiopia

The potential role of exotic tree plantations in facilitating successional processes on degraded areas was evaluated in southern Ethiopia by comparing seedling characteristics, transpiration and photo-synthetic performance of Podocarpus falcatus seedlings in Eucalyptus plantation, Pinus plantation, adjacent natural forest and clear-felled plantation site. P. falcatus seedlings exhibited differences in architecture between Eucalyptus and Pinus plantations. They had higher leaf area, shorter internode length and greater number of lateral branches in Eucalyptus plantation. At similar vapor pressure deficit (VPD), P. falcatus transpired much less than E. saligna, especially at higher VPDs. Analysis of fluorescence parameters in the leaves showed no significant differences in the level of dark-adapted and light-adapted fluorescence yield (Fv /Fm and ΔF/Fm, respectively), electron transport rate (ETR) and nonphotochemical quenching (NPQ) among seedlings grown inside plantations and adjacent natural forest, indicating similar photosynthetic performance. Nevertheless, there was evidence of photoinhibition in P. falcatus in the clear-felled site which had low fluorescence yield but high values of NPQ as protection from photoamage. The light response curves of ETR, NPQ and ΔF/F m , showed similar light saturation behavior among the seedlings grown inside plantations and natural forest and suggested a sequence of light-adapted to shade-adapted behavior in Natural forest > Eucalyptus plantation > Pinus plantation. The results show the structural flexibility, better water-use and adaptability of P. falcatus in its use of the understory environment of plantation species.     

Pathogenicity of ophiostomatoid fungi on Picea abies in Slovenia

Pathogenic fungi can survive and develop in living plants, often causing diseases in the host. Some theories speculate that pathogenic ophiostomatoid fungi provide benefits to its vectors – bark beetles – by overcoming the tree's defence mechanisms. This study reports the results of an experiment in south‐eastern Europe in which mature and seedling Norway spruce trees were artificially inoculated with various ophiostomatoid fungi. The aim of the experiment was to determine the relative virulence of ophiostomatoid fungi by assessing the ability of the fungi to stimulate host tree defence mechanisms through inoculation experiments. Experiments were performed by inoculation of Picea abies in seedling and mature trees. The following fungi were used in low‐density and seedling inoculations: Ophiostoma ainoae, O. brunneo‐ciliatum, Grosmannia cucullata and an unidentified Leptographium sp., O. bicolor, O. fuscum, O. piceae, G. penicillata and G. piceiperda. Endoconidiophora polonica was used in mass and seedling inoculations. Various characteristics such as host vitality, blue stain, lesion and resin outflow were measured before and after the trees were felled. E. polonica caused blue stain, induced large lesions and killed some of the mature trees and seedlings, confirming earlier reports that it is a strong wound pathogen. Only E. polonica, Leptographium sp. and O. ainoae caused blue stains in the sapwood of inoculated seedlings. In low‐density inoculations, G. piceiperda induced intense necrosis and had higher values for all the characteristics monitored. Some of the other ophiostomatoid fungi showed a moderate level of pathogenicity. Fungi with the capacity to stimulate a host defence mechanism could play a role in the establishment of bark beetle populations.     

树木枝干溃疡病菌致病力分化研究

应用人工接种的方法,对不同地理区域、不同寄主上的24个Botryosphaeria dothidea菌株、6个B.berengeriana菌株在不同抗性杨树上接种后的致病力分化进行了研究.结果显示:接种后是否形成愈伤组织是树木抗性的表现,也是真菌致病力大小的表现;B.dothidea菌株存在致病力分化现象,表现为强、中、弱3种类型,中等致病力的菌株是优势类群;以杨树为接种材料,24个B.dothidea菌株的致病力呈现一定的地理分化现象,致病力从强到弱依次为:华东(华南)>西北>华北(华中)>东北,即菌株致病力从我国南方向北方有逐渐变小的趋势;苹果轮纹病菌和干腐病菌的致病力也呈现出南方菌株大于北方菌株的特点.从溃疡病菌株的致病力出发分析了5种杨树的抗病性,其结果与以往的观察结果相一致.     

Ophiostomatoid fungi associated with root‐feeding bark beetles on Scots pine in Poland

Ophiostomatoid fungi are known to be associated with various species of bark beetles. However, information about fungal associates of root‐feeding bark beetles in Europe is still fragmentary. For this reason, the fungal associates of Hylastes ater, H. opacus and Hylurgus ligniperda on Pinus sylvestris were isolated and identified. A total of 743 fungal isolates were collected and separated into 10 morphological groups. Analyses of ITS rDNA and partial β‐tubulin gene sequences confirmed that these groups represented distinct species. The 10 species included a total of 13 associations between fungi and bark beetles that had not been recorded previously. All of the bark beetles examined were frequently associated with ophiostomatoid fungi. The fungal diversity and relative abundance of species were very similar in the three species of root‐feeding bark beetles. The most commonly encountered associates of these beetles were Grosmannia radiaticola, Leptographium lundbergii, L. procerum and L. truncatum. Insect infestation data furthermore suggest that Hylastes spp. and Hg. ligniperda are also important vectors of the fungal pathogen Sphaeropsis sapinea.     

Root‐colonizing ophiostomatoid fungi associated with dying and dead young Scots pine in Poland

Ophiostomatoid fungi are carried by various bark beetles. However, very little is known about the role of these fungi in conifer roots. We studied ophiostomatoid fungi in roots of dying and dead Pinus sylvestris trees and tested the potential phytotoxicity of some isolates using a sensitive bioassay with Lepidium sativum in Poland. Fungi were identified based on their morphology and DNA sequencing. Three ophiostomatoid fungi, Leptographium procerum, Sporothrix inflata and Ophiostoma pallidulum, were isolated from the roots. The most abundant soil‐borne fungus, S. inflata, and relatively rare O. pallidulum were isolated for the first time from roots of dying and dead pine trees. The frequency of S. inflata and O. pallidulum correlated with tree decline. The fungi were isolated more frequently from roots of dead than dying trees. Sporothrix inflata and O. pallidulum slightly reduced the stem and root growth of L. sativum. Leptographium procerum reduced more significantly root than stem growth. This species reduced root elongation 32–54% after 10–17 days of incubation.     

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.     

Pathogenic wood‐decaying fungi in China

Wood‐decaying fungi on living trees in China were surveyed over the last 12 years. In all, 102 potentially pathogenic Basidiomycetes were found in natural forests, forest plantations, parks and gardens, and among them 20 species were recorded for the first time on living trees in China. The host(s), occurrence, type of damage, type of decay and distribution of each species in China are given. Most of these wood‐destroying fungi are polypores in the Aphyllophorales, and the majority were found in temperate and boreal forests. Of all the species detected, 88 species are known to cause white rot, and 14 cause brown rot; 25 species are considered as common, 44 occasional, and 33 rare.     

In vitro interactions in artificial and wood‐based media between fungi colonizing stumps of Sitka spruce

Interspecific interactions between fungi that colonize stumps of Picea sitchensis in Scotland were tested in dual cultures on Norkrans agar, spruce sawdust and in autoclaved blocks cut from roots. Isolates were ranked according to competitive ability on the different media, based on their ability to overgrow competitors or to form deadlock interactions. On the defined medium, Phaeolus schweinitzii was the species most able to overgrow competitors, followed by Stereum sanguinolentum and Heterobasidion annosum; Resinicium bicolor was the least able to overgrow competitors on this medium. By contrast, R. bicolor was the most competitive on spruce sawdust medium. Deadlocking interactions were formed most often in dual cultures on Norkrans agar. Observation of hyphal interactions on Norkrans agar under the microscope identified several different response types including growth of thin hyphae compared to control cultures, hyphal coiling, vacuolation of hyphae, hyphal lysis of one competitor and deposition of crystals in the agar. Hymenomycetes caused varying amounts of decay in autoclaved root blocks. Resinicium bicolor was able to replace other species in most co‐inoculations. Stereum sanguinolentum appeared to be the least competitive species in root block inoculations, being replaced by Melanotus proteus and R. bicolor, although interactions with H. annosum varied widely. These results indicate that substrate has a marked effect on interspecific fungal interactions, with wood‐based, particularly intact woody tissues closely matching competitive behaviour displayed in the field.     

In vitro antagonism of Trichoderma and naturally occurring fungi from elms against Ophiostoma novo‐ulmi

The antagonist ability of four fungal endophytes isolated from elms (Trichoderma atroviride GF13, Diaporthe eres GF2, Diplodia ulmi GF5 and Alternaria alternata GF6) and two fungi from a collection (Trichoderma harzianum and Trichoderma virens) was evaluated against six strains of Ophiostoma novo‐ulmi (1, 10, 19, 32, 43 and O‐Va), the causal agent of Dutch elm disease. Dual culture tests were established in PDA and MA culture media to test the interactions between pathogenic fungi and potential antagonist ones. The fungi showed varying levels of antagonist capacity against O. novo‐ulmi. Among them, T. atroviride and T. harzianum had the greatest inhibiting effect (around 50%). Culture filtrates and previously inoculated cellophane discs from T. harzianum and T. atroviride also caused growth inhibition of O. novo‐ulmi, with less intensity in strains 10 and 19. The control mechanisms involved in the process were competition, mycoparasitism and the production of soluble and diffusible antifungal metabolites. The new isolate T. atroviride GF13 was efficient as antagonist. It showed antagonist ability against non‐elm‐derived fungi such as Verticillium, Fusarium, Aspergillus, Rhizopus and Penicillium. These results suggest interesting application possibilities in the biological control of Dutch elm disease.     

Association of seed‐borne Fusarium species on Pinus ponderosa with germination and seedling viability in Argentina

Pinus ponderosa seedlots from eight seed orchards in Western Patagonia, Argentina, were evaluated for the presence of Fusarium species, and isolates obtained were subjected to pathogenicity tests. The following species were isolated: Fusariumoxysporum, Fusarium equiseti, Fusarium verticillioides, Fusarium incarnatum, Fusarium acuminatum, Fusarium graminearum and Fusarium proliferatum. With the exception of F. proliferatum and F. graminearum all Fusarium species were recovered from P. ponderosa seedlings with symptoms of damping‐off. Five Fusarium species were re‐isolated from necrotic roots of seedlings showing chlorosis (F. oxysporum, F. equiseti, F. incarnatum, F. acuminatum and F. verticillioides). High damping‐off losses occurred in Fusarium‐contaminated seed of P. ponderosa and low disease incidence occurred on less contaminated seed. The role of seed‐borne inoculum in disease spread within forest nurseries is discussed.     

Pathogenicity of fungi isolated from Quercus suber in Catalonia (NE Spain)

Summary Thirty‐four fungal species isolated from cork oak (Quercus suber) in Catalonia (NE Spain) during 1992–95 were tested for pathogenicity either in stem, leaf or root inoculations. Eleven species were found to be pathogenic on stem: Biscogniauxia mediterranea, Botryosphaeria stevensii, Diatrype cf. stigma, Endothia gyrosa, Fusarium solani, Graphium sp., Ophiostoma quercus, Phomopsis sp., Phytophthora cinnamomi, Sporendocladia bactrospora and an unidentified Coelomycete. Three fungi showed pathogenic effects on leaves: Dendrophoma myriadea, Lembosia quercina and Phomopsis quercella. No clear pathogenic effects were detected in the root inoculation experiment. Trunk pathogens were differentiated into two groups according to the effects induced in the inoculated plants; B. stevensii, Phomopsis sp. and P. cinnamomi caused the death of the inoculated plants and induced the formation of large cankers and vascular necroses. The other pathogenic species also produced severe cankers and vascular lesions, but no significant mortality was detected. Water stress increased the lesions caused by B. mediterranea and Phomopsis sp., but limited those of P. cinnamomi and the rest of the inoculated fungi. However, water stress did not significantly affect the damage caused by B. stevensii, which was the most virulent of the species tested. Leaf pathogens only showed their effects if the leaf cuticle was previously damaged. Lembosia quercina caused small dark lesions whereas D. myriadea and P. quercella produced large necrotic areas in well‐watered plants. The lesions caused by the last two fungi were reduced by water stress.     

Effect of phenolic compounds on the in vitro growth of two fungi associated with Ips typographus

The susceptibility of Ophiostoma piceaperdum (n = 17) and O. bicolor isolates (n = 7) to catechin and resveratrol has been investigated with growth bioassays on malt‐agar medium. While catechin (10^?3 m ) did not have any significant influence, resveratrol (10^?3 m ) inhibited the growth of the two species, particularly the growth of O. piceaperdum isolates. We did not observe any synergistic inhibitory effect between the two phenolic compounds. Significant differences either in fungal growth on malt agar medium or susceptibility to resveratrol occurred among isolates for both species. Our results corroborate previous observations of high genetic and virulence variability among isolates of ophiostomatoid fungi and indicate that further studies should take into account the intraspecific variability of the fungal species of this group.     

Intraspecies variation of mature Pinus taeda in response to root‐infecting ophiostomatoid fungi

Seedling screening studies have shown intraspecies variation in susceptibility of Pinus taeda (loblolly pine) to Leptographium terebrantis and Grosmannia huntii, the causal agents of root infection in Pinus species. However, it is critical to understand the susceptibility of mature P. taeda trees. Roots of mature P. taeda families determined as susceptible and tolerant to L. terebrantis and G. huntii by previous seedling screening trials were artificially inoculated with the same fungal isolates. Dark necrotic lesion and vascular occlusion were recorded 8 weeks later. Families previously considered as susceptible had relatively longer lesions and occlusions. The variation in susceptibility to the two fungi remained the same as exhibited by families in the seedling trial. These results suggest intraspecies variation in relative susceptibility of P. taeda to L. terebrantis and G. huntii remains similar regardless of the age of the tree.     

Mountain pine beetle‐associated blue‐stain fungi in lodgepole × jack pine hybrids near Grande Prairie,Alberta (Canada)

The mountain pine beetle (MPB), the most serious pest of lodgepole pine in mountainous western Canada, spread northeastward into lodgepole × jack pine hybrids in the boreal forest of Alberta in 2006. The MPB vectors three species of blue‐stain fungi, which contribute to the success of the beetles. These fungi were isolated from MPB larvae and galleries in several lodgepole × jack pine stands in the Grande Prairie region of northwestern Alberta in autumn 2006 and winter and spring 2007. Fungi were recovered from more than 95% of gallery systems. The three fungi were similarly prevalent but Ophiostoma montium was the most frequently isolated fungus at each sampling point, isolated from 72% to 90% of gallery systems compared with 63% to 78% for Grosmannia clavigera, and 61% to 86% for Leptographium longiclavatum. Ophiostoma montium and G. clavigera were isolated from more larvae than gallery samples, with the opposite true for L. longiclavatum. Most gallery systems contained multiple fungi with three fungi per gallery system being more common in autumn and winter and two fungi more common in the spring. The combination of G. clavigera and L. longiclavatum was less common among gallery systems with two fungi than either of the pairwise combinations containing O. montium. Fungal prevalence was the same above and below snow level. The prevalence of the three fungi did not differ significantly among stands sampled in the spring but stands with more G. clavigera tended to have less L. longiclavatum. The winter of 2006–2007 was colder than average throughout Alberta with temperatures below ?30°C in November, January and February, and all three fungi were present after the cold winter while most larvae had died, suggesting that overwintering mortality of the fungi will not limit persistence and spread of MPB in the boreal forest.     

Pathogenicity of entomopathogenic fungi to <Emphasis Type="Italic">Thaumetopoea pityocampa</Emphasis> (Schiff.) (Lepidoptera: Thaumatopoeidae) larvae in laboratory conditions

Pathogenicity of 13 strains of entomopathogenic fungi belonging to genera Paecilomyces, Tolypocladium, Beauveria, Metarhizium, and Lecanicillium (=Verticillium) was evaluated on fourth instar larvae of Thaumetopoea pityocampa (Schiff.) (Lep.: Thaumatopoeidae). Larvae were treated individually by dipping into conidial suspension with the concentration of 1.0 × 10⁵ conidia ml⁻¹. All the tested isolates resulted in various levels of mortality (16–100%), and eleven of them were able to grow and sporulate on dead larvae. Mortality caused by three isolates of P. fumosoroseus, one isolate of Beauveria bassiana and one isolate of Metarhizium anisopliae were significantly higher than that of the other treatments. Three isolates of P. fumosoroseus, one isolate of P. farinosus, and one isolate of M. anisopliae showed mycelial growth and sporulation on significantly more cadavers compared to other treatments. Dose–mortality test was conducted with P. fumosoroseus (isolate no: 2679), which appeared to be the most promising isolate among the tested fungi, by spraying five concentrations of its conidia (1.0 × 10⁶, 1.5 × 10⁶, 7.5 × 10⁶, 1.0 × 10⁷ and 1.5 × 10⁷ conidia ml⁻¹). The estimation of the LC₅₀ (95% confidence limits) was 3.4 × 10⁶ (9.3 × 10⁵ − 6.9 × 10⁶) conidia ml⁻¹. This study showed that P. fumosoroseus (isolate no: 2679) could be a good candidate as a microbial control agent against T. pityocampa in forest environment.     

Ecology of wood‐inhabiting fungi in northern forests of Iran

Wood‐decay fungi are important in forest ecosystems and play an important role in nutrient and carbon recycling. The scope of this research was to identify wood‐inhabiting and wood‐decay macrofungi and determine their ecology. We sampled standing trees and dead wood of oak and hornbeam in the northern forests of Iran, specifically in Mazandaran and Golestan provinces. We assessed the influence of field slope, elevation, site direction and the height of fungal sporocarp position on the tree. The species of wood‐inhabiting and wood‐decay fungi belonged to 11 families: Polyporaceae; Stereaceae; Ganodermataceae; Physalacriaceae; Agaricaceae; Xylariaceae; Pluteaceae; Coprinaceae; Strophariaceae; Pleurotaceae; and Hydnaceae. The most common fungal species identified belonged to the Polyporaceae and Ganodermataceae and predominantly grew on trees growing on a slope of 20%, at an altitude of 700–900 m, 0–3 m from the ground a southwesterly site direction and the northern sides of both tree species. These results indicated that field slope was the most important factor in determining fungal sporocarp abundance.     

Effects of oleoresins and monoterpenes on in vitro growth of fungi associated with pine decline in the Southern United States

As a means of exploring pine resistance to root disease and declines, the effects of host plant secondary metabolites on the growth of root colonizing fungi associated with three diseases/declines of southern pines – loblolly pine decline, littleleaf disease and annosum root rot were tested. The associated fungi –Leptographium huntii, L. serpens, L. terebrantis, L. procerum, Heterobasidion annosum and Phytophthora cinnamomi– were grown in saturated atmospheres or in direct contact with, pure monoterpenes and crude oleoresin collected from the four southern pines (Pinus taeda, P. eschinata, P. palustris and P. elliotti) for 7 day. Fungal growth was measured at 3, 5 and 7 day. Root‐infecting fungi differed significantly in sensitivity to crude oleoresin and pure monoterpenes. All fungi tested were inhibited, to some extent, by the resins tested. H. annosum and P. cinnamomi were strongly inhibited by all the monoterpenes tested. The ophiostomatoid fungi were significantly less affected by the compounds tested. L. huntii and L. serpens were less inhibited by monoterpenes than either L. terebrantis or L. procerum. These fungal growth studies show that the kind and amount of secondary metabolite produced by the host plant have a profound effect on tree pathogens. Alterations of tree physiology may have implications for defenses against tree pathogens as well as to the ecology and management of forest ecosystems. Difference in incidence of root disease observed in the field may be explained by the ability of the fungus to tolerate these host defense mechanisms.