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1.
Leptoglossus occidentalis, an insect native to North America, was inadvertently introduced into Italy about 1999. The insect damages the cones of conifer trees, especially Pinus pinea (Italian Stone pine). Pinus pinea is also affected by Diplodia pinea, a fungus native to Italy, which is becoming an increasing threat because pine trees are becoming more susceptible to it as a consequence of global warming. Because the insect and the fungus both have the pine cones as a common habitat, a possible interaction between them has been postulated. The aim of this study was to ascertain whether L. occidentalis and D. pinea interact on P. pinea cones. The interaction was studied using real‐time PCR on a group of naturally infected insects collected from a forest, and a group raised in the laboratory and artificially inoculated with D. pinea conidia. Molecular analysis showed that D. pinea DNA occurred on both naturally infected and inoculated insects, but with significant differences between the two groups. The rapid and sensitive molecular technique made it possible to detect D. pinea DNA on the bodies of the insects, and to show that the native D. pinea occurred on the exotic insect.  相似文献   

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In this research, we examined decay patterns occurring in Quercus castaneifolia wood under natural conditions compared with controlled decay in vivo. Pleurotus ostreatus‐infected oak wood was obtained from the Sari forests in the north of Iran. The species causing decay was verified as P. ostreatus using rDNA‐ITS sequencing of pure cultures from infected sapwood. In addition to P. ostreatus, two wood‐inhabiting Ascomycota, Trichoderma harzianum and T. lixii, were present. Mass loss in oak sapwood samples exposed to P. ostreatus for 60 days was around 10 per cent. Samples were prepared from both naturally decayed wood and wood decayed under controlled conditions and examined using microscopy. P. ostreatus was found to produce a simultaneous white‐rot decay pattern in both conditions.  相似文献   

4.
An increasing decline and mortality of cork oak trees have been recently observed in central Italy and Sardinia Island. Following surveys conducted in three declining cork oak forests, a Phytophthora species was consistently isolated from soil samples collected from trees displaying different level of decline. Based on morphological features, growth rates at different temperatures and analysis of DNA sequences of the ITS region, all isolates were identified as Phytophthora cinnamomi Rands. This pathogen caused large brownish lesions on inoculated freshly cut branches of cork oak. It was re‐isolated from all infected tissues. These findings represent the first report of P. cinnamomi on cork oak trees in Italy.  相似文献   

5.
The epidemic outbreak in northern Europe of Neonectria neomacrospora, the causal agent of dieback in Abies spp., led the European and Mediterranean Plant Protection Organization (EPPO) to include the pathogen on its alert list in 2017. Effective monitoring of this pathogen calls for a rapid and sensitive method of identification and quantification. A probe‐based real‐time PCR (qPCR) assay based on the β‐tubulin gene was developed for the detection and quantification of N. neomacrospora in infected wood samples, and directly for ascospores. This study presents the first published species–specific molecular detection assay for N. neomacrospora. The analytical specificity was validated on taxonomically closely related fungal species as well as on 18 fungal species associated with the host (Abies sp.). The analytical sensitivity was tested on naturally infected wood, on purified pathogen DNA in a matrix of host DNA and on N. neomacrospora ascospores for detection of airborne inoculum. The latter was tested both with a DNA extraction step prior to qPCR and without DNA extraction by direct qPCR on collected ascospores. The assay was specific to N. neomacrospora, with a sensitivity of 130 fg purified DNA, or 10 ascospores by direct qPCR. Omitting DNA extraction and amplifying directly on unpurified ascospores improved assay sensitivity significantly.  相似文献   

6.
Ceratocystis platani was detected at several locations in the European side of Istanbul, causing severe dieback and mortality, mainly on Platanus × acerifolia imported from Italy approximately 160 years ago, but also on Platanus orientalis. In the work reported here, the causal agent of the dieback and mortality was identified based on morphological characteristics in culture and internal transcribed spacer (ITS) sequencing, confirming the presence of C. platani in Turkey. All analysed C. platani isolates from Istanbul shared the same ITS sequence with European C. platani strains available in the NCBI, but differed from other species previously ascribed to the Ceratocystis fimbriata species complex. Pathogenicity of C. platani was proven by inoculation onto 25‐cm‐long and 4–9 mm diam. P. orientalis twigs and the causal agent reisolated from symptomatic tissues. In a survey of 976 Platanus in streets and parks of Istanbul, 26.5% of trees showed symptoms of C. platani infections, whilst 5.6% were dead. The proportion of symptomatic trees was higher where management interventions, such as topping or pruning and felling infected trees had been carried out.  相似文献   

7.
The anamorphic fungus Cryptostroma corticale (Xylariaceae) causing sooty bark disease (SBD) of Acer pseudoplatanus has been reported within the past decade in several European countries. In the last decade, severe drought conditions during summer also resulted in an outbreak of SBD in park areas in Prague (Czech Republic). Trees with the typical syndromes were recorded in four localities in Prague. Some trees were also located in the alluvial plain of the Vltava River with groundwater available all year long. We suppose that the trees were stressed by full water saturation for several months after flood in 2012 and that SBD was triggered by anoxia and subsequent phytopththora root rot. Sterile fungi isolated from one tree and one lesion were identified as C. corticale based on the similarity of their ITS rDNA sequences with strains of C. corticale in a public collection. Furthermore, C. corticale was placed in the family Xylariaceae within the genus Biscogniauxia based on phylogenetic analysis of four genes (ITS nrDNA, actin, RPB2 and β‐tubulin). The closest relatives are B. bartholomei and Graphostroma platystoma.  相似文献   

8.
Sissoo (Dalbergia sissoo), commonly known as shisham, is amongst the finest woods of South Asia, but ‘wilt’ disease has caused a rapid decline in this species. The cause of the disease remains uncertain. The aim of this study is to identify the causal agent of the disease and characterize isolates made from diseased trees, based on genomic data and variations in virulence. Samples of infected roots, stems and the ooze exuded from infected trees were obtained from plants showing symptoms in different geographical regions of India for the isolation of microorganisms. Isolates were used to inoculate healthy plants. Based on the morphological characteristics, genus‐ and species‐specific PCR, and in silico analysis of 5.8S rDNA‐ITS regions, of the 38 fungal isolates, 24 and 14 were identified as Fusarium solani and Fusarium sp., respectively. In a pathotyping study, eighteen F. solani isolates, isolated from roots and stem parts of symptomatic plants, induced typical wilt symptoms when inoculated through soil and roots on D. sissoo seedlings of 1–15 months in age. The population of F. solani was the highest in infected roots and the lowest in parts of stems, gradually decreasing with height, and was isolated constantly up to approximately 40% height of the seedling. F. solani isolates used in inoculations were successfully re‐isolated from the rhizosphere, infected roots and wilted stems, as confirmed using isolate‐specific DNA fingerprints. Molecular phylogenies based on rDNA‐ITS sequences showed that the 38 isolates fell into 2 groups. Group I comprised of F. solani isolates from D. sissoo and F. solani sequences in the NCBI GenBank database, whereas group II included Fusarium isolates other than F. solani. These results are helpful in developing integrated control measures for this highly variable pathogen and to establish a base for future population studies.  相似文献   

9.
[目的]鉴定引起新疆巩留县杏树果实斑点病病原菌,研究造成杏树果实病害的原因,为当地杏树防治工作提供依据。[方法]采用常规组织分离法分离得到罹病杏真菌菌株,利用传统形态学观察和分子系统学分析相结合的方式对所分离出菌株进行分类鉴定及其致病性检测。[结果]杏果病斑处的病原菌在显微镜下观察到分生孢子形态与经PDA培养基培养后观察到分生孢子形态均与Thyrostroma carpophilum(Lév.) B.Sutton所产生分生孢子一致;将分离获得的3株真菌的rDNA-ITS片段测序后与NCBI参考序列进行多重序列比对的结果显示,其序列与T.carpophilum一致性为100%;在基于ITS基因序列构建的系统发育树中,3株菌与T.carpophilum聚在同一分支。在接种了T.carpophilum后,杏果实和叶片均产生明显病斑并且从其所产生的病斑上再次分离到所接菌,满足柯赫氏法则。[结论]从新疆巩留县杏果实病斑处分离获得的3株真菌,经鉴定为引起杏穿孔病的病原菌T.carpophilum。这是该菌首次在该地区发现并报道。  相似文献   

10.
During the winter of 2012, a leaf spot disease was observed on Euphrates poplar (Populus euphratica) in the forest areas of Khuzestan province, south‐western Iran, causing significant damage in the Karun's riverside forests. Symptoms consisted of necrotic dark brown, circular to oval, 5‐ to 10‐mm spots on both surfaces of the leaves. A fungus having distinct dictyospores similar to those produced by Alternaria spp. was observed. The morphological characteristics, as well as the phylogenetic analysis of the internal transcribed spacer (ITS1‐5,8S‐ITS2) region, confirmed the identity of the strains belonging to the species Alternaria alternata. Pathogenicity tests were conducted on the alive leaves of P. euphratica on the young branches, as well as on the detached leaves in Petri dishes, through inoculation with spore suspension. Target spot symptoms similar to those observed in naturally infected leaves were developed on the inoculated leaves seven to 10 days after inoculation in both the inoculation procedures. A. alternata was consistently re‐isolated from the spots. Interestingly, similar symptoms were observed 7 days after detached leaf treatment with droplets of 15‐day‐old fungal culture filtrate, suggesting the production of pathotoxic compounds by the fungus. To our knowledge, this is the first report of A. alternata causing leaf spot on Euphrates poplar in Iran.  相似文献   

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12.
Heterobasidion annosum (Fr.) Bref. sensu lato is an important fungal parasite of coniferous trees throughout the temperate regions of the world. Approximately 15% of Heterobasidion isolates are infected by dsRNA viruses, which are considered as obligately intracellular and transmit vertically into both basidiospores and asexual conidia. Insects such as H. abietis and its larvae feeding on wood colonized by Heterobasidion fungi may carry Heterobasidion conidia and hyphae. In this study, we used H. abietis as a model species to reveal whether Heterobasidion viruses resist in their fungal host passing through the digestive tract of insects that may potentially serve as disseminators of fungal propagules. Pinus sylvestris branches were inoculated by three different strains of Heterobasidion parviporum, hosting taxonomically diverse virus species: Heterobasidion RNA virus 2, HetRV4 and HetRV6. Then, the inoculated branches were fed to H. abietis insects, and beetle excrements were investigated for Heterobasidion infections. All the inoculated fungal strains survived passage through the alimentary tract of the insects (survival rate 25–80%). The passage rates of the viruses inside their hosts varied considerably, ranging from 0 to 67%. Two different mycoviruses, HetRV2 and HetRV6, survived the intestinal passage of their fungal host, while the virus species HetRV4 was detected among none of the five H. parviporum isolates retrieved from insect faeces. The relative stability of fungi harbouring viruses suggests that if viruses are to be used for biological control against Heterobasidion species, it is likely that insect feeding does not considerably decrease the virus effect, but might instead enhance short‐range dispersal of the viral biocontrol agent.  相似文献   

13.
Diplodia corticola is one of the most aggressive fungal pathogens of Quercus species and is involved in the decline of Mediterranean cork oak forests and Californian oaks. Information regarding variation in virulence between strains is scarce. We hypothesize that D. corticola strains differ in virulence and consequently induce different symptoms in infected plants. To test this, infection assays were carried out on Quercus suber half‐sib seedlings with seven strains of D. corticola. Visual symptoms of infection (external lesions, leaf wilting, exudation and others) were recorded in parallel with physiological and biochemical parameters. All strains were able to cause lesions but at differing levels of aggressiveness. We show that internal lesion length did not correlate directly with strain aggressiveness and this agrees with physiological parameters that should be taken into account to infer about strain pathogenicity. Infection by all strains induced an overall negative impact on the net photosynthetic rate and an increase in the oxidative stress status of plants; however, significant differences were found when the effects of different strains were compared. Results also suggest that being under optimum growth conditions, prior to and during infection, allowed plants to respond to the pathogen. At the end of the experiment, some strains of D. corticola established a latent pathogen‐like relationship with cork oak. This is the first study to show that D. corticola virulence is strain‐dependent.  相似文献   

14.
The aim of this study was to assess genetic diversity and spatial distribution of 123 strains of the root‐rot pathogen Neonectria macrodidyma isolated from fine living roots of bare‐root nursery cultivated Pinus sylvestris. We found that ITS rDNA sequences were 100% identical for all strains. Arbitrary primed PCR fragment analysis showed the presence of two distinct N. macrodidyma types (Type 1 and Type 2), which included respectively 11 and 14 genotypes composed of 1–29 strains. Results of this study indicate that N. macrodidyma is commonly associated with fine living roots of pine seedlings, is largely disseminated by asexual means of local genotypes and has even distribution in the forest nursery soils. In living roots, N. macrodidyma is likely present as dormant propagules but under favourable conditions, it may develop rapidly and have a significant negative effect on plant health and productivity.  相似文献   

15.
Diplodia pinea causes shoot blight and collar rot diseases of pines in forest tree nurseries and sporulates on colonized seedling needles and stems. In late summer 2005, pycnidia of D. pinea were observed on shoots that had been excised by top pruning red pine seedlings earlier that summer during the third season of growth. This observation prompted surveys to determine the incidence and abundance of D. pinea conidia on excised shoots. At each of two nurseries, excised shoots were collected from the seedling canopy and adjacent alleyway soil in two subplots in each of five beds (plots). Excised shoots from both nurseries bore pycnidia with conidia of D. pinea. A water washing and filtration technique was used to quantify D. pinea conidia extracted from these shoots. Excised shoots collected from the seedling canopy yielded more D. pinea conidia than shoots collected from adjacent alleyway soil. Collection and removal of excised shoots resulting from top pruning of pine nursery seedlings should be considered as a means of reducing inoculum in areas where D. pinea is present.  相似文献   

16.
Acacia hybrid (Acacia mangium × Acacia auriculiformis) clones are widely planted in Vietnam with a total of approximately 400,000 ha to meet the demand for pulpwood, sawn timber and wood chip exports. Silvicultural techniques such as pruning and thinning have been applied to improve productivity and sawlog quality of Acacia hybrid plantations. However, those techniques may also create opportunities for wood decay fungi to enter the Acacia hybrid stems through wounds and cause stem defects that reduce sawlog quality and the value of the plantation. The presence of fungal decay agents in Acacia hybrid trees was examined in two Vietnamese plantations. In July 2011, just prior to a second thinning, discoloured wood samples were taken from a three‐year‐old Acacia hybrid plantation at Phan Truong Hai for the isolation of fungi. In July 2012, approximately 18 months after pruning and thinning treatments, discoloured wood samples were taken from a three‐year‐old Acacia hybrid plantation at Nghia Trung for the isolation of fungi. DNA sequencing of the rDNA ITS identified the isolates. In May 2015, approximately 4 years after thinning and fertilizer treatments, discoloured and decayed wood samples were taken from the above (7‐year‐old) Acacia hybrid plantation at Phan Truong Hai for fungal identification. DNA was extracted directly from discoloured and decayed wood samples and fungal rDNA ITS amplicons sequenced on a Roche 454 sequencer. The results showed that silvicultural treatments did not affect the fungal communities associated with discoloured and decayed wood of Acacia hybrid plantation at Phan Truong Hai. A total of 135 fungal species or OTUs (operational taxonomic units) were identified, including 82 members of Ascomycota and 52 Basidiomycota.  相似文献   

17.
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.  相似文献   

18.
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.  相似文献   

19.
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.  相似文献   

20.
In 2013, the death of needles of dwarf Alberta spruce (Picea glauca ‘Conica’) seedlings was observed in an arboretum in Slovenia. Needles on infected P. glauca ‘Conica’ twigs were covered with thick, yellowish brown hyphal cushions. The outgrowing mycelium was white and had completely overgrown some needles and connected them to each other. To definitively identify the fungus affecting P. glauca ‘Conica’, morphological examinations of isolated pure cultures were performed, and the internal transcribed spacer (ITS) region of the rDNA was sequenced. Based on morphological and molecular phylogenetic analyses together with the performed pathogenicity trial, we identified the causal agent of the observed symptoms as Rhizoctonia butinii – a recently described species. This report connects this fungus to a new locality and a new host plant.  相似文献   

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