Needle blight of pine caused by two species of Dothistroma in Hungary

Dothistroma septosporum and D. pini cause the serious pine needle disease, Dothistroma needle blight (DNB). Of these, D. septosporum has a global distribution, while D. pini is known only from the USA, Russia, Ukraine and France. During a study of D. septosporum isolates from Pinus nigra in Hungary, microsatellite markers revealed the presence of a second Dothistroma species. The aim of this study was to identify the DNB pathogens occurring in Hungary using four different molecular techniques. These included sequencing of the rDNA ITS region, a species‐specific ITS‐RFLP, mating type primers and a diagnostic microsatellite marker, Doth_A. Results showed that both D. septosporum and D. pini occur in Hungary and that both DNB pathogens were present on the same trees and in some cases, the same needles. Mating types MAT1 and MAT2 of D. septosporum were shown to be present in Hungary, but only the MAT2 of D. pini was found. In addition, examination of needles with DNB symptoms from Russia revealed the presence of D. pini on Pinus pallasiana and P. nigra as well as on a new host, Pinus mugo. The molecular markers applied in this study were sufficiently robust to identify and differentiate between the two DNB pathogens, both in culture and directly from needles. They will consequently be useful to document the geographical range and to monitor the spread of D. septosporum and D. pini in future studies.     

New host and country records of the Dothistroma needle blight pathogens from Europe and Asia

Dothistroma needle blight (DNB) is a serious disease of pines (Pinus spp.), with a worldwide distribution. It is caused by the ascomycete fungi Dothistroma septosporum (teleomorph: Mycosphaerella pini) and Dothistroma pini (teleomorph unknown). Recently, DNB was found on Pinus peuce in Austria, Pinus pallasiana in Ukraine and the European part of south-western Russia, as well as on Pinus radiata and Pinus wallichiana in Bhutan. Based on DNA sequence comparisons of the internal transcribed spacer and β-tubulin gene regions, isolates from Austria and Bhutan were identified as D. septosporum, while isolates from Ukraine and south-western Russia were identified as D. pini. Additional isolates studied from Pinus mugo in Hungary confirmed the presence of D. septosporum in this country. The record of D. septosporum on exotic P. peuce in Austria represents a new host report of this needle blight pathogen in Europe. Likewise, DNB and the associated pathogen, D. septosporum are reported from Bhutan, eastern Himalayas, for the first time. In addition, D. pini was found in two European countries and on a new host, P. pallasiana. These European records represent the only reports of D. pini from outside the north-central USA. Morphological examination of selected specimens from different hosts and countries showed that D. septosporum and D. pini overlap in the length of their conidia, while the width is slightly wider in D. pini than in D. septosporum. The differences in conidial width are so small, however, that identification of the two Dothistroma species solely based on morphology is virtually impossible. The new host and country records provided here are consistent with the continuing trend of reports of the DNB pathogens from new hosts and new geographical areas during the last two decades, particularly in the northern hemisphere.     

Dothistroma Needle Blight in Slovenia is caused by two cryptic species: Dothistroma pini and Dothistroma septosporum

Dothistroma Needle Blight (DNB) has been known in Slovenia since 1971, but the disease symptoms have intensified in recent years. With this study, the DNB symptoms in Slovenia are for the first time connected to both phytopathogenic Dothistroma species: D. pini and D. septosporum. Based on the ITS‐rDNA comparisons, the Slovenian isolates of D. pini group with the D. pini isolates from France, Hungary and the USA and not with the D. pini isolates from Russia and Ukraine. Both mating types MAT1 and MAT2 of D. septosporum are present in Slovenia, while for D. pini, only MAT2 was found. The current widespread occurrence of D. pini in native forest stands indicates the prolonged presence of this species in Slovenia.     

The hosts and geographic range of Dothistroma needle blight in Slovakia

The occurrence and distribution of Dothistroma needle blight (DNB) were studied in 2014–2017 around Slovakia. A total of 84 localities, both native and planted, were investigated, and the presence of DNB was confirmed in 73 of them. In all positive locations, symptoms typical of DNB were observed and the Dothistroma species was confirmed using species‐specific primers either from fungal cultures or directly from needles. Both Dothistroma species—D. septosporum and D. pini—were identified. Both species occurred together in 29 locations, only D. septosporum in 42 and only D. pini in two locations. The host range of D. septosporum included 10 pine species and two spruce species. The host range of D. pini comprised the same number of pine hosts but only one spruce species. Five pine hosts, P. aristata, P. coulteri, P. densiflora, P. jeffreyi, P. × schwerinii, and one spruce host P. abies are new hosts species of D. pini. P. densiflora and Picea pungens have earlier been reported to be susceptible for DNB. In this study, D. septosporum was found from both tree species.     

Dothistromin biosynthesis genes allow inter‐ and intraspecific differentiation between Dothistroma pine needle blight fungi

Dothistroma septosporum and D. pini are the causal agents of Dothistroma needle blight (DNB) of Pinus spp. in natural forests and plantations. The main aim of this study was to develop molecular diagnostic procedures to distinguish between isolates within D. septosporum, for use in biosecurity and forest health surveillance programmes. This is of particular interest for New Zealand where the population is clonal and introduction of a new isolate of the opposite mating type could have serious consequences. Areas of diversity in the dothistromin toxin gene clusters were identified in D. septosporum (51 isolates) and D. pini (6 isolates) and used as the basis of two types of diagnostic tests. PCR‐restriction fragment length polymorphism (RFLP) of part of the dothistromin polyketide synthase gene (pksA) enabled distinction between two groups of D. septosporum isolates (A and B) as well as distinguishing D. septosporum and D. pini. The intergenic region between the epoA and avfA genes allowed further resolution between some of the A group isolates in RFLP assays. These regions were analysed further to develop a rapid real‐time PCR method for diagnosis by high‐resolution melting (HRM) curve analysis. The pksA gene enabled rapid discrimination between D. septosporum and D. pini, whilst the epoA–avfA region distinguished the New Zealand isolate from most other isolates in the collection, including some isolates from DNB epidemics in Canada and Europe. Although this study is focused on differences between the New Zealand isolate and other global isolates, this type of diagnostic system could be used more generally for high‐throughput screening of D. septosporum isolates.     

Dothistroma spp. in Western Ukraine and Georgia

Dothistroma needle blight (DNB) is among the most serious foliar diseases affecting Pinus spp. globally. Infected needles were collected from potential host species in four locations in western Ukraine and in four locations in eastern Georgia during spring–summer 2015 to update the knowledge on pathogen distribution in these countries. Dothistroma spp. were detected using isolation, sequencing and species‐specific priming (SSPP) PCR. Two new hosts for Dothistroma spp. were recorded in western Ukraine: D. septosporum on Pinus nigra var. australica and D. pini on P. nigra var. mollet. D. septosporum was found on 15‐year‐old P. strobus in western Ukraine. New hosts for D. septosporum were recorded in Georgia on 5‐ to 10‐year‐old naturally regenerated P. sylvestris var. hamata and on 40‐ to 50‐year‐old P. ponderosa trees. D. pini was found for the first time in Georgia on 30‐ to 40‐year‐old P. nigra trees. The work confirmed the presence of both D. septosporum and D. pini in western Ukraine and Georgia, and demonstrated new hosts for both Dothistroma species.     

Modelling of the spread of Dothistroma septosporum in Europe

Dothistroma needle blight (DNB), a disease affecting several pine species, is currently generating great concern in Europe. Caused by Dothistroma pini and Dothistroma septosporum, DNB affects pine needles and causes premature defoliation, which results in growth reduction and, in extreme cases, mortality. The disease has increased in importance in Europe over the last 20 years, with an increase in the number of observations of DNB in regions with large areas of Pinus sylvestris in northern Europe. This article presents a cell‐based spatiotemporal model for predicting the likelihood and intensity of the future spread of D. septosporum in Europe. Here, “spread” includes both invasion of new regions and infection of healthy stands within already‐colonized regions. Predicted spread depends on the availability of host species, climatic suitability of different regions to D. septosporum and dispersal of sexual and asexual spores from infected trees to surrounding forests via water splash, mist and wind. Long‐distance spread through transport of infected seedlings is also included in the model. Simulations of spread until 2007 and 2015 were used to validate the model. These simulations produced similar patterns of spread to those observed in Europe. Simulations for 2030 suggested that additional and new outbreaks are likely to occur in Scotland, southern Norway, southern and central Sweden, northern parts of Germany and Poland, Estonia, Latvia and south‐west Finland. Preventing the delivery of infected seedlings would be an effective method for reducing the spread of D. septosporum in the Nordic countries, Scotland and Ireland, the Baltic countries, and parts of Germany, Poland and Belarus. In these states, prevention of transport of infected seedlings can reduce the probability of additional spread by 15%–40%.     

Fungicide sensitivity of Dothistroma septosporum isolates in the UK

Dothistroma septosporum, a causal agent of Dothistroma needle blight (DNB), is a damaging fungal pathogen of pines that has recently started to affect native Scots pine woodlands in the UK. In addition to silvicultural methods, fungicide spraying of forest nursery stock can help prevent the spread of DNB. However, the effectiveness of modern single‐site fungicides against D. septosporum and the risk of fungicide resistance evolution remain largely unknown. In this project, we aimed to establish sensitivity profiles of D. septosporum to some widely used single‐site fungicide classes in vitro, and to determine whether fungicide resistance is already present, as this could increase the spread of D. septosporum genotypes on planting stock in native woodlands. For this purpose, we compared isolates of D. septosporum, originating from pine stands unexposed to fungicides, with isolates from nursery outbreaks, for sensitivity to a range of commonly applied fungicides. Most of the fungicides we tested were effective in vitro and we observed no significant shifts in sensitivity in forest nurseries. Although further tests in planta are required to confirm effectiveness of single‐site fungicides against D. septosporum, our results suggest that they can be successfully used in DNB control, although appropriate measures to prevent the evolution of fungicide resistance are strongly recommended.     

Dothistroma pini—Eine Gefahr fürPinus sylvestris?

Zusammenfassung Dothistroma pini wurde auf einer ganzen Reihe von Kiefernarten festgestellt. Sporenl?nge und das gelegentliche Auftreten der HauptfruchtformScirrhia pini belegen das Vorhandensein der var.linearis. Konidien wurden von M?rz bis in den November gefunden, besonders zahlreich von April bis Juni. Erstmals, festgestellt wurde das Pathogen aufPinus cembra, P. aristata, P. koraiensis undP. tabuliformis, allerdings ist anzunehmen, da? die Infektion dieser Arten nur unter starkem Infektionsdruck stattfindet und die wirtschaftliche Bedeutung des Pilzes in diesen F?llen sehr begrenzt ist. Gleiches gilt offensichtlich auch fürPinus sylvestris. Diese Kiefernart wird nach unseren Beobachtungen auch bei engem Kontakt mit sehr stark infizierten Schwarzkiefern nur unbedeutend oder überhaupt nicht befallen. In diese Richtung deuten auch die ersten Ergebnisse eines Infektionsversuches, bei demP. sylvestris symptomfrei blieb, w?hrendP. nigra z. T. sehr starke Symptome einer Infektion zeigte. Eine nennenswerte Sch?digung vonP. sylvestris konnte bei Freilandbeobachtungen in keinem Falle festgestellt werden. Gegenteilige Hinweise in der Literatur sind auf Verwechselungen (des Pathogens oder des Wirtes) zurückzuführen.

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Dothistroma pini—a danger for Pinus sylvestris?

Summary Dothistroma pini was found on several different pine species. The length of the conidia and the existence of the teleomorph, which sometimes could be noticed, prove the presence of the var.linearis of the fungus. Conidia were detected between March and November, in high quantities especially from April to June. For the first time, the pathogen has been mentioned onPinus cembra, P. aristata, P. koraiensis andP. tabuliformis. It must be assumed, however, that these species only become infected under high infection pressure. That means, that the economic importance of the pathogen is very limited in these cases. Obviously the same is true withPinus sylvestris. After our observations this pine species was little or not at all infested even when in direct contact with heavily attackedP. nigra-plantations. The result of an infection experiment, in whichP. sylvestris did not show any symptoms whereasP. nigra was partly heavily attacked, points in the same direction. Damages toPinus sylvestris worth mentioning could not be found during observations in the field. Opposite indications from the literature can be explained by mistaking pathogens or host species.

     

Detection of pine needle diseases caused by Dothistroma septosporum,Dothistroma pini and Lecanosticta acicola using different methodologies

Dothistroma and Lecanosticta needle blight are among the most damaging foliage diseases in plantations and natural pine stands worldwide, and are therefore listed as quarantine organisms in numerous countries. The aim of this study was to validate different methodologies used for the detection of both diseases. Based on multiplex quantitative PCR (qPCR), simultaneous detection and discrimination of the three pathogens Dothistroma septosporum, Dothistroma pini and Lecanosticta acicola were possible in symptomatic needles. Additionally, the same set of needles was analysed for the presence of all three pathogens using novel end‐point PCR assays followed by enzymatic digestion of PCR products. Results were compared with the qPCR data, and for validation, collected needles were screened morphologically for the presence of typical fruiting bodies and spores. Differences in detection sensitivity were found between the detection tools. The qPCR‐based detection allowed for specific and efficient identification and quantification of all three pathogens simultaneously. At the same time, conventional PCR assays, which target the multicopy ITS region, showed increased detection sensitivity and can be conducted without the use of expensive infrastructure and reagents.     

Invasive brown spot needle blight caused by Lecanosticta acicola in Estonia

Lecanosticta acicola (Thüm.) Syd., a serious foliage pathogen of pines in many regions of the world, is an emerging invasive species in northern Europe. After the first record of L. acicola on Pinus ponderosa Dougl. ex Laws. in northern Estonia in 2008, monitoring was started to investigate the spread and host range of the fungus in the country. By the beginning of 2015, L. acicola was also recorded on P. uncinata Mill. ex Mirb., P. mugo Turra and on P. mugo var. pumilio (Haenke) Zenari, being the northernmost records of the fungus in Europe. So far, the single native pine species Pinus sylvestris L. has not been found to be infected. Molecular analysis proved infection of L. acicola on pines in five different localities of Estonia: in Tallinn (mainly in the Botanic Garden, northern Estonia), in Tori and Kärdla (western Estonia), in Vasula and Kärevere (east-central Estonia). The sexual state (teleomorph) of the fungus was not found, but the existence of the both mating types, MAT1-1 and MAT1-2, was confirmed.     

First molecular detection of Lecanosticta acicola from Poland on Pinus mugo

The presence of quarantine pathogen Lecanosticta acicola, the causal agent of brown spot needle blight, was confirmed using molecular methods for the first time in northern part of Poland on the coastal area adjacent to the Baltic Sea. This area includes sandy beaches, where Pinus mugo and P. sylvestris were planted. Symptomatic needles were collected in 2017 from 20 P. mugo trees growing in one stand in Ustka. Typical symptoms of brown spot needle blight infection, including dead needle tips and central zones with yellow or reddish brown, circular spots in green tissue, were observed on all samples. Only, the asexual stage of L. acicola was obtained during this work. The pathogen species identity was confirmed using classical morphological methods (microscopic examination of the infected needles), real‐time and species‐specific priming (SSPP) PCR, and ITS sequencing. Analysis of mating‐type (MAT) genes showed the presence of both mating types in northern Poland.     

Artificial inoculation and susceptibility of Pinus armandii to Dothistroma septosporum

Dothistroma needle blight (DNB) is a serious needle disease of conifers that primarily affects pine species (Pinus spp.). Dothistroma septosporum is one of the DNB pathogens that has a diverse range of host species excluding Pinus armandii. In 15 inoculated P. armandii seedlings, D. septosporum acervuli were observed in 43 infected needles of ten seedlings with a mean disease severity of 1.11% at 25 weeks after inoculations, demonstrating the potential of D. septosporum to cause symptoms on the needles of P. armandii via artificial inoculation. The disease severity of P. armandii was similar to the positive control, Pinus nigra (median 0.75 for P. armandii to 0.70 for P. nigra), thus, P. armandii acts under artificial conditions as a susceptible host species.     

Production of a polyclonal antibody to Phellinus pini and examination of its potential use in diagnostic assays

In order to differentiate among Phellinus pini, Inonotus tomentosus and Inonotus circinatus a polyclonal antibody was raised to a N-terminal part of 25-kDa P. pini-specific protein. The specificity of the polyclonal antibody produced against a synthetic N-terminal peptide of this protein was investigated for diagnostic purposes using Western immunoblot, indirect enzyme-linked immunosorbent assay (ELISA), and inhibition ELISA techniques. The N-terminal synthetic peptide, used as the immunogen, was found to be more than 80% pure by reverse-phase high-performance liquid chromatography. Following immunization, antisera were collected at three different time intervals. The antibody molecules were purified from the crude antisera using immunoaffinity gel chromatography. Following one-dimensional sodium dodecylsulfate-polyacrylamide gel electrophoresis, Western immunoblot analysis showed that the P. pini I polyclonal-antibody detected the immunogen, the 25-kDa protein, in all but one of the P. pini isolates examined, but in none of the isolates of the nontarget species I. tomentosus and I. circinatus. Nevertheless, cross-reactivity was a problem because the P. pini I polyclonal-antibody also recognized bands at other molecular weights in nearly all of the isolates of the other species tested. With the indirect ELISA the P. pini isolates tended to have higher affinity for the polyclonal antibody than the nontarget species, but some cross-reactivity did occur. Inhibition ELISAs, performed over a range of soluble antigen concentrations (1.56–400 ng/100 μl), failed to show a clear distinction between P. pini and the two Inonotus spp. The low level of cross-reactivity observed for I. tomentosus isolate 52 (9%) was also apparent in the indirect ELISA analysis. All three assays indicated that P. pini isolate 41 was the most antigenic. Despite cross-reactivity, the antibody is useful in Western immunoblots for the diagnosis of most P. pini isolates.     

Dothistroma (red‐band) needle blight of pines and the dothistromin toxin: a review

Dothistroma (red‐band) needle blight has been a problem in plantations of exotic pines in the southern hemisphere for many decades. The prevalence of this disease is currently increasing in the northern hemisphere and is now affecting trees in their native ranges. The fungal pathogen Mycosphaerella pini with its anamorph Dothistroma pini, which is responsible for the disease, produces a toxin, dothistromin, that is closely related to the potent carcinogen, aflatoxin. Understandably this has provoked concern about possible effects on the health of forestry workers. This review gives a broad coverage of literature on both disease and toxin. The fungus has a complicated taxonomy with many synonyms and in most countries only the anamorph is found. It is a necrotrophic pathogen that kills needle tissue and completes its life cycle in the lesion thus formed. Dispersal of the disease is normally by rain splash of conidiospores but there is evidence that long range dispersal has occurred by transport of contaminated plant tissue and by wind/cloud dispersal of spores in air currents. The severity of disease is affected by humidity, temperature and light. There is variation in susceptibility of different Pinus species and some achieve increased resistance with age. The current method of control in southern hemisphere plantation forests is through spraying with copper fungicides and, with P. radiata, increased disease resistance has been achieved through a breeding programme. The dothistromin toxin is a difuroanthraquinone and is similar in structure to the aflatoxin precursor versicolorin B. Part of a gene cluster encoding dothistromin biosynthetic genes has been cloned and this has confirmed parallels between the dothistromin and aflatoxin biosynthetic pathways. Dothistromin produces damaging oxygen radicals by reductive oxygen activation rather than by photosensitization, but is also thought to exert its toxic effects on specific cellular targets. Studies have shown that dothistromin is a weak mutagen and clastogen and is therefore a potential carcinogen. Although the risks to forest workers are considered very low it is prudent to avoid unnecessary exposure during periods when dothistromin levels are likely to be at their peak.     

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.     

Chloroplast DNA polymorphism of an exotic P. mugo Turra population introduced to seaside spit of Kursiu Nerija in Lithuania

The aim of this study was to elucidate the introduction history of P. mugo in the unique landscape of the Lithuanian seaside spit of Kursiu Nerija by assessing its genetic structure and the genetic diversity. The individuals were sampled in 12 populations within an area of 3 km × 50 km along the Lithuanian part of Kursiu Nerija. P. mugo was introduced over 200 years ago to prevent sand erosion by establishing a forest cover. Chloroplast DNA polymorphism of 220 individuals of P. mugo together with 18 P. sylvestris and 11 putative P. sylvestris × P. mugo hybrids was assessed by the aid of five microsatellite markers. The standard intra-population diversity indexes were calculated. The intra-specific variation between distinct morphotypes as well as the population differentiation within the most spread P. mugo ssp. rotundata morphotype was assessed based on the haplotype frequencies by hierarchical AMOVA, G_ST/R_ST test, UPGMA clustering and PCA methods. The genetic diversity of P. mugo in Kursiu Nerija was high (H_e = 0.95; 83 different haplotypes). All except one of the P. mugo populations sampled contained a notable share of private haplotypes. AMOVA revealed high intra-specific diversity but low differentiation between the P. mugo populations. Most of the haplotypic variance was within populations. The UPGMA clustering produced groups more corresponding to the sub-species morphotypes than the geography of the populations. There was no geographical pattern of reduction in genetic diversity towards the younger plantations. A strong candidate for a species-specific DNA marker was found. After several events of introduction, the genetic diversity of P. mugo in Kursiu Nerija is very high and is structured based on the sub-species morphotypes rather than geography. The high frequency of shared and notable frequency of private haplotypes in most of the populations indicate that the major part of the P. mugo material originates from a number of geographically and genetically related sources, which more likely are introductions from abroad that the local collections. The high frequency of private haplotypes in the northernmost populations leaves a possibility for minor introductions from other genetically distinct sources. The absence of private haplotypes in one of the sampled populations indicates the use of local seed collections. The large number of shared haplotypes provides a strong evidence for a geneflow among the P. mugo taxa.     

Early fungal community succession following crown fire in Pinus mugo stands and surface fire in Pinus sylvestris stands

The early post-fire development of mycobiota following a crown fire in mountain pine plantations and a surface fire in Scots pine plantations, and in the corresponding unburnt stands in the coastal sand dunes of the Curonian Spit in western Lithuania was investigated. Species numbers in unburnt Pinus mugo and Pinus sylvestris stands showed annual fluctuation, but in the burnt sites, the numbers of fungi increased yearly, especially in the crown fire plots. Both burnt stand types—P. mugo and P. sylvestris—showed strongly significant (two-way ANOSIM; R = 1, p < 0.05) differences in species composition; the differences between unburnt sites were clearly expressed but less significant (R = 0.86, p < 0.05). Fungal species composition of burnt P. mugo and P. sylvestris sites was qualitatively different from that of corresponding unburnt sites (two-way ANOSIM; R ≥ 0.75, p < 0.05). The chronosequence of mycobiota in surface fire burns was less clearly defined than in crown fire sites, reflecting the greater patchiness of impacts of the surface fire. Although both fire types were detrimental or at least damaging to all functional groups of fungi (saprobic on soil and forest litter, wood-inhabiting, biotrophic, and mycorrhizal and lichenized fungi), their recovery and appearance (fructification) patterns varied between the groups and among the burn types. The end of the early post-fire fungal succession (cessation of sporocarp production of pyrophilous fungi) was recorded 3 years after the fire for both crown and surface fire types, which is earlier than reported by other authors. Rare or threatened fungal species that are dependent on fire regime were not recorded during the study.     

Dothistroma pini Hulbary: A New Disease on Pine in Britain

Dothistroma pini Hulbary, not hitherto reported in Britain,is described. Since 1954, and possibly before this date, ithas caused serious defoliation of nursery pine stock, particularlyof Pinus nigra and P. ponderosa. It has been reported only fromtwo nurseries both in the Wareham area, Dorset, but the reasonfor this extremely limited distribution is unknown. Attacksare worst in wet summers. Spore inoculations on potted plantsgave severe infection.     

Inoculation of young pine seedlings with Peridermium pini from north-east Scotland

Seedlings of Pinus sylvestris, P. nigra ssp. laricio, and P. mugo vars mughus, rostrata and pumilio were inoculated at their cotyledon stage with aeciospores of Peridermium pini from north-east Scotland. Infection was determined by the presence of both mycelia and haustoria in free-hand or paraffin sections. In the second growing season, infected seedlings showed symptoms such as stem-swelling, development of spermogonia and aecia, and death. In a second experiment, young P. sylvestris seedlings from seven seed sources collected in Great Britain were inoculated and the infection were examined after 6 weeks. In the samples of 50 seedlings from each source, 30–70% seedlings were infected.