First report of Sphaeropsis canker on cypress in South Africa

In this study a Sphaeropsis sp. is recorded as causing cankers on cypress in South Africa. These cankers are either found alone or on trees which are infected with the well-known cypress canker pathogen, Seiridium unicorne. When the two fungi occurred together, lesions could not be distinguished from each other, although the greater number of lesions were caused by the Sphaeropsis sp. The fungus appears to be identical to that described as Sphaeropsis sapinea f.sp. cupressi, in Israel, which is a fungus very different from the pine pathogen, Sphaeropsis sapinea. Pathogenicity tests showed that the Sphaeropsis sp. is significantly more pathogenic to Cupressus lusitanica than to Pinus roxburgii and Pinus elliottii. S. sapinea was pathogenic only to the Pinus spp. tested, and not to C. lusitanica. In contrast, the cypress pathogen Seiridium unicorne, was pathogenic to C. lusitanica and also to the two Pinus spp. tested.     

Mycoflora associated with slash-pine seeds from cones collected at seed orchards and cone-processing facilities in the south-eastern USA1

A diverse mycoflora was associated with slash-pine seeds from cones obtained at seed orchards and cone-processing facilities in the southeastern USA. Potentially pathogenic fungi, including Lasiodiplodia tbeobromae, Sphaeropsis sapinea, and Fusarium spp., were isolated from seeds of cones obtained at all locations. Fusarium proliferatum and F. semitectum were the Fusarium spp. most often associated with seeds; F. subglutinans was rarely isolated. Many other species of fungi were associated with seeds from all locations. Incidence of fungus-damaged seeds ranged from 2 to 11% among collection sites. Disease incidence was found to increase during cone storage. Types of fungi and the incidence of fungus-damaged seeds were similar for damaged/discoloured cones and apparently healthy cones. The potential for slash-pine seeds to become colonized by pathogenic fungi, such as S. sapinea and the various Fusarium spp., could have important implications for the establishment of these fungi in forest-tree nurseries, as well as their introduction into foreign countries.     

Persistence of some pine pathogens in coarse woody debris and cones in a Pinus pinea forest

The persistence of Sphaeropsis sapinea, Leptographium serpens and Heterobasidion annosum s.s. in artificially inoculated pine branch pieces (S. sapinea and L. serpens) and wood blocks (L. serpens and H. annosum s.s.) was investigated in order to discuss the alternative of leaving coarse woody debris in stands of Italian stone pine (Pinus pinea). Also, natural colonization by S. sapinea of pine cones of different ages was assessed. Methods used for inoculating branch pieces and wood blocks were highly effective for all fungi. Type of a forest stand in which branch pieces and wood blocks have been incubated did not affect the persistence of the pathogens in the inoculated samples. For branch pieces, the success of re-isolation of L. serpens dropped as the sample incubation time increased, while S. sapinea was always successfully (100%) re-isolated (even 12 months after the inoculation). L. serpens and H. annosum s.s. were re-isolated from most of the buried wood blocks (from more than 95% samples) up to 3 months following the inoculation. Of the observed P. pinea cones (in most cases, more than 2 years old), 74% were naturally infected byS. sapinea. All three investigated pathogens were able to survive in dead plant tissues for long periods of time (at least for several (3–12) months). The persistence of these pine-pathogenic species in dead plant material questions the feasibility of leaving coarse woody debris in managed Italian stone pine forests meant for landscape conservation and leisure activities.     

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.     

Association of Sphaeropsis sapinea with insect‐damaged red pine shoots and cones

The association of the shoot blight and canker pathogen Sphaeropsis sapinea with red pine (Pinus resinosa) shoots and cones damaged by insects (especially Dioryctria sp.) was investigated. Samples from a single plantation approximately 35 years old, in Sauk Co., Wisconsin and also from three plantations, between approximately 40 and 50 years old, located in an area of pine shoot moth activity in the preceding year in Adams Co., Wisconsin were visually examined. Samples were arbitrarily collected from trees felled in the first plantation in May. Pycnidia of S. sapinea and insect damage were observed on 56 of 91 (62%) of closed cones and 17 of 165 (7%) of previous year's shoots. In the absence of insect damage, pycnidia of the pathogen were identified only on eight of 91 (9%) closed cones and never on previous year's shoots. In each of the other three plantations, 10 trees were located at intervals along transects in mid‐June; one branch from the lower half of the crown per tree was pruned off, and both current and previous year's shoots were examined. Insect damage and S. sapinea pycnidia were too rare on current year's shoots to draw any conclusions. Insect damage occurred on 20–40% of over 2000 previous year's shoots that were examined, but pycnidia of the pathogen were identified on only about 5%. Although infrequent, S. sapinea was identified in association with insect‐damaged previous year's shoots from these three plantations three times more frequently than those without insect damage. Random amplified polymorphic DNA (RAPD) markers from eight randomly selected isolates were consistent with the A group of S. sapinea, which can be aggressive on red pine. This ability to exploit insect‐damaged shoots may facilitate long‐term persistence of S. sapinea at low disease incidence and severity. The potential role of insect wounds as infection courts and insects as vectors of this important pathogen of pines deserves further study.     

The aggressiveness of Sphaeropsis sapinea on Austrian pine varies with isolate group and site of infection

The aggressiveness of the A and B isolate groups (morphotypes) of Sphaeropsis sapinea on shoot tips and lower stems of Austrian pine (Pinus nigra) was compared. The distinct differences in aggressiveness between the two S. sapinea groups, previously reported for other conifers, are confirmed for Austrian pine. However, the relative aggressiveness of the B group isolate varied by site of infection. Although the B group isolate of S. sapinea was not aggressive on shoot tips of Austrian pine, it was aggressive on stems of this host.     

The ecology of fungal cankers on Cupressus macrocarpa in southern England

Bark isolations from cankers on Cupressus macrocarpa in southern England yielded Seiridium cardinale, Pestalotiopsis funerea and Pestalotiopsis monochaetioides. The recovery pattern of the three fungi indicated that only S. cardinale was a primary pathogen; the two Pestalotiopsis species being secondary. Pathogenicity data obtained following the inoculation of C. macrocarpa branches with mycelium of the three fungi confirmed this view. No evidence of vegetative incompatibility was found in S. cardinale, but five vegetative-compatibility (vc) groups were found in P. funerea and six in P. monochaetioides. Different vc groups of the two Pestalatiopsis species were often found in the same S. cardinale lesion, suggesting that a number of separate colonization events were involved. It is possible that this sometimes leads to complete replacement of the pathogen.     

Genotypic diversity and distribution of Sphaeropsis sapinea within Pinus radiata trees from northern Spain

Sphaeropsis sapinea is an important latent pathogen of Pinus spp., outbreaks of which have a considerable impact on plantations. This study considers the population diversity and distribution of S. sapinea in northern Spain at different spatial scales from single plantations to a wide area covered by Pinus radiata trees. Estimation of genotypic diversity is an important component of the analysis of the genetic structure of plant pathogen populations. Ten simple sequence repeat (SSR) markers were used, together with vegetative compatibility tests, to study the genetic diversity among S. sapinea isolates. Polymorphism analysis at SSR loci is a simple and direct approach for estimating the genetic diversity of S. sapinea isolates. From a total of 86 isolates collected from four different areas, 14 microsatellite haplotypes and 13 vegetative compatibility groups (VCGs) were identified. The percentage of maximum genotypic diversity, based on Stoddart and Taylor's index, for microsatellites of the northern Spain population ranged from 14.6% to 38.1% and from 8.0% to 29.4% for VCGs. Analysis of these markers and vegetative compatibility groups confirmed that S. sapinea reproduces mainly asexually due to its reduced genotypic diversity in spatially close populations. Isolates of S. sapinea from northern Spain populations were predominantly monomorphic at the tested SSR loci. Vegetative compatibility groups also indicate a low level of genetic variability in these samples, which appear to be clonal.     

Pathogenicity of A and B morphotypes of Sphaeropsis sapinea confirmed on American larch (tamarack) and European larch

Fungi identified as Sphaeropsis sapinea (or synonyms) have been reported from Larix. Lack of pathogenicity tests, confusion about the identity of isolates mentioned in previous literature, and existence of distinct populations (A and B morphotypes) of the pathogen indicated the need to evaluate the ability of S. sapinea to cause disease of larch. Elongating snoot tips of Larix decidua, Larix laricina, and known hosts of the pathogen, Pinus banksiana and Pinus resinosa, were inoculated with water-agar plugs colonized with an A or a B isolate, or sterile plugs (controls). Each of five replications included five seedlings for each treatment-species combination. After 6 weeks, no symptoms had developed on control seedlings, but two-way analyses of variance revealed significant effects of isolate morpnotype and host on both incidence and severity of disease (values of p ≤ 0.01). The A isolate killed almost all shoot tips, but the B isolate killed from no P. resinosa shoots to 56% of L. laricina shoots. The average length of shoot killed by the A isolate was also greater than that killed by the B isolate. Response to the B isolate again varied among species, with greater average lengths of shoot killed on the larches (compared with the pines). Both A and B morphotypes of 5. sapinea should be considered among the fungi encountered on L. decidua and L. laricina.     

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.     

Variation among New Zealand isolates of Sphaeropsis sapinea

Sphaeropsis sapinea is a pathogen of many coniferous species and causes significant losses to the plantation forestry industries of many countries. New Zealand isolates of S. sapinea were examined for colony and morphological characteristics, and pathogenicity to Pinus radiata seedlings. Considerable variation was observed in colony growth form and colour, and in the growth rates on four agar media. The conidial dimensions, carbon and nitrogen utilization, and pathogenicity also varied among the isolates. The variation observed was such that the isolates could not be placed within previously described distinct morphotypes and the present results concur with previous studies cautioning the use of morphological criteria alone to partition the species. These results contrast markedly with previous reports and seemingly indicate a change in the New Zealand S. sapinea population. This difference may reflect the re‐introduction or spread of additional isolates or a shift in the structure of the resident population.     

Biocontrol of decay in seasoning utility poles. I. Growth rate and colonizing ability of bluestain and decay fungi in vivo and in vitro

Seasoning (air drying) of utility poles for 6–12 months is essential before preservative treatment can be achieved. However, during seasoning, pine sapwood is often colonized by decay fungi, thereby compromising the performance and service life of the poles. This study investigated the potential of bluestain fungi to act as short‐term biocontrol agents against decay during seasoning. An important attribute for biocontrol is rapid growth, so growth rates of common bluestain (Ceratocystis coerulescens, Ophiostoma minus, Ophiostoma piceae, Ophiostoma piliferum, Sphaeropsis sapinea) and decay fungi (Heterobasidion annosum, Phlebiopsis gigantea, Stereum sanguinolentum) were compared on agar medium and pine in logs at various temperatures. On agar, the growth temperature optimum of most bluestain fungi and all the decay fungi was ～25°C, with little growth at ≤5°C or above 32.5°C. Overall, the fastest growing were S. sapinea and O. minus. In logs, the most effective colonizers were S. sapinea and O. minus with pathogenic abilities that made them well fitted to colonize the sapwood of freshly felled pine. Within these species, certain isolates produced much larger lesions in phloem and the sapwood tangential plane than all the decay fungi. Notably, there was significant variation in colonizing ability between different isolates within a species, emphasizing the need for testing a range of isolates when selecting a potential biocontrol agent.     

The pine pathogen Diplodia sapinea is associated with the death of large Douglas fir trees

Climate change poses severe pressures to European conifer forests. Using non-native tree species, such as Douglas fir (Pseudotsuga menziesii), is one proposed strategy to circumvent adverse effects for forest management. However, novel forest health risks can impair the cultivation of non-native trees. In 2022, we observed large Douglas fir trees (approximately 40–50 years old, diameter at breast height (dbh) 21–41 cm) that had recently died in spring or summer 2022 in three forest stands in Eastern Austria. Intensive resin flow, blue-staining of the sapwood and the absence of bark- and wood-boring insects indicated a fungal infection. Isolations from blue-stained sapwood of the dead trees consistently yielded cultures of the opportunistic pathogen Diplodia sapinea. In a greenhouse wound inoculation experiment, seven D. sapinea isolates obtained from Douglas fir caused phloem necrosis, blue-staining of sapwood and mortality and thus displayed pathogenicity towards seedlings of both Ps. menziesii and its common host, Scots pine (Pinus sylvestris). Diplodia sapinea produced significantly longer areas of blue-stain as well as higher and faster mortality in Ps. menziesii compared to P. sylvestris. We conclude that D. sapinea substantially contributed to the death of seven of the 13 examined large Douglas fir trees. While this fungus has been described as a pathogen of young Douglas fir trees before, this is the first report that it can potentially kill large individuals of this conifer species under drought conditions. Thus, our results indicate that D. sapinea could represent a severe threat to the cultivation of Ps. menziesii in European forestry.     

First report of Mycosphaerella dearnessii in Switzerland

Brown-spot needle blight caused by Mycosphaerella dearnessii [syn. Scirrbia acicola; anamorphic state: Lecanosticta acicola (syn. Septoria acicola)] was observed for the first time in Switzerland. So far, the disease seems to be confined to planted Pinus mugo and Pinus uncinata near Zurich. Pinus sylvestris adjacent to diseased P. uncinata did not show any disease symptoms.     

An effective medium for isolating Sphaeropsis sapinea from asymptomatic pines

The pathogen Sphaeropsis sapinea can persist in stems and branches of asymptomatic pines and can later induce disease when triggered by host stress. Several experiments were conducted to test if:(i) medium amended with tannic acid (TA) can increase the frequency of cultural detection of this shoot blight and canker pathogen from asymptomatic red pine (Pinus resinosa) stems, and (ii) S. sapinea can persist in asymptomatic red pine in the field following artificial inoculation. TA (0.5% w/v) in 2% (w/v) water agar proved to be the best medium for isolation of S. sapinea among a larger number of tested media. The addition of TA had little or no effect on the growth of two group A and two group B isolates of S. sapinea. However, when TA was added, 11 other fast‐growing fungal isolates from stems/branches of red or jack pines (P. banksiana) were inhibited and grew more slowly (p < 0.05) than both S. sapinea groups. The TA‐amended medium improved cultural detection of S. sapinea from 2‐year‐old, asymptomatic red pine nursery seedlings compared with two other methods used for the cultural detection of S. sapinea (32%vs. 8.5% and 18% recovery; p < 0.001 and p = 0.031, respectively). A field test using the TA‐amended media established that S. sapinea can persist asymptomatically in red pine trees for at least 1 year. This medium significantly reduces the frequency of false‐negatives from asymptomatic field material.     

Induction of phenylalanine ammonia lyase activity and control of Sphaeropsis sapinea infection in Pinus radiata by 5-chlorosalicylic acid

Phenylalanine ammonia lyase (PAL) activity increased rapidly in Pinus radiata seedlings after treatment with 2 mm 5-chlorosalicylic acid (5CSA) and peaked after 48 h at four-times the level of the control. Elevation of PAF is often associated with the induction of host resistance mechanisms. Thus, effects of 5CSA on subsequent infection of P. radiata by Sphaeropsis sapinea was investigated. Only one out of 16 seedlings treated with 2 mm 5CSA, 4 days before inoculation, developed deadtop compared to 14 of the controls. The 5CSA was directly inhibitory at this concentration and reduced growth rate of S. sapinea on agar by 27%. However, antifungal activity does not fully account for the observed disease control afforded by 5CSA since 0.5 mm and 0.25 mm 5CSA, which did not inhibit growth of S. sapinea, significantly reduced the incidence of deadtop. Furthermore, 1 mm 5CSA protected seedlings against fresh inoculation challenge for up to 32 days after application. Such durable resistance is unlikely to be attributable to the direct effects of the applied compound and may suggest the involvement of inducible host resistance mechanisms.     

Experimentelle Infizierung einiger anБereuropäischerPinus-Arten durch die KiefernnadelgallmückeThecodiplosis brachyntera (Schwägr.)(Diptera,Cecidomyiidae)

Zusammenfassung Die KiefernnadelgallmückeThecodiplosis brachyntera (Schwägr.), die in Europa anPinus sylvestris L.,P. mugo Turra undP. nigra Arnold lebt, wurde im Arboretum Bolevec in Westböhmen auch an der nordamerikanischen KiefernartPinus resinosa Aiton und den ostasiatischen KiefernartenPinus tabuliformis Carriere undP. yunnanensis Franchet festgestellt.Experimentell gelang es durch die Kiefernnadelgallmücke vonPinus mugo subsp.mughus (Scop.) Domin, auch die obengenannten außereuropäischen Kiefernarten und als Kontrolle die europäischen ArtenPinus sylvestris L. undPinus mugo subsp.uncinata (Ant.) Domin zu infizieren. Experimentelle Infizierung war erfolglos an den Kiefernarten:Pinus cembra subsp.alpina (Rikli),P. strobus L.,P. rigida Miller,P. radiata D. Don.,P. echinata Miller,P. contorta Douglas undP. thunbergii Parlatore.Die Ursache der Nichtinfizierung der untersuchten Kiefernarten kann folgenden Ursprung haben: Unterschiede in dem Chemismus, besonders in der Qualität und Quantität der Terpene im Innern der Nadeln, oder: Unterschiede in der Gestalt der Nadeln, besonders die Nichtanwesenheit der Nadelscheide, die den Basalteil der Nadeln umhüllt und deshalb die Entwicklung der Eier und Larven nicht ermöglicht, oder aber: es waren morphologisch die Nadeln in der Zeit der Infizierung für die Eiablage nicht geeignet — entweder noch mit Schutzschuppen bedeckt, oder schon zu groß entwickelt.Die befallenen Nadeln der außereuropäischen Arten waren verkürzt, in dem Basalteile grö ßtenteils einige mm zusammengewachsen. Die Lavalkammer befand sich in der Mehrzahl der Fälle in der Nadelbasis.Nach den gewonnenen Ergebnissen erscheint es notwendig, eine taxonomische Revision der 5 von 11 nordamerikanischen Kiefernarten beschriebenen Gallmückenarten durchzuführen.

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Summary The Needle shortening gall midgeThecodiplosis brachyntera (Schwägr.) occurring in Europe on the pinesPinus sylvestris L.,P. mugo Turra andP. nigra Arnold was found in an Arboretum in Bolevec in Western Bohemia on the North American speciesP. resinosa Aiton and on the East Asian speciesP. tabuliformis Carriere andP. yunnanensis Franchet.The following non-European species:Pinus resinosa Aiton,P. tabuliformis Carriere andP. yunnanensis Franchet and European species:P. sylvestris L. andP. mugo subsp.uncinata (Ant.) Domin were experimentally infested withThecodiplosis brachyntera (Schwägr.) fromPinus mugo subsp.mughus (Scop.) Domin. The experimental infestation was not successful in the case ofPinus cembra subsp.alpina (Rikli),P. strobus L.,P. rigida Miller,P. radiata D. Don.,P. echinata Miller,P. contorta Douglas andP. thunbergii Parlatore.The failure to infest some pine species might have three causes: the chemical compositions of the pine needles differed; the sheath shielding the basal part of brachyblast was missing; the needles of some pine species were not in the right state for oviposition and development of larvae at the time of infestation — they were either too mature or completely covered with scales.Infested needles of the non-European species were considerably shortened compared with non-infested ones. Their basal part usually coalesced for several mm and in most cases the larval chamber was situated right in the base of the needle.Results mentioned above showed the necessity of revision of the taxonomic position of 5 gall midge species developing in the bases of 11 North-American pine species.

     

Sapstain fungi on <Emphasis Type="Italic">Pinus radiata</Emphasis> logs – from New Zealand Forest to export in Japan

Supply of clear, unstained logs from an export country is important economically as well as in reducing the biosecurity risk to an importing country such as Japan. Although conditions found within the holds of ships containing logs are thought to be ideal for rapid colonization of sapstain fungi, no research has been conducted. Research-focussed log export trials were designed to determine the extent of sapstain colonisation at specific points in the processing of logs from harvesting to arrival at the export destination. Two trials were established, in the austral summer and winter, in which mature Pinus radiata logs were harvested in New Zealand and shipped to export ports in Japan. Data loggers placed above and below deck of the ships recorded microclimatic conditions. Nine species of sapstain fungi were isolated from logs during the summer trial; the most common species isolated were Ophiostoma floccosum, Ophiostoma querci, and Ophiostoma setosum. In contrast, ten sapstain species were detected during the winter trial, with Sphaeropsis sapinea, O. querci, O. floccosum, and O. setosum most commonly isolated. This research was the first successful attempt at measuring visual sapstain development and isolating sapstain fungi from the time of harvesting to arrival at an export destination.