Endophytische Pilze in Nadeln von gesunden und geschädigten Fichten (Picea abies [L.] Karsten)

Endophytic fungi in needles of healthy-looking and diseased Norway spruce (Picea abies [L.] Karsten). Endophytic fungi were isolated from surface-sterilized, 5 to 6 year old needles of healthy-looking and diseased trees. Two fungal species could be isolated frequently: Lophodermium piceae and Tiarosporella parca. At any site neither L. piceae nor T. parca could be isolated more frequently from needles of diseased trees. The possible types of relationship between these two fungi and Norway spruce are discussed.     

Histology of Diplodia pinea in diseased and latently infected Pinus nigra shoots

Diplodia tip blight is a serious disease of >30 conifer species worldwide. Symptoms are particularly severe on non‐native, two‐needled Pinus species, and typically include stunted, necrotic needles and shoots and a general decline of the tree. Latent Diplodia pinea infections occur in current‐year shoots of some symptomless pines, and in some apparently healthy current‐year shoots of diseased pines. Latent infections also occur in symptomless terminal buds. A histological approach was used to investigate the nature of latent infections in shoot stems and terminal buds of landscape Pinus nigra. Fungal colonization was compared in healthy, diseased and latently infected tissues. A nested‐PCR technique that is specific for D. pinea was used to differentiate latently infected tissues from uninfected ones. Latent D. pinea infections were localized in the outer stem cortex, usually in the vicinity of needle scales at leaf axes. In contrast, pathogenic D. pinea infections were characterized by fungal colonization throughout the shoot stem tissues, even very early in symptom development. The presence of necrophylatic periderms in two of the latently infected samples suggests that host defences play a role in the production and maintenance of latent stem infections. Latent infections of terminal buds appeared to originate from the distal bud scales of axillary buds in the terminal bud cluster, and not from the subtending shoot. Fungal tissues were never observed inside asymptomatic, PCR‐negative shoots.     

Needle nutrient status of Sirococcus shoot blight‐diseased and healthy Norway spruces

In this study the needle nutrient status of Sirococcus shoot blight‐diseased and healthy Norway spruces (Picea abies) was investigated. Element contents were determined in the current year and 3‐year‐old needles collected in December 2000 from 72 mature trees randomly distributed in the investigated stand. Half of the trees were severely affected by Sirococcus conigenus and had a reduced crown whereas the other trees were unaffected and vigorous. Compared with the uninfected trees, contents of Mg, Ca, P and Mn were significantly lower in the current year and 3‐year‐old needles of diseased trees. Furthermore, analyses of the nutrient contents of needles from severely affected trees revealed insufficient Mg and Ca supply and enhanced N/Mg and N/Ca ratios, when compared with threshold values. The results of this study suggest that imbalanced tree nutrition either has a significant impact on the expression of Sirococcus disease symptoms or fungal infection itself is affecting needle nutrient contents.     

Rapid living crown reduction caused by Gremmeniella abietina affects foliar nutrient concentrations of Scots pine

The foliar chemistry of diseased and healthy trees was studied one growing season after severe reduction in living crown caused by Gremmeniella abietina in four young Scots pine (Pinus sylvestris) stands. Sample trees were chosen pairwise on the basis of the living crown length: a diseased tree with about 50% live crown reduction and a healthy tree in each pair. Fifteen elements were determined in the youngest healthy needles on the lateral top shoots of each sample tree. Diseased trees had higher foliar boron, manganese and sodium concentrations and lower magnesium, iron, zinc, copper, potassium, nitrogen and sulphur concentrations compared to the healthy trees. Foliar calcium, aluminium, phosphorus, carbon and hydrogen concentrations did not differ between the diseased and healthy trees, except for P and A1 in two of the stands when the stands were analysed separately. Significant correlations between the needle element concentrations and crown ratio (length of the living crown/tree height) were found especially for B (increasing B with decreasing crown ratio) and for Mg, Fe and Zn (decreasing concentrations with decreasing crown ratio). The effect of G. abietina-induced living crown reduction on tree nutrient status and the role of these mineral nutrients in the susceptibility are discussed.     

Seasonal patterns of colonization of Norway spruce needles by Lophodermium piceae

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

Effects of different ecological and silvicultural factors on beetle catches in the Turkish fir (<Emphasis Type="Italic">Abies bornmülleriana</Emphasis> Mattf.) ecosystems

To study the bark beetle attacks on Turkish fir stands and to determine the principle site and stand factors influencing beetle attacks, trap log method was used in the Western Black Sea Region of Turkey. The study was carried out in ten different locations prior to flying time in February 2003. On each experimental plot, three healthy fir trees with 30–50 cm diameter and 200 m apart were cut down with a chainsaw. From each sampling tree, needles were sub-sampled for determination of specific leaf-area and macro-nutrient concentrations. In addition, on each experimental plot, crown closure, basal-area, and stand density were measured. To determine the bulk density and nutrient concentrations of the soil, samples were taken around trees on each plot. The relationship between measured stand variables, the number of beetle species and beetle populations were evaluated using correlation and stepwise multiple regression procedure. Eight different species belonging to four different families from Coleoptera were identified in examining the trap logs. Six of these eight species (Pissodes piceae Illig., Pissodes notatus Fabr., Cryphalus piceae Ratz., Pityophthorus pityographus Ratz., Xyloterus lineatus Oliv., Pityokteines curvidens Germ.) were identified as harmful beetles (HB). The other two (Thanasimus formicarius L. and Rhizophagus dispar Payk.) were identified as predator species. The analysis showed that the total number of HB increased as the site’s slope and needle specific leaf area increased. However, as needle N concentrations increased, HB population decreased. Increased Ca concentration in fir needle reduced total insect attack.     

First report of Lecanosticta acicola on non‐native Pinus mugo in southern Sweden

In 2017, severe symptoms of brown spot needle blight, similar to those caused by Lecanosticta acicola, were observed on needles of non‐native Pinus mugo var. Hesse planted in an arboretum in southern Sweden. Microscopic characterization and molecular diagnostics of isolates obtained from diseased needles confirmed the identity of L. acicola. This is the first report of the quarantine pathogen L. acicola in Sweden. Subsequent surveys are urgently needed to assess the presence and extent of the L. acicola invasion in Scots pine (Pinus sylvestris) stands in Sweden and other Scandinavian countries.     

The impact of cronartium flaccidum on the growth of pinus sylvestris

The blister rust of two‐needle pines in Europe is caused by the rust fungus, Cronartium flaccidum (Alb. Schw.) Wint. There are two races of the fungus: One host‐alternating and another pine‐to‐pine race. The latter race is considered to be more common in northern Sweden. The impact of this rust on growth of Scots pine (Pinus sylvestris L.) was investigated in three selected stands in northern Sweden. Radial stem increment was reduced 40–70% by severe attacks and 20–40% by minor attacks. The reduction of stem volume growth was probably greater. The faster growing trees seemd to be more severely attacked. Trees growing on poor soils appeared to lose almost as much growth capacity from a minor attack as from a severe attack.     

Mycobiota in needles of Abies alba with and without symptoms of Herpotrichia needle browning

The ascomycete Nematostoma parasiticum (syn.: Herpotrichia parasitica) is commonly perceived as the causal agent of the so‐called Herpotrichia needle browning in silver fir (Abies alba). However, its fruitbodies are rarely present on symptomatic needles, which are also colonized by many presumably saprotrophic fungi. We compared the internal colonization of healthy and symptomatic needles on two sites in Poland. In addition, the endophytic mycobiota in needles of various age was recorded on two other sites without disease symptoms. Fungi were isolated from 95.6% of the dead needles and from 62.9% of the living needles on symptomatic trees, whereas on healthy trees, only 45.0% of the needles were colonized internally. Colonization frequency increased with needle age. From a total of 2017 isolates, 116 fungal taxa were identified. Frequency of many species was influenced by needle type. Anthostomella formosa, Gloeosporidiella sp., Hypoxylon fragiforme, Xylaria hypoxylon and X. polymorpha were the most common fungi isolated from living needles. In symptomless living needles, fungi occurred significantly more often in the basal than in the apical parts. In dead needles, the most common fungi were Alternaria alternata, Paraconiothyrium sporulosum, Fusarium sp., Mollisia cinerea, Rhizoctonia sp., Rhizosphaera oudemansii, Thysanophora penicillioides, Xylaria hypoxylon and X. polymorpha. Rhizoctonia sp. was the most frequently isolated fungus in dead needles (23.4%) but occurred rarely also in living needles (0.3–1.1%). The supposed pathogen N. parasiticum was detected only sporadically (at most in 0.6% of the needles). Our findings demonstrate the need for understanding the role of Rhizoctonia sp. in Herpotrichia needle browning disease aetiology.     

Lophodermium piceae and Rhizosphaera kalkhoffii in fallen needles of Norway spruce (Picea abies)

Falling needles collected from under individual 70-year-old Norway spruce (Picea abies) trees for about four years were checked for fungal fruitbodies. The most common fungi were Lophodermium piceae and Rhizosphaera kalkhoffii. Maximum frequencies of fruitbodies occurred in late autumn for L. piceae (ca. 80% of the needles), in summer for R. kalkhoffii (ca. 60%) and in winter for the third most common fungus, Tiarosporella parca (ca. 10%). The frequencies of needles with fruiting fungi varied greatly within and between years. This is the first report of T. parca from Sweden.     

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

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

Life cycle of Lophodermium piceae in Norway spruce needles

The spruce needle inhabiting ascomycete Lophodermium piceae can live symptomless for several years, before it completes its life cycle within less than one year as detailed studies have shown.     

Epicuticular waxes,peroxidase activity and contents of chlorophyll,phaeophytin and carotenoids of needles from Pinus syivestris and picea abies as indicators of air pollution in the region of Stockholm

To trace any possible effects of air pollution stress and injuries on trees around a city centre, needles of Scots pine (Pinus Syivestris L.) and Norway spruce (Picea abies (L.) Karst.) from sites of different distances and directions from Stockholm were analysed. Comparisons were made between needles of different ages, a half to three and a half years old (C ‐ C + 3) for pine up to six and a half years old (C ‐ C + 6) for spruce. Epicuticular waxes, studied by scanning electron microscopy (SEM), showed age‐dependent but site‐independent changes. Different age‐patterns were found in the pine and spruce needles but no dust coverings or lesions were detected, irrespective of the sites. A proportionally increasing peroxidase activity (EC 1.11.1.7) with increasing needle‐age up to the third year was observed in both species. No certain indication of site‐effects, expressed by an unproportional increase of the enzyme activity with increasing needle‐age could be found. Chlorophyll a and b, phaeophytin a and b and carotenoid analyses on the corresponding materials only indicated disturbances in trees from some scattered sampling plots. Thus, neither could the pigment analyses reveal any signs of a general pollution stress in pine and spruce trees from the area in question.     

The relationship between Swiss needle cast symptom severity and level of Phaeocryptopus gaeumannii colonization in coastal Douglas‐fir (Pseudotsuga menziesii var. menziesii)*

This study examined 108 15‐year‐old Douglas‐fir (Pseudotsuga menziesii var. menziesii) trees to investigate whether trees exhibiting less severe Swiss needle cast (SNC) symptoms were more resistant (had less fungal colonization) or more tolerant (maintained healthy foliage under similar infection levels). Trees were sampled from six open pollinated families that were categorized into three disease severity groups (two families for each group; mild, moderate and severe disease symptoms). The amount of retained foliage and level of discoloration were visually assessed on trees in the field. Fungal colonization (as determined by proportion of stomata occluded with pseudothecia and by amount of Phaeocryptopus gaeumannii DNA in sampled needles) was measured on 1‐ and 2‐year‐old needles in the laboratory. Trees in the different disease severity groups were similar with respect to amount of fungus in their needles, yet the trees in the mild symptom group retained higher proportions of needles and maintained greener foliage. The relationship between amount of P. gaeumannii in needles and SNC symptom severity was statistically significant (p < 0.05) for amount of fungal DNA in 1‐year‐old needles and average needle retention (NR) over the last four growing seasons. Average NR decreased with increased amount of pathogen DNA in the mild disease symptom families. This relationship was reversed in the severe disease symptom group and there was no relationship in the moderate disease symptom group. Because the amount of P. gaeumannii DNA in foliage did not differ significantly among the groups, differences in symptom severity were attributed to tolerance, not resistance. Visual scoring of individual trees for average NR over the past four growing seasons could be used to effectively assess for SNC tolerance in Douglas‐fir.     

Necrotic spots induced by simulated acid rain on needles of Abies balsamea saplings

Simulated acid rain (SAR) of pH 2.6, 3.6 and 5.6 (control) was applied to Abies balsamea (L.) Mill. saplings at 2-week intervals for 3 years. Ambient precipitation was excluded from the trees during this period. Light to dark brown necrotic spots along the midribs of needles, 0.1 to 0.5mm in diameter and sometimes having a white center, were associated with the most acidic (pH 2.6) SAR treatment. These spots appeared over a significantly (p = 0.05) larger area of the foliage in the pH 2.6 treatment than in controls (pH 5.6). Needles that received SAR of pH 3.6 had intermediate numbers of spots. The spots persisted for at least 1²/₃ years after cessation of SAR application. Needle retention, needle lengths, and the occurrence of epicormic shoots and the needle rust caused by Uredinopsis longimucronata Faull were not significantly (p = 0.05) affected by the SAR treatments. Needle retention tended to be lessened by the most acidic (pH 2.6) SAR treatment. The occurrence of Lophodermium piceae (Fuckel) Höhnel needle cast was significantly lower in the middle tree sections of the pH 5.6 and 3.6 treatments than in the pH 2.6, drought and ambient treatments. Damage by naturally occurring insects was negligible, and was not related to the SAR treatments.     

Genetic diversity of Lophodermium piceae in South Finland

Lophodermium piceae, the dominant needle endophyte of Norway spruce (Picea abies), is known to be extremely diverse. This work aimed to test the possible occurrence of cryptic species within the morphological species L. piceae. Genetic variation in 36 South Finnish L. piceae isolates originating from six localities was investigated by comparing DNA sequences of three genetic markers. One of the markers was the internal transcribed sequence (ITS) of the ribosomal DNA and the other two (LP1 and LP2) were based on sequence characterized amplified regions designed for L. piceae. The LP2 marker could be detected only from isolates of L. piceae but not from 20 other ascomycete species tested. This sequence, therefore, is considered as a species‐specific marker for L. piceae. For comparison, ITS sequences of isolates representing two other Lophodermium species, L. pinastri and L. seditiosum, were also investigated. In a neighbor‐joining analysis of ITS sequences all L. piceae isolates fell into one cluster, which was clearly separate from those of L. pinastri and L. seditiosum. Dendrograms of the three markers were incongruent indicating that the L. piceae population examined consisted of a single phylogenetic species. No geographical differentiation was observed. Our results confirm that L. piceae is a genetically highly diverse endophytic species.     

Effects of Atlas cedar (<Emphasis Type="Italic">Cedrus atlantica</Emphasis>) defoliation on performance of the pine processionary moth (<Emphasis Type="Italic">Thaumetopoea pityocampa</Emphasis>)

The pine processionary moth (Thaumetopoea pityocampa) is an important defoliating insect of native cedar stands in Northern Africa. In 2002 and 2003, we studied the size of needles of Atlas cedar trees that had been defoliated or not in the previous year, and compared the oviposition preference and larval performance of processionary moth on these two types of trees in cedar stands of central Algeria. Needle length and their number per needle cluster were twice as high on non-defoliated trees than on defoliated trees. There were about ten times fewer egg batches on defoliated than on non-defoliated trees in both study years. Young larvae raised in the laboratory on the foliage of previously defoliated trees were smaller compared to those fed on the foliage from healthy trees. Larval colonies transferred to trees in the field showed two times higher mortality on defoliated trees. It was concluded that the decline in population density in defoliated stands often observed after outbreaks results both from the fact that the female moths avoid laying their eggs on defoliated trees and the lower performance of larvae which later feed on the new-grown needles.     

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.     

Detection of Diplodia pinea in asymptomatic pine shoots and its relation to the Normalized Insolation index

Diplodia pinea (syn. Sphaeropsis sapinea), a common pathogenic fungus, causes considerable damage in Italy, particularly to pine stands in which trees are subjected to environmental stress. The occurrence of D. pinea in symptomless Pinus nigra shoots was investigated and related to the amount of radiation received by the trees growing on a site in a year, expressed as the Normalized Insolation index (NIi). Twenty‐seven pines were selected from nine locations in Trentino (northern Italy). For each pine the incidence of the fungus in apparently healthy shoots was determined by both culturing on an agar medium and application of real‐time PCR. The incidence of D. pinea determined by culturing samples taken from asymptomatic trees was 59% (16 of 27 trees), compared with 85% found using real‐time PCR (23 of 27 trees). Detection of the pathogen in healthy pine tissue was positively correlated (p < 0.05) with the NIi values, using both detection methods.     

Photosynthesis in Norway spruce seedlings infected by the needle rust Chrysomyxa rhododendri

Chrysomyxa rhododendri (DC.) De Bary is a needle rust with a host shift between Rhododendron sp. and Norway spruce (Picea abies (L.) Karst.), penetrating only the new developing flushes of the conifer. Because little is known about its effects on trees, we investigated several parameters related to photosynthesis in artificially infected 3-year-old Norway spruce seedlings. The potential efficiency of photosystem II (PSII; derived from chlorophyll fluorescence measurements) was reduced in infected current-year needles as soon as disease symptoms were visible, about three weeks after inoculation. Two weeks later, photosynthetic O(2) evolution (P(max)) of infected needles was less than 20% of control needles, whereas respiratory O(2) uptake (R(D)) was about three times higher than that of control needles. Nonstructural carbohydrate concentrations were about 60% of control values in all parts of the shoots of infected trees. Photosynthetic inhibition was associated with marked decreases in chlorophyll concentration and chlorophyll a/b ratio but only a small reduction in carotenoid concentration. In infected trees, P(max) of noninfected 1-year-old and 2-year-old needles was 50 and 80% higher than in the corresponding age class of needles of control trees. Estimation of potential daily net dry mass production, based on P(max), R(D), specific leaf area, carbon content and needle biomass, indicated that seedlings infected once were able to produce 60%, and those infected twice only 25%, of the dry mass of controls. We conclude that afforestation and regeneration of Norway spruce is seriously impaired in regions where seedlings are frequently attacked by Chrysomyxa.