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.     

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.     

Quantification of Phaeocryptopus gaeumannii colonization in Douglas‐fir needles by ergosterol analysis

Current assessments of infection levels of Phaeocryptopus gaeumannii, the incitant pathogen of Swiss needle cast disease on Pseudotsuga menziesii, typically rely on surveys of abundance of fruit bodies on diseased needles. The relationship between this measure and internal fungal colonization is unknown. In this article, a series of experiments to determine whether ergosterol can be used to quantify P. gaeumannii internal colonization within Douglas‐fir needles is reported. It was found that ergosterol content in seven commonly occurring Douglas‐fir foliar fungi is proportionally related to biomass, and in P. gaeumannii this relationship is not affected by age of the culture. Furthermore, at four sites tested, P. gaeumannii was the most common fungus species isolated from Douglas‐fir needles, accounting for approximately 50% of the isolations. Ergosterol content in these needles was best related to P. gaeumannii despite the presence of other fungi. The strong relationship between ergosterol and P. gaeumannii is attributed to its greater contribution to total fungal biomass compared with all other fungi present within Douglas‐fir needles.     

A method for quantifying changes in the epicuticular wax structure of Norway spruce needles

A method for quantifying changes in the epicuticular wax structure of spruce needles (Pieea abies) has been developed for the scanning electron microscope. This evaluation system, based on a five-point scale, classifies only the epistomatal waxes of current-year needles. Two criteria, the different crystal wax forms and the amount of changed wax as a percentage of the total stomatal area have been used for classification. Statistically significant results (accuracy below ± 10% at 5% error level) were obtained under the following conditions. Mixed needle samples were taken from two adjacent trees per site. These were in a dominant and predominant social position in the stand. The needles were collected from the 7^th whorl of sun crowns exposed to all wind directions. The wax quality was determined by evaluating three stomata per needle. One stoma was investigated at the needle base, one m the middle and the third at the apex of the needle. 24 needles were evaluated per site. SEM-Investigations of needle waxes of 25 Austrian forest sites with no local emission source (background sites) showed, that the mean wax quality ranged from 1.6 to 4.1, according to this classification method. Stepwise multiple regression analyses with site characteristics and element contents of the needles (nutrient elements, heavy metals) confirmed correlations between the wax quality and the manganese- and sulphur-content of the needles and also the average degree of crown defoliation.     

Über Sukzession und pathogene Eigenschaften Kiefernnadeln bewohnender Pilze

Succession and pathogenic properties of fungi inhabiting pine needles . Isolates from healthy and dying pine needles of various age permit conclusions about the relationship of some needle fungi to their host. Of the six most prevalent fungi, only Lopbodermium seditiosum has a pathogenic nature.     

The biology of Lophodermella conjuncta Darker on Corsican pine needles

The biology of Lophodermella conjuncta on Corsican pine needles was studied. Results of a spore exclusion experiment and histological examination of needles indicated that infection of young current years needles occurred and that the fungus first colonized only the endodermis. Infection remained latent until symptoms developed, from hyphae penetrating the mesophyll from the endodermis, in July of the year following infection. Ascocarps were produced on second year needles by mid-August. Aseospores were trapped throughout the year.Infected needles were cast prematurely during the third season. The phenomenon of latent infection of pine needles by some pine needle fungi is discussed.     

Colonization and lignin decomposition of pine needle litter by Lophodermium pinastri

The colonization and lignin decomposition of pine (Pinus thunbergii and Pinus densiflora) needle litter by an endophytic fungus, Lophodermium pinastri, was examined with field observations, a field experiment and laboratory experiments. In pine needles collected from the field, needle mass per length and lignin content were lower in needle portions bearing Lophodermium fruiting bodies than in the remaining needle portions, whereas total carbohydrate content was not different between them. Total and live hyphal lengths were greater in needle portions bearing Lophodermium fruiting bodies than in the remaining needle portions. Lophodermium fruiting bodies were not formed on sterilized needles after a 6‐month incubation on the forest floor, whereas they formed on 20% of non‐sterilized needles, indicating that this fungus can only colonize live needles on the branch. In pure culture decomposition tests, mass loss of lignin was detected for several isolates of L. pinastri, but was variable among isolates and between the needles of two pine species. A comparison of the rDNA‐internal transcribed spacer sequences between L. pinastri isolates of P. thunbergii and P. densiflora indicated that the two groups were phylogenetically separated. This study is the first to show that L. pinastri has the ability to decompose lignin in dead pine needles.     

Colonization of beech leaves by two endophytic fungi in northern Japan

Leaves of Japanese beech (Fagus crenata) were collected monthly during the vegetation period at five sites in the Tohoku district in Japan to isolate endophytic fungi. Leaves were also collected only once at two additional sites. Two endophytic fungi were dominant, a Discula species and a sterile mycelium. This result strongly suggests that these two fungi are generally associated with leaves of the Japanese beech at different sites. At most sites the isolation frequency of Discula sp. was greatest in June and gradually decreased from July to October whereas the isolation frequency of the sterile mycelium increased during the vegetation period and remained at a high isolation frequency in October. Spores of Discula sp. were released for a very short time in late May, just after the disappearance of the snow cover on the forest floor. These spores may be important for the infection of newly sprouting leaves.     

Diversity and identification of fungi associated with needles of Pinus radiata in Tasmania

Needle fungi have been extensively studied in conifers but rarely in Pinus radiata. Previous studies of P. radiata have been based on fungal isolation and not direct PCR detection from needles. This research was a component of a study examining factors linked to spring needle cast (SNC) in Tasmanian P. radiata plantations and aimed to identify as many as possible of the fungal species commonly associated with P. radiata needles in Tasmanian plantations. Needle samples were collected from 13 sites representative of the range of sites in which P. radiata is grown in Tasmania. Fungi were detected by a direct PCR approach and identified using barcode sequences from a reference collection of isolates from Tasmania, mainland Australia and New Zealand as well as sequences from the international nucleotide sequence databases. The total number of molecular operational taxonomic units (MOTUs) was 152, with 127 detected by direct PCR and sequencing, and only 35 operational taxonomic units (OTUs) isolated by culture-based methods. Dothideomycetes was the most diverse class detected in this study, with many MOTUs detected by direct PCR and not isolated. Leotiomycetes was the second-most diverse class and Sordariomycetes third, with several OTUs frequently isolated but rarely or not at all detected by direct PCR. DNA sequence data facilitated the discrimination and identification of OTUs, but some effort was required to avoid confusion caused by poorly identified isolates in public DNA databases. This is the most comprehensive study yet of fungi associated with pine needles in Australia and highlights the hitherto unrecognised diversity of Teratosphaeriaceae in P. radiata.     

Fungi infecting Pinus sylvestris needles of various ages

The studies were carried out in three 8–11 year old stands of Pinus sylvestris in Southwestern Poland. They included the isolation of fungi from infection spots which were sampled after 1, 1, 5 and 2, 5 years on living needles developed in 1978 on lower and upper branches of tree crowns. There were 28 fungal species isolated from such needles. An attempt was made to evaluate the role of the 5 most numerous species in causing needle cast of Pinus sylvestris.     

油松毛虫产卵、取食偏嗜行为及油松针叶内含物测定

[目的]为明确油松毛虫在油松上的产卵和取食部位以及可能机理,为未来害虫发生预测模拟提供基础信息数据。[方法]通过室内模拟试验,统计油松毛虫在盆栽油松幼苗不同部位上的产卵量和取食不同油松针叶的偏嗜情况及死亡率;测定油松针叶内含物含量并与油松毛虫偏嗜行为进行了相关性分析。[结果]在一年生油松针叶上平均产卵块数最多(3.10±0.28),显著高于二年生针叶和三年生针叶;取食一年生油松针叶的油松毛虫数量最多(13.40±0.91)、死亡率最低(13.60%±4.87%),显著优于取食二年生油松针叶和三年生油松针叶的幼虫。油松毛虫幼虫取食选择、成虫产卵选择和幼虫体长与油松针叶内含水量成显著正相关(P0.01);与寄主总酚含量(P0.01)和碳氮比(P0.05)成显著负相关。[结论]油松毛虫对一年生油松针叶具有明显的产卵和取食偏嗜行为。一年生油松针叶内含水量最高、总酚含量最低、有机碳含量最低可能是造成寄主偏嗜行为的原因。     

The influence of soil scarification on the turn‐over rate of slash needles and nutrient release

The effect of soil scarification on decomposition of green Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) needles and mineral nutrient release was investigated. The treatments studied were ploughing, disk trenching, mound on inverted humus and untreated control. The study was conducted on two clear‐felled sites with different ground moisture conditions, located in south Sweden. The needles decomposed and released their contents of N and Ca considerably faster on scarified areas than on untreated ones. For P, Mg and especially K, only minor differences between scarified and untreated areas were established. At the wet site, needle decomposition and nutrient mineralization were generally affected to the same extent by all three scarification methods studied. At the dry to fresh site, mounding and ploughing enhanced needle decomposition and nutrient mineralization more than disk trenching. The effect of different scarification methods on the long‐term production capacity of soils is discussed.     

Environmental Effects on Norway Spruce Needle Wax

In spring, tubular wax structures were observed on the needle surfaces of new Norway spruce needles and also on previous-year needles. In young plants, as in mature trees, the fusing and eroding of the wax tubes on needles of increasing age had taken place in relation to their exposure, particularly to wind and precipitation. The quantity of needle wax and certain fractions of the extracted wax differed in mature trees at two field sites and in young clonal plants. No difference in wax quantity and only small differences in wax composition occurred among young plants which had undergone different treatments (fumigation with O³, SO², ambient air, filtered air, or planting at the sites). Environmental effects on wax structure, wax quantity and the presence of certain compounds in the wax were identified.     

Study on biological characteristics ofHendersornia acicola

The morphological and cultural characteristics ofHendersonia acicola Munch & Tub., isolated from the needles ofPinus sylvestris var.mongolica Litvin, were described. The fungi was cultured on three kinds of media and the speed of mycelial growth and the required time to produce conidia were different. On the medium of needle decoction agar, the fungi grew fast and produced conidia after 5 days culturing. On PDA came second and it needed 7 days to produce conidia. On the medium of maize flour agar, it grew slowly and produces conidia after 27 days incubating. The fungi cannot produce conidia under darkness on all three kinds of media. The conidia can germinate at 8°C–28°C. The optimum conditions for conidia to germinate are 25°C, RH 100% and pH 4 and under these conditions it only takes four hours for conidia to germinate. The conidia germination is not affected by light     

Morphological and molecular characterization of Fusarium oxysporum and Fusarium avenaceum causing a novel needle blight of Picea asperata in China

Picea asperata is a unique tree species in China. It has important economic and ecological values. In this study, the disease symptoms resembling needle blight, a new disease of P. asperata (10- to 15-year-old), were detected in a forest area in Tianshui City, Gansu Province, China, in August 2020. Affected needles turned yellow, the colour became darker at the junction of infected and healthy areas, the needles wilted and even fell off the tree. Through pathogenicity tests, the two pure fungi strains isolated from diseased needles caused distinct needle blight symptoms on P. asperata, with symptoms similar to those observed in the forest. It was observed that they had sickle-shaped conidia. Subsequently, multilocus phylogenetic analysis of partial sequences of the internal transcribed spacer region (ITS), translation elongation factor 1-α (TEF-1α) and RNA polymerase II second major subunit (RPB2) genes was performed. They were found closely clustered with Fusarium oxysporum and F. avenaceum, respectively. Based on morphological and molecular biological determinations, the two pathogens were identified as F. oxysporum and F. avenaceum. To our knowledge, this is the first report of needle blight caused by F. oxysporum and F. avenaceum on P. asperata in China.     

Breakdown and macroinvertebrate colonization of needle and leaf litter in conifer plantation streams in Shikoku,southwestern Japan

Breakdown and macroinvertebrate colonization of conifer needles (Cryptomeria japonica) and deciduous broadleaves (Euptelea polyandra) were investigated using litter bags in two study sites in streams flowing through a conifer plantation of C. japonica in Shikoku, southwestern Japan (one site with conifer canopy and another with mixed conifer and broadleaved canopy). Breakdown rates and macroinvertebrate densities were compared between litter species (conifer needle vs broadleaf) and between the two sites (conifer vs mixed canopy) to determine (1) whether breakdown rate of broadleaves is higher than conifer needles, (2) whether macroinvertebrates prefer broadleaves to conifer needles, and (3) whether the difference in riparian canopy is reflected in macroinvertebrate abundance. The results indicated that breakdown rates of broadleaves were higher than those of conifer needles, suggesting poorer quality of the latter as food for macroinvertebrates. Differences in macroinvertebrate density between needles and broadleaves were generally consistent with those in breakdown rates: broadleaves tended to have higher densities than needles, suggesting that conifer needles were not preferred by macroinvertebrates. However, total macroinvertebrate density in the conifer site was not significantly different from that in the mixed site, although the dominant shredder taxon differed (conifer site: gammarids; mixed site: lepidostomatids). Although conifer needles are low-quality food for macroinvertebrates, this may offer some advantages. Conifer needles remain on the streambed for longer periods owing to their lower breakdown rates, being a constantly available resource. In addition, accumulations of conifer litter may effectively trap and retain particulate organic matter.     

Lophodermium pinastri and an unknown species of Teratosphaeriaceae are associated with needle cast in a Pinus radiata selection trial

Spring needle cast (SNC) in Tasmania is defined by the early casting of pine needles and appears in 6‐ to 7‐year‐old pine plantations after canopy closure. The severity of SNC can vary significantly among trees growing within the same plantation. Fungal communities were surveyed in a Pinus radiata family trial, using DNA extraction from needles, PCR with fungal‐specific primers and sequencing to detect the presence of fungal species. Samples were taken at the same time as the trees were scored for disease severity. Trees with contrasting levels of SNC disease severity have significantly different needle fungal communities, but family pedigree and different ages of needle are not clearly distinguished by their fungal communities. All common fungal pathogens previously implicated in causing SNC were identified from the study, but of these, only Lophodermium pinastri was correlated with high levels of disease. Several species of Teratosphaeriaceae were detected, and one of these was also strongly associated with needle cast.     

Reflectance features of water stressed Larix gmelinii needles

In recent years, Xing’an larch (Larix gmelinii) has been seriously infected by pests and drought. In order to improve the accuracy of monitoring the damage to larch by remote sensing (RS) and to predict the health of the larch, we studied the reflectance features of larch needles under different water conditions at the needle level by using the LIBERTY (Leaf Incorporating Biochemistry Exhibiting Reflectance and Transmittance Yields) model. Before applying the LIBERTY model, we recalibrated it for the needles of L. gmelinii based on ten field-measured spectral curves. After recalibration, LIBERTY can accurately model the needle reflectance spectra of L. gmelinii. Based on the recalibrated LIBERTY model, we extracted and analyzed the sensitive bands to needle water content by simulating the needle reflectance spectra under different drought conditions. Then, we established mathematical equations between the spectral indices (MSI, NDWI, and GVMI) and needle water content. Results show that the variations of larch needle water content can significantly change the needle spectra at the near-infrared and short-wave infrared bands. The higher the water content is, the higher the absorption peak is. We believe that our study will provide the theoretical basis and an optional method to investigate the forest water stress using multi-spectral or hyper-spectral remote sensing data.     

Relation Between Defoliation and Litterfall in Some Danish Picea Abies and Fagus Sylvatica Stands

Assessing defoliation and measuring litterfall are two different ways of estimating the shedding of needles and leaves from the forest canopy. Both variables can be said to reflect the crown condition, but the two methods have rarely been compared. In this study the visual observations of defoliation of individual trees were compared with sampled litterfall data for the two main tree species in Denmark, Fagus sylvatica and Picea abies. Defoliation assessments and litterfall measurements were performed in seven level II plots. Six of the stands were 40 yrs old and the remaining stand was 81 yrs old. Both a positive and a negative correlation, which were not significant, were observed between defoliation and total leaf litterfall on the beech sites. Similarly, no significant correlation was observed between defoliation and the yearly needle litterfall on the younger Norway spruce plots. However, the defoliation and the yearly needle litterfall at the old stand at Klosterhede were positively correlated on a 10% significance level. A positive correlation was also apparent at the younger stands between the defoliation and the needle litterfall from the period April-July of the same year in which defoliation was assessed. The absence of a clear connection between the two assessments is discussed. The two assessments are apparently of widely different origin, which makes comparison difficult.     

Presence of fungi in Scots pine needles found to correlate with air quality as measured by bioindicators in northern Spain

Atmospheric pollution has increased worldwide during the last decades affecting forest ecosystems. Thermal power plants give off high levels of contaminants, which can damage forest health. Bioindicators can be helpful tools used for evaluating environment changes by giving an accurate measure of the extent of pollution. The focus of this study was to analyse fungal mycoflora in Pinus sylvestris stands near a thermal power plant and the possible correlation between the presence of the fungi and the air quality as measured by lichens and mosses. Fungi associated with pine needles were selected as subjects of this study because of their phytopathological importance and association with the pine trees' decline. Samples for this study were selected from eight plots in close proximity to a thermal power station in northern Spain. Symptoms of decline were previously observed in all the plots before sampling was performed. Lichens were used as bioindicators of environmental quality in two ways, first using the Index of Atmospheric Purity and second through categorization of lichen species based on their sensitivity. In addition, botanical quality was measured regarding the presence or absence of mosses. With two seasonal samplings (autumn and spring) and the use of four types of pine needle plant material (green needles, dried needles, half–green, half‐dried needles and fallen needles), both endophytes and epiphytes present in the needles were isolated and observed. Thirty fungal species in total were identified of the 1095 isolates obtained. Furthermore, significant differences in fungal isolates were observed between seasons and among the different plant material. Results showed that both environmental air quality and botanical quality were negatively correlated with the relative isolation frequency of fungi. The higher number of isolates was attributed to a possible infection produced by fungi, which could be a leading factor in the trees' decline.