Environmental factors and host genetic variation shape the fungal endophyte communities within needles of Scots pine (Pinus sylvestris)

To determine the role of environmental and host genetic factors in shaping fungal endophyte communities we used culturing and metabarcoding techniques to quantify fungal taxa within healthy Scots pine (Pinus sylvestris) needles in a 7-y old provenance-progeny trial replicated at three sites. Both methods revealed a community of ascomycete and basidiomycete taxa dominated by the needle pathogen Lophodermium seditiosum. Differences in fungal endophyte taxon composition and diversity indices were highly significant among trial sites. Within two sites, fungal endophyte communities varied significantly among provenances. Furthermore, the communities differed significantly among maternal families within provenances in 11/15 and 7/15 comparisons involving culture and metabarcoding data respectively. We conclude that both environmental and host genetic variation shape the fungal endophyte community of P. sylvestris needles.     

Lichen-rich pinewood,Cladonia ciliata-Pinus sylvestris community in north-eastern Scotland

Summary

Ground dominated by Cladonia lichens of the sub-genus Cladina was found locally on pinewood floors on sandy or gravelly soils in north-eastern Scotland. Most sites are under mature or old planted Scots pine, Pinus sylvestris, but some in younger plantations and a few in native pinewood. Nearly all are in areas of fairly dry climate, and all are in open situations with exposure to light and wind. The vegetation, prior to 1990 not a well recognized feature of Scottish pinewoods, is distinguished in this paper as a separate community, the Cladonia ciliata — Pinus sylvestris Community. It is part of the class Vaccinio-Piceetea and resembles the lichen-rich coniferous forests in the circumpolar boreal zone, but in Scotland occurs at the extreme climatic limit of this type of vegetation. Many stands are vulnerable to trampling and land-use change, and some have little protection against such environmental vicissitudes.     

Relationship between foliar composition of red-pine and jack-pine seedlings and vulnerability to Lophodermium needle-cast disease

Summary The Mn and Al content of needles from two-year-old jack pine (Pinus banksiana) seedlings was found to be more than twice that of red pine (Pinus resinosa). It is speculated that the high Mn and Al content of jack pine seedlings may impart some degree of resistance to needle cast disease caused byLophodermium pinastri. Contribution from the Soils Dept., Univ. of Wis., Madison, in cooperation with and supported in part by the Wisconsin Dept. of Natural Resources. Publication approved by the Director of the Wisconsin Agricultural Experiment Station.     

Karyologische Untersuchungen an keimenden Ascosporen und Hyphenzellen von Lophodermium pinastri (Schrad.) Chev.

Zusammenfassung Die vorliegenden Untersuchungen sollen die karyologischen Verhältnisse in keimenden Ascosporen und Hyphenzellen von Lophodermium pinastri klären. Die Kernuntersuchungen erfolgten an fixiertem und gefärbtem Material (HCl-Giemsa).Bevor sich bei den Ascosporen Keimhyphen bilden, setzen Kernteilungen sowie die Bildung von Querwänden ein. 2 hauptsächliche Keimformen können unterschieden werden. L. pinastri ist ein Organismus mit mehrkernigen Hyphenzellen (im Mittel 3–6 Kerne/Zelle). Die Terminalzelle enthält im Mittel 7–30 Zellkerne. Zwischen den Kernzahlen der Terminal-und der ersten Folgezelle besteht eine schwache positive Korrelation.Die 4 untersuchten Stämme unterscheiden sich in ihren Kernzahlen hoch signifikant. Die mittlere Kernzahl je Hyphenzelle kann durch veränderte Umweltbedingungen (z. B. Nährboden) variieren.Die Ergebnisse stellen eine cytologische Bestätigung der zu beobachtenden morphologischen und physiologischen Variabilität von L. pinastri dar.

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Karyological studies of germinating ascospores and hyphal cells in Lophodermium pinastri (Schrad.) Chev.

Summary The studies clarify the karyological constitution in germinating ascopores and hyphal cells of Lophodermium pinastri. For the nuclear studies fixed and stained material (HCl-Giemsa) was used. Before the development of ascospore germ tubes, nuclear divisions occur and septa are formed. 2 main types of germinating ascospores could be distinguished. L. pinastri is an organism with multinucleate hyphal cells (average of 3–6 nuclei/cell). The hyphal tip cell contains an average number of 7–30 nuclei. There is a weak positive correlation between the number of nuclei in the tip cell and in the second cell following.The 4 strains studied differ highly significantly in their numbers of nuclei. The average number of nuclei/cell can be influenced by changed environmental conditions.The results confirm cytologically the morphological and physiological variability observed in L. pinastri.

     

Fungi in Living Symptomless Needles of Pinus sylvestris with Respect to Some Observed Disease Processes

The mycoflora in living symptomless needles of Pinus sylvestris was investigated in three 5–8 year-old plantations in Southern Poland from 1987 to 1990. The needles from trees with symptoms of the autumn needle cast and from trees without such symptoms were treated separately. In total 960 of the current year and 720 of the previous year needles were collected. Fungi were isolated from 80.1 % of needles and 3671 cultures belonging to 86 fungal taxa were obtained. Seven species of fungi showed the rate of infection above 5%: Anthostomella formosa, Lophodermium seditiosum, Cydaneusma minus, Cenangium ferruginosum, Lophodermium pinastri, Sclerophoma pythiophila and Anthostomella pedemontana. The frequency of infection of needles by these species depended on needle age, season of the year, calendar year and site. The spread of fungi in needles was not too high. They were found in 35.8 % of needle sections (isolations were made from 10,080 sections). The needle base, middle and top were infected with more or less similar frequency, but the differences were considerable for the individual fungal species. Anthostomella formosa and A. pedemontana generally occurred slightly more frequently on needles of trees without symptoms of the autumn needle cast, while the other four common species ( Cenangium ferruginosum, Cyclaneusma minus, Lophodermium pinastri, Lophodermium seditiosum ) generally occurred more frequently on needles of trees showing such symptoms. There were considerable differences in this respect among plantations investigated.     

Development and seasonal variations of Lophodermium populations on Pinus thunbergii needle litter

A quantitative method was developed to describe Lophodermium (Rhytismataceae, Ascomycetes) populations on Pinus thunbergii needle litter, and seasonal patterns of population variation were examined based on this method. We focused on the formation of black zone lines across needles and the production of Lophodermium ascocarps on needle portions delimitated by zone lines. The study was carried out at a soil erosion control site on a mountain slope and at a coastal sand dune site. Most Lophodermium spp. on needle litter were identified as L. pinastri according to morphological characteristics of ascocarps and ascospores and DNA analysis. Zone lines were produced on needles when isolates of L. pinastri were in contact with other isolates of L. pinastri or with isolates of other species in dual culture tests. This observation provided a rationale to consider that individual colonies with ascocarps and delimitated by zone lines were occupied by a single Lophodermium isolate. Frequency of occurrence of Lophodermium colonies, total colony length, and mean colony number per needle were higher at the coastal sand dune site than at the soil erosion control site. Total colony length and mean colony number also varied with season. Mean colony length and mean ascocarp number per colony were not different between sites or seasons.     

Dehydrin stress proteins in <Emphasis Type="Italic">Pinus sylvestris</Emphasis> L. needles under conditions of extreme climate of Yakutia

This is the first study to investigate stress proteins dehydrins with the use of specific antibodies in the Scots pine (Pinus sylvestris L.) needles and their changes in the annual cycle under extreme climate of Yakutia. No pronounced polymorphism of major dehydrins (14–15 and 66 kDa) has been found during the winter dormancy period of P. sylvestris. A clear correlation between the seasonal variations in dehydrins and changes in the water content in needles was revealed. Consistently high levels of dehydrins was retained throughout the period of low negative temperatures. It is assumed that dehydrins can participate in the formation of P. sylvestris L. resistance to the permafrost conditions.     

Toxic effects of high copper content on physiological processes in <Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.

The aim of this study was to determine the impact of increased copper contents on selected physiological processes in oneyear-old Pinus sylvestris L. needles from a former German timber storage area in Warcino Forest District, a subject to an environmental quality survey. Samples were collected from the area with the high copper content in the soil. The control area was a nearby pine tree stand showing unimpeded growth. The significant growth inhibition was found in dwarf shoots and whole needles, increased water content, and reduced dry mass were also observed. The chlorophyll content was lowered, while 20% higher electrolyte leakage was found. Chlorophyll a fluorescence indicated only higher values of the nonphotochemical quenching in P. sylvestris from the Cu-site. Significant differences were shown in the rate of gas exchange measured by changes in carbon dioxide or oxygen concentration. The intensity of photosynthesis in needles of P. sylvestris from the Cu-site measured by CO₂ uptake was considerably higher than that of oxygen production. The rate of respiration in the needles from the Cu-site measured by the amount of released CO₂ was higher only by 15%, while according to O₂ consumed, the rate increased by 30% in relation to the control. Our results suggest that the copper accumulation in P. sylvestris needles affected the morphology and physiology of the studied organs.     

Fungal succession on pine needles in Germany

The mycofloral succession on the needles ofPinus sylvestris was investigated in Tübingen, southwest Germany. Dead needles attached to the branches (D-type), those caught on branches (C-type) and three types of fallen needles, i.e., freshly fallen (L-type), slightly discolored (OL-type) and almost black needles (F-type) were examined for their fungal flora. Common primary saprophytes were rich on the dead needles on the tree, and on the L-type needles. They were replaced by successive species that contained the well-known species preferring pine needles as their substratum, such asVerticicladium trifidum orSympodiella acicola. Their ecological niches in pine leaf litter and their distribution patterns from a biogeographical viewpoint were discussed.     

<Emphasis Type="Italic">Lophodermium</Emphasis> on needles of conifers from Yunnan Province,China

Twelve Lophodermium species on needles of conifers are known in Yunnan, the eastern Himalayas in China. Recently, additional specimens were collected and identified. Two new species, Lophodermium dilutum on Pinus armandii and Lophodermium tsugae on Tsuga dumosa, were described and illustrated, and six species, L. autumnale, L. confluens, L. himalayense, L. kumaunicum, L. laricinum, and L. nitens, were recorded for the first time for Yunnan. A key to the Lophodermium species on needles of conifers for Yunnan is provided.     

Influence of atmospheric pollution on the morphological parameters of the assimilation apparatus of pine and spruce in the basin of the Northern Dvina

The dynamics of damage and litter of the needles of pine Pinus sylvestris (L.) and spruce Picea obovata Ledeb. × P. abies (L.) Karst. in the basin of the Northern Dvina under atmospheric pollution was studied. Correlations of morphometric parameters of the assimilation apparatus with the distance to the source of emissions and concentrations of pollutants in the air, snow, and soil and heavy metals in needles were revealed.     

Pinewood Nematode,Bursaphelenchus xylophilus,Associated with Red Pine, Pinus resinosa,in Western Maryland

Red pines Pinus resinosa in Garrett and Allegany counties, Maryland, were examined during 1982-84 to determine distribution of the pinewood nematode, Bursaphelenchus xylophilus, within and among trees. Approximately 25-year-old (younger) and 47-year-old (older) trees were subdivided into the following categories: 1) trees with mostly green needles; 2) trees with mostly reddish-brown needles; 3) trees lacking needles but with bark intact; 4) trees lacking both needles and bark; and 5) trees with chlorotic, bleached-green needles. Bursaphelenchus xylophilus was found infecting 68% of younger red pines and 77% of older red pines. Nematodes were not evenly distributed in trees within any given tree decadence category or in trees of the same age. Nematodes were recovered from 20% of wood samples from trunks and primary and secondary branches in younger pines and from 15 % of older red pines. On the basis of tree decadence category, the highest incidence of infection in younger trees (31%) was in bleached-green needled trees (category 5), whereas in older trees the highest infection (25%) occurred in green needled trees (category 1). At both sites trunks were infected more often than branches.     

Effect of provenance on free amino acid and chemical composition of Scots pine needles

Free amino acid (16 amino acids) and chemical composition (N, P, K, Mg, Ca, S, Fe, Mn, Cu, Zn) of Scots pine (Pinus sylvestris L.) needles were compared between six provenances in three different experimental areas. The main free amino acids in the needles were in the sequence of quantity; glutamic acid, glutamine, arginine and γ-aminobutyric acid. There were no significant differences in the concentrations of phenylalanine, γ-aminobutyric acid, methionine, proline and threonine in pine needles between the sites or between the provenances. Significant differences in the foliar concentrations of alanine and leucine were found between the sites and in the foliar concentrations of isoleucine, glutamine, glycine and tyrosine between the provenances. The concentrations of aspartic acid, glutamic acid, arginine and lysine were significantly affected by the sites and the provenances. The foliar nutrients, except copper, had statistically significant differences, both between the sites and between the provenances. Calcium did not differ between the provenances. This revised version was published online in June 2006 with corrections to the Cover Date.     

Assay and Electrophoresis of Superoxide Dismutase from Red Spruce (Picea rubens Sarg.), Loblolly Pine (Pinus taeda L.), and Scotch Pine (Pinus sylvestris L.) : A Method for Biomonitoring

This paper reports a method for extracting the antioxidant enzyme superoxide dismutase (SOD) from the needles of red spruce (Picea rubens Sarg.), loblolly pine (Pinus taeda L.), and scotch pine (Pinus sylvestris L.) with high efficiency and free from interfering compounds. The extraction employs phosphate buffer with polyvinylpolypyrrolidone and Triton X-100 followed by dialysis overnight. The isozymes of SOD in each species were separated electrophoretically and tested for their sensitivity to KCN and H₂O₂. An isozyme resistant to these inhibitors was found in the spruce but not the pine needles. The isozymes from the spruce needles were examined for individual responses to aging and H₂O₂ inhibition. Four of the five CuZn isozymes in spruce were found to have increased significantly but equally by October of their first year and two of those four isozymes were found to be more sensitive to H₂O₂. The response of the SOD isozymes in loblolly pine seedlings to O₃ was also examined and the isozymes were found to be induced equally. Because the SOD activity in the young pine needles was too low to electrophorese, the SOD activity from the pines in the O₃ experiment had to be partially purified using CHCl₃ and ethanol, then concentrated.     

Effects in s.e. Queensland during 1967–72 of insects introduced to controlLantana camara

Seasonal fluctuations in populations of the introduced speciesTeleonemia scrupulosa Stål and damage by this insect to common taxa ofLantana camara, were observed at several field sites and experimental plots. At 2 plots effects ofT. scrupulosa populations on untreated plants were compared with plants protected by application of insecticides. On untreated plants damage varied seasonally and between taxa ofL. camara. In general attack byT. scrupulosa reduced fruiting and caused severe short term foliage damage. Combined with severe stress from some other cause heavy attack may result in long term damage or death to plants. However Common PinkL. camara was more resistant to damage than were the other taxa studied. At the time of these experiments other insect species were at very low population levels and did not have observable effects onL. camara.     

Identification of Naemacyclus minor hyphae within needle tissues of Pinus sylvestris by immunoelectron microscopy

Cultural investigations revealed that Naemacyclus minor, Lophodermium seditiosum and Cenangium ferruginosum were the most frequent colonizers of asymptomatic and symptomatic Pinus sylvestris needles. Since ultrastructural observations showed that morphological features were not suitable to differentiate hyphae of N. minor from hyphae of other isolates, the on-section immunogold labelling technique was applied in combination with an anti-N. minor specific immunoserum. The specificity of this serum was tested against culture hyphae of all isolates. Anti-N. minor specific immunoserum was then used to identify N. minor hyphae in thin sections of green P. sylvestris needles. The infection loci identified were restricted to small tissue areas located in the vicinity of stomata. In the hypodermis, hyphae and endocell-containing hyphae were located within the lumina of host cells but outside the protoplast. The growth of hyphae from cell to cell occurred through pits. The hyphae spreat into the mesophyll intercellularly and continued with the intracellular colonization of moribund and dead mesophyll cells in a later stage of infection. The observed host-parasite interactions at cellular and ultrastructural level are discussed in connection with the still controversial interpretation of the pathogenicity of N. minor.     

The growth and nutrients status of conifers on ash-treated cutaway peatland

Key message

Use of wood ash or a mixture of wood and oil shale ashes increases the concentrations of P and K in the assimilation organs of conifers and stimulates tree growth.

Abstract

The effect of fertilization with wood ash (10 and 15 t ha^?1) and a mixture of wood ash (10 t ha^?1) and oil shale ash (8 t ha^?1) on the growth (height, root collar diameter, biomass, biomass production) and nutrient concentrations in subsoil and needles of young Pinus sylvestris and Picea abies plants on the Puhatu (Northeast Estonia) cutaway peatland in the first 2 years were studied. After the second growing year differences in the average height growth of P. abies and P. sylvestris were statistically significantly higher on ash-treated plots than on the control plots (p < 0.05), being respectively 1.4–1.6 and 1.5–1.7 times greater than height growth of the control trees. The best results on root collar diameter were observed on mixture ash treatments: the root collars were 1.9 (P. abies) and 2.2 (P. sylvestris) times larger than of the control trees. The biomass of the two conifer species and the biomass production of P. sylvestris in 2012 was the greatest on the mixture ash treatments. Five months after fertilization with ashes the concentrations of P, K, Ca and Mg were higher on the treated plots than on the control plot. Although the concentrations of P and K in P. sylvestris needles rose after the treatment with ash, seedlings suffered from P and K deficiency. The concentrations of P and K in P. abies needles were on optimum. The P/N and the K/N ratios in needles were also improved compared to control trees needles.     

Chlorophyll Synthesis in Dark-Grown Pine Primary Needles

The pigment content of dark-grown primary needles of Pinus jeffreyi L. and Pinus sylvestris L. was determined by high-performance liquid chromatography. The state of protochlorophyllide a and of chlorophylls during dark growth were analyzed by in situ 77 K fluorescence spectroscopy. Both measurements unambiguously demonstrated that pine primary needles are able to synthesize chlorophyll in the dark. Norflurazon strongly inhibited both carotenoid and chlorophyll synthesis. Needles of plants treated with this inhibitor had low chlorophyll content, contained only traces of xanthophylls, and accumulated carotenoid precursors. The first form of chlorophyll detected in young pine needles grown in darkness had an emission maximum at 678 nm. Chlorophyll-protein complexes with in situ spectroscopic properties similar to those of fully green needles (685, 695, and 735 nm) later accumulated in untreated plants, whereas in norflurazon-treated plants the photosystem I emission at 735 nm was completely lacking. To better characterize the light-dependent chlorophyll biosynthetic pathway in pine needles, the 77 K fluorescence properties of in situ protochlorophyllide a spectral forms were studied. Photoactive and nonphotoactive protochlorophyllide a forms with emission properties similar to those reported for dark-grown angiosperms were found, but excitation spectra were substantially red shifted. Because of their lower chlorophyll content, norflurazon-treated plants were used to study the protochlorophyllide a photoreduction process triggered by one light flash. The first stable chlorophyllide photoproduct was a chlorophyllide a form emitting at 688 nm as in angiosperms. Further chlorophyllide a shifts usually observed in angiosperms were not detected. The rapid regeneration of photoactive protochlorophyllide a from nonphotoactive protochlorophyllide after one flash was demonstrated.     

Pinus uncinata Ramond taxonomy based on needle characters

 11 needle characters of Pinus uncinata Ramond from the Spanish Pyrenees, P. mugo Turra from the Tatra Mountains, and P. sylvestris L. from N.E. Poland were analysed. It was shown that P. uncinata is much more closely related to P. mugo than to P. sylvestris. Nevertheless P. uncinata remains distinct from both in a number of the characters examined. The species differs from P. mugo especially in the length of the needles and in the distance between the vascular bundles. It differs from P. sylvestris in such characters as the number of resin canals, thickness of epidermis cells and the distance between the vascular bundles. Width of epidermis cells was similar in all taxa studied. Received May 31, 2000 Accepted February 13, 2001