Separation of allelopathy and resource competition by the boreal dwarf shrub Empetrum hermaphroditum Hagerup

An experimental technique was used to separate and evaluate the magnitude of allelopathic interference relative to resource competition by the boreal dwarf shrub Empetrum hermaphroditum Hagerup (Empetraceae). To test for resource competition and allelopathy, respectively, Scots pine (Pinus sylvestris L.) seedlings were grown in both the greenhouse and in the field over a 3 year period, in four different treatments within E. hermaphroditum vegetation: (1) PVC tubes were used to reduce effects of interspecific below-ground competition; (2) activated carbon was spread on the soil to adsorb toxins leached from E. hermaphroditum litter and green leaves, thus reducing effects of allelopathic interference; (3) E. hermaphroditum vegetation was left untreated to evaluate inhibiting effects when both allelopathy and resource competition were present; (4) PVC tubes, placed in E. hermaphroditum vegetation spread with activated carbon were used to determine growth of seedlings when both allelopathy and resource competition were reduced. Scots pine seedlings grown in untreated vegetation (with both root competition and allelopathy present) had the lowest shoot length and dry weight; seedlings with both allelopathy and root competition reduced (activated carbon in tube) were the largest. Reducing either root competition alone (tube treatment) or allelopathy alone (carbon treatment) produced seedlings of intermediate size, but reduced competition had a greater effect than reduced allelopathy (although, in the greenhouse, significantly so only for root biomass). In the greenhouse experiment, biomass production of seedlings grown free of both interactions (carbon in tube) was greater than the simple sum of the growth response to the individual interactions (tube treatment and carbon treatment, respectively). Larger shoot:root ratios were also found when pine seedlings were grown without tubes (i.e. when resource competition was occurring). In the field, the removal of allelopathy (carbon treatments) increased shoot:root ratio when compared to the removal of resource competition. The study showed that two different interference mechanisms of E. hermaphroditum can be separated and quantified, and that below-ground competition and allelopathy by E. hermaphroditum are both important factors retarding growth of Scots pine.     

The role of cattle in maintaining plant species diversity in wet dune valleys

The succession of species-rich wetland vegetation in dune valleys into species-poor dwarf shrub vegetation was followed by means of permanent vegetation plots, in which the cover of vascular plant, moss and lichen species were recorded over a period of up to 33 years. Low density cattle grazing is an effective substitute for rabbits in stalling this succession, thus preserving the local plant species diversity. The influence of direct rabbit grazing pressure on the vegetation was studied in exclosures. The differences were significant, but not long-lasting after the exclosures had been opened again to the rabbits. Cladina lichen-rich Empetrum-heathland in the dunes is a stage in a probably cyclic succession that might be triggered by grazing. The total vascular plant, moss and lichen diversity of the dune ecosystem can be maintained by a combination of extensive cattle grazing and a regular but limited re-creation of pioneer situations.     

Facilitation of Nothofagus antarctica (Fagaceae) seedlings by the prostrate shrub Empetrum rubrum (Empetraceae) on glacial moraines in Patagonia

Abstract Although positive interactions among plants are believed to be common in primary succession, they have rarely been demonstrated on glacial moraines. In Patagonia, the prostrate shrub Empetrum rubrum is dominant in early succession in recently deglaciated valleys, eventually being replaced by the small tree Nothofagus antarctica. This study experimentally evaluates the effects of E. rubrum shrubs on density, biomass, growth and survival of N. antarctica seedlings. The initial density of the seedlings was significantly higher under the canopy of Empetrum than in open areas. A removal experiment was performed to evaluate the influence of Empetrum on survival and growth of seedlings that germinated naturally beneath the canopy of this species. The shrub cover was removed from half of a sample of randomly selected seedlings, and left intact above the remainder. One year later, seedlings with an intact cover of E. rubrum showed significantly higher survival, growth and leaf number than seedlings which had been exposed. The results suggest that the presence of E. rubrum facilitates the establishment of N. antarctica in post‐glacial succession, mainly as a result of more favourable microclimatic conditions under the shrub canopy.     

Ecosystem disturbance reduces the allelopathic effects of Empetrum hermaphroditum humus on tundra plants

Question: Do naturally occurring ecosystem disturbances interfere with allelopathic effects and alleviate growing conditions for plants typical of more productive ecosystem states? Location: An unproductive tundra ecosystem in Northern Fennoscandia. Methods: We designed a factorial phytotron experiment to monitor the growth of seedlings of two herbaceous species in undisturbed and disturbed humus of the dominant dwarf shrub Empetrum hermaphroditum, which releases allelopathic substances that accumulate in soil over time. Herbivore defecation and wildfire are two major ecosystem disturbances likely to alleviate effects of these allelopathic substances. Humus and seeds of herbaceous species were sampled from Empetrum‐dominated heaths. Disturbance effects on humus were simulated by adding reindeer faeces and by low‐severity burning. Results: The concentration of the allelopathic substance batatasin‐III in Empetrum humus was 0.32±0.16 (SE) μg g^?1, which is similar to previous reports from boreal areas. Reindeer faeces addition and burning treatments to Empetrum humus caused three‐ and five‐fold increases in growth of the herbaceous species Avenella flexuosa and Solidago virgaurea respectively. Changes in soil chemistry caused by the disturbance, i.e. increased pH and nutrient availability, likely interfered with allelopathic effects and improved conditions for growth. Conclusions: Humus from Empetrum‐dominated tundra areas is infertile for seedlings of local herbaceous plants. Simulated herbivore defecation and wildfire can alleviate this infertility. Based on our results, we discuss the potential role of these naturally occurring disturbances in switching vegetation composition from Empetrum to herbaceous dominance, i.e. from an unproductive to a more productive tundra ecosystem.     

Abundance,diversity, and vitality of mycorrhizae of Scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) in lignite recultivation sites

Scots pine (Pinus sylvestris L.) stands cover large areas in the Lusatian and the Middle German lignite mining districts. Due to adverse chemical substrate conditions, the root systems of the trees are restricted to the ameliorated top-spoil and the organic forest floor layers. To investigate functioning of fine root systems under the prevailing site factors, we studied mycorrhizal colonization rate and frequency as well as mycorrhizal diversity, vitality and growth phases in Scots pine ecosystems along a chronosequence in both mining districts. Mycorrhizal rate was close to 100% in both districts. Mycorrhizal abundance was higher in the organic forest floor layer than the mineral soil layer. In total, 25 morphotypes were recorded. Diversity differed between the districts. The mycorrhizae of Amphinema byssoides, Tuber puberulum, Pinirhiza discolor, Pinirhiza cf. bicolorata and E-type were present in both mining areas. These morphotypes are typical of nutrient-rich soils with high pH values. Compared with the undisturbed sites, vitality of mycorrhizae was very high at the test sites on spoil substrate, correlating with the high growth dynamics of mycorrhizae at recultivation sites. A relatively high carbon flow to the mycorrhizal root systems at these sites seems likely. Thus, mycorrhizal root systems are able to cope with the ameliorated top-spoil and the organic layer. The main reason for the adaptation is the large number of ectomycorrhizal fungal species available in this area where Pinus sylvestris is indigenous.     

Ectomycorrhizal community structure of different genotypes of Scots pine under forest nursery conditions

In this paper, we report the effect of Scots pine genotypes on ectomycorrhizal (ECM) community and growth, survival, and foliar nutrient composition of 2-year-old seedlings grown in forest bare-root nursery conditions in Lithuania. The Scots pine seeds originated from five stands from Latvia (P1), Lithuania (P2 and P3), Belarus (P4), and Poland (P5). Based on molecular identification, seven ECM fungal taxa were identified: Suillus luteus and Suillus variegatus (within the Suilloid type), Wilcoxina mikolae, Tuber sp., Thelephora terrestris, Cenococcum geophilum, and Russuloid type. The fungal species richness varied between five and seven morphotypes, depending on seed origin. The average species richness and relative abundance of most ECM morphotypes differed significantly depending on pine origin. The most essential finding of our study is the shift in dominance from an ascomycetous fungus like W. mikolae in P2 and P4 seedlings to basidiomycetous Suilloid species like S. luteus and S. variegatus in P1 and P5 seedlings. Significant differences between Scots pine origin were also found in seedling height, root dry weight, survival, and concentration of C, K, Ca, and Mg in the needles. The Spearman rank correlation coefficient revealed that survival and nutritional status of pine seedlings were positively correlated with abundance of Suilloid mycorrhizas and negatively linked with W. mikolae abundance. However, stepwise multiple regression analysis showed that only survival and magnesium content in pine needles were significantly correlated with abundance of ECM fungi, and Suilloid mycorrhizas were a main significant predictor. Our results may have implications for understanding the physiological and genetic relationship between the host tree and fungi and should be considered in management decisions in forestry and ECM fungus inoculation programs.     

Endobacteria in some ectomycorrhiza of Scots pine (Pinus sylvestris)

The diversity of cultivable endobacteria associated with four different ectomycorrhizal morphotypes (Suillus flavidus, Suillus variegatus, Russula paludosa and Russula sp.) of Scots pine (Pinus sylvestris) was analysed by restriction fragment length polymorphism profiling of PCR-amplified rDNA intergenic spacer regions and by sequence analyses of 16S rRNA genes. Ectomycorrhizal root tip surface-sterilization methods were developed and assessed for their efficiencies. Bacterial communities from surface-sterilized ectomycorrhizal root tips were different from those of ectomycorrhizal root tips without surface-sterilization for all the morphotypes studied. Endobacteria belonging to the genera Pseudomonas, Burkholderia and Bacillus were isolated from more than one ectomycorrhizal morphotype, whereas species of Rahnella, Janthinobacterium and Rhodococcus were only isolated from the single morphotypes of S. variegatus, R. paludosa and Russula sp., respectively. Some of the isolated endobacteria utilized fungal sugars more readily than typical plant sugars in carbon utilization assays.     

Ectomycorrhizal development on Scots pine (Pinus sylvestris L.) seedlings in different soils

In the present study ectomycorrhizal development of Laccaria bicolor, Rhizopogon luteolus and Suillus bovinus associated with Scots pine (Pinus sylvestris) seedings was studied as affected by primary stand humus, secondary stand humus, podsolic sandy soil or peat in perspex growth chambers. After 9 weeks, ectomycorrhizal development with S. bovinus was significantly greater in peat and primary stand humus than in secondary stand humus or podsolic sandy soil. Ectomycorrhizal development with R. luteolus in secondary stand humus was higher than in primary stand humus. Degree of ectomycorrhizal development of L. bicolor, R. lutuelus and S. bovinus on Scots pine was related to potassium concentration, organic matter content and pH of the soils suggesting that chemical composition of the soils affects ectomycorrhizal development.     

Effects of aluminium on growth and nutrient uptake of small Picea abies and Pinus sylvestris plants

Summary The effects of aluminium concentrations between 0.2 and 30 mM at pH 3.8 ±0.2 on small plants of Norway spruce [(Picea abies (L.) Karst], Scots pine (Pinus sylvestris L.), and Scots pine infected with the ectomycorrhizal fungus Suillus bovinus (L. ex Fr.) O. Kuntze were investigated. The plants were grown at maximum relative growth rate (R_G % day^–1) with free access but very low external concentrations of nutrients. Steady-state conditions with respect to relative growth rate (R_G) and internal nutrient concentrations were achieved before addition of aluminium, which was added as AlCl₃ and/or Al(NO₃)₃. There were reductions in rg at aluminium concentrations of 0.3 mM in spruce, 6 mM in pine and 10 mM in ectomycorrhizal pine, i. e. at aluminium concentrations considerably higher than those normally occurring in the top layer of the mineral soil where most fine roots are found. Nutrient uptake rate per unit root growth rate was calculated for different nutrient elements. The uptake rate of calcium and magnesium was reduced at aluminium concentrations of 0.2 mM (spruce), 1 mM (pine) and 3 mM (ectomycorrhizal pine), without influencing R_g. The results question the validity of the hypothesis of aluminium toxicity to forest tree species at low external concentrations.     

The threatened plant intermediate wintergreen (<Emphasis Type="Italic">Pyrola media</Emphasis>) associates with a wide range of biotrophic fungi in native Scottish pine woods

The plant intermediate wintergreen (Pyrola media, Ericaceae) is in need of conservation action in Scotland. Although widespread, it is locally distributed in dwarf shrub heath and more commonly in Scots pine (Pinus sylvestris) woodlands. A recent study on the mycorrhizal status of Pyrola suggested that they associate with a restricted range of ectomycorrhizal (ECM) fungi. Here, we examined the hypothesis that specialisation by P. media for fungi usually associated with Scots pine is a factor in promoting its occurrence in this habitat. The fungal community associated with the roots of P. media growing in a Scots pine forest was determined by morphotyping, polymerase chain reaction, cloning and sequencing. Molecular identification found 49 taxa representing ecto- and ericoid mycorrhizal fungi, dark septate endophytes, saprotrophs, and fungi of unknown trophic status. The majority of the taxa (67.4%) were Basidiomycota, with 24.4% known to be ECM fungi specific to Pinus sp. or conifers. However, a wide range of other mycorrhizal fungi with varying degrees of host specificity were also found, including taxa usually associated with deciduous hosts. In conclusion, the broad range of mycorrhizal fungi recovered from the roots of P. media suggests that specialization is not a major factor in determining its distribution.     

Ectomycorrhizal root growth in scots pine stands in response to manipulation of litter and humus layers

 The effect on ectomycorrhizal root growth in a nitrogen-enriched planted stand of Scots pine (Pinus sylvestris L.) on podzolic sandy soil to manipulation of litter and humus layers (removal, doubling and control treatments) was examined, and compared to ectomycorrhizal root growth in an untreated naturally established Scots pine stand on nutrient-poor non-podzolic sandy soil. Half a year after manipulation of litter and humus layers in the planted stand, ingrowth-cores to a depth of 60 cm were installed in both stands. Scots pine roots were sampled four times during two growing seasons. Ectomycorrhizal roots were found at all sampled soil depths to 60 cm in all plots. Root growth and ectomycorrhizal development were greater in the naturally established stand than in all plots in the planted stand. Numbers of ectomycorrhizal root tips in the litter and humus removal plots were generally higher than in the control plots in the planted stand until May 1992. Doubling litter and humus did not significantly affect root length or the numbers of ectomycorrhizal root tips. The N _dissolved , NH₄ ⁺ and NO₃ ^– concentrations and the organic matter content in the upper 5 cm of the mineral soil in the planted stand on podzolic sandy soil were generally higher and the pH significantly lower than in the naturally established stand on non-podzolic sandy soil. Root growth and ectomycorrhizal development in the secondary stand may have been negatively affected by the chemical composition of the podzolic sandy soil. Accepted: 19 March 1997     

Partial restoration of fungal and plant species diversity by removal of litter and humus layers in stands of Scots pine in the Netherlands

A field experiment was set up in five mature Scots pine (Pinus sylvestris) stands in the Netherlands in order to study the effects of removal of litter and humus on ectomycorrhizal fungi and vegetation of higher plants, bryophytes, and lichens. Lime was also applied as a countermeasure to the removal of surplus cations. Application of lime did not have significant effects on mycorrhizal fungi. Removal of ectorganic material increased species richness of ectomycorrhizal fungi. The number of fruit bodies was also higher in the sod-cut plots, but the difference compared with the controls was not significant in all years. A major effect of removal of the ectorganic layer was a reversion of ectomycorrhizal succession to earlier stages. Liming increased the number and cover of ruderal, nitrophilous plant species. Sod-cutting caused a reversion of the green vegetation to earlier successional stages.     

Limiting factors for seedling emergence and establishment of the invasive non-native Rosa rugosa in a coastal dune system

The relative importance of seed, habitat and microhabitat limitation has rarely been investigated for invasive non-native species, although this is critical for their effective management and for predicting future range expansion. Rosa rugosa is an abundant non-native shrub in coastal habitats of NW Europe; it is common along the Baltic coast but more scarce in exposed dunes of the North Sea. To investigate whether invasion of exposed dunes is limited by seed, habitat and microhabitat limitation, seedling emergence and establishment were examined in a factorial sowing, transplant and disturbance experiment. Twenty plots were randomly placed in each of five dune habitats (white dune, Empetrum dune, grey dune, outer dune heath, inner dune heath), and studied over 2 years. Seedling emergence in control subplots was zero in all habitats, whereas 96% and 98% of the undisturbed and disturbed seeded subplots produced seedlings. Disturbance had a positive effect on emergence and subsequent survival in white dune, outer and inner dune heath. Seedling survival and growth, and growth of transplanted seedlings, were markedly lower in grey dune. These findings indicate that establishment of R. rugosa is seed-limited in coastal dune habitats, and that the species is able to establish in both active and fixed dunes once seeds have arrived. Although differential seedling emergence and growth indicate that habitats differ in their degree of invasion susceptibility by R. rugosa, the positive influence of small-scale disturbance suggests microhabitat limitation in some dune habitats as well. Dune management should aim to reduce seed production and dispersal of R. rugosa in near-natural sites, and anthropogenic changes of habitat dynamics should be prevented.     

Ectomycorrhizal responses to organic and inorganic nitrogen sources when associating with two host species

While it is established that increasing atmospheric inorganic nitrogen (N) deposition reduces ectomycorrhizal fungal biomass and shifts the relative abundances of fungal species, little is known about effects of organic N deposition. The effects of organic and inorganic N deposition on ectomycorrhizal fungi may differ because responses to inorganic N deposition may reflect C-limitation. To compare the effects of organic and inorganic N additions on ectomycorrhizal fungi, and to assess whether host species may influence the response of ectomycorrhizal fungi to N additions, we conducted an N addition experiment at a field site in the New Jersey pine barrens. Seedlings of two host species, Quercus velutina (black oak) and Pinus rigida (pitch pine), were planted at the base of randomly-selected mature pitch pine trees. Nitrogen was added as glutamic acid, ammonium, or nitrate at a rate equivalent to 227.5 kg ha⁻¹ y⁻¹ for eight weeks, to achieve a total application of 35 kg ha⁻¹ during the 10-week study period. Organic and inorganic N additions differed in their effects on total ectomycorrhizal root tip abundance across hosts, and these effects differed for individual morphotypes between oak and pine seedlings. Mycorrhizal root tip abundance across hosts was 90 % higher on seedlings receiving organic N compared to seedlings in the control treatment, while abundances were similar among seedlings receiving the inorganic N treatments and seedlings in the control. On oak, 33–83 % of the most-common morphotypes exhibited increased root tip abundances in response to the three forms of N, relative to the control. On pine, 33–66 % of the most-common morphotypes exhibited decreased root tip abundance in response to inorganic N, while responses to organic N were mixed. Plant chemistry and regression analyses suggested that, on oak seedlings, mycorrhizal colonization increased in response to N limitation. In contrast, pine root and shoot N and C contents did not vary in response to any form of N added, and mycorrhizal root tip abundance was not associated with seedling N or C status, indicating that pine received sufficient N. These results suggest that in situ organic and inorganic N additions differentially affect ectomycorrhizal root tip abundance and that ectomycorrhizal fungal responses to N addition may be mediated by host tree species.     

Growth and Mycorrhizal Community Structure of Pinus sylvestris Seedlings following the Addition of Forest Litter

We report the effects of pine and oak litter on species composition and diversity of mycorrhizal fungi colonizing 2-year-old Pinus sylvestris L. seedlings grown in a bare-root nursery in Lithuania. A layer of pine or oak litter was placed on the surface of the nursery bed soil to mimic natural litter cover. Oak litter amendment appeared to be most favorable for seedling survival, with a 73% survival rate, in contrast to the untreated mineral bed soil (44%). The concentrations of total N, P, K, Ca, and Mg were higher in oak growth medium than in pine growth medium. Relative to the control (pH 6.1), the pH was lower in pine growth medium (5.8) and higher in oak growth medium (6.3). There were also twofold and threefold increases in the C content of growth medium with the addition of pine and oak litter, respectively. Among seven mycorrhizal morphotypes, eight different mycorrhizal taxa were identified: Suillus luteus, Suillus variegatus, Wilcoxina mikolae, a Tuber sp., a Tomentella sp., Cenococcum geophilum, Amphinema byssoides, and one unidentified ectomycorrhizal symbiont. Forest litter addition affected the relative abundance of mycorrhizal symbionts more than their overall representation. This was more pronounced for pine litter than for oak litter, with 40% and 25% increases in the abundance of suilloid mycorrhizae, respectively. Our findings provide preliminary evidence that changes in the supply of organic matter through litter manipulation may have far-reaching effects on the chemistry of soil, thus influencing the growth and survival of Scots pine seedlings and their mycorrhizal communities.     

An assessment of below-ground ectomycorrhizal diversity of Abies alba miller in central Italy

ABSTRACT

In the framework of an ongoing study on the mycorrhizal associations of silver fir (Abies alba Mill., Pinaceae), we investigated the below-ground diversity of ectomycorrhizal communities in ten field sites located in five distinct natural A. alba woods, situated in the central part of the Apennine chain (Abruzzo region, Italy). Based on macro- and microscopic features, a total of 48 morphologically distinct ectomycorrhizal types have been classified on mature trees of A. alba, 37 of which are reported here for the first time. Ectomycorrhizal morphotypes were partially characterized, and their main features described; in many cases, the relevant fungal symbiont was identified at the level of species or genus. The number of distinguishable morphotypes per site was, with few exceptions, rather homogeneous, ranging from (5) 8 to 13 (20). Comparison of morphotype occurrence revealed that only few types were encountered in five or more sampled sites, whereas the vast majority of types was less frequent. The present study revealed a considerably high species diversity of the ectomycorrhizal symbionts of A. alba in a quite restricted area, thus raising interesting questions as to the ectomycorrhizal potential of this host tree over its entire, large natural range.     

Recent 14C-labelled assimilate allocation to Scots pine seedling root and mycorrhizosphere compartments developed on reconstructed podzol humus,E- and B- mineral horizons

In northern boreal forests, podzolic soils prevail that comprise of a distinct upper organic humus/mor (O) horizon that is supported by underlying eluvial (E) and illuvial (B) mineral horizons. The dominant tree species, Scots pine (Pinus sylvestris L.), is known to be highly dependent on root symbiosis with ectomycorrhizal fungi that develop in constituent podzol horizons for growth in these nutrient limited soils. The aim of this microcosm-based study was a quantification of photosynthetically fixed ¹⁴C allocation, following standard pulse-feeding of 7-month-old Scots pine seedling shoots, to respective root and mycorrhizosphere compartments that developed in the reconstructed podzol (O, E and B) profile. Biomass of roots and mycorrhizas decreased with increasing soil depth but no soil origin, control forest vs. clear-cut area, related differences were observed. Similarly, no major soil origin- or podzol horizon-related differences in categorised ectomycorrhizal morphotypes and number of mycorrhizas, in relation to pooled root and mycorrhiza biomass, were detected. However, the total recovery of ¹⁴C-label was significantly higher in clear-cut soil microcosms compared to control counterparts. A significant finding was equivalent ¹⁴C-carbon allocation to roots and ectomycorrhizas in all three major, organic and mineral, podzol profile horizons studied. These carbon allocation data provide additional support for direct (or indirect) roles of roots and symbiotic mycorrhizal fungi in mineral weathering and biodegradation of organic ligands that are central for plant acquisition of growth limiting nutrients and the podzolization process in boreal forest ecosystems.     

Use of Shrubs as Nurse Plants: A New Technique for Reforestation in Mediterranean Mountains

Common techniques currently used for afforestation in the Mediterranean basin consider the pre‐existing vegetation (mainly shrubs) as a source of competition for trees, and consequently it is generally eliminated before planting. Nevertheless, it has been demonstrated that woody plants can facilitate the establishment of understory seedlings in environments that, like the Mediterranean area, are characterized by a pronounced dry season. In this study, we experimentally analyze the usefulness of shrubs as nurse plants for afforestation of two native conifers, Pinus sylvestris L. (Scots pine) and Pinus nigra Arnold (black pine). Two‐year‐old seedlings were planted in four microhabitats: (1) open interspaces without vegetation (which is the usual method used in afforestation programs), (2) under individuals of Salvia lavandulifolia, (3) under the north side of spiny shrubs, and (4) under the south side of spiny shrubs. Pine survival was remarkably higher when planted under individuals of the shrub S. lavandulifolia (54.8% for Scots pine, 81.9% for black pine) compared with open areas (21.5% for Scots pine, 56.8% for black pine; chi square, p < 0.05). The survival of both pines was also higher when planted on the north side of spiny shrubs, although the survival on the south side was similar to that found in open areas. In addition, pine growth was not inhibited when planted in association with shrubs. This pattern appears to result from the combination of abiotic conditions imposed by the presence of a nurse shrub, which leads to improvement in seedling water status and therefore reduced summer mortality by drought. The results show that the use of shrubs as nurse plants is a technique that offers both economic and ecological advantages, in terms of savings in labor and plant material and reduced and even negligible impact on the pre‐existing vegetation.     

Lack of regeneration and climatic vulnerability to fire of Scots pine may induce vegetation shifts at the southern edge of its distribution

Aim Forest ecosystems dominated by fire‐sensitive species could suffer shifts in composition under altered crown fire regimes mediated by climate change. The aims of this study were to: (1) study the spatio‐temporal patterns and the climatic distribution of fires in Scots pine (Pinus sylvestris) forests during the last 31 years in north‐eastern Spain, (2) evaluate the climatic vulnerability to fire of these forests in Spain, (3) analyse the regeneration of Scots pine after fire, and (4) predict the mid‐term maintenance or replacement of Scots pine in burned areas. Location Catalonia (north‐eastern Spain): the southern distribution limit of Scots pine. Methods We characterized the spatio‐temporal and the climatic distribution of fires that occurred in Catalonia between 1979 and 2009. We used a generalized linear model to characterize the climatic vulnerability to fire of Scots pine in the whole of Spain. We assessed the regeneration of the species after crown fires in nine burned areas in Catalonia. The resulting data were integrated into a stochastic matrix model to predict the mid‐term maintenance or replacement of Scots pine in burned areas. Results During the last three decades, Scots pine forests distributed in dry sites were most affected by fire. Our assessment of the vulnerability to fire of Scots pine forests in Spain as a whole, based on climatic and topographical variables, showed that 32% of these forests are vulnerable to fire, and that this proportion could increase to 66% under a conservative climate change scenario. Field data showed almost no regeneration of Scots pine after crown fires, and a limited capacity to recolonize from unburned edges, even in relatively old fires, with 90% of recruits located in the first 25 m from the edge. This process could be delayed by the elapsed time for new recruits to achieve reproductive maturity, which we estimated to be c. 15 years. Finally, our matrix model predicted the replacement of burned Scots pine forests by oak (Quercus sp.) forests, shrublands or mixed resprouter forests. Main conclusions Increased vulnerability to fire of Scots pine forests under future, warmer conditions may result in vegetation shifts at the southern edge of the distribution of the species.     

Pisolithus tinctorius promotes germination and forms mycorrhizal structures in Scots pine somatic embryos in vitro

The results of the present study show that inoculation with the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker and Couch potentially enhances the germination of Scots pine (Pinus sylvestris L.) somatic embryos in vitro. Stimulation by Pisolithus tinctorius was only observed in the absence of direct contact between the symbionts; mature embryos were not sufficiently robust for balanced interaction with the fungus on half-strength DCR medium. Subsequently, on MMN medium with a reduced sugar concentration, direct contact between somatic embryo-derived plants and the fungus resulted in in vitro formation of mycorrhiza. Ex vitro inoculation also improved adaptation of the somatic embryo-derived plants, even though mycorrhizal structures were not observed. The reactions to Pisolithus tinctorius varied between different Scots pine cell lines both in vitro and ex vitro.