Root architecture might account for contrasting establishment success of <Emphasis Type="Italic">Pseudotsuga menziesii</Emphasis> var. <Emphasis Type="Italic">menziesii</Emphasis> and <Emphasis Type="Italic">Pinus sylvestris</Emphasis> in Central Europe under dry conditions

Key message

Pinus sylvestris seedlings quickly expand their roots to deeper soil layers while Pseudotsuga menziesii concentrates its root system in the topsoil, thereby running the risk of desiccation during long dry spells, as indicated by lower survival after simulated summer drought.

Context

Pseudotsuga menziesii (Douglas-fir) is regarded as a promising species to maintain the productivity of Central European lowland forests given the projected increase of long dry spells.

Aims

Will the species be able to regenerate from seed and spread outside plantations in a drier temperate Europe?

Methods

We measured the relative growth rate, biomass allocation, root architecture, and phenotypic plasticity of Pseudotsuga menziesii seedlings sown in a common garden and grown under current precipitation and prolonged drought, respectively. The species’ competitive ability with respect to Pinus sylvestris L., the most drought-tolerant native conifer in Central Europe, was assessed during three growing seasons.

Results

Pinus sylvestris seedlings had higher relative growth rates than did Pseudotsuga menziesii seedlings, first in terms of aboveground biomass and later in terms of shoot height. This resulted in heavier and taller seedlings after three growing seasons under both moist and dry conditions. Shorter vertical roots corresponded with lower survival of Pseudotsuga menziesii seedlings under dry conditions.

Conclusion

Fast root proliferation allows Pinus sylvestris seedlings to reach deeper water pools that are less rapidly depleted during transient drought. By contrast, the shallow root system might put Pseudotsuga menziesii seedlings at the risk of desiccation during prolonged dry spells.

     

Changes in soil properties under <Emphasis Type="Italic">Eucalyptus</Emphasis> relative to <Emphasis Type="Italic">Pinus massoniana</Emphasis> and natural broadleaved forests in South China

The ecological effects of eucalypt plantations (EPs) have garnered increasing attention. To understand their effect on soil quality at a landscape scale, and to determine whether soil quality parameters differ due to different stand types, we evaluated soil characteristics in twenty-one groups of EPs, Pinus massoniana Lamb. plantations (PMPs) and natural broadleaved forests (NBFs) across Guangdong Province, China. Both the physical characteristics of soil hydrology and the properties of soil nutrients in A and B horizons were determined. Results showed that, compared to NBFs, EPs and PMPs produced a shallower litter layer, reduced canopy density, higher soil bulk density, significantly lower total porosity, non-capillary porosity, total water volume, and hygroscopic water in the A horizon (P < 0.05). Moreover, total N, available K, and soil organic carbon (SOC) in EPs and PMPs were significantly lower than in NBFs. EPs and PMPs did not differ significantly in N, P or K content, but PMPs had significantly lower SOC and boron in the A horizon than EPs. Low pH and poor capacity to buffer acidification generally occurred in all cover types. Both EPs and PMPs showed a decline in soil properties relative to NBFs, but EPs and PMPs exhibited no significant difference. These results indicate that actions are needed to ameliorate the potential negative effects on soil quality in forestry plantations.     

Spatial pattern of beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.) and oak (<Emphasis Type="Italic">Quercus pubescens</Emphasis> Mill.) seedlings in natural pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) woodlands

Spatial pattern of recruitment is an important factor influencing population dynamics of plant communities. The spatial pattern is determined by seed dispersal and by the spatial variability of germination and initial survival. In the process of forest expansion following farmland abandonment, mid- and late-successional species are often dispersed in pioneer forests by birds. This could result in an aggregated spatial pattern of seeds that could influence the dynamics of these species in mixed pioneer forests. In the sub-Mediterranean area, mid- and late-successional species such as Quercus pubescens (downy oak) and Fagus sylvatica (European beech) are expected to replace pioneer Pinus species. Using a point sampling method we demonstrated that beech and oak seedlings (height <2 m) have a clumped distribution in the understorey of pine. This could result from an aggregated dispersal by jays (dispersal effect) or from preferential recruitment in particular habitats (habitat effect). To test these hypotheses we proposed a statistical analysis of spatial patterns of regeneration of beech and oak. Ground cover variables (i.e. cover by rock outcrops, herbs, box shrubs, mosses or pine) did not differ significantly around seedlings as compared with random sample plots. Likewise, clumped seedlings had growth similar to isolated seedlings, thus refuting the hypothesis of preferential location in the most favourable microsites. Aggregated dispersal seems to be involved in the process of regeneration. Since beech and oak seedlings have contrasting ecological demands, we discuss the implication of this pattern for the replacement dynamics of pine by these species.     

The colonisation of Scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) by <Emphasis Type="Italic">Tomicus minor</Emphasis> Hartig in southern Poland: modelling and monitoring

Tomicus minor Hartig (Col., Curculionidae, Scolytinae), occurring on Pinus sylvestris L., is a species which demonstrates high reproductive capability on weakened stands, accelerating the process of forest death. In protected areas, T. minor is regarded as a sensitive bioindicator that reacts to decline in the health and vitality of forests. Although there have been many publications concerning T. minor, no precise method has yet been given for estimating its population so as to enable the monitoring of forest vitality and assessment of the role played by T. minor in the forest ecosystem. The aim of the present work is to develop a statistical method for estimating populations of T. minor, requiring minimum work and interference with the forest ecosystem and permitting the computation of estimation errors. Research was carried out in the years 1992–2011 in pine stands aged over 80 years, growing in a variety of habitats and situated at varying distances from sawmill timber yards. Attack density of T. minor was measured on trap logs made from uninfested living trees. The population of T. minor on the trap logs was described using a multiple linear regression model with two explanatory variables. Among the features investigated, the T. minor population was found to depend significantly on the number of egg galleries on the fifth metre of the trap log counted from the thinner end (p < 0.001) and on the diameter of the trap log in bark at the thinner end (p < 0.05). The model explains approximately 85% (R ² = 0.8564) of the variation in the total number of T. minor egg galleries on the trap logs. The numbering of units beginning from the thinner end of the log enabled increased precision in determining the model parameter resulting from the concentration of egg galleries on certain units of the log. In all validated plots, the mean real and model values for the number of T. minor egg galleries on the trap logs are similar (p > 0.5), confirming the high accuracy of the developed model.     

Regeneration dynamics of mixed stands of <Emphasis Type="Italic">Pinus pinaster</Emphasis> Ait. and <Emphasis Type="Italic">Pinus pinea</Emphasis> L. in Central Spain

The dynamics of mixed stands are more complex and less studied than those of monospecific stands. The objective of this work was to analyze the variables involved in seedling occurrence and seedling survival in mixed stands of Pinus pinaster and P. pinea in Mediterranean areas. From 2011 to 2016, regeneration of both species was monitored at two sites located in Central Spain. We installed 72 regeneration plots where seedling dynamics were monitored. All the trees in the study areas were measured and mapped. Additionally, we took hemispherical photographs in each regeneration plot. The average density of P. pinea seedlings over the study period was almost 20 times larger than that of P. pinaster. Our results indicate that the seedlings of both species grow under moderate light conditions. In addition, we found that the occurrence of seedlings of both species was related to the structure of the stand. P. pinea seedlings grew where the density and size of P. pinaster trees were low and where P. pinea trees provided moderately sheltered conditions, whereas the number of P. pinaster seedlings was related to under intermediate densities of P. pinaster trees. Furthermore, seedling survival was positively associated with age of the seedlings and negatively with the August average maximum temperature. The temporal continuity of mixed stands of P. pinea and P. pinaster in the study area is compromised by the observed lack of regeneration of P. pinaster.     

Species proportions by area in mixtures of Scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) and European beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.)

Scots pine (Pinus sylvestris L.) and European beech (Fagus sylvatica L.) dominate many of the European forest stands. Also, mixtures of European beech and Scots pine more or less occur over all European countries, but have been scarcely investigated. The area occupied by each species is of high relevance, especially for growth evaluation and comparison of different species in mixed and monospecific stands. Thus, we studied different methods to describe species proportions and their definition as proportion by area. 25 triplets consisting of mixed and monospecific stands were established across Europe ranging from Lithuania to Spain in northern to southern direction and from Bulgaria to Belgium in eastern to western direction. On stand level, the conclusive method for estimating the species proportion as a fraction of the stand area relates the observed density (tree number or basal area) to its potential. This stand-level estimation makes use of the potential from comparable neighboring monospecific stands or from maximum density lines derived from other data, e.g. forest inventories or permanent observations plots. At tree level, the fraction of the stand area occupied by a species can be derived from the proportions of their crown projection area or of their leaf area. The estimates of the potentials obtained from neighboring monospecific stands, especially in older stands, were poorer than those from the maximum density line depending on the Martonne aridity index. Therefore, the stand-level method in combination with the Martonne aridity index for potential densities can be highly recommended. The species’ proportions estimated with this method are best approximated by the proportions of the species’ leaf areas. In forest practice, the most commonly applied method is an ocular estimation of the proportions by crown projection area. Even though the proportions of pine were calculated here by measuring crown projection areas in the field, we found this method to underestimate the proportion by 25% compared to the stand-level approach.     

Effects of <Emphasis Type="Italic">Eucalypt</Emphasis> and <Emphasis Type="Italic">Acacia</Emphasis> plantations on soil water in Sumatra

Indonesia’s pulp and paper industry needs a large area of sustainably grown plantations to support its continued development. Acacia mangium has been the key species underpinning the pulp and paper industries in Sumatra, however increased disease pressure on A. mangium is expected to require large-scale conversion of Acacia plantations to Eucalyptus in the near future. The effect of such a large scale change in plantation species on soil moisture, for both tree production, and catchment hydrology is unknown. In this study we sought to characterize the impacts of plantation species (Acacia or Eucalyptus) and nitrogen management, on soil moisture, soil water depletion and depth to groundwater under stands of Acacia mangium and Eucalyptus pellita over the first 2–3 years after establishment. The study was conducted in experiments at four sites in Sumatra, Indonesia. Soil moisture and soil water depletion were not influenced by plantation species or fertilizer treatment. Soil moisture content and soil water depletion were strongly influenced by shallow groundwater at two of the four sites, however depth to groundwater did not influence stem growth. Results from the field trials cautiously suggest that large scale conversion of Acacia mangium to Eucalypt species in these regions is unlikely to result in increased moisture stress, nor is conversion of plantation species likely to lead to substantial differences in catchment hydrology. This study demonstrated the importance of conducting multi-site studies when investigating biophysical relationships in forest/plantation systems.     

Factors determining enzyme activities in soils under <Emphasis Type="Italic">Pinus halepensis</Emphasis> and <Emphasis Type="Italic">Pinus sylvestris</Emphasis> plantations in Spain: a basis for establishing sustainable forest management strategies

Key message

Water availability and soil pH seem to be major constraints for enzyme activities in calcareous soils under Pinus halepensis and acidic soils under Pinus sylvestris plantations respectively. Proposals for improving enzyme activities may include the promotion of broadleaf species to increase soil pH and the modulation of stand density or the implementation of soil preparation techniques to facilitate water infiltration.

Context

Soil enzymes play a key role in nutrient turnover in forest ecosystems, as they are responsible for the transformation of organic matter into available nutrients for plants. Enzyme activities are commonly influenced by temperature, humidity, nutrient availability, pH, and organic matter content.

Aims

To assess the differences between enzyme activities in calcareous soils below Pinus halepensis and acidic soils below Pinus sylvestris plantations in Spain and to trace those differences back to edapho-climatic parameters to answer the questions: Which environmental factors drive enzyme activities in these soils? How can forest management improve them?

Methods

The differences in climatic, soil physical, chemical, and biochemical parameters and the correlations between these parameters and enzyme activities in soils were assessed.

Results

Low pH and high level of phenols in acidic soils under Pinus sylvestris and water deficit in calcareous soils under Pinus halepensis plantations appeared to be the most limiting factors for enzyme activities.

Conclusion

Options such as the promotion of native broadleaf species in the Pinus sylvestris stands and the modulation of Pinus halepensis stand density or the implementation of soil preparation techniques may improve enzyme activities and, therefore, nutrient availability.

     

Comparison of postfire mortality in endemic Corsican black pine (<Emphasis Type="Italic">Pinus nigra</Emphasis> ssp. <Emphasis Type="Italic">laricio</Emphasis>) and its direct competitor (<Emphasis Type="Italic">Pinus pinaster</Emphasis>)

• Introduction   

Laricio pine (Pinus nigra J.F. Arn. ssp. laricio (Poiret) Maire var Corsicana Hyl.) is a form of black pine endemic to Corsica, that may now be under threat due to current fire regimes and competition with maritime pine (Pinus pinaster Aiton).     

Fertility variation in endemic populations of Ehrami black pine (<Emphasis Type="Italic">Pinus nigra</Emphasis> Arnold. subsp. <Emphasis Type="Italic">pallasiana</Emphasis> var. <Emphasis Type="Italic">pyramidata</Emphasis>)

Fertility variation and effective number of parents were estimated in the 100 individuals from each of three plantation populations (P1, P2 and P3) of the endangered Pinus nigra. subsp. pallasiana var. pyramidata established in the Isparta–Golcuk district in southern Turkey in 1990. Potential relations between cone yield and growth characters (height, diameter at breast height, diameter at base and crown diameter) were also investigated. Large differences were found among populations and within populations for cone production and growth characters. Mean cone production was eight (ranging from 6.5 in P2 to 9.1 in P1) for the three populations. The 10 most productive individuals for cones produced 32% of the total cones in P1, 39% in P2 and 34% in P3; 21, 16 and 6 individuals, respectively did not produce any cones in the populations. According to the correlation analyses, the relation between cone yield and growth characters changed for the populations, while height, diameter at breast height and crown diameter were positively significantly correlated (r = 0.182, 0.135 and 0.209; p ≤ 0.05) with cone production for the three populations. Fertility variation was 1.81 in total of three populations, ranging from 1.72 in P3 to 3.45 in P1). Effective number of parents was 142 (55.2 of number of parents censused; 29%, P1; 49.8%, P2; 58.3%, P3).     

Osmoregulators in <Emphasis Type="Italic">Hymenaea courbaril</Emphasis> and <Emphasis Type="Italic">Hymenaea stigonocarpa</Emphasis> under water stress and rehydration

The objective of this work was to evaluate the effect of different water deficiency and rehydration levels on the concentrations of osmoregulators in two plant species (Hymenaea courbaril and H. Stigonocarpa) in the Amazon. We adopted a 2 × 5 × 5 factorial system, referring to 2 species (H. courbaril and H. stigonocarpa) and 5 stages of hydration and rehydration. The five hydration and rehydration stages were established in: (1) Control treatment E0; (2) Plants with 13 days of stress after incubation—E13; (3) Plants with 26 days of stress E26; (4) The plants that were established after 26 days after incubation and rehydrated for two days (RD2); (5) rehydrated for two days (RD4). The plants that were established after 26 days after incubation and rehydrated for four days. The experiment totaled fifty young plants with five replicates. Biochemical measurements were performed at the beginning of the experiment (E0) at 13 (E13) and 26 (E26) days after the water stress, in which the plants were rehydrated, repeating the analyses after two (RD2) and four (RD4) days. Both species increased the sucrose concentration by 18%, with a decrease of 52% in starch content. The RD4 time presented the highest mean starch concentration (0.19 mmol g^?1 of the residue for H. courbaril and 0.27 mmol g^?1 of residue for H. stigonocarpa). Increased proline concentrations were recorded for controls until RD2 for both species. For glycine betaine, the highest increases in treatments E26 and RD2 were observed for the H. courbaril species. Our rehydration period was not sufficient for total recovery of pre-stress concentrations of all studied solutes.     

Dynamics of ecosystem services in <Emphasis Type="Italic">Pinus sylvestris</Emphasis> stands under different managements and site quality classes

Timber and biodiversity are considered two antagonistic ecosystem services (ES), largely influenced by silviculture and site ecological conditions. In order to address the trade-offs between these two ES over time, we performed a retrospective study at compartment level in the Pinus sylvestris forests of the Spanish Central Mountain System. Archival data from Management Plans for eight forests with contrasting silvicultural systems (uniform shelterwood system, group shelterwood system, irregular shelterwood system,) and three different site quality classes were analysed. Timber production was assessed through stand volume, harvested timber volume and a stand volume index. Biodiversity was examined through structural diversity (Gini index, Shannon tree size diversity index, Simpson’s reciprocal index and evenness index, all applied to diameter classes) and the abundance of large living trees. For all silvicultural systems investigated, stand volume and harvested timber volume have grown since the beginning of the management plans (beginning of the twentieth century in some forests). The largest yields of timber corresponded to the best quality sites with more intensive silvicultural treatments (uniform and group shelterwood systems). The uniform shelterwood system showed lower figures for structural diversity, though not always significant. The best site qualities maintained notable structural diversity values, even under the most intensive management system. The application of the different management systems over decades has revealed a synergy between timber production and structural diversity, particularly in those systems maintaining more than one age class, although results are conditioned by forest harvesting history. The interaction between historical silvicultural treatment and site quality has been identified as an important source of information to understand forest dynamics and functioning of ES provision.     

Fine root biomass and morphology of <Emphasis Type="Italic">Pinus densiflora</Emphasis> under competitive stress by <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>

The fine root (diameter ≤2.0 mm) biomass and morphology of Japanese red pines (Pinus densiflora) grown under different aboveground conditions (i.e., high and low competitive environments) were examined in a pine–cypress mixed forest. All P. densiflora subject trees were about 40 years old, and the aboveground condition (i.e., size) of red pines appeared to be influenced by the surrounding Japanese cypress (Chamaecyparis obtusa). Smaller P. densiflora exhibited lower fine root biomasses, shorter root lengths, and lower root tip densities, but longer specific root lengths and higher specific root tip densities relative to larger pines. These results suggest that P. densiflora may adjust the morphological traits of fine roots to the different conditions in biomass allocation to fine roots of individuals with different aboveground growth.     

Recessive inheritance of morphological characteristics of utsukushimatsu,<Emphasis Type="Italic">Pinus densiflora</Emphasis> f.<Emphasis Type="Italic">umbraculifera</Emphasis>

Pinus densiflora f.umbraculifera, commonly known as utsukushimatsu, is a distinctively shaped form of Japanese red pine whose growth is restricted to a forest stand in Shiga Prefecture, Japan. The inheritance mode of morphological characteristics of utsukushimatsu was studied to preserve the genetic resource of this pine. As previously reported, F₁ trees grown from open-pollinated seeds harvested from trees inhabiting the native stand showed two phenotypes: one resembling utsukushimatsu, which produces multiple trunks, and the other resembling normalP. densiflora, which produces one or a few trunks. In the present study, controlled pollination was carried out using F₁ and normalP. densiflora trees. Segregation ratios of the two phenotypes observed in the F₂ population showed that the morphological characteristics of utsukushimatsu are inherited recessively. This suggests that the mutation of one gene or a few closely linked genes controls the morphological characteristics of utsukushimatsu. Since multiple trunk formation of utsukushimatsu might be related to a loss of lateral bud inhibition, it follows that a simple gene mutation breaks apical dominance inP. densiflora.     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

Variation in cone and seed characters in clonal seed orchards of <Emphasis Type="Italic">Pinus sylvestris</Emphasis>

Cone and seed characters were observed on top, middle and bottom portions of tree crown in 3 ramets/clone in three Turkish Pinus sylvestris seed orchards. Broad sense heritability (clonal repeatability), and correlations among characters were estimated. Around one quarter of the seed production occurred in the top portion, half in the middle and one quarter in the bottom portion of the crown for all orchards. The percentage of filled seeds varied little with the crown position, indicating more or less similar levels of selfing in the bottom of the crown as in the top. The seed weight was typically 11 mg. Differences were found for studied cone and seed characters among orchards and crown positions. Variation among grafts within clone was higher than among clones for most characters. The heritability was on average below 0.5 (e.g., cone diameter, number of filled seed per cone) and rarely rose above that (e.g., cone form, length/diameter; percentage of filled seed) for any individual characters. The coefficient of variation within clones was often higher than among clones. Thus, non-genetic factors seem often more important for the variation in performance of grafts than their genetic constitution. Cone form (length/diameter) was the character where the clone influence was the strongest. Cone number and cone dry weight showed significant correlations with seed characters (numbers of total and filled seed, percentage and weight of filled seeds). Significant correlation was found between seed characters.     

Production of <Emphasis Type="Italic">Pinus halepensis</Emphasis> seedlings inoculated with the edible fungus <Emphasis Type="Italic">Lactarius deliciosus</Emphasis> under nursery conditions

Inoculation with edible fungi bestows an added value on mycorrhized plants since production of mushrooms as a secondary crop can be an economically-valuable resource in forestry management. In order to establish a protocol for controlled mycorrhization of Pinus halepensis with the edible fungus Lactarius deliciosus, several factors such as fertilisation (35, 60 or 120 mg total N and 13.5, 27 or 54 mg total P), potting substrate (sphagnum peat or sphagnum peat/vermiculite) and fungal inoculum (mycelial slurry produced in a bioreactor, alginate beads or peat/vermiculite) were assessed. The most effective inoculum was mycelial slurry at a dose of 10 ml/plant. The two potting substrates assessed were compatible with mycorrhiza formation. The greatest number of mycorrhized seedlings was obtained with a moderate level of N (35 mg/plant) or P fertilisation (27 mg/plant). Inoculation did not produce a consistent growth effect on P. halepensis, but seedlings met the quality requirements, making them suitable for planting out.     

Antifeedant activities of secondary metabolites from <Emphasis Type="Italic">Ajuga nipponensis</Emphasis> against adult of striped flea beetles, <Emphasis Type="Italic">Phyllotreta striolata</Emphasis>

Antifeedant activities of the isolated chemical compounds from Ajuga nipponensis, were studied against adult of striped leaf beetles. The methanol, petroleum ether, ethyl acetate and n-butanol extracts at 1.0 mg/ml, were used in this study. All four extracts exhibited more than 65 antifeedant index at 24 h and ethyl acetate extract showed significant activity against striped leaf beetles with 83.12 antifeedant index. Six compounds and one fraction were isolated by chromatography and their structures were identified by NMR, MS and FTIR spectra. At 2.0 mg/ml for 24 h the three compounds 20-hydroxyecdysone, acacetin and apigenin showed considerable activities with antifeedant indexes 59.29, 51.22 and 61.55, respectively. In contrast to this the antifeedant indexes of acacetin and apigenin, were sharply reduced as the time extended and that of 20-hydroxyecdysone remained unchanged. In addition, the synergistic effects of two mixtures of secondary metabolites, were studied and no sharp difference was observed.     

Phenotypic plasticity blurs ecotypic divergence in the response of <Emphasis Type="Italic">Quercus coccifera</Emphasis> and <Emphasis Type="Italic">Pinus halepensis</Emphasis> to water stress

The Mediterranean evergreen woody plants Quercus coccifera and Pinus halepensis grow in a range of environments where selection by drought, heat and high irradiance can drive genetic and phenotypic differentiation of populations. However, the role of these stresses in filtering out maladaptive genotypes remains unknown. We hypothesize that this filtering is an important process for woody Mediterranean species due to their low phenotypic plasticity reported in previous studies. We have studied the response of saplings of Q. coccifera and P. halepensis, originating from two contrasting populations (a rock outcrop and a garrigue formation), to water stress. Isozyme characterization of genetic diversity was done to determine whether populations were genetically distinct. Water response analysis was based on water relations, gas exchange, chlorophyll a fluorescence, pigment content, antioxidant status and morphological and structural parameters. Ecotypic differentiation was found for both Q. coccifera and P. halepensis populations, with a higher population isozyme similarity and a higher frequency of dominance of a few genotypes at the rock outcrop in both the species. P. halepensis exhibited small but significant differences between populations for plastic responses to water, with lower phenotypic plasticity in saplings from the rock outcrop. Although it was not found in Q. coccifera, this pattern suggests that ecotypic differentiation rendering stress-tolerant ecotypes involves a decreased plasticity. Phenotypic plasticity was not high but it explained over 75% of the total variability among individual plants. Thus, and although evidence for ecotypic divergence was found in both the species, saplings were plastic enough to blur ecotypic differentiation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

An inoculation experiment of Japanese <Emphasis Type="Italic">Bursaphelenchus</Emphasis> nematodes on Japanese black and red pine, <Emphasis Type="Italic">Pinus thunbergii</Emphasis> and <Emphasis Type="Italic">P. densiflora</Emphasis>

The pinewood nematode (PWN) Bursaphelenchus xylophilus is an invasive pathogen that was introduced from North America to Asian countries and Portugal and is devastating native pine forests. Some native European and Asian Bursaphelenchus nematodes also have weak to moderate pathogenicity to native pine species. To evaluate the potential risk of native Bursaphelenchus species, we inoculated ten Japanese Bursaphelenchus species into native pine species (the dominant forest species) in Japan, and evaluated their pathogenicity using mortality and tracheal tissue damage as indices. Inoculation was conducted on August 3, 2007, and the symptoms were observed every 2 weeks until February 1, 2008. None of the inoculated trees, excluding the pathogenic PWN inoculated control, showed external disease symptoms; however, four species [a less pathogenic PWN isolate, B. luxuriosae, Bursaphelenchus sp. NK215 (undescribed), and NK224 (undescribed)] caused tracheal tissue damage in inoculated seedlings and showed weak pathogenicity. Therefore, we conclude that there are some potentially pathogenic native species of nematodes distributed in Japan. Interestingly, two of these weakly pathogenic species, B. luxuriosae and NK215, are not associated with Pinaceae trees, suggesting that nematode pathogenicity may be a pre-adaptive character. More experimental studies under different conditions are necessary to accurately evaluate the potential risk of these pathogens.