Absence of geographical structure of morphological variation in <Emphasis Type="Italic">Juniperus oxycedrus</Emphasis> L. subsp. <Emphasis Type="Italic">oxycedrus</Emphasis> in the Balkan Peninsula

We examined leaf and mature seed cone variation of Juniperus oxycedrus L. subsp. oxycedrus in 12 natural populations across the species range in the Balkan Peninsula. We measured 10 morphological traits from a minimum of 100 leaves in each of 190 individuals, and two morphological traits from 30–50 seed cones in each of 94 females. High phenotypic variation was found, but no geographical structure or cline across populations was detected for any of the studied traits. Mean values of comparable leaf and cone morphological traits did not differ considerably from values reported elsewhere. Gender dimorphism in leaf morphology was detected, but it was not distributed uniformly throughout the studied area. An ANOVA model with both nested and crossed effects revealed that the largest proportion of the total variation was, as expected, contained within populations, partly as among-tree variation (18–47%, depending on the trait) and partly as within-tree variation (33–77%), which was remarkably high. Gender dimorphism explained only 0–3% of the total variation. Differences among populations (2–23%) were significant for all studied traits except one; however, PCA showed no clear geographical differentiation of the studied populations. This lack of phylogeographical structure may be the consequence of repeatedly occurring colonisation-retreat scenarios and suggests the existence of several small refugial populations scattered over a large part of the Balkan Peninsula in the Pleistocene. Further research including palaeobotanical and molecular genetic studies will be needed to better understand the forces that shaped current variation patterns of J. oxycedrus L. subsp. oxycedrus in the Balkan Peninsula.     

Post-fire attractiveness of maritime pines (<Emphasis Type="Italic">Pinus pinaster</Emphasis> Ait.) to xylophagous insects

The spectrum and dynamics of xylophagous insects infesting Pinus pinaster after a fire event were studied from November 2007 to December 2008 at Sierra de Outes (Galicia, NW Spain). Insects were collected within and outside the fire area. Relationships among species status, species abundance, tree parameters and tree injury were assessed. Mortality of injured trees was also recorded. Insect colonization built up consistently during the first year after the fire, although the pests did not colonize unburned trees in nearby areas. Thirteen insect species from six xylophagous families were collected. Tomicus spp., Buprestis novenmaculata, Anobium punctatum and Pissodes castaneus were the first species detected. Tomicus spp. (29%) and Ips sexdentatus (23%) were the dominant taxa. Bole char height and soil burn severity were positively correlated with insect presence. Trees with short stem diameter and thinner bark were also preferred. B. novenmaculata presence was positively correlated with crown scorch. Loss of cambium hydration was negatively related to the occurrence of pests, likely because sapwood desiccation and loss of nutrient impede brood development. About 33.3% of the fire-scorched pines died. The results recommend the removal of injured and dying pines after fire, to avoid the rise of pest population threatening recovering trees in the burned areas.     

Effect of understory vegetation management through liming and sewage sludge fertilisation on soil fertility and <Emphasis Type="Italic">Pinus radiata</Emphasis> D. Don growth after reforestation

Tree–understory competition is one of the most important aspects that control tree growth after reforestation. The relationship between trees and the understory can be modified by improving acidic soils with lime and by fertilisation. This experiment aims to evaluate the effect of soil improvements on the pasture–tree relationship by liming and fertilisation on different dates in a Pinus radiata-reforested area. Both lime and sewage sludge improved soil fertility by increasing Ca and reducing Al in the soil. Initially, tree development was reduced by lime, which improved the establishment of competitive grasses. Tree growth in limed treatments did not initially respond to sludge inputs, likely because both tree and grass roots shared the same soil depth layer. Three years after establishment, the use of high doses of sewage sludge in limed plots caused a growth rate similar to the best treatments of unlimed plots, which grew with a poorly sown grass establishment. After 2 years of the experiment, the presence of Erica woody shrub diminished tree development. High doses of sewage sludge with lime, as well as high doses of sewage sludge without lime, applied in April and low doses of sewage sludge without lime added in early February improved tree growth. From a practical point of view, lime and sewage sludge dose close to 100 kg total N ha⁻¹ should be recommended if a silvopastoral system is established, as it enhances pasture production and tree growth.     

Logistic regression models for human-caused wildfire risk estimation: analysing the effect of the spatial accuracy in fire occurrence data

About 90% of the wildland fires occurred in Southern Europe are caused by human activities. In spite of these figures, the human factor hardly ever appears in the definition of operational fire risk systems due to the difficulty of characterising it. This paper describes two spatially explicit models that predict the probability of fire occurrence due to human causes for their integration into a comprehensive fire risk–mapping methodology. A logistic regression technique at 1 × 1 km grid resolution has been used to obtain these models in the region of Madrid, a highly populated area in the centre of Spain. Socio-economic data were used as predictive variables to spatially represent anthropogenic factors related to fire risk. Historical fire occurrence from 2000 to 2005 was used as the response variable. In order to analyse the effects of the spatial accuracy of the response variable on the model performance (significant variables and classification accuracy), two different models were defined. In the first model, fire ignition points (x, y coordinates) were used as response variable. This model was compared with another one (Kernel model) where the response variable was the density of ignition points and was obtained through a kernel density interpolation technique from fire ignition points randomly located within a 10 × 10 km grid, which is the standard spatial reference unit established by the Spanish Ministry of Environment, Rural and Marine Affairs to report fire location in the national official statistics. Validation of both models was accomplished using an independent set of fire ignition points (years 2006–2007). For the validation, we used the area under the curve (AUC) obtained by a receiver-operating system. The first model performs slightly better with a value of AUC of 0.70 as opposed to 0.67 for the Kernel model. Wildland–urban interface was selected by both models with high relative importance.     

Susceptibility of the Mexican bean beetle <Emphasis Type="Italic">Epilachna varivestis</Emphasis> Mulsant (Coleoptera: Coccinellidae) to endemic isolates of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae)

Entomopathogenic fungi represent excellent candidates for biological insecticides. Among the many entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) is one of the most promising species. Here, we report the results of infection studies performed using four B. bassiana native isolates (Bb37, Bb38, Bb40, and Bb45) compared with the commercial product Mycotrol^® (strain GHA) on adult Mexican bean beetle Epilachna varivestis Mulsant (Coleoptera: Coccinellidae), one of the most important pests of bean production in Mexico. First, single concentration (1 × 10⁸ conidia mL^?1) assays were carried out using all isolates applied using an immersion method. Examinations at 10 days after inoculation indicated that isolates Bb37, Bb38, and Bb40 significantly reduced adult survival, causing 60–75% mortality compared to the commercial strain GHA (33%) and control (29%). Second, using two selected isolates (Bb37 and Bb40), the mean lethal concentration values (LC₅₀) were estimated for third instar larvae and adult E. varivestis. The LC₅₀ values for Bb37 and Bb40 isolates decreased 5.26-fold and 3.19-fold, respectively, in third instars compared with adults. However, such difference for Bb40 isolate was not significant. Finally, an experiment, to compare the median survival time (MST) values between third instar and adult E. varivestis, was conducted using Bb37 and Bb40 at 1 × 10⁹ conidia mL^?1. MST values were significantly reduced against larvae (87 and 100 h) compared with adults (130 and 134 h) by Bb37 and Bb40, respectively. We conclude that native isolates of B. bassiana represent an important alternative for the control of E. varivestis. However, to demonstrate its effectiveness under field conditions, detailed studies are needed.     

Ecology and distribution of Armillaria species in Norway

The occurence of Armillaria species was assessed in Norway, enabling the northern‐most distribution of this genus to be determined in Europe. Four Armillaria species were found in Norway. Armillaria borealis was the most common species occurring on woody vegetation to the permafrost zone (ca. 69°N). Armillaria cepistipes was present in southern and central Norway, but was not found further than 66°N. Armillaria solidipes and Armillaria gallica were rare, found at only one locality each; 59°40′ and 59°32′, respectively. Armillaria species were found on 14 hosts, but there was no significant difference between occurrence of A. borealis and A. cepistipes on declining and dead trees. Phylogenetic analyses separated each species into separate clades. All isolates of A. borealis, except one, and most isolates of A. solidipes were in separate clades. However, a subclade within the A. borealis clade was formed of two A. ostoyae and one A. borealis isolates. Two small A. cepistipes genets were found in a declining oak stand.     

Resveratrol oligomers isolated from Carex species inhibit growth of human colon tumorigenic cells mediated by cell cycle arrest

Research has shown that members of the Carex genus produce biologically active stilbenoids including resveratrol oligomers. This is of great interest to the nutraceutical industry given that resveratrol, a constituent of grape and red wine, has attracted immense research attention due to its potential human health benefits. In the current study, five resveratrol oligomers (isolated from Carex folliculata and Carex gynandra ), along with resveratrol, were evaluated for antiproliferative effects against human colon cancer (HCT-116, HT-29, Caco-2) and normal human colon (CCD-18Co) cells. The resveratrol oligomers included one dimer, two trimers, and two tetramers: pallidol (1); α-viniferin (2) and trans-miyabenol C (3); and kobophenols A (4) and B (5), respectively. Although not cytotoxic, the resveratrol oligomers (1-5), as well as resveratrol, inhibited growth of the human colon cancer cells. Among the six stilbenoids, α-viniferin (2) was most active against the colon cancer cells with IC(50) values of 6-32 μM (>2-fold compared to normal colon cells). Moreover, α-viniferin (at 20 μM) did not induce apoptosis but arrested cell cycle (in the S-phase) for the colon cancer but not the normal colon cells. This study adds to the growing body of knowledge supporting the anticancer effects of resveratrol and its oligomers. Furthermore, Carex species should be investigated for their nutraceutical potential given that they produce biologically active stilbenoids such as α-viniferin.     

Radical scavenging capacity and antimutagenic properties of purified proteins from Solanum betaceum fruits and Solanum tuberosum tubers

In this study, antioxidant activities in free-radical-mediated oxidative systems and the genotoxic/antigenotoxic effects of two proteins with molecular mass around 17 kDa, purified from Solanum betaceum fruits (cyphomine) and Solanum tuberosum tubers (solamarine), were investigated. Both proteins inhibited uric acid formation with IC(50) values between 55 and 60 μg/mL, and both proteins were able to reduce oxidative damage by scavenging hydroxyl radicals and superoxide anion in a dose-dependent manner. Furthermore, the DPPH? reduction assay showed SC(50) values of 55-73 μg/mL. Cyphomine and solamarine were able to retain their antioxidant activity after heat treatment at 80 °C for 15 min. Allium cepa and Salmonella /microsome assays showed no genotoxic and mutagenic effects. Solamarine showed an antimutagenic effect against a direct mutagen (4-nitro-o-phenylenediamine). Consequently, the present study showed that the investigated proteins are promising ingredients for the development of functional foods with a beneficial impact on human health and an important source for the production of bioactive peptides.