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.     

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.     

Field-based artificial crossings indicate partial compatibility of reciprocal crosses between <Emphasis Type="Italic">Pinus sylvestris</Emphasis> and <Emphasis Type="Italic">Pinus mugo</Emphasis> and unexpected chloroplast DNA inheritance

Crossability relationships between Scots pine (Pinus sylvestris L.) and mountain dwarf pine (Pinus mugo Turra) was studied, using artificial pollination approach. Partial compatibility of the reciprocal crossings of these species was proved experimentally, validating the idea of a spontaneous formation of their hybrid swarms under natural conditions. The hybrids were validated using organellar DNA markers and nuclear DNA microsatellites. Based on the percentage of filled seeds, the interspecific crossings were less efficient than the intraspecific cross-pollinations of P. sylvestris and P. mugo individuals. Both species were found to intercross readily with individuals of their putative hybrid swarm, P. mugo exhibiting a higher hybridological affinity towards putatively hybrid individuals than P. sylvestris. Validation of the hybrids confirmed the paternal inheritance of chloroplast DNA (cpDNA) in the combination P. sylvestris × P. mugo only. Surprisingly, in the reciprocal crossing P. mugo × P. sylvestris, maternal inheritance of cpDNA was revealed. Obtained results offer a new insight into the direction and intensity of gene flow within the hybrid swarms of Scots pine and mountain dwarf pine.     

Adaptive Responses of Morphological Forms of the Pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) under Stressful Conditions of the Northern Taiga (in the Northern Dvina Basin)

The variability of physiological and biochemical indicators and radial growth of different forms of Pinus sylvestris L. f. (var.) sulfuranthera Kozubow and f. (var.) erythranthera Sanio under flooding conditions has been studied. It is shown that the dynamics of photosynthetic pigments (chlorophylls and carotenoids), proline, proteins, ascorbic acid, peroxidase activity, and the light-harvesting complex in the pine needles depend on meteorologic factors and the phenophase. Forms with different colors of anthers differ in seasonal dynamics of the content of stressful metabolites and age variability of the radial growth of wood. Our results indicate that different forms of Pinus sylvestris L. have nuances in adaptation to stress conditions.     

The composition of fatty acids in the endosperm and embryo lipids of <Emphasis Type="Italic">Pinus sibirica</Emphasis> and <Emphasis Type="Italic">P. sylvestris</Emphasis> seeds

The fatty acid (FA) composition of storage lipids in the seed endosperms and embryos of two pine species, Pinus sibirica and P. sylvestris, and possible biosynthetic pathways of these acids were studied by the GLC method. Linoleic acid predominated in the embryo and endosperm lipids of both P. sibirica (43.5 and 42.6%) and P. sylvestris (44.8 and 46.8%); this was evidently determined by a high expression of the gene encoding stearoyl-Δ9 acyl-lipid desaturase and the fad2 gene encoding microsomal ω6 acyl-lipid desaturase. P. sibirica lipids of the embryo and endosperm contained more oleic acid (22.0 and 24.0%, respectively) than corresponding P. sylvestris lipids (18.7 and 14%). Storage lipids of conifer seeds contain Δ5-unsaturated FAs: taxoleic (18:2Δ5,9), ephedrenic (18:2Δ5,11), pinoleenic (18:3Δ5,9,12), skiadonic (18:3Δ5,11, 14), and coniferonic (18:4Δ5,9,12,15). In the endosperm and embryos of P. sylvestris, the content of pinolenic acid was higher (22.1 and 19.6%) than in P. sibirica seeds (19.1 and 18.6%).     

Characterization of a <Emphasis Type="Italic">Pinus sylvestris</Emphasis> thaumatin-like protein gene and determination of antimicrobial activity of the in vitro expressed protein

Thaumatin-like proteins (TLPs) are pathogenesis-related proteins, which are involved in plant defense responses to pathogen infection. Expression of the Pinus sylvestris L. TLP gene is up-regulated by methyl jasmonate treatment and inoculation with Heterobasidion annosum. A full-length Pinus taeda TLP gene sequence was used to design PCR primers for amplification of the full-length TLP gene from P. sylvestris. A putative 705-bp open reading frame of TLP gene was cloned into Escherichia coli cells, and then subcloned into the overexpression vector pET100 using BL21 Star expression bacteria. Optimization of the expression of recombinant TLP was achieved by decreasing both expression temperature and IPTG concentration. The purified 24.6-kDa TLP shows antimicrobial activity against 12 fungal species.     

Structure of the <Emphasis Type="Italic">TADHN</Emphasis> gene for dehydrin-like protein of soft wheat and activation of its expression by ABA and 24-epibrassinolide

The structure of the cloned fragment of wheat (Triticum aestivum L.) TADHN gene encoding dehydrin-like protein was examined. A comparative analysis of nucleotide and deduced amino acid sequences revealed a high homology of this fragment with sequences of the barley dhn8 gene and wheat wcor gene family. In deduced amino acid sequence of the TADHN fragment, a 15-residue region EKKGFLEKIKEKLPG was found, which corresponded to a highly conserved K-segment of dehydrins. Wheat seedling treatment with 3.7 μM ABA and 0.4 μM 24-epibrassinolide exerted similar stimulatory effects on expression of the TADHN gene, which indicates the involvement of dehydrins in the protective action of these phytohormones in wheat plants.     

Dehydrins in Buds of Main Birch Species under Conditions of Karelia

The composition and seasonal dynamics of stress proteins-dehydrins in the buds of the main birch species (downy birch (Betula pubescens Ehrh.), silver birch (B. pendula Roth)) and its varieties (Karelian birch (B. pendula var. carelica (Mercklin) Hämet-Ahti)), growing in northwest Russia (on the example of the Republic of Karelia) were investigated for the first time. It was shown that the level of low-molecular dehydrins, mainly with a molecular mass of 17 kD, is subjected to major seasonal changes, regardless of the specific features of the birch. The maximal level of 17 kD dehydrin was formed during the autumn preparation of plants to dormancy and was persistently preserved during the cold period of the year. The content of medium-molecular weight dehydrins of 66–69 kD was almost at the same level all year round. Significant inter-and intraspecific polymorphism of the major dehydrins of 17 and 66–69 kD in the buds of downy birch, silver birch, and Karelian birch during dormancy was not found. The significant similarity in the composition of total proteins and dehydrins, as well as the uniform nature of their seasonal changes, mainly 17 kD dehydrin, indicates the phylogenetic proximity and similar mechanisms of adaptation of the main species of the genus Betula L. to the temperate continental climate of Karelia.     

Biochemical Adaptation of Scots Pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) to Technogenic Pollution

We have investigated changes in the complex of biochemical parameters reflecting the disturbance of the homeostatic state and the activity of the adaptive reactions in Pinus sylvestris L. under the conditions of technogenic pollution. Disorders of the physiological state of trees growing under the effect of the excessive accumulation of pollutant elements result in changes in a number of parameters: a decrease in the ratio of protein and nonprotein nitrogen fractions in the needles by 52%; a decrease in the total phosphorus and its acid-soluble fraction by 40 and 63%, respectively; a decrease in carbon concentration by 31%; and a decrease in the amount of chlorophyll a, chlorophyll b, and carotenoids based on the weight of one needle by 23, 40, and 42%, respectively. The activation of protective reactions of pine is proven by an increase in the amount of ascorbic acid in needles by 48%; in the amount of water-soluble phenolic compounds by 29%; in the ratio of Chl. a/Chl. b by 35%; in the ratio of green pigments to yellow by 40%; and in the level of water- and alcohol-soluble proteins 40 and 30%, respectively. The highest activity of biochemical protection components was recorded in trees growing near the Shelekhovsky industrial center, the emissions of which contain a large amount of fluorides and polycyclic aromatic hydrocarbons (PAHs).     

Maternal inheritance of chloroplast DNA in <Emphasis Type="Italic">Pinus mugo</Emphasis> Turra: a case study of <Emphasis Type="Italic">Pinus mugo</Emphasis> × <Emphasis Type="Italic">Pinus sylvestris</Emphasis> crossing

The opposite modes of chloroplast DNA (cpDNA) inheritance were found to operate in the reciprocal crossings of Scots pine (Pinus sylvestris L.) and mountain dwarf pine (Pinus mugo Turra). The crossings were found to be partially compatible. In P. sylvestris × P. mugo crossing, the paternal transmission of cpDNA to the offspring takes place corroborating the generally acknowledged concept of the paternal cpDNA inheritance in gymnosperms. On the contrary, in P. mugo × P. sylvestris crossing the seed progeny exhibited P. mugo haplotype of the mother tree deviating conspicuously from the above concept. In the open pollination offspring of the putatively hybrid individuals of the Scots and mountain dwarf pines, a biparental inheritance of cpDNA was revealed in mother tree with P. mugo haplotype indicating a loosened control of the maternal inheritance of cpDNA in the putative hybrids. Implications and impacts of this finding for further studies are discussed.     

Efficacy of botanical extracts from Brazilian savannah against <Emphasis Type="Italic">Diabrotica speciosa</Emphasis> and associated bacteria

Botanical extracts are a plentiful resource of molecules with different biological activities, such as insecticides and antimicrobial pesticides. In this context, the aim of this work was to evaluate the efficacy of botanical extracts from the Brazilian savannah against Diabrotica speciosa and bacterial strains isolated from the gut of this insect under aseptic conditions. The bacterial isolates were identified by genomic and proteomic approaches, and bioassayed against eighteen botanical extracts in vitro. The best results of bacterial inhibitions were obtained for the extracts of Casearia sylvestris and Psidium laruotteanum. Fractions of C. sylvestris and P. laruotteanum, quantitatively evaluated by chromatographic analyses, showed a relationship between the bactericidal activity and phytochemical profile. In vivo assays showed that P. laruotteanum was also effective for the control of D. speciosa. Those results show that selected natural products can have both antimicrobial and insecticidal activities.     

New and little-known crickets of the subfamily Phalangopsinae (Orthoptera,Gryllidae). 10. The genus <Emphasis Type="Italic">Parendacustes</Emphasis> (Part 1)

The composition of the genus Parendacustes Chop. and of its two subgenera is briefly discussed. Nine new taxa of the nominotypical subgenus are described: P. berezini sp. n., P. arachne sp. n., P. pahangi borealis subsp. n., P. sylvestris sp. n., P. tkatshevae sp. n., P. tawau sp. n., P. tiomani sp. n., P. ectoparameralis sp. n., and P. magnispeculum sp. n. New data on the distribution of P. p. pahangi Gor. and considerations on the generic position of P. latifrons Chop. and P. lifouensis Desutter-Grandcolas are also provided.     

A scientific note on the evidence for the production and use of wax by a captive female <Emphasis Type="Italic">Bombus</Emphasis><Emphasis Type="Italic">sylvestris</Emphasis>

The presence of wax glands in the socially parasitic subgenus Bombus (Psithyrus) was established only recently (Sramkova and Ayasse, 2008), but the uses for the produced wax are unknown. In this investigation nest-searching Bombus sylvestris females were collected and kept in darkness, in conditions of controlled temperature and relative humidity, using rearing conditions for non-parasitic bumblebees. The results show that in Bombus sylvestris not only wax glands are present but they also can produce wax which can be used as a building material.     

Comparative studies of leaf surface chemical biosynthesis in different tobacco cultivars

To understand the differences in the biosynthesis of leaf surface chemicals and their influence on aphid preference for different tobacco cultivars (Nicotiana tabacum), we analyzed the secretory characteristics of glandular trichomes of four commercial cultivars, K326 (flue-cured), Beinhart 1000-1 (cigar), Basma YNOTBS1 (oriental), and Dabai 1 (burley), and their parental species, Nicotiana sylvestris and Nicotiana tomentosiformis. Trichome-type observation showed that K326 and N. sylvestris have three kinds of glandular trichomes (non-glandular, long stalked glandular, and short stalked glandular trichomes), whereas Beinhart 1000-1, Basma YNOTBS1, Dabai 1, and N. tomentosiformis had two kinds of glandular trichomes (long and short stalked glandular trichomes). The gas chromatography–mass spectrometry profiles of leaf exudates indicated that N. tomentosiformis synthesized only labdanoids; N. sylvestris, K326 and Dabai 1 synthesized only cembranoids; and Beinhart 1000-1 and Basma YNOTBS1 synthesized cembranoids and labdanoids. Gene expression pattern analysis revealed that the labdanoid synthesis-related genes NtABS and NtCPS2 were expressed in N. tomentosiformis, Beinhart 1000-1, and Basma YNOTBS1, whereas the cembranoid synthesis-related genes NtCYC and NtCYP71D16 were expressed in N. sylvestris and all four commercial cultivars. Evolutionary analysis indicated that NtCYC and NtCYP71D16 might be phylogenetically originated from N. sylvestris, whereas NtABS and NtCPS2 expressed in Basma YNOTBS1 and Beinhart 1000-1 might be derived from N. tomentosiformis. In addition, aphid attraction (number of aphids) was significantly and positively correlated with the total glandular secretion (r² =?0.9425, P?≤?0.05), and it was significantly and positively correlated with amount of CBT-diol (r²?=?0.9224; P?≤?0.05). These results provide new insights into the biosynthesis of diterpenoids and biotic stress resistance in tobacco.     

A multitrophic interaction: consequences of grazing on decomposition of needles of <Emphasis Type="Italic">Pinus pinea</Emphasis> L. by <Emphasis Type="Italic">Ommatoiulus sabulosus</Emphasis> L.

The aim of this study was to assess how the physical (needle fragmentation by trampling) and chemical (nutritional enrichment from faeces) changes brought about by grazing influence the consumption of needles of Pinus pinea L. by Ommatoiulus sabulosus L. Adult individuals of O. sabulosus were introduced into trays with pine needles subjected to four treatments: (i) intact needles, (ii) trampled needles, (iii) intact needles fertilized with liquid manure, and (iv) trampled needles fertilized with liquid manure. After 30 days, litter decomposition was determined as a function of mass loss over time. Biometric and chemical characteristics of the needles were also measured. The pine needle decomposition rate was significantly higher in the presence of O. sabulosus (20–40%) than in its absence, presenting in addition a significantly higher rate in the treatments enriched with nitrogen. Average needle length was significantly shorter in the trampled treatments (6.89 ± 0.50 cm) than in those with intact needles (11.95 ± 0.69 cm). With regard to leaf mass per area and resistance to breakage, no significant differences were found. The fertilized treatments presented significantly higher N content (50% higher) than those in the non-fertilized ones with a significantly lower C:N ratio. The results of the present study indicate that goat grazing (which produces both trampling and fertilization) favoured the consumption of pine needles by O. sabulosus, which thus influences, along with the microbial community, the decomposition rate of these needles and thereby promotes incorporation of N into the system.     

Males and females of <Emphasis Type="Italic">Juniperus communis</Emphasis> L. and <Emphasis Type="Italic">Taxus baccata</Emphasis> L. show different seasonal patterns of nitrogen and carbon content in needles

Genders of dioecious species often show secondary sexual dimorphisms (not directly related to the sex organs), which may be related to reproductive demand for resources during the year. Our working hypothesis stated that phenology influences yearly sex-specific pattern of foliar nitrogen concentration in dioecious species. The concentration of carbon (C) and nitrogen (N) of last year’s needles (on part of twigs where strobili are bearing) was measured in 1-month intervals from March to November in Taxus baccata L. and Juniperus communis L. separately for males and females. Seasonal C concentration of needles was unrelated to gender, probably due to storage elsewhere in the plants. Needles of J. communis females had higher N concentration in March and April but this dropped to the same level as males after flowering and vegetative growth began. This suggests that females of J. communis, a species of N-poor habitats, have a long-term strategy of N storage. Nitrogen concentration was not different between the sexes during the rest of the growing season. In T. baccata, N concentration was higher in males throughout the analysed period, but the highest differences were in the period of intensive shoot elongation and radial growth. The high demand for N in the period of intensive growth and female cone maturation may result in restrictions in the vegetative growth of females. The results indicate that the time (phenology) and species-specific strategy significantly affect the concentration of N in females and males.     

Antifungal and Biocontrol Evaluation of Four <Emphasis Type="Italic">Lysobacter</Emphasis> Strains Against Clubroot Disease

The effect of crude extract (Ce), seed coating agent (SCA) and whole bacterial broth culture (WBC) of Lysobacter strains was evaluated against the causal agent of clubroot formation in Cruciferous vegetables. The ability of four Lysobacter strains (L. antibioticus 6-B-1, L. antibioticus 6-T-4, L. antibioticus 13-B-1 and L. capsici ZST1-2) inhibited Plasmodiophora brassicae of resting spores and disease. Application of WBC of four Lysobacter strains inhibited clubroot disease, indicating that the disease suppression was due to antifungal compounds produced by the biocontrol bacterium in the culture. Development of clubroot on Chinese cabbage was inhibited when the WBC and SCA were applied before P. brassicae inoculation. Crude extract (Ce) of culture filtrate was effective in arresting the germination of resting spores of P. brassicae on slides. However, Lysobacter strains differed in their biocontrol effects, the strain L. capsci ZST1-2 recorded a high level of disease limiting effect.     

Morphology and molecules reveal the alien <Emphasis Type="Italic">Posthodiplostomum centrarchi</Emphasis> Hoffman, 1958 as the third species of <Emphasis Type="Italic">Posthodiplostomum</Emphasis> Dubois, 1936 (Digenea: Diplostomidae) in Europe

Metacercariae of two species of Posthodiplostomum Dubois, 1936 (Digenea: Diplostomidae) were subjected to morphological and molecular studies: P. brevicaudatum (von Nordmann, 1832) from Gasterosteus aculeatus (L.) (Gasterosteiformes: Gasterosteidae), Bulgaria (morphology, cox1 and ITS1-5.8S-ITS2) and Perca fluviatilis L. (Perciformes: Percidae), Czech Republic (morphology, cox1, ITS1-5.8S-ITS2 and 28S); and P. centrarchi Hoffman, 1958 from Lepomis gibbosus (L.) (Perciformes: Centrarchidae), Bulgaria (morphology, cox1 and ITS1-5.8S-ITS2) and Slovakia (cox1 and ITS1-5.8S-ITS2). In addition, cercariae of P. cuticola (von Nordmann, 1832) from Planorbis planorbis (L.) (Mollusca: Planorbidae), Lithuania (morphology and cox1) and metacercariae of Ornithodiplostomum scardinii (Schulman in Dubinin, 1952) from Scardinius erythrophthalmus (L.) (Cypriniformes: Cyprinidae), Czech Republic, were examined (morphology, cox1, ITS1-5.8S-ITS2 and 28S). These represent the first molecular data for species of Posthodiplostomum and Ornithodiplostomum Dubois, 1936 from the Palaearctic. Phylogenetic analyses based on cox1 and ITS1-5.8S-ITS2, using O. scardinii as the outgroup and including the three newly-sequenced Posthodiplostomum spp. from Europe and eight published unidentified (presumably species-level) lineages of Posthodiplostomum from Canada confirmed the distinct status of the three European species (contrary to the generally accepted opinion that only P. brevicaudatum and P. cuticola occur in the Palaearctic). The subspecies Posthodiplostomum minimum centrarchi Hoffmann, 1958, originally described from North America, is elevated to the species level as Posthodiplostomum centrarchi Hoffman, 1958. The undescribed “Posthodiplostomum sp. 3” of Locke et al. (2010) from centrarchid fishes in Canada has identical sequences with the European isolates of P. centrarchi and is recognised as belonging to the same species. The latter parasite, occurring in the alien pumpkinseed sunfish Lepomis gibbosus in Europe, is also supposed to be alien for this continent. It is speculated that it colonised Europe long ago and is currently widespread (recorded in Bulgaria, Slovakia and Spain); based on the cox1 sequence of an adult digenean isolate from the Ebro Delta, Spain, only the grey heron (Ardea cinerea L.) (Ciconiiformes: Ardeidae) is known to be its definitive host in Europe.     

Acaricidal active fractions from acetone extract of <Emphasis Type="Italic">Aloe vera</Emphasis> L. against <Emphasis Type="Italic">Tetranychus cinnabarinus</Emphasis> and <Emphasis Type="Italic">Panonychus citri</Emphasis>

This study aimed to isolate acaricidal active fractions from acetone extract of Aloe vera L. and investigate the toxicity of these fractions against Tetranychus cinnabarinus (T. cinnabarinus) and Panonychus citri (P. citri). Acetone extract of A. vera L. was isolated by immersing in acetone for 72 h, and diverse fractions were fractionated by column chromatography. The acaricidal activity of each fractions was evaluated by corrected mortality of T. cinnabarinus through slide-dip bioassay. The 8th and 13th fractions of acetone extract with good acaricidal activity were indentified by LC/MS, and the toxicity of these two fractions to T. cinnabarinus and P. citri was identified by regression analysis. Acetone extract of A. vera L. exhibited obvious acaricidal activity, from which a total of 18 fractions were isolated. The 8th and 13th fractions with strong acaricidal activity against T. cinnabarinus were identified to be 3-O-alpha-d-mannopyranosyl-d-mannopyranose (OAMM) and aloe emodin. When compared with spirodiclofen, both OAMM and aloe emodin exhibited higher toxicity to T. cinnabarinus, while only OAMM exhibited a higher toxicity to P. citri (P < 0.05). OAMM and aloe emodin isolated from acetone extract of A. vera L. exhibited obvious acaricidal activities against T. cinnabarinus and P. citri.