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.     

Quality of citizen science data and its consequences for the conservation of skipper butterflies (Hesperiidae) in Flanders (northern Belgium)

Citizen science projects have become important data sources for ecologists. However, opportunistic data are not only characterized by spatial and temporal biases, but probably also contain species identification errors, especially concerning morphologically similar species. Such misidentifications may result in wrongly estimated distribution ranges and trends, and thus in inadequate conservation measures. We illustrate this issue with three skipper butterflies (Hesperiidae) in Flanders (northern Belgium) using photographs uploaded with observations in data portals. Ochlodes sylvanus and Thymelicus lineola records had relatively low identification error rates (1 and 11?%, respectively), but the majority (59?%) of Thymelicus sylvestris records turned out to be misidentified. Using verified records only allowed us to model their distribution more accurately, especially for T. sylvestris whose actual distribution area had hitherto been strongly overestimated. An additional field study on T. sylvestris confirmed the species distribution model output as the species was almost completely restricted to sites with verified records and was largely absent from sites with unverified records. The preference of T. sylvestris for unimproved grasslands was confirmed by the negative correlation between its model-predicted presence and elevated nitrogen and ammonia levels. Thus, quality control of citizen science data is of major importance to improve the knowledge of species distribution ranges, biotope preferences and other limiting factors. This, in turn, will help to better assess species conservation statuses and to suggest more appropriate management and mitigation measures.     

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.     

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.     

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

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

Heterosis,transmission genetics,and selection for increased growth rate in a <Emphasis Type="Italic">N. tabacum</Emphasis> × synthetic tobacco cross

Cultivated tobacco (Nicotiana tabacum L.) is a classic amphidiploid, and hybrids between this cultivated species and closely related diploid Nicotiana relatives often exhibit heterotic effects for growth rate and yield. Crosses between N. tabacum and synthetic tobaccos, 4x(Nicotiana sylvestris × Nicotiana otophora) or 4x(N. sylvestris × Nicotiana tomentosiformis), may provide superior routes for genome-wide introgression from diploid relatives and allow increased potential to capitalize on heterotic effects in tobacco. Significant levels of mid-parent heterosis were observed for yield and growth rate in F₁ hybrids between synthetic tobaccos and a standard tobacco cultivar. Microsatellite marker genotyping of an F₂ population derived from a K326 × [4x(N. sylvestris × N. otophora)] cross was carried out to preliminarily investigate the relative importance of different types of gene action on observed heterosis in the original interspecific cross. Results suggested a role for both partial dominance and overdominance. Marker genotyping also indicated an overall reduced level of recombination in the N. tabacum × synthetic tobacco cross relative to a N. tabacum × N. tabacum cross but no evidence of genomic regions with severely restricted levels of recombination. Results suggest that populations derived from N. tabacum × synthetic tobacco crosses may be more efficient for introgressing germplasm from diploid relatives, as compared to populations derived from crosses between N. tabacum and diploid forms where preferential pairing between N. tabacum homologues can reduce the potential for introgression of alien chromatin. Such materials may be useful as sources of favorable alleles influencing quantitative characters in tobacco.     

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.     

Cork-warts on leaves of Lauraceae: confirming a suspicion

Some studies have reported the presence of leaf cork-warts in several families of plants; however, in Lauraceae, these structures have not been confirmed. Therefore, based on studies of leaf anatomy, we have herein documented the presence of leaf cork-warts in Mezilaurus species of Lauraceae. To accomplish this, we investigated fifteen species from all six genera of the Mezilaurus group, including Anaueria, Chlorocardium, Clinostemon, Mezilaurus, Sextonia, and Williamodendron. Analyses of leaf anatomy were performed through optical and scanning electron microscopy. Our results revealed cork-warts in six species of the genus Mezilaurus: M. crassiramea, M. navalium, M. revolutifolia, M. subcordata, M. synandra, and M. vanderwerffii. In the investigated species M. crassiramea and M. navalium, cork-warts originated from the stomatal complex, from epidermal cells covering oil cells located in mesophyll, and from epidermal cells around trichomes. Therefore, cork-warts could be considered an additional taxonomic character to delimit Mezilaurus species.     

Searching chromosomal landmarks in Indian lentils through EMA-based Giemsa staining method

Lentil is one of the oldest protein-rich food crop with only one cultivated and six wild species. India is one important cultivator, producer and consumer of lentils and possesses a large number of germplasms. All species of lentil show 2n?=?14 chromosomes. The primary objective of the present paper is to search chromosomal landmarks through enzymatic maceration and air drying (EMA)-based Giemsa staining method in five Indian lentil species not reported elsewhere at a time. Additionally, gametic chromosome analysis, tendril formation and seed morphology have been studied to ascertain interspecific relationships in lentils. Chromosome analysis in Lens culinaris, Lens orientalis and Lens odemensis revealed that they contain intercalary sat chromosome and similar karyotypic formula, while Lens nigricans and Lens lamottei showed presence of terminal sat chromosomes not reported earlier. This distinct morphological feature in L. nigricans and L. lamottei may be considered as chromosomal landmark. Meiotic analysis showed n?=?7 bivalents in L. culinaris, L. nigricans and L. lamottei. No tendril formation was observed in L. culinaris, L. orientalis and L. odemensis while L. nigricans and L. lamottei developed very prominent tendrils. Based on chromosomal analysis, tendril formation and seed morphology, the five lentil species can be separated into two distinct groups. The outcome of this research may enrich conventional and biotechnological breeding programmes in lentil and may facilitate an easy and alternative method for identification of interspecific hybrids.     

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%).     

Toxicity of some goldenrods

This study was performed to determine the biological activity in mice of eight species of Solidago (goldenrod) and to determine some of the chemical groups present in these species. Biologically active substances were present in all of the species tested. Tests of six species were positive for alkaloids. All but one of the species contained demonstrable saponins. All of the species contained tannins, whereas six or seven species were positive for flavonoids. Quaternary bases were not found in any of the species tested. A comparison of the biological activity in mice of these species of Solidago, extracts of which were injected intraperitoneally, showed their descending order of activity to be: (1)S. flexicaulis L., (2)S. hispida Muhl., (3)S. juncea Ait., (4)S. serotina Ait., (5)S. canadensis L., (6)S. rugosa Ait., (7)S. uliginosa Nutt., and (8)S. squarrosa Muhl.     

Stratification of the ant species (Hymenoptera,Formicidae) in the urban broadleaf woodlands of the city of Kiev

Investigations were carried out in June–August 2012 and 2013 in broadleaf forests and planted stands of the northern red oak Quercus rubra in Kiev. The ant visitation rates of 3–6 most abundant plant species within each vegetation layer were analyzed. In all, 16 species of ants were found in the broadleaf forest (Quercus robur + Acer spp. + Carpinus betulus), with the dominance of three species: Formica rufa, Lasius fuliginosus, and L. emarginatus. Eight ant species occurred in Q. rubra stands with the undergrowth of Acer campestre and A. platanoides; the dominant species were Lasius fuliginosus and L. emarginatus. The northern red oak stands with conspecific undergrowth had only four ant species with no dominants among them. Plants with the highest ant visitation rates in the herbage layer of the broadleaf forest were the invasive Impatiens parviflora and the native Aegopodium podagraria. Ants were rare or absent on the remaining herbs: Impatiens noli-tangere, Carex sylvatica, Stellaria holostea, and Galium odoratum. The herbage layer was mostly visited by influents, the dominants being represented only by Lasius emarginatus. Within the shrub layer, ants most frequently visited the undergrowth of the maple Acer platanoides, often harboring colonies of the aphid Periphyllus lyropictus; the highest visitation rates were recorded in the dominants L. emarginatus and L. fuliginosus. The arboreal layer had the highest visitation rate and was most often visited by the dominants Lasius fuliginosus, L. emarginatus, and F. rufa. The common oak Q. robur was the most visited tree in broadleaf forests. In addition to providing ants with food (colonies of the aphids Lachnus roboris and Stomaphis quercus, and also phytophagous insects and other invertebrates), these trees were used for nesting by the dominants L. fuliginosus and L. emarginatus. The presence of aphid colonies was also typical of ripe trees and undergrowth of the maple A. platanoides, the second-visited tree in broadleaf forests. The common hornbeam Carpinus betulus was the least frequently visited, though some ant species, usually L. emarginatus, nested in the trunks and branches of old trees. The vertical distribution of ants in broadleaf forests and red oak stands formed two clusters: the influents occupied the lower (herbage and shrub) vegetation layers, while the subdominants and dominants occupied the upper (shrub and arboreal) ones. The smallest number of ant species (1 or 2) in all the vegetation layers was recorded in the areas with F. rufa; areas with L. fuliginosus had twice as many species; the greatest numbers of ant species were found in the areas with L. emarginatus and in those without dominants. This pattern may result from different territoriality of the dominants: the strongest in F. rufa (defending the whole territory) and the weaker in the other two species (defending only a part of the territory). Plants of all the vegetation layers varied in their attractiveness to ants. As a result, the layers had a mosaic structure in their vertical (between-layer) and horizontal (between different plant species within one layer) arrangement. From 60 to 100% of plants of certain species were visited by ants while plants of other species were not visited at all. The main reason for visiting plants by ants was the presence of aphid colonies.     

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.     

Intestinal <Emphasis Type="Italic">Lactobacillus</Emphasis> community structure and its correlation with diet of Southern Chinese elderly subjects

This study aimed to investigate the relationship between the intestinal Lactobacillus species and diet of elderly subjects in a longevity area in Southern China. Healthy elderly subjects ranging from 80 to 99 years old were respectively selected from the regions of Bama and Nanning, Guangxi, China. The nested polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) technology was used to analyze the intestinal Lactobacillus community structure. Results showed that Weissella confusa, L. mucosae, L. crispatus, L. salivarius, and L. delbrueckii were the representative Lactobacillus of elderly subjects. Among them, L. crispatus and L. delbrueckii were the dominant Lactobacillus of all species. In comparison to Nanning elderly subjects, the detection frequencies of W. confusa and L. salivarius were significantly increased in Bama elderly subjects (P < 0.01), whereas L. mucosae was significantly decreased (P < 0.01). Interestingly, it was also found that there were 4 kinds of representative Lactobacillus, which were significantly correlated with dietary fiber. W. confusa (P < 0.01) and L. salivarius (P < 0.05) were significantly positively correlated with the intake of dietary fiber, while L. mucosae (P < 0.01) and L. crispatus (P < 0.05) were significantly negatively correlated with the intake of dietary fiber, respectively. Results confirmed that different diets had obvious effects on the intestinal Lactobacillus community structure of elderly subjects in Southern China, which may provide a certain theoretical basis for the elderly’s healthy food strategic design and probiotics product development.     

Spatial change of reservoir nitrite-dependent methane-oxidizing microorganisms

Nitrite-dependent anaerobic methane oxidation (n-damo), catalyzed by microorganisms affiliated with bacterial phylum NC10, can have an important contribution to the reduction of the methane emission from anoxic freshwater sediment to the atmosphere. However, information on the variation of sediment n-damo organisms in reservoirs is still lacking. The present study monitored the spatial change of sediment n-damo organisms in the oligotrophic freshwater Xinfengjiang Reservoir (South China). Sediment samples were obtained from six different sampling locations and two sediment depths (0–5 cm, 5–10 cm). Sediment n-damo bacterial abundance was found to vary with sampling location and layer depth, which was likely influenced by pH and nitrogen level. The presence of the n-damo pmoA gene was found in all these samples. A remarkable shift occurred in the diversity and composition of sediment n-damo pmoA gene sequences. A variety of distinctively different n-damo pmoA clusters existed in reservoir sediments. The pmoA sequences affiliated with Candidatus Methylomirabilis oxyfera formed the largest group, while a significant proportion of the obtained n-damo pmoA gene sequences showed no close relationship to those from any known NC10 species. In addition, the present n-damo process was found in reservoir sediment, which could be enhanced by nitrite nitrogen amendment.     

Shade tolerance of <Emphasis Type="Italic">Ailanthus altissima</Emphasis> revisited: novel insights from southern Switzerland

The tree of heaven (Ailanthus altissima (Mill.) Swingle) is considered to be an early-successional, gap-obligate pioneer species with vigorous height growth, low shade tolerance, early fecundity and large seed production. It is a highly invasive species in many temperate and Mediterranean ecosystems outside its natural range, especially after disturbance. Due to its low shade tolerance, the potential of A. altissima to colonise undisturbed forests is thought to be low. In this study we analysed the potential of juvenile A. altissima to grow and survive in sweet chestnut (Castanea sativa Mill.) forests in southern Switzerland. We used hemispherical photography to assess the light conditions of 204 individuals of A. altissima (31 % generative, 69 % vegetative) aged between 1 and 7 years (median: 3 years) in six sites. Generative (seed-borne) and vegetative (clonal ramet) offspring of A. altissima are able to grow in light conditions well below the requirements of shade-intolerant tree species such as European larch (Larix decidua Mill.) and Scots pine (Pinus sylvestris L.). The relatively low light conditions found to be sufficient for the growth and survival of generative regeneration of A. altissima suggest a higher shade tolerance for this species than previously stated, at least for early regeneration. Consequently, the colonisation frontier of A. altissima should be intensively monitored in both forest openings but also in closed canopy forests in the vicinity of seed-bearing A. altissima.     

Production of gold nanoparticles by biogenesis using bacteria

Diazotrophic cyanobacteria Anabaena sp. PCC 7120, four Nostoc strains, and two Azotobacter species (A. vinelandii and A. chroococcum) were found to produce gold nanoparticles (GNP) under nitrogen fixation conditions. GNP biogenesis occurred at AuHCl₄ concentrations from 0.1 to 1 mM. In the cultures of unicellular cyanobacteria Synechococcus sp. PCC 7942 and Synechocystis sp. PCC 6803 incapable of nitrogen fixation, no GNP were formed at the same concentrations of gold salts. The plasmon resonance band peak was located at 552 nm. This position is characteristic of spherical GNP 10 to 30 nm in size. Small amounts of GNP were also formed in the culture liquid supernatants of the tested nitrogen-fixing bacteria at AuHCl₄ concentrations from 0.25 to 0.5 mM.     

Cytological Variability in <Emphasis Type="Italic">Artemisia</Emphasis> L. inhabiting North-West Himalayas: B Chromosomes in <Emphasis Type="Italic">Artemisia gmelini</Emphasis> Weber ex Stechm

Present work is a part of our studies on the cytological details of some species of Artemisia L. inhabiting Ladakh region of Jammu and Kashmir state. Of the six populations investigated for three different species (Artemisia sieversiana Ehrh. ex Willd., A. tournefortiana L. and A. gmelinii Weber ex Stechm.), some variations were noticed only for Artemisia gmelinii. The species occurring in Leh region at altitudinal range of 3992 masl is diploid with 2n = 18 (n = 9). In one of the population of the species, sprawling in Hemis region (4009 masl), В chromosome was found to be present in addition to the diploid chromosome complement. Comparison of the different morphological and reproductive features revealed that the population with В chromosome had reduced vigour.     

A review of oribatid mites of the family Liacaridae (Acariformes,Oribatida) from the Caucasus

The paper summarizes the data on oribatid mites of the family Liacaridae and their distribution in the Caucasus. Two new species, Liacarus subiasi from Azerbaijan (differing from the known species in the shape of cuspids and short lamellae) and Dorycranosus musaevi from Daghestan (differing from closely related D. splendens (Coggi, 1898) and D. punctulatus Miheli?i?, 1956 in the presence of the smooth notogaster, wide distal lamellar cuspids, and also in the absence of inner cuspid tooth and the presence of longer ventral setae), are described. The species status of D. punctulatus is reestablished. Liacarus lencoranicus Krivolutsky, 1967 and L. nitidulus Krivolutsky, 1967 are synonymized with L. coracinus C.L. Koch, 1898. D. zachvatkini Kuliev, 1962 (= D. ibericus Dzaparidze, 1973) is redescribed. Data on species variability of Adoristes poppei (Oudemans, 1906) and A. ovatus Koch, 1849 are given.