Embryological peculiarities of interspecific hybridization in <Emphasis Type="Italic">Pinus sibirica</Emphasis>

Cytoembryological research of the ovules in experiments with interspecific hybridization of Pinus sibirica (pollination be the pollen of P. koraiensis, P. armandii, P. parviflora, P. strobus, P. hokkaidensis, P. wallichiana, P. monticola, and P. сembra) revealed that the development of megagametophytes occurred in them by the usual scenario and resulted in the formation of mature archegonia. Pollen successfully germinated on the nucellus of ovules. However, disturbances were observed in the process of male gametophyte development, and pollen tubes on the nucellus were not visible by the period of archegonia maturation. Fertilization was usually absent. The development of embryonic channel is determined by egg cell maturity. The only exception was the variant of the controlled pollination of Pinus sibirica × P. сembra, in which the embryo has been formed.     

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%).     

Nucleotide polymorphisms of candidate genes of adaptive significance in the ural populations of <Emphasis Type="Italic">Larix sibirica</Emphasis> Ledeb.

The objectives of conservation and sustainable forest management require in depth study of genomes of woody plants and definition of their intraspecific genetic diversity. In recent years, an approach was developed based on the study of “candidate genes” that can potentially be involved in the formation of adaptive traits. In this study, we investigated nucleotide polymorphism of several adaptive candidate genes in the populations of Siberian larch (Larix sibirica Ledeb.) in the Urals. Representatives of this genus are among the most valuable and widely distributed forest tree species in Russia. From ten selected gene loci in the genome of L. sibirica, we isolated and investigated three loci, one of which (ABA-WDS) was sequenced in L. sibirica for the first time. The total length of the analyzed sequence in each individual amounted to 2865 bp. The length of locus alignment was from 360 bp to 1395 bp. In total, we identified 200 polymorphic positions. The most conservative is locus 4CL1-363, and the most polymorphic is locus sSPcDFD040B03103-274. The studied populations of L. sibirica are characterized by a high level of nucleotide polymorphism in comparison with other species and genuses (Picea, Pinus, Pseudotsuga, Abies) conifers plants (Hd = 0.896; π = 0.007; θ_W = 0.015). The most selectively neutral polymorphism (D _T =–0.997) was attributed to locus 4CL1-363, and polymorphism with high probability of adaptability (D _T =–1.807) was determined for the ABA-WDS locus. We identified 54 SNP markers, only five of which were nonsynonymous (9.26%) replacements. The average frequency of SNPs in the three studied loci of L. sibirica was one SNP in 53 bp. We detected unique SNP markers for eight populations, which could potentially be used to identify populations. Populations that are characterized by the highest number of unique SNP markers can be recommended for selection in order to preserve the gene pool of the species.     

Investigation of the photosynthetic activity of bark phelloderm of arboreous plants using the fluorescent method

Seasonal changes in the characteristics of chlorophyll fluorescence were studied in the bark of several species of trees originating in various climatic zones: Siberian cedar (Pinus sibirica), larch (Larix sibirica), eastern arborvitae (Thuja occidentalis), pendent white birch (Betula pendula), wild black cherry (Padus virginiana), horse chestnut (Aesculus hippocastanum), red oak (Quercus rubra), Manchurian catalpa (Catalpa bungei), linden (Tilia cordata), goat willow (Salix caprea), Amur cherry (Padus maackii), and apple Korichnaya (Malus domestrica B.). Tree bark has a sufficient amount of chlorophyll for measuring the parameters of chlorophyll fluorescence throughout the year. The relative yield of the variable fluorescence of chlorophyll (F _v/F _m) can be used to assess seasonal changes in the physiological state of various trees.     

Variability of metric and qualitative traits of <Emphasis Type="Italic">Nitraria</Emphasis> species in relation to ecological and climatic conditions of Siberian habitats

The metric traits of shrubs, leaves, seeds, and flower parts have been studied in 19 Siberian populations of Nitraria sibirica Pall. and N. schoberi L. According to the obtained data, N. schoberi is characterized by larger shrubs (150–179 cm in height), leaves (15–30 mm in length, 5.7–8.8 mm in width), and seeds (up to 10 mm in length) and has rather wide (~2.5 mm) corolla petals. The size of anthers exceeds that of N. sibirica 1.2–1.5 times. The height of N. sibirica shrubs does not exceed 80 cm; the length of lamina, seed, and petals of corolla is 8–10, 3.7–7.4, and ~3.5 mm, respectively; and the size of anthers is 0.5–1 mm. Irrespective of habitat conditions, the stability of metric traits typical for both species has been shown to be rather high. The only dependence on climatic parameters is observed for the height of N. sibirica, which forms creeping forms in continental regions, especially in the mountains of the Altai Republic. Vegetative organs (laminas and young shoots) of N. sibirica are always more pubescent than those of N. shoberi.     

Activities and properties of phosphorylases of turbellarias <Emphasis Type="Italic">Phagocata sibirica</Emphasis> and cestodes <Emphasis Type="Italic">Bothriocephalus scorpii</Emphasis>

Activities and properties of phosphorylases of cytosol and mitochondrial fractions are studied in free-living turbellarias Phagocata sibirica and cestodes Bothriocephalus scorpii. The phosphorylase activities in P. sibirica and B. scorpii differ significantly both in form and in total activity of this enzyme. Dependence of the phosphorylase reaction rate on substrate concentration is studied. The high activity of phosphorylase as compared with that of hexokinase suggests glycogen to be the main energy source of the studied flatworms. Action of various effectors on activities of the cytosol and mitochondrial phosphorylases has been studied.     

Quantum Speciation of <Emphasis Type="Italic">Yersinia pestis</Emphasis> Plague Microbe in a Heteroimmune Environment: In the Populations of Hibernating Tarbagan Marmots (<Emphasis Type="Italic">Marmota sibirica</Emphasis>)

This review is devoted to the ecological mechanism for the transformation of the population of the psychrophilic saprozoobiont pseudotuberculosis microbe Yersinia pseudotuberculosis O:1b into the population of the obligate blood plague microbe Yersinia pestis in the host–parasite system of the marmot flea (Marmota sibirica–Oropsylla silantiewi). The correspondence of this mechanism to the evolutionary principle of quantum speciation has been shown. The most significant population–genetic microbial transformations took place in the populations of hibernating marmots. The main factor of rapid speciation was the heterothermic and, accordingly, heteroimmune state of host marmots during hibernation. During winter awakenings, the body temperature of marmots increases in an S-shaped manner from 5 to 37°C within a short period of time (from dozens of minutes to several hours). A drastic acceleration of metabolic and immune processes occurs in the temperature range of 20–30°C. Rapid adaptation to the “explosive” increase in the immune activity of the primary host (tarbagan marmot) during its regular winter awakenings was the essence of the process of Y. pestis speciation.     

Correlations between the ages of<Emphasis Type="Italic">Alnus</Emphasis> host species and the genetic diversity of associated endosymbiotic<Emphasis Type="Italic">Frankia</Emphasis> strains from nodules

Nodule samples were collected from four alder species:Alnus nepalensis, A. sibirica, A. tinctoria andA. mandshurica growing in different environments on Gaoligong Mountains, Yunnan Province of Southwest China and on Changbai Mountains, Jilin Province of Northeast China. PCR-RFLP analysis of the IGS betweennifD andnifK genes was directly applied to unculturedFrankia strains in the nodules. A total of 21 restriction patterns were obtained. TheFrankia population in the nodules ofA. nepalensis had the highest genetic diversity among all fourFrankia populations; by contrast, the population in the nodules ofA. mandshurica had the lowest degree of divergence; the ones in the nodules ofA. sibirica andA. tinctoria were intermediate. A dendrogram, which was constructed based on the genetic distance between the restriction patterns, indicated thatFrankia strains fromA. sibirica andA. tinctoria had a close genetic relationship.Frankia strains fromA. nepalensis might be the ancestor ofFrankia strains infecting otherAlnus species. From these results and the inference of the ages ofAlnus host species, it is deduced that there was a co-evolution betweenAlnus and its microsymbiontFrankia in China.     

<Emphasis Type="Italic">Pinus contorta</Emphasis> invasion into treeless steppe reduces species richness and alters species traits of the local community

Pinus contorta, one of the most invasive tree species in the world, has been proposed as a model species for improving our understanding of invasion ecology. In this study, we assessed the impact of P. contorta invasions on the species richness, diversity and species traits of a resident treeless steppe community. In a Pinus contorta invasion gradient (Patagonia, Chile), we surveyed vegetation from high canopy closure pine invasion to treeless steppe, and computed species richness, diversity and Sørensen similarity indexes. For all species, we determined functional trait values from the literature, data bases, and personal observations. Species richness and diversity were related to canopy cover (a proxy for invasion intensity) using generalized linear mixed-effects models. Changes in species traits due to canopy cover were analyzed using RLQ ordination analysis and the fourth-corner analysis. We found that Pinus contorta canopy cover significantly reduced the number of native species by 70 %, implying a strong effect on species exclusion. A few native species, however, prevail in the novel conditions (e.g. Baccharis magellanica, Acaena integerrima). Species traits changed significantly with increasing pine canopy cover, where P. contorta promoted the existence of traits related to shade-tolerance and conservative reproductive strategies. We conclude that the negative impacts of Pinus contorta into the treeless steppe, including a reduction in the number of species and the shifting to traits adapted to tolerate shade and associated with conservative reproductive strategies, can have severe implications for the conservation of biodiversity and ecosystem functioning where it invades.     

Invasion of Siberian Pine Populations in Mountain Tundra in the Northern Urals

A new method for determining the upper forest border (UFB) as the border of the “root-closed” stand and regrowth as its potential vanguard is proposed. The mass dispersal of Pinus sibirica populations by the nutcracker (Nucifraga caryocatactes) in the UFB ecotone in the Northern Urals (Pavdinskii Kamen’ and Tretii Bugor Mountain) from the middle-mountain taiga to the mountain forest tundra and tundra at a distance reaching 1 km is found. It is done based on the original reconstruction method of the dynamics of population number in P. sibirica annual seedlings and on an analysis of their relations with the dynamics of the seed bearing and population number of the nutcracker. The trustworthy consortive relations of the number of Pinus sibirica generations in the mountain forested tundra and tundra zones, with its seed bearing and nutcracker number in the previous year in the middle-mountain taiga zone, are found. A significant increase, 3.7 times, in the number of seedling generations during the last 49 years is found in connection with the increase in the summer air temperature in the subarctic to 2.0°C. A hypothesis of the formation of the united genetic metapopulation of P. sibirica in the forest–tundra mountain ecotone zone as a consequence of its seed ornitochory from the highly different populations is formulated. A forecast of the mosaic formation of the vanguard “thin forests” in 20–25 years and the root-closed curtains of the forest in 40–50 years in the mountain tundra of the Northern Urals at the current rate of the rise in temperature is made.     

Variance in Female Reproductive Success Differentially Impacts Effective Population Size in the Short-Nosed Fruit Bat, <Emphasis Type="Italic">Cynopterus sphinx</Emphasis>

Effective population size (N _e) quantifies the effects of micro-evolutionary processes and the rate of loss of genetic diversity in a population. Several demographic and mating parameters reduce N _e. Theoretical studies elucidate the impacts of various demographic and mating system parameters on N _e, while empirical studies illustrate realized N _e for species with differing life histories and mating systems. However, effect of intra-specific variation in mating system on effective size remains largely unexplored. In this paper we investigated the effect of promiscuous and polygynous mating on N _e in two wild populations of the short-nosed fruit bat, Cynopterus sphinx. N _e/N (ratio of effective population size to census size) was lower than unity in both populations, and much lower for the polygynous population compared to promiscuous population. Elasticity analyses reveal that N _e/N was sensitive to deviations in the sex ratio. Variance in female reproductive success had a higher impact on N _e compared to variance in male reproductive success in the promiscuous population. However, for the polygynous population, impact of variance in male reproductive success on N _e was higher than that of variance in female reproductive success. Our results suggest that depending on mating system, different populations of the same species could have alternate evolutionary trajectories. The rate of loss of genetic diversity would be lower for the promiscuous population compared to the polygynous population. Our study is the first to highlight which parameters would most significantly impact population specific N _e under different mating systems.     

Phylogenetic reconstructions in the genus <Emphasis Type="Italic">Capra</Emphasis> (Bovidae,Artiodactyla) based on the mitochondrial DNA analysis

Mitochondrial genome fragments were examined in all species of the genus Capra (Bovidae, Artiodactyla). Phylogenetic analysis was carried out using 59 cytochrome b gene sequences (392 bp), and 22 sequences of the mtDNA variable fragment (402 bp). In the control region, two unique deletions were revealed. One of the deletions was found only in Capra cylindricornis (17 bp), while another one grouped C. caucasica with C. aegagrus (1 bp). The group of Caucasian wild goats splits into two clades, and furthermore, the sequences of C. caucasica demonstrate remarkable similarity to the sequences of C. aegagrus, while C. cylindricornis seems to have evolved independently for a long period of time. It was demonstrated that C. pyrenaica and C. ibex were extremely close to one another. Capra sibirica formed an outer group relative to the other species, and according to our data, was the most ancient species of the genus. On the contrary, genetic distance separating C. falconeri (the most independent species of the genus related to its morphology) from the other species is small.     

Middle Cambrian trilobites from the vicinity of Dolgii Mys Mountain (Khakassia,Batenevsky Ridge)

The trilobites Oryctocarella sibirica (Tomash.) and Lancastria plana (Tomash.) from the basal Middle Cambrian in the vicinity of Dolgii Mys Mountain (Batenevsky Ridge, Khakassia) are described. These trilobites were previously described by V.D. Tomashpolskaya, but the original collection was subsequently lost. Repeated finds allowed a detailed study of these species. It is established that relatively small specimens of Oryctocarella sibirica are more similar to Ovatoryctocara granulata. In addition, these species have much in common with the genus Arthricocephalus in morphological characters. New material of Lancastria plana is described in detail. These trilobites were originally assigned to the genus Paraoryctocephalops, although according to the data of Blacker and Peel (1997), they should be transfered to the genus Lancastria.     

Reproductive Biology of <Emphasis Type="Italic">Leptocybe invasa</Emphasis> Fisher &amp; La Salle (Hymenoptera: Eulophidae)

Leptocybe invasa Fisher & La Salle (Hymenoptera: Eulophidae) is an invasive pest in Eucalyptus plantations around the world. The successful colonization of L. invasa is possibly related to its reproductive biology. The objective of this study was to examine the reproductive biology of L. invasa. In Guangxi Province, the sex ratio (proportion of female, 0.99) of L. invasa was female-dominant throughout the year based on natural and artificial infestation. This result was similar to the ratios observed for other geographic populations in China, including those in Fujian (0.99), Guangdong (0.98), Hainan (0.95), Jiangxi (0.96), and Sichuan (0.99). The offspring sex ratio favored females. A large number of females emerged from the galls produced by females, with few males found. Galls on the petioles and midribs of Eucalyptus plants could be caused by newly emerged females with mature eggs. The lengths of the ovariole, spermatheca, common oviduct, and reproductive glands did not differ among L. invasa females, but their lateral oviducts showed differences from 0 to 42 h after emergence, indicating that this insect is proovigenic. These results could explain why L. invasa populations can rapidly increase in invaded areas.     

Functional Conservation of an <Emphasis Type="Italic">AGAMOUS</Emphasis> Orthologous Gene Controlling Reproductive Organ Development in the Gymnosperm Species <Emphasis Type="Italic">Taxus chinensis</Emphasis> var. <Emphasis Type="Italic">mairei</Emphasis>

Arabidopsis AGAMOUS (AG) has roles in specifying reproductive organ (stamens and carpels) identity, floral meristem determinacy, and repression of A-function. To investigate possible roles of AG orthologous genes in gymnosperm species and evolution of C function, we isolated and identified AG orthologous gene TcAG from Taxus chinensis var. mairei (family Taxaceae, order Coniferales), a member of the last divergant lineage from higher Conifer that sisters to Gnetales. Sequence alignment and phylogenetic analysis grouped TcAG into the gymnosperm AG lineage. TcAG was expressed in both developing male and female cones, but there was no expression in juvenile leaves. Ectopic expression of TcAG in an Arabidopsis ag mutant produced flowers with the third whorl petaloid stamen and fourth whorl normal carpel, but failed to convert first whorl sepals into carpeloid organs and second whorl petals into stamenoid organs. A 35S::TcAG transgenic Arabidopsis ag mutant had very early flowering, and produced a misshapen inflorescence with a shortened floral axis. Our results suggest that establishment of the complete C-function occurred gradually during AG lineage evolution even in gymnosperms.     

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.     

Autecology of a new species of carnivorous marsupial,the endangered black-tailed dusky antechinus (<Emphasis Type="Italic">Antechinus arktos</Emphasis>), compared to a sympatric congener,the brown antechinus (<Emphasis Type="Italic">Antechinus stuartii</Emphasis>)

The carnivorous marsupial Antechinus are one of the few mammal genera known to exhibit the phenomenon of semelparous reproduction, where all males die at the end of a frenetic annual mating period. The genus Antechinus has recently been revised, seeing four new species named and one existing subspecies raised to species status. Here, we present the first ecological assessment of one new species, the endangered black-tailed dusky antechinus (Antechinus arktos), based on a 2-year mark-recapture study of two proximate sites within the cloud forest of Springbrook National Park, south-east Queensland, Australia. We also present comparative ecological data from a sympatric congener, Antechinus stuartii. In total, 103 A. arktos (49 male; 54 female) and 2125 A. stuartii (1229 male; 896 female) captures were made over 16,630 trap nights at the two sites. The occurrence and synchrony of reproductive events observed in A. arktos closely parallel the semelparous reproductive strategy exhibited by all congeners studied to date. The A. arktos populations mate during mid-September, with birth of up to six young occurring during mid-October. The low abundance and fecundity, together with a limited and apparently retracting distribution, suggest A. arktos is under threat of climate-induced extinction in the coming decades.     

The fitness of the cotton mealybug <Emphasis Type="Italic">Phenacoccus solenopsis</Emphasis> (Tinsley) is reduced after feeding on virus-infected <Emphasis Type="Italic">Hibiscus rosa</Emphasis>-<Emphasis Type="Italic">sinensis</Emphasis>

The cotton mealybug Phenacoccus solenopsis (Tinsley) and Cotton leaf curl Multan virus (CLCuMV), serious threats to economic crops and garden plants, have invaded southern China and widely infected Hibiscus rosa-sinensis. Whether an inter-species connection has facilitated the invasion process is unclear. In this study the interaction between P. solenopsis and H. rosa-sinensis infected with CLCuMV was investigated in the laboratory. We observed that 1st and 2nd instar nymphs of P. solenopsis preferred to feed on healthy H. rosa-sinensis leaves, whereas 3rd instar nymphs and female adults had no preference between healthy and virus-infected H. rosa-sinensis leaves. The developmental time of each P. solenopsis developmental stage increased significantly after feeding on infected H. rosa-sinensis leaves (p < 0.05). In particular, the development time for 2nd instar female and male nymphs and 3rd instar female nymphs increased by approximately twofold. The generation time of female mealybugs increased from 25.84 d on healthy H. rosa-sinensis to 32.12 d when feeding on CLCuMV-infected H. rosa-sinensis, and the survival rate decreased from 71.04 % on healthy H. rosa-sinensis to 5.80 % on infected plants. Nymph survival was most affected by feeding on infected plants. Additionally, the fecundity of female mealybugs feeding on infected H. rosa-sinensis decreased by 47.8 %. Thus, feeding on CLCuMV-infected H. rosa-sinensis significantly decreased the biological fitness and invading and colonizing abilities of P. solenopsis.