Comparative study of chemical composition and antitumor activity of aqueous-ethanol extracts of brown algae <Emphasis Type="Italic">Laminaria cichorioides,Costaria costata</Emphasis>, and <Emphasis Type="Italic">Fucus evanescens</Emphasis>

The chemical composition of water-ethanol extracts of the brown algae Laminaria cichorioides, Costaria costata, and Fucus evanescens were studied. The extracts contained mannitol, iodine, micro elements, free amino acids, glycolipids, polyunsaturated fatty acids, fucosterine, and polyphenols. The extracts were distinguished by high contents of mannitol and iodine (L. cichorioides), lipophilic matter (C. costata), and polyphenol compounds (F. evanescens). All the extracts under study inhibited the growth of DLD-1 and HT-29 human intestine tumor cells. The strongest inhibitory effect was exerted by the extract of F. evanescens at a concentration of 50 μg/ml. The extracts can be recommended for the production of fucosterol, phlorotannins, chlorophyll derivatives, mannitol, and compositions for medical and veterinary application.     

Influence of abiotic factors on the structure of the population of the brown alga <Emphasis Type="Italic">Fucus vesiculosus</Emphasis> in East Murman (Barents Sea)

The peculiarities of the structure of the population of the brown alga Fucus vesiculosus in East Murman at different combinations of abiotic factors were studied. Salinity below 10‰ and high surf activity reduced the average life span of the plants (to 1 year) and caused a significant predominance of female specimens (60–90% of the fertile specimens). The proportion of female specimens increased with age. The greatest average age (3 branchings) and the highest survival of F. vesiculosus occurred in the wave-protected parts of bays. The distribution of F. vesiculosus was related to water movement intensity, salinity, and substrate type. The size-weight characteristics of F. vesiculosus thalli were decreased during maximum salinity fluctuations. The optimal conditions for this species were those existing on a weakly protected shore with short-term fluctuations of salinity (24–25‰).     

Symbiotic performance,nitrogen flux and growth of lima bean (<Emphasis Type="Italic">Phaseolus lunatus</Emphasis> L.) varieties inoculated with different indigenous strains of rhizobia

Nitrate uptake and nitrogen inclusion into amino acids were studied in the intact thallus and isolated bionts of the lichen Parmelia sulcata with the aid of mass spectroscopic tracing of heavy isotope ¹⁵N. The isolated photobiont, the green algae Trebouxia sp. did not take up nitrate, whereas the mycobiont and intact thalli were enriched in ¹⁵N when incubated with Na¹⁵NO₃. Pulse feeding experiments with intact thalli followed by separation of photobiont showed that the labelled nitrate was originally assimilated by the mycobiont and only after that was detected in the photobiont. The isolated mycobiont after pulse labeling excreted labeled compounds into the incubation medium. Amino acids were detected in the exudate. The quantities of two amino acids considerably exceeded those of the others. One was identified as alanine, the other could not yet be identified with certainty. Both of these high-quantity compounds were also much more enriched in ¹⁵N than the others. These two compounds are proposed to be the transport forms of nitrogen within the Parmelia sulcata thallus.     

Distributional and ecophysiological study on the Antarctic lichens species pair <Emphasis Type="Italic">Usnea antarctica</Emphasis>/<Emphasis Type="Italic">Usnea aurantiaco</Emphasis>-<Emphasis Type="Italic">atra</Emphasis>

The factors that control lichen distribution in Antarctica are still not well understood, and in this investigation we focused on the distribution, local and continental, and gas exchange of a species pair, closely related lichens with differing reproductive strategies, Usnea aurantiaco-atra (fertile) and Usnea antarctica (sterile, sorediate). The local distributions of these species were recorded along an altitudinal gradient of nearly 300 m at South Bay, Livingston Island, and microclimate was also recorded over 1 year. The photosynthetic responses to temperature, light and thallus water content were determined under controlled conditions in the laboratory. The species were almost identical in their photosynthetic profiles. Locally, on Livingston Island, U. antarctica was confined to low altitude sites which were warmer and drier, whilst U. aurantiaco-atra was present at all altitudes. This contrasts with its distribution across Antarctica where U. antarctica grows 9° latitude further south than U. aurantiaco-atra. Temperature appears not to be the main controller of distribution in these species, but dryness of habitat, which will influence length of activity periods, may be important.     

The effect of inoculation with <Emphasis Type="Italic">Rhizobium leguminosarum</Emphasis> on the contents of cytoplasmic protein and free amino acids in the roots of pea seedlings

The changes in the contents of protein and free amino acids in pea plants inoculated with Rhizobium leguminosarum were studied taking into account the susceptibility of roots to root nodule bacteria. The content of cytoplasmic protein during infection increased in the actively growing root zone (0–5 mm) and decreased in the root zones susceptible to rhizobia (5–20 mm from the root tip). The quantitative composition of free amino acids changed essentially upon inoculation of pea seedlings with R. leguminosarum.     

Three new species and one new combination of <Emphasis Type="Italic">Gypsoplaca</Emphasis> (lichenized Ascomycota) from the Hengduan Mountains in China

Three new species are described as follows: Gypsoplaca alpina H. X. Shi and Li S. Wang, G. bullata H. X. Shi and Li S. Wang, and G. rosulata H. X. Shi, Li S. Wang, and Timdal sp. nov. Gypsoplaca alpina is characterized by a small thallus, upper cortex with very thick gelatinized layer, ascocarp warty with a cerebriform ridging, and by growing on soil in alpine meadows. Gypsoplaca bullata characterized by having bullate ascocarp, uneven to bullate thallus. Gypsoplaca rosulata is characterized by rosulate thallus and 4–6-spored asci. The new combination Gypsoplaca blastidiata (Zhurb) H. X. Shi and Li S. Wang comb. & stat. nov. is proposed. A phylogenetic tree is established, based on combined sequences of ITS and nrLSU, and all species formed monophyletic clades. Two main chemical compounds, brialmontin and sesterterpenes, are reported for the first time from Gypsoplaca by analysis of NMR and MS data. Triterpenoids are absent in Chinese Gypsoplaca specimens.     

The biogeochemical background and trace metal accumulation by brown algae of the genus <Emphasis Type="Italic">Fucus</Emphasis> in coastal waters of the Sea of Japan,the Sea of Okhotsk,and the White Sea

The ranges of background concentrations of Fe, Mn, Zn, Cu, and Cd in Fucus algae from the Sea of Japan, the Sea of Okhotsk, and the White Sea were determined. The lower background threshold of element concentration was calculated as Me₁₅–2MAD₁₅, the upper background threshold was determined as Me+2MAD. The upper background threshold of the metal contents in Fucus algae can be used as the maximum permissible regional concentration in assessing pollution levels of marine habitats. Comparison of the Me₁₅–2MAD₁₅ values showed that Fucus algae of the White Sea had increased Mn and decreased Cd concentrations compared to those in Pacific algae. The concentrations of Zn were higher in Fucus algae of the Sea of Japan than in macrophytes from the White Sea and the Sea of Okhotsk. The background concentration range of Fe in Fucus algae of the Sea of Japan was much narrower compared to that in their counterparts from the White Sea and the Sea of Okhotsk. The background metal concentrations in Fucus algae are specific to a sea region; their variations are connected not only with the trace element contents in abiotic components of the environment, but also with the functioning of coastal ecosystems.     

Pharmacognostic study of aerial parts of Baikal skullcap (<Emphasis Type="Italic">Scutellaria baicalensis</Emphasis> Georgi)

A pharmacognostic study of aerial parts of Scutellaria baicalensis Georgi (Lamiaceae family) has been carried out. A merchandising analysis has been performed to determine the basic quality parameters of S. baicalensis. A chemical composition analysis has revealed the presence of organic acids, free carbohydrates, polysaccharides, amino acids, alkaloids, flavonoids, phenolic acids, anthocyanins, tannins, photosynthetic pigments, lipids, and triterpene compounds. Ethyl acetate and ethanol fractions, extracted from the aerial parts of S. baikalensis, have shown a significant antiradical activity.     

The characterization of lectins from the thripartite lichen <Emphasis Type="Italic">Peltigera aphthosa</Emphasis> (L.) Willd.

Agglutinin from Pemtigera aphthosa (L.) Willd. was isolated and its characteristics were obtained. It is a glycoprotein lectin with D-galactose specificity. Lectin concentration decreases from the apical parts of the thallus to its basal region. Finer differences in the agglutinating activity of different parts of the thallus reflect physiological differences in certain areas and their role in the functioning of lichen symbiosis.     

Distribution of representatives of the genus <Emphasis Type="Italic">Neoabbottiella</Emphasis> (Rhodophyta: Halymeniales) in the Russian Far Eastern Seas and taxonomic status of <Emphasis Type="Italic">N. valentinae</Emphasis> N. Klochkova et Pisareva, 2013

The new information on distribution of the red algae Neoabbottiella in the Russian Far East changes the concept of the areal of these algae and their preferred temperatures. A comparison of N. valentinae N. Klochkova et Pisareva with Schizymenia dubyi f. palmata Yamada collected by Y. Yamada from Urup Island and stored with its type and paratypes in the collection of Hokkaido University Museum (Sapporo, Japan) showed their conspecificity. Based on the data, N. valentinae was recognized as a synonym of Schizymenia dubyi f. palmata. Since this form does not possess the main generic feature of the genus Schizymenia and at the same time has well-expressed generic features of Neoabbottiella, the form S. dubyi f. palmata is transferred to this genus with the name N. palmata (Yamada) N. Klochkova et Pisareva.     

Metatrancriptomic analysis from the Hepatopancreas of adult white leg shrimp (<Emphasis Type="Italic">Litopenaeus vannamei)</Emphasis>

The amoeba, Mayorella viridis contains several hundred symbiotic green algae in its cytoplasm. Transmission electron microscopy revealed strong resemblance between symbiotic algae from M. viridis the symbiotic Chlorella sp. in the perialgal vacuoles of Paramecium bursaria and other ciliates. Although it is thought that the M. viridis and symbiotic algae could be model organisms for studying endosymbiosis between protists and green algae, few cell biological observations of the endosymbiosis between M. viridis and their symbiotic algae have been published. In this study, we characterized the specificity of endosymbiotic relationships between green algae and their hosts. Initially, we established stable cultures of M. viridis in KCM medium by feeding with Chlorogonium capillatum. Microscopic analyses showed that chloroplasts of symbiotic algae in M. viridis occupy approximately half of the algal cells, whereas those in P. bursaria occupy entire algal cells. The symbiotic algae in P. bursaria contain several small spherical vacuoles. The labeling of actin filaments using Acti-stain? 488 Fluorescent Phalloidin revealed no relationship between host actin filaments and symbiotic algal localization, although the host mitochondria were localized around symbiotic algae. Symbiotic algae from M. viridis could infect algae-free P. bursaria but could not support P. bursaria growth without feeding, whereas the original symbiotic algae of P. bursaria supported its growth without feeding. These data indicated the specificity of endosymbiotic algae relationships in M. viridis and P. bursaria.     

The nepionic stage of <Emphasis Type="Italic">Solenomeris</Emphasis> Douvillé, 1924 (Acervulinidae,Foraminiferida): new observations from the uppermost Maastrichtian-early Danian of Austria (Kambühel Formation,Northern Calcareous Alps)

In the shallow-water limestone succession of the Kambühel Formation (upper Maastrichtian to upper Thanetian, Northern Calcareous Alps), the benthic-vagile to encrusting-sessile acervulinid foraminifer Solenomeris Douvillé was identified in upper Maastrichtian and, more commonly, lower Danian rocks. We report on the free nepionic (juvenile) stage of this foraminifer, its relation to the adult encrusting stage, and competitive interaction with coralline algae. Whereas morphology and chamber arrangement of the free nepionts clearly assign the Alpine specimens to Solenomeris, the chamber shape in axial sections differs from the Eocene type-species S. ogormani, accounting for classification in open nomenclature. Nepiont chamber shape can be used as a further specific criterion besides size of adult chambers and, perhaps, growth habit. A two-stage concept of ontogenetic development is presented. Solenomeris flourished in early Danian high-stress environments (perhaps related to cooler waters and/or elevated nutrient level) under conditions less favorable for encrusting corallines that competed for substrate. Absence of Solenomeris in upper Danian limestones and in upper Thanetian coralgal limestones that form the top of the Kambühel Formation may be related to recovery of coralline algae in shallow-marine environments of low to moderate nutrient level.     

Cloning,characterization and function analysis of <Emphasis Type="Italic">DAX1</Emphasis> in Chinese loach (<Emphasis Type="Italic">Paramisgurnus dabryanus</Emphasis>)

The mechanisms of sex determination and differentiation have not been elucidated in most fish species. In this study, the full-length cDNAs of DAX1 was cloned and characterized in aquaculture fish Chinese loach (Paramisgurnus dabryanus), designated as Pd-DAX1. The cDNA sequence of Pd-DAX1 was 1261 bp, including 795 bp open reading frame (ORF) encoding 264 amino acids. Pd-DAX1 shares highly identical sequence with DAX1 homologues from different species. The expression profiles of Pd-DAX1 in different developmental stages and diverse adult tissues were analyzed by quantitative real-time RT-PCR and in situ hybridization (ISH). Pd-DAX1 was continuously expressed during embryogenesis, with the extensive distribution in the development of the central nervous system. Tissue distribution analysis revealed that Pd-DAX1 expressed widely in adult tissues, with the highest expression level found in testis, moderate level in ovary, showing a sex-dimorphic expression pattern. Pd-DAX1 mainly located in spermatogonia cells, spermatocytes, primary oocytes and previtellogenic oocyte cells, implying that Pd-DAX1 may involve in gametogenesis. These preliminary findings suggest that Pd-DAX1 gene is highly conserved during vertebrate evolution and involved in a wide range of developmental processes including embryogenesis, central nervous system development and gonad development.     

Changes in leaf epicuticular wax,gas exchange and biochemistry metabolism between <Emphasis Type="Italic">Jatropha mollissima</Emphasis> and <Emphasis Type="Italic">Jatropha curcas</Emphasis> under semi-arid conditions

Jatropha curcas and Jatropha mollissima plants were evaluated under conditions of high (HSM) and low (LSM) soil moisture in a semi-arid environment, as changes in the content and concentration of epicuticular wax and the leaf metabolism which could have a relationship with drought tolerance. Besides epicuticular wax, gas exchange, antioxidant system and biochemical parameters of the photosynthetic metabolism were measured. The epicuticular wax content increased only in J. mollissima leaves 95 % under LSM, when compared with HSM conditions. Therefore, J. curcas invested less in the production of long-chain n-alkanes than did J. mollissima under LSM conditions. J. mollissima plants showed the highest CO₂ assimilation rate during the HSM period compared to J. curcas. Both species showed high stability in some leaf biochemistry products, highlighting the highest sugar content, free amino acids, total soluble protein, and photosynthetic pigments in the leaves of J. mollissima plants under both of the soil moisture conditions. Moreover, the stability and performance of the different parameters, such as morphologic variables, seem to allow J. mollissima plants to tolerate semi-arid conditions.     

Taste attractiveness of free amino acids for juveniles of Persian sturgeon <Emphasis Type="Italic">Acipenser persicus</Emphasis>

Taste preferences towards 20 free amino acids (L-isomers, 0.1–0.0001 M) were determined in juveniles of Persian sturgeon Acipenser persicus. It was found that most amino acids (16) had a positive effect on extraoral gustatory reception (increased the frequency of catching artificial pellets by fishes). The most efficient amino acids were the following: threonine, histidine, arginine, asparagine, phenylalanine, cysteine, glutamine, and glycine. Fifteen amino acids were efficient for intraoral gustatory receptors: serine, arginine, cysteine, histidine, alanine, and some others. The presence of these amino acids in pellets increased the consumption. Amino acids decreasing the frequency of catching or consumption of pellets were not found. Highly significant positive correlation was found between the amino acid ranges mediated by the extraoral and intraoral gustatory reception. The fishes demonstrated maximum sensitivity to aspartic acid, 0.01 and 0.001 M, respectively, for intraoral and extraoral gustatory systems. The threshold concentrations of arginine and glycine were higher for the intraoral gustatory system (0.01 M) than for the extraoral one (0.1 M). The comparison of Persian sturgeon with sturgeon fishes (Russian sturgeon A. gueldenstaedtii, Siberian sturgeon A, baerii, and starred sturgeon A. stellatus) studied earlier confirmed high species specificity of intraoral taste preferences in representatives of Acipenser genus. Species specificity of extraoral taste ranges was less pronounced.     

Cloning and functional characterization of the <Emphasis Type="Italic">SmNCED3</Emphasis> in <Emphasis Type="Italic">Salvia miltiorrhiza</Emphasis>

As one of the most important phytohormones, the abscisic acid (ABA) is often used to breed stress-tolerant crop lines with both higher yields and active ingredient contents. In higher plants, the 9-cis-epoxycarotenoid dioxygenase (NCED) has been found to be a regulatory enzyme involved in ABA biosynthesis. In research, the novel gene SmNCED3 was isolated from S. miltiorrhiza. The open reading frame of SmNCED3 was 1725-bp, and it was encoding 574 amino acids with a calculated molecular mass of 63,822 kDa, which was verified by the expression of SmNCED3 in E. coli. The deduced SmNCED3 amino acid sequence had high sequence homology with NCED sequences from other plants and contained a putative chloroplast transit targeting signal peptide at its N terminus. Phylogenetic analysis demonstrated that SmNCED3 had a closer affinity to NCED3 in Arabidopsis thaliana (AtNCED3). The 1732-bp 5′ flanking sequence of SmNCED3 was also cloned. It contained several phytohormone response elements, biotic or abiotic stress-related elements, and plant development-related elements. Real-time PCR revealed that SmNCED3 was highly expressed in leaves, and was strongly induced by exogenous ABA. A subcellular localization experiment indicated that SmNCED3 was located in chloroplast stroma, chloroplast membranes, and thylakoid membranes. The overexpression of SmNCED3 promoted ABA accumulation. These results indicated that SmNCED3 might be a rate-limiting gene regulating ABA biosynthesis, and improving abiotic stresses tolerance and active ingredient contents in plants.     

Molecular characterization of <Emphasis Type="Italic">bmyC</Emphasis> gene of the mosquito pupicidal bacteria, <Emphasis Type="Italic">Bacillus amyloliquefaciens</Emphasis> (VCRC B483) and in silico analysis of bacillomycin D synthetase C protein

A strain of Bacillus amyloliquefaciens (VCRC B483) exhibiting mosquito pupicidal, keratinase and antimicrobial activities was isolated from mangrove forest ecosystem of Andaman and Nicobar Islands. Molecular characterization of the strain showed the presence of lipopeptide encoding bmyC gene. Phylogenetic tree based on protein sequence of this gene exhibited homology with mycosubtilin synthetase of Bacilus atropheus and Iturin synthetase of Bacillus subtilis and B. amyloliquefaciens. This is the first report on the evolutionary conservation of amino acids concerned with the function and structure of bmyC protein of B. amyloliquefaciens. The presence of valine at the 1197th position in our strain was found to be unique and different from the existing strains of B. subtilis and B. amyloliquefaciens. Molecular modelling studies revealed significant changes in the structure of epimerization domain of the bmyC protein with A1197V variation. Crude metabolite of this strain exhibited antifungal activity against Fusarium sp. and Carvularia sp.     

Comparative Analysis of the Amino Acid Spectrum of Blood Plasma in Chiroptera (<Emphasis Type="Italic">Vespertilio murinus</Emphasis> L., 1758 and <Emphasis Type="Italic">Myotis dasycneme</Emphasis> B., 1825) in the Fauna of the Ural Mountains

The article presents the results of a new comparative analysis of free amino acids in the blood plasma of representatives of insectivorous Chiroptera (Mammalia: Vespertilionidae) in the fauna of the Ural Mountains: the pond bat (Myotis dasycneme Boie, 1825) and the parti-colored bat (Vespertilio murinus Linnaeus, 1758). This is the first study to show the species variability of free amino acids in resident and migratory species of bats from different ecosystems of the Ural Region.     

Molecular Characterization of a Newly Identified Subfamily Member of Penaeidin from two Penaeid Shrimps, <Emphasis Type="Italic">Fenneropenaeus indicus</Emphasis> and <Emphasis Type="Italic">Metapenaeus monoceros</Emphasis>

Penaeidins are a major group of antimicrobial peptides found in penaeid shrimps. This study reports a new isoform of penaeidin from the hemocytes of Indian white shrimp, Fenneropenaeus indicus (Fi-PEN, JX657680), and the pink shrimp, Metapenaeus monoceros (Mm-PEN, KF275674). Mm-PEN is also the first antimicrobial peptide to be identified from M. monoceros. The complete coding sequences of the newly identified Fi-PEN and Mm-PEN consisted of an ORF of 338 bp encoding 71 amino acids with a predicted molecular weight of 5.66 kDa and a pI of 9.38. The penaeidins had its characteristic signal peptide region (19 amino acids), which was followed by a mature peptide with a proline-rich domain (24 amino acids) at the N-terminal region and a cysteine-rich domain (28 amino acids) at the C-terminal region, designating it to penaeidin-3 subgroup. Structural analysis revealed an alpha-helix in its secondary structure and an extended structure at the proline-rich domain. The newly identified penaeidin isoform showed maximum similarity of 63 % to a penaeidin-3 isoform of P. monodon, which further proves it to be a new isoform. Phylogenetic analysis showed that it possessed similar evolutionary status like other penaeidins, which has subsequently diverged at different phases of evolution. The wide distribution of penaeidins in penaeid shrimps indicates the importance of these AMPs in the innate immunity.