Diurnal Dynamics of Activity of Peptidases at Early Ontogenesis of Bleak <Emphasis Type="Italic">Alburnus alburnus</Emphasis> (L.) from Coastal Shallows of the Rybinsk Reservoir

The diurnal dynamics of the activity of peptidases in fish larvae by the example of bleak Alburnus alburnus (L.) as the dominant species in the open and protected shallows of the Rybinsk Reservoir has been studied for the first time. Two peaks of enzyme activity in the intestinal mucosa of fish larvae are revealed: morning and night. The data on species composition and daily dynamics of number of fish larvae in the shallows of different types are presented.     

Sexual Dimorphism in the Pelvis of <Emphasis Type=Italic>Microcebus</Emphasis>

Pelvic sexual dimorphism occurs in many anthropoid species and is often attributed to obstetric selection on female pelvic morphology. Few studies of pelvic dimorphism have included strepsirrhine taxa, which typically have relatively smaller infants than those of anthropoids. Because smaller female primates give birth to relatively larger infants, it is possible that the pelves of Microcebus, the smallest extant primate genus, will show some evidence of selection on obstetric adequacy. A comparison of adult female and neonatal body masses indicates that individual neonatal Microcebus are relatively large compared to adult female body mass, even though members of the taxon frequently produce twins. I examined variation in the bony pelvis within a sample of Microcebus. I measured specimens from a single locality, which probably represent 1 population. I measured 8 pelvic and 3 femoral variables to investigate skeletal size and pelvic size and shape dimorphism. Females significantly exceed males in absolute values of sacral width, pelvic height, pubic length, and distances from the pubic symphysis to the ischial tuberosity and points on the sacrum. Measurements of the femur are not significantly greater in females, suggesting that the pelvic differences are not due to skeletal size dimorphism. Significant pelvic shape or ratio differences, calculated via the geometric mean of 5 variables as the denominator, included greater relative pubic length and sacral width in females. Hence selection for obstetric adequacy may occur in the extremely small-bodied Microcebus.     

High-density AFLP map of nonbrittle rachis 1 (<Emphasis Type=Italic>btr1</Emphasis>) and 2 (<Emphasis Type=Italic>btr2</Emphasis>) genes in barley (<Emphasis Type=Italic>Hordeum vulgare</Emphasis> L.)

Wild relatives of barley disperse their seeds at maturity by means of their brittle rachis. In cultivated barley, brittleness of the rachis was lost during domestication. Nonbrittle rachis of occidental barley lines is controlled by a single gene (btr1) on chromosome 3H. However, nonbrittle rachis of oriental barley lines is controlled by a major gene (btr2) on chromosome 3H and two quantitative trait loci on chromosomes 5HL and 7H. This result suggests multiple mutations of the genes involved in the formation of brittle rachis in oriental lines. The btr1 and btr2 loci did not recombine in the mapping population analyzed. This result agrees with the theory of tight linkage between the two loci. A high-density amplified fragment-length polymorphism (AFLP) map of the btr1/btr2 region was constructed, providing an average density of 0.08 cM/locus. A phylogenetic tree based on the AFLPs showed clear separation of occidental and oriental barley lines. Thus, barley consists of at least two lineages as far as revealed by molecular markers linked to nonbrittle rachis genes.Electronic Supplementary Material Supplementary material is available for this article at An erratum to this article can be found at     

Efficient separation and purification of allophycocyanin from <Emphasis Type=Italic>Spirulina</Emphasis> (<Emphasis Type=Italic>Arthrospira</Emphasis>) <Emphasis Type=Italic>platensis</Emphasis>

Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0). In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A ₆₅₀/A ₂₈₀ of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g⁻¹ wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from 5.6 to 4.0. The absorbance ratio A ₆₅₀/A ₂₈₀ of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g⁻¹ wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE.     

A new name in <Emphasis Type=Italic>Pavetta</Emphasis> L. (<Emphasis Type=Italic>Rubiaceae</Emphasis>)

Summary  The name Pavetta modesta (Hiern) S. E. Dawson is a later homonym of P. modesta Bremek. Pavetta crystalensis is proposed as a new name.     

Dynamics of energy metabolism in ontogenesis of striped shield bug (<Emphasis Type=Italic>Graphosoma lineatum</Emphasis> L.) and cabbage moth (<Emphasis Type=Italic>Mamestra brassicae</Emphasis> L.)

Dynamics of growth and oxygen consumption during ontogenesis of insects with direct (striped shield bug Graphosoma lineatum L.) and indirect (cabbage moth Mamestra brassicae L.) development have been compared. The correlation between a character of energy metabolism alteration and peculiarities of development of the insects has been shown. Cyclic decrease of oxygen consumption during molt and sharp dropping during metamorphosis have been observed in insects with indirect development. The decrease of oxygen consumption has been observed in insects with direct development only during molts. The coefficient a of allometric dependence of oxygen consumption on body weight of imago for cabbage moth was two times higher than that for striped shield bug.     

Cryopreservation of <Emphasis Type=Italic>Robinia</Emphasis><Emphasis Type=Italic>pseudoacacia</Emphasis>

Cryopreservation of Robinia pseudoacacia explants by vitrification achieved 78% survival following the stepwise preculture of shoot tips in (0.3 + 0.5 + 0.7 M) sucrose with a 80 min incubation in PVS2; compared to 87% survival after desiccation of explants to 30% water content, following 3 days alginate bead (with glycerol and sucrose treatments) preculture in 0.7 M sucrose.     

Seasonal changes in water source of four families of loblolly pine (<Emphasis Type=Italic>Pinus</Emphasis><Emphasis Type=Italic>taeda</Emphasis> L.)

Source water utilized by four families of loblolly pine (Pinus taeda L.) was assessed by comparing the H isotope composition ('D) of xylem sap and of soil water from four depths (0-20 cm, 20-40 cm, 1.2 m, and 2.1 m) across 1 year. Soil water 'D values varied with soil moisture content in the well-drained, sandy site and at each of the four soil depths. In September and November 1997 and May through November 1998, xylem sap 'D values closely matched the soil water 'D values of the upper soil horizons (0-20 and 0-40 cm) and, therefore, reflected significant water uptake from upper regions of the soil profile. In March 1998, xylem sap 'D values closely matched the soil water 'D values of the 1.2 m soil depth and, therefore, indicated that trees during this portion of the growing season were obtaining their water from deep in the soil profile. Analysis of source water use with a two-ended mixing model in the 3 months of collection that exhibited a range of soil water 'D values across the soil profile confirmed that trees utilized different sources of water depending upon season of the year. In September 1997 and November 1998, source water uptake was primarily from the upper soil profile while in March 1998, source water uptake was from deep in the soil profile. With few exceptions, we did not find striking differences in source water use between drought-hardy families and those that were locally adapted.     

Functional characterization of the powdery mildew susceptibility gene <Emphasis Type=Italic>SmMLO1</Emphasis> in eggplant (<Emphasis Type=Italic>Solanum melongena</Emphasis> L.)

Eggplant (Solanum melongena L.) is one of the most important vegetables among the Solanaceae and can be a host to fungal species causing powdery mildew (PM) disease. Specific homologs of the plant Mildew Locus O (MLO) gene family are PM susceptibility factors, as their loss of function results in a recessive form of resistance known as mlo resistance. In a previous work, we isolated the eggplant MLO homolog SmMLO1. SmMLO1 is closely related to MLO susceptibility genes characterized in other plant species. However, it displays a peculiar non-synonymous substitution that leads to a T → M amino acid change at protein position 422, in correspondence of the MLO calmodulin-binding domain. In this study, we performed the functional characterization of SmMLO1. Transgenic overexpression of SmMLO1 in a tomato mlo mutant compromised resistance to the tomato PM pathogen Oidium neolycopersici, thus indicating that SmMLO1 is a PM susceptibility factor in eggplant. PM susceptibility was also restored by the transgenic expression of a synthetic gene, named s-SmMLO1, encoding a protein identical to SmMLO1, except for the presence of T at position 422. This indicates that the T → M polymorphism does not affect the protein role as PM susceptibility factor. Overall, the results of this work are of interest for the functional characterization of MLO proteins and the introduction of PM resistance in eggplant using reverse genetics.     

The surface topography of <Emphasis Type=Italic>Eudiplozoon nipponicum</Emphasis> (Monogenea) developmental stages parasitizing carp (<Emphasis Type=Italic>Cyprinus carpio</Emphasis> L.)

Using scanning electron microscopy (SEM), the external morphology of all developmental stages (egg, oncomiracidium, diporpa, just fused juvenile and adult) of the parasite, Eudiplozoon nipponicum (Monogenea, Diplozoidae), from the gills of carp was studied. During the ontogeny, the tegument, tegumentary and sensory structures are subsequently developed. The tegument of free swimming oncomiracidium occurs in two types — the ciliated and non-ciliated with numerous uniciliated sensory structures. An attachment apparatus starts to form during the oncomiracidium stage. Further developmental stages are adapted to the environment of the gills. Tegumentary folds become more apparent later in development and assist to the parasite’s attachment. In connection with its reproductive strategy, the two morphological structures of diporpa (ventral sucker and dorsal papilla) appear to play important role. On the gills, two individuals need to meet and these structures mediate the fusion between two diporpae. The hindbody of adult parasite is highly modified for attachment. The haptor, folds and lobular extensions are most developed. The forebody is flexible and able to interact with host gill tissue via the mouth and associated mouth structures. The process of food intake of the parasite was discussed.     

Morphological Variability of Roach <Emphasis Type=Italic>Rutilus rutilus</Emphasis> (L.) from the Volga Reach of Rybinsk Reservoir

A comparative analysis of literature data and our own results on the morphology of roach from the Volga Reach (Rybinsk Reservoir) in different years is conducted. Recently, (from 2007 to 2010), the morphological characteristics of the roach have been similar to those observed between 1976 and 1978. The roach is characterized by an intermediate position between the herbivorous and malacophagous morphs, but it is more similar to the herbivorous morph.     

Attenuation of fibrosis <Emphasis Type=Italic>in vitro</Emphasis> and <Emphasis Type=Italic>in vivo</Emphasis> with <Emphasis Type=Italic>SPARC</Emphasis> siRNA

Introduction  

SPARC is a matricellular protein, which, along with other extracellular matrix components including collagens, is commonly over-expressed in fibrotic diseases. The purpose of this study was to examine whether inhibition of SPARC can regulate collagen expression in vitro and in vivo, and subsequently attenuate fibrotic stimulation by bleomycin in mouse skin and lungs.     

<Emphasis Type=Italic>Agrobacterium</Emphasis>-mediated transformation of cotton (<Emphasis Type=Italic>Gossypium hirsutum</Emphasis> L.) via vacuum infiltration

AnAgrobacterium-mediated gene transfer system with recovery of putative transformants was developed for cotton (Gossypium hirsutum L.) cv. Cocker-312. Two-month-old hypocotyl-derived embryogenic calli were infected through agroinfiltration for 10 min at 27 psi in a suspension ofAgrobacterium tumefaciens strain GV3101 carrying tDNA with theGUS gene, encoding β-glucuronidase (GUS), and the neomycin phosphotransferase II (nptII) gene as a kanamycin-resistant plant-selectable marker. Six days after the histochemicalGUS assay was done, 46.6% and 20%GUS activity was noted with the vacuum-infiltration and commonAgrobacterium-mediated transformation methods, respectively. The transformed embryogenic calli were cultured on selection medium (100 mg/L and 50 mg/L kanamycin for 2 wk and 10 wk, respectively) for 3 mo. The putative transgenic plants were developed via somatic embryogenesis (25 mg/L kanamycin). In 4 independent experiments, up to 28.23% transformation efficiency was achieved. PCR amplification and Southern blot analysis fo the transformants were used to confirm the integration of the transgenes. Thus far, this is the only procedure available for cotton that can successfully be used to generate cotton transformants.     

Biochemical Characterization of Cytokinin Oxidases/Dehydrogenases from <Emphasis Type=Italic>Arabidopsis thaliana</Emphasis> Expressed in <Emphasis Type=Italic>Nicotiana tabacum</Emphasis> L.

Transgenic tobacco plants overexpressing single Arabidopsis thaliana cytokinin dehydrogenase (CKX, EC 1.5.99.12) genes AtCKX1, AtCKX2, AtCKX3, AtCKX4, AtCKX5, AtCKX6, and AtCKX7 under the control of a constitutive 35S promoter were tested for CKX-enzymatic activity with varying pH, electron acceptors, and substrates. This comparative analysis showed that out of these, only AtCKX2 and AtCKX4 were highly active enzymes in reaction with isoprenoid cytokinins (N ⁶ -(2-isopentenyl)adenine (iP), zeatin (Z)) and their ribosides using the artificial electron acceptors 2,6-dichlorophenol indophenol (DCPIP) or 2,3-dimethoxy-5-methyl-1,4-benzoquinone (Q₀). Turnover rates of these cytokinins by four other AtCKX isoforms (AtCKX1, AtCKX3, AtCKX5, and AtCKX7) were substantially lower, whereas activity of AtCKX6 was almost undetectable. The isoenzymes AtCKX1 and AtCKX7 showed significant preference for cytokinin glycosides, especially N ⁶ -(2-isopentenyl)adenine 9-glucoside, under weakly acidic conditions. All enzymes preferentially cleave isoprenoid cytokinins in the presence of an electron acceptor, but aromatic cytokinins are not resistant and are degraded with lower reaction rates as well. Cytokinin nucleotides, considered as resistant to CKX attack until now, were found to be potent substrates for some of the CKX isoforms. Substrate specificity of AtCKXs is discussed in this study with respect to the structure of the CKX active site. Further biochemical characterization of the AtCKX1, AtCKX2, AtCKX4 and AtCKX7 enzymes showed pH-dependent activity profiles.     

Molecular phylogeny of the <Emphasis Type=Italic>Sinocyclocheilus</Emphasis> (Cypriniformes: Cyprinidae) fishes in northwest part of Guangxi,China

The northwestern part of Guangxi is a typical karst landform. There are about ten species of the genus Sinocyclocheilus distributed in the region. To investigate the phylogenetic relationships and resolve the taxonomic status of these species, we determined nucleotide sequences of complete mitochondrial cytochrome b gene of 11 ingroup species and one outgroup species Barbodes laticeps. Phylogenetic trees were reconstructed using maximum parsimony and maximum likelihood analyses. Phylogenetic results showed that these species cluster as one clade with strong support. The 95% credibility interval (CI) of the divergence time between the species in the northwest part of Guangxi and other species in the genus Sinocyclocheilus is 8.28–12.40Mya. For the karst environment, these species show marked, recent genetic differentiation. From the overall consideration of morphology and molecular data Sinocyclocheilus tianeensis should be a valid species.     

<Emphasis Type=Italic>In vitro</Emphasis> organogenesis of <Emphasis Type=Italic>Passiflora alata</Emphasis>

Passiflora alata in vitro organogenesis was studied based on explant type, culture medium composition, and incubation conditions. The results indicated that the morphogenic process occurred more efficiently when hypocotyl segment-derived explants were cultured in media supplemented with cytokinin and AgNO₃ incubated under a 16-h photoperiod. The shoot bud elongation and plant development were obtained by transferring the material to MSM culture medium supplemented with GA₃ and incubated in flasks with vented lids. Histological analyses of the process revealed that the difficulties in obtaining plants could be related to the development of protuberances and leaf primordia structures, which did not contain shoot apical meristem. Roots developed easily by transferring elongated shoots to 1/2 MSM culture medium. Plant acclimatization occurred successfully, and somaclonal variation was not visually detected. The efficiency of this organogenesis protocol will be evaluated for genetic transformation of this species to obtain transgenic plants expressing genes that can influence the resistance to Cowpea aphid borne mosaic virus.     

Presence of <Emphasis Type=Italic>C. albidus</Emphasis>, <Emphasis Type=Italic>C. laurentii</Emphasis> and <Emphasis Type=Italic>C. uniguttulatus</Emphasis> in Crop and Droppings of Pigeon Lofts (<Emphasis Type=Italic>Columba livia</Emphasis>)

Columba livia is an important reservoir and carrier of Cryptococcus neoformans, Cryptococcus uniguttulatus, Cryptococcus laurentii and Cryptococcus albidus. Upper digestive tract of this species is also known as a habitat for Cryptococcus neoformans. Given the increasing clinical interest of this microorganism, 331 swabs from crop and 174 dropping samples from pigeon lofts in Grand Canary Island have been studied. The obtained results show an extensive presence samples 81 positive (24.47%) of Cryptococcus spp. in analysed crops: 32 (9.66%) for C. neoformans, 24 (7.2%) for C. uniguttulatus, 23 (6.9%) for C. albidus and 2 (0.6%) for C. laurentii. In the same way, Cryptococcus spp was also isolated in 82 (47.13%), dropping samples: C. neoformans in 59 (33.9%), C. uniguttulatus, in 9 (5.17%), C. laurentii in 8 (4.59%) and C. albidus in 6 (3.44%) of the investigated samples, respectively. The cryptococcosis produced by species of cryptococci other than C. neoformans has become more important during the last decade, supporting the study on the role of pigeon in the epidemiology of this disease.     

Chromosomes are eliminated in the symmetric fusion between <Emphasis Type=Italic>Arabidopsis </Emphasis><Emphasis Type=Italic>thaliana</Emphasis> L. and <Emphasis Type=Italic>Bupleurum scorzonerifolium</Emphasis> Willd.

In order to investigate chromosome elimination in symmetric somatic hybridization between Bupleurum scorzonerifolium and Arabidopsis thaliana, protoplasts were isolated from suspension cultures of both A. thaliana and B. scorzonerifolium parents. Biparental protoplasts were mixed at a rate of 1.5:1 and fused with PEG-method. After protoplast fusion, the products were cultured in the P5 liquid medium for microcallus formation. Single cell lines formed from microcalli after subculturing on the MB1 (Xia and Chen, Plant Sci 120:197–203, 1996) solid medium. The putative somatic hybrid cell lines were identified by cytological and molecular analysis. Of the 132 somatic cell lines generated, 16 were identified as somatic hybrids, with the phenotypes resembled B. scorzonerifolium parent. These hybrids showed a complete set of B. scorzonerifolium chromosome and 0–2 small chromosome(s) of A. thaliana. A few of them showed nuclear and cytoplasmic SSR fragments of A. thaliana. These hybrid cell lines could differentiate to green spots, buds/leaves through complementation of regeneration ability. The chromosomes elimination of A. thaliana was discussed. Wang Minqin and Zhao Junsheng contributed equally to the work.