Sequence-level analysis of the diploidization process in the triplicated FLOWERING LOCUS C region of Brassica rapa

Strong evidence exists for polyploidy having occurred during the evolution of the tribe Brassiceae. We show evidence for the dynamic and ongoing diploidization process by comparative analysis of the sequences of four paralogous Brassica rapa BAC clones and the homologous 124-kb segment of Arabidopsis thaliana chromosome 5. We estimated the times since divergence of the paralogous and homologous lineages. The three paralogous subgenomes of B. rapa triplicated 13 to 17 million years ago (MYA), very soon after the Arabidopsis and Brassica divergence occurred at 17 to 18 MYA. In addition, a pair of BACs represents a more recent segmental duplication, which occurred approximately 0.8 MYA, and provides an exception to the general expectation of three paralogous segments within the B. rapa genome. The Brassica genome segments show extensive interspersed gene loss relative to the inferred structure of the ancestral genome, whereas the Arabidopsis genome segment appears little changed. Representatives of all 32 genes in the Arabidopsis genome segment are represented in Brassica, but the hexaploid complement of 96 has been reduced to 54 in the three subgenomes, with compression of the genomic region lengths they occupy to between 52 and 110 kb. The gene content of the recently duplicated B. rapa genome segments is identical, but intergenic sequences differ.     

An Insect Multiligand Recognition Protein Functions as an Opsonin for the Phagocytosis of Microorganisms

We characterize a novel pathogen recognition protein obtained from the lepidopteran Galleria mellonella. This protein recognizes Escherichia coli, Micrococcus luteus, and Candida albicans via specific binding to lipopolysaccharides, lipoteichoic acid, and β-1,3-glucan, respectively. As a multiligand receptor capable of coping with a broad variety of invading pathogens, it is constitutively produced in the fat body, midgut, and integument but not in the hemocytes and is secreted into the hemolymph. The protein was confirmed to be relevant to cellular immune response and to further function as an opsonin that promotes the uptake of invading microorganisms into hemocytes. Our data reveal that the mechanism by which a multiligand receptor recognizes microorganisms contributes substantially to their phagocytosis by hemocytes. A better understanding of an opsonin with the required repertoire for detecting diverse invaders might provide us with critical insights into the mechanisms underlying insect phagocytosis.     

Proteome Analysis of Cellular Response of Pseudomonas putida KT2440 to Tetracycline Stress

Tetracycline-induced proteome of Pseudomonas putida KT2440 was analyzed by 2-D gel electrophoresis and matrix-assisted laser desorption ionization–time of flight/mass spectrum (NALDI-TOF/MS) in order to understand cellular response to tetracycline. Of the proteins upregulated in a culture medium containing subinhibitory concentration of tetracycline (50 μg/mL), we identified 38 proteins from cytosol and precipitated fractions by peptide mass fingerprinting and mass spectrum/mass spectrum analysis. Various amino acids ABC transporters, a ribose ABC transporter, and a sulfate ABC transporter were found to be upregulated. Protein synthesis-related proteins, stress proteins, energy metabolic enzymes, and unknown proteins were also strongly induced. Of the identified upregulated proteins, several proteins (isocitrate lyase, branched-chain amino acid ABC transporter, superoxide dismutase, etc.) were also upregulated under phenol-induced stress condition. These results demonstrate that tetracycline at a high concentration induced comprehensive stress in P. putida KT2440 and the global induction of proteins related to bacteria survival. Proteome analysis was found to be a useful tool for the elucidation of antibiotic-induced proteins in the present study.     

Toward unraveling the structure of Brassica rapa genome

Genomic research in any organism encompasses understanding structure of the target genome and genes, their function, and evolution. Brassica rapa , which is phylogenetically related to Arabidopsis thaliana , is an important species with respect to its uses as vegetable, oil, and fodder. The availability of suitable genetic and genomic resources is a prerequisite to undertake genomic research in B. rapa . We have developed reference mapping populations of Chinese cabbage ( B. rapa ssp. pekinensis ) comprising 78 doubled haploid lines and over 250 recombinant inbred lines. Two Bacterial Artificial Chromosome (BAC) libraries, generated by restriction enzymes Hin dIII (KBrH) and Bam HI (KBrB), comprise 56 592 and 50 688 clones, respectively. We have also constructed 22 cDNA libraries from different plant tissues consisting of 104 914 clones with an average length of 575 bp. Initial BAC-end sequence analysis of 1473 clones of the KBrH library led us to understand the structure of B. rapa genome with respect to extent of genic sequences and their annotation, and relative abundance of different types of repetitive DNAs. Full-length sequence analysis of BAC clones revealed extensive triplication of B. rapa DNA segments coupled with variable gene losses within the segments. The formulation of the 'Multinational Brassica Genome Project' has laid the foundation to sequence the complete genome of B. rapa ssp. pekinensis by the international Brassica research community. It has been proposed to undertake BAC-to-BAC sequencing of genetically mapped seed BACs. In recent years, development of bioinformatics tools in Brassica has given a boost to structural genomics research in Brassica species. The research undertaken with the availability of various genomic resources in the public domain has added to our understanding of the structure of B. rapa .     

Isolation and characterization of a novel lipase from a metagenomic library of tidal flat sediments: evidence for a new family of bacterial lipases

We cloned lipG, which encoded a lipolytic enzyme, from a Korean tidal flat metagenomic library. LipG was related to six putative lipases previously identified only in bacterial genome sequences. These enzymes comprise a new family. We partially characterized LipG, providing the first experimental data for a member of this family.     

Comparative seed morphology of Boronia and related genera (Boroniinae: Rutaceae) and its systematic implications

Comparative seed morphology of Boronia Sm., Boronella Baill., Neobyrnesia J. A. Armstr. and Zieria Sm., 31 taxa in total, in Rutaceae is studied. The seeds are ventrally ellipsoid to ovoid (0.93–4.42 × 0.50–2.31 × 0.95–2.70 mm). Colour is brown to black. An elaiosome which is related to myrmechocory was observed in Boronella, Boronia, Neobyrnesia and Zieria. Six testa surface pattern types are identified: Type I (Boronia sect. Boronia) – smooth, and cells not barely discernable; Type II (Boronella) – smooth and cells more or less rectangular and often cells elongated; Type III (Boronia sects. Boronia, Cyanothamnus, Imbricatae and Valvatae, Neobyrnesia) – colliculate with four subtypes by presence, absence of ridges or ridge characters; Type IV (Boronia sect. Alatae) – digit form; Type V (Boronia sect. Algidae) – alveolate; Type VI (Zieria) – ridged, the interridge area formed by both cells that form the ridges and cells that do not form the ridges. Seed morphology, especially testa surface patterns, was partially congruent with the current classification of Boronia, with most sections having apomorphies, and provided additional support for the placement of Boronella near Boronia sect. Boronia. Hilum (surrounded by labiose margin or not) and elaiosome shape also offer useful diagnostic characters for Boronia at infrageneric levels. Neobyrnesia and Zieria both had unique testa surface patterns. The seed morphological data contribute to our understanding of systematic relationship in the subtribe Boroniinae.     

Shewanella seohaensis sp. nov., isolated from a tidal flat sediment

A Gram-negative, motile and rod-shaped bacterial strain, designated S7-3^T, was isolated from a tidal flat sediment at Saemankum on the western coast of Korea. Phylogenetic analyses based on 16S rRNA gene and gyrB sequences showed that strain S7-3^T belonged to the genus Shewanella, clustering with Shewanella decolorationis S12^T. Strain S7-3^T exhibited 98.8 % 16S rRNA gene sequence similarity and 96.8 % gyrB sequence similarity to S. decolorationis S12^T, respectively. The 16S rRNA gene sequence similarity values between strain S7-3^T and other members of the genus Shewanella were in the range of 93.0–98.0 %. Strain S7-3^T contained simultaneously both menaquinones (MK) and ubiquinones (Q); the predominant menaquinone was MK-7 and the predominant ubiquinones were Q-7 and Q-8. The fatty acid profiles of strain S7-3^T and S. decolorationis JCM 21555^T were similar; major components were C_17:1 ω8c, iso-C_15:0 and iso-C_15:0 2-OH and/or C_16:1 ω7c. The DNA G+C content of strain S7-3^T was 51.8 mol% and its mean DNA–DNA relatedness value with S. decolorationis JCM 21555^T was 43 %. Differential phenotypic properties of strain S7-3^T, together with the phylogenetic and genetic distinctiveness, revealed that this strain is distinguishable from recognized Shewanella species. On the basis of the data presented, strain S7-3^T is considered to represent a novel Shewanella species, for which the name Shewanella seohaensis sp. nov. is proposed. The type strain is S7-3^T (=KCTC 23556^T = CCUG 60900^T).     

Ohioensin F suppresses TNF-α-induced adhesion molecule expression by inactivation of the MAPK, Akt and NF-κB pathways in vascular smooth muscle cells

AimsThe expression of cell adhesion molecules on vascular smooth muscle cells is central to leukocyte recruitment and progression of atherosclerotic disease. Ohioensin F, a chemical compound of the Antarctic moss Polyerichastrum alpinum, exhibited inhibitory activity against protein tyrosine phosphatase 1B and antioxidant activity. However, published scientific information regarding other biological activities and pharmacological function of ohioensin F is scarce. In the present study, we aimed to examine the in vitro effects of ohioensin F on the ability to suppress TNF-α-induced adhesion molecule expression in vascular smooth muscle cells (VSMCs).Main methodsThe inhibitory effect of ohioensin F on TNF-α-induced upregulation in expression of adhesion molecules was investigated by enzyme-linked immunosorbent assay, cell adhesion assay, RT-PCR, western blot analysis, immunofluorescence, and transfection and reporter assay, respectively.Key findingsPretreatment of VSMCs with ohioensin F at nontoxic concentrations of 0.1–10 μg/ml dose-dependently inhibited TNF-α-induced expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1). In addition, ohioensin F suppressed adhesion of THP-1 monocytes to TNF-α-stimulated VSMCs. Ohioensin F reduced TNF-α-induced production of intracellular reactive oxygen species (ROS) and phosphorylation of p38, ERK, JNK and Akt. Finally, ohioensin F inhibited TNF-α-induced CAM mRNA expression and NK-κB translocation.SignificanceThese results suggest a new mechanism of ohioensin F's anti-inflammatory action, owing to the negative regulation of TNF-α-induced adhesion molecule expression, monocyte adhesion and ROS production in vascular smooth muscle cells. Our finding also supports ohioensin F as a potential pharmacological, anti-inflammatory molecule that has a protective effect on the atherosclerotic lesion.