Arthrobacter bambusae sp. nov., isolated from soil of a bamboo grove.

A Gram-stain-positive, aerobic, motile by gliding, rod-shaped bacterial strain, THG-GM18(T), was isolated from soil of a bamboo grove. Strain THG-GM18(T) was able to grow in the presence of up to 6.0 % (w/v) NaCl, at 4-37 °C and at pH 7.0-10.0 in R2A medium. Based on 16S rRNA gene sequence similarity, strain THG-GM18(T) was closely related to species of the genus Arthrobacter. The most closely related strains to strain THG-GM18(T) are Arthrobacter ramosus CCM 1646(T) (98.5 % similarity), Arthrobacter nitroguajacolicus G2-1(T) (98.4 %), Arthrobacter nicotinovorans DSM 420(T) (98.2 %), Arthrobacter aurescens DSM 20116(T) (98.1 %) and Arthrobacter chlorophenolicus A6(T) (98.0 %). Strain THG-GM18(T) possessed chemotaxonomic properties consistent with those of members of the genus Arthrobacter, such as peptidoglycan type A3α (l-Lys-l-Ala-l-Thr-l-Ala), MK-9 as major menaquinone and anteiso- and iso-branched compounds (anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0) as major cellular fatty acids. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, an unidentified phosphoglycolipid, unidentified phospholipids, unidentified aminolipids, an unidentified glycolipid and unidentified lipids. The G+C content of the genomic DNA was 61.0 mol%. The DNA-DNA relatedness values between strain THG-GM18(T) and its closest phylogenetic neighbours were below 26.0 %. The results of physiological and biochemical tests allowed the differentiation of strain THG-GM18(T) from species of the genus Arthrobacter with validly published names. Arthrobacter bambusae sp. nov. is the proposed name, and the type strain is THG-GM18(T) ( = KACC 17531(T) = JCM 19335(T)).

Chryseobacterium gwangjuense sp. nov., isolated from soil.

A Gram-stain-negative, strictly aerobic, non-motile, rod-shaped bacterial strain, THG-A18(T), was isolated from soil of Gwangju province in South Korea. Strain THG-A18(T) grew optimally at 25-30 °C, at pH 7.0-8.0 and in the absence of NaCl. Strain THG-A18(T) displayed ß-glucosidase activity, which enabled it to convert ginsenoside Rb1 to Rd. According to 16S rRNA gene sequence analysis, strain THG-A18(T) was shown to belong to the genus Chryseobacterium. The closest phylogenetic neighbours were Chryseobacterium ginsenosidimutans THG 15(T) (97.9 % 16S rRNA gene sequence similariity), C. defluvii B2(T) (97.7 %), C. daeguense K105(T) (97.6 %), C. taiwanense BCRC 17412(T) (97.5 %), C. indoltheticum LMG 4025(T) (97.4 %), C. gregarium P 461/12(T) (97.4 %) and C. lathyri RBA2-6(T) (97.3 %), but DNA-DNA relatedness values between these strains and strain THG-A18(T) were below 41.9 %. The G+C content of the genomic DNA was 36.4 mol%. The major respiratory quinone (MK-6) and fatty acids [iso-C15 : 0, iso-C17 : 0 3-OH, summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) and summed feature 9 (comprising iso-C17 : 1ω9c and/or 10-methyl C16 : 0)] supported the affiliation of strain THG-A18(T) with the genus Chryseobacterium. The polar lipids of strain THG-A18(T) were phosphatidylethanolamine, four unidentified aminolipids and seven unidentified lipids. A number of physiological and biochemical tests allowed phenotypic differentiation of strain THG-A18(T) from recognized species of the genus Chryseobacterium. The name Chryseobacterium gwangjuense sp. nov. is proposed, with THG-A18(T) ( = KACC 16227(T) = LMG 26579(T)) as the type strain.

Flavobacterium kyungheensis sp. nov., isolated from soil of a ginseng field.

A Gram-staining negative, strictly aerobic, motile by gliding, non-spore-forming, pale yellow pigmented and rod-shaped bacterium designated strain THG-107(T) was isolated from soil of a ginseng field on Ganghwa Island in the Republic of Korea and its taxonomic position was investigated by using a polyphasic study. Growth of strain THG-107(T) was found to occur at 4-37 °C (optimum, 20-30 °C), at pH 5.5-10 (optimum, pH 7.0) and in the presence of 0-1 % (w/v) NaCl (optimum, absence) on R2A agar. On the basis of 16S rRNA gene sequence similarity, strain THG-107(T) was shown to belong to the family Flavobacteriaceae and was related to Flavobacterium denitrificans ED5(T) (99.1 % similarity). The G+C content of the genomic DNA was determined to be 34.2 mol%. These results are consistent with characteristics of members of the genus Flavobacterium. The only isoprenoid quinone detected in strain THG-107(T) was menaquinone-6 (MK-6) and the major polyamine was identified as homospermidine (82.9 %). The major polar lipid detected was phosphatidylethanolamine and the major cellular fatty acids were identified as iso-C15:0 (26.3 %), iso-C17:0 3OH (12.6 %) and summed feature 3 (comprising C16:1 ω7c and/or C16:1 ω6c; 11.6 %). Flexirubin-type pigments were found to be present. Strain THG-107(T) has ß-glucosidase activity to convert ginsenosides Rb1 and Rd into Gyp17 and F2. DNA-DNA hybridization with F. denitrificans ED5(T) was 52 %. Strain THG-107(T) could be distinguished from F. denitrificans ED5(T) and the other species of the genus Flavobacterium by its phylogenetic and genetic distinctiveness and by several phenotypic properties. Therefore, strain THG-107(T) is considered to represent a novel species in the genus Flavobacterium, for which the name Flavobacterium kyungheensis sp. nov. is proposed (type strain THG-107(T) = KACC 16219(T) = LMG 26575(T)).

Small RNA regulation of ovule development in the cotton plant, G. hirsutum L.

BACKGROUND: The involvement of small RNAs in cotton fiber development is under explored. The objective of this work was to directly clone, annotate, and analyze small RNAs of developing ovules to reveal the candidate small interfering RNA/microRNAs involved in cotton ovule and fiber development. RESULTS: We cloned small RNA sequences from 0-10 days post anthesis (DPA) developing cotton ovules. A total of 6691 individual colonies were sequenced from 11 ovule small RNA libraries that yielded 2482 candidate small RNAs with a total of 583 unique sequence signatures. The majority (362, 62.1%) of these 583 sequences were 24 nt long with an additional 145 sequences (24.9%) in the 21 nt to 23 nt size range. Among all small RNA sequence signatures only three mirBase-confirmed plant microRNAs (miR172, miR390 and ath-miR853-like) were identified and only two miRNA-containing clones were recovered beyond 4 DPA. Further, among all of the small RNA sequences obtained from the small RNA pools in developing ovules, only 15 groups of sequences were observed in more than one DPA period. Of these, only five were present in more than two DPA periods. Two of these were miR-172 and miR-390 and a third was identified as 5.8S rRNA sequence. Thus, the vast majority of sequence signatures were expressed in only one DPA period and this included nearly all of the 24 nt sequences. Finally, we observed a distinct DPA-specific expression pattern among our clones based upon sequence abundance. Sequences occurring only once were far more likely to be seen in the 0 to 2 DPA periods while those occurring five or more times were the majority in later periods. CONCLUSION: This initial survey of small RNA sequences present in developing ovules in cotton indicates that fiber development is under complex small RNA regulation. Taken together, the results of this initial small RNA screen of developing cotton ovules is most consistent with a model, proposed by Baulcombe, that there are networks of small RNAs that are induced in a cascade fashion by the action of miRNAs and that the nature of these cascades can change from tissue to tissue and developmental stage to developmental stage.

Alveolar Bone Protective Effect of Hiziki Extracts on the Progression of Periodontitis.

The purpose of this study was to evaluate the effects of hiziki extract on alveolar bone loss, inflammation, and osteo-biomarker expression in hPDL cells (10, 50, 100 µg/ml final concentrations in culture medium) and on a ligature-induced periodontitis rat model (50, 100, 200 mg/kg with oral administration). Hiziki extract increased alkaline phosphatase activity and mineralized nodule formation in hPDL cell. In western blot analysis, hiziki extract resulted in increased expression of osteoblast markers, including transforming growth factor beta (TGF-ß), SMAD anchor for receptor activation (SARA) and runt-related transcription factor 2 (RUNX2) in hDPL cells. Additionally, expression of osteoclast markers and inflammatory cytokines was inhibited, which were receptor activator of NF-κB (RANK), RANK receptor (RANKL) and nuclear factor of activated T cells, cytoplasmic 1 (NFATc1). Hiziki extract also prevented alveolar bone loss in a ligature-induced periodontitis rat model through reducing the distance between cementoenamel junction and alveolar bone crest (CBJ-ABC) and furcation involvement. These findings suggested that hiziki extract has prophylactic potential for the prevention of periodontitis through anti-inflammation and, anti-bone resorption effects and the inhibition of alveolar bone destruction.

The inductive effect of ginsenoside F2 on hair growth by altering the WNT signal pathway in telogen mouse skin.

This study was conducted to confirm the possibility of using minor ginseng saponin F2 by oral administration on hair anagen induction effects. The signaling pathway and anagen induction effect of ginsenoside F2 were investigated and compared with finasteride on the effect of hair growth induction. The cell-based MTT assay results indicated that the proliferation rates of HHDPC and HaCaT treated with F2 significantly increased by 30% compared with the finasteride-treated group. A western blot study showed that the expression of ß-catenin Lef-1 and DKK-1 increased by 140, 200% and decreased by 40% in the F2-treated group, respectively compared to that of finasteride-treated group. C57BL/6 mice were subjected to the same treatments. The hair growth promotion rates were compared with groups treated with finasteride, which was 20% higher in the F2-treated group. Tissue histological analysis results showed the number of hair follicles, thickness of the epidermis, and follicles of the anagen phase which increased in the F2-treated group, compared with the finasteride-treated groups. Moreover, the effect of F2 on hair growth was confirmed through the immunofluorescence (IF) methods indicating the expression aspect of Wnt signal pathway-related factors in the tissue of C57BL/6 mouse. Our results considered the expression increase in ß-catenin, Lef-1 which was suggested as a major factor related to the development and growth of hair follicle and the decrease in DKK-1 when entering catagen by F2. As the data showed, F2 might be a potential new therapeutic source for anagen induction and hair growth through the Wnt signal pathway.

Solid-phase colorimetric method for the quantification of fucoidan.

We described the simple, selective, and rapid method for determination of fucoidans using methylene blue staining of sulfated polysaccharides, immobilized into filter paper and consequent optic density (at A (663) nm) measurement of the eluted dye from filter paper. This solid-phase method allows selective determination of 1-20 µg fucoidan in presence of potentially interfering compounds (alginic acid, DNA, salts, proteins, and detergents). Further, we demonstrated the alternative way of using image processing software for fucoidan quantification without extraction of methylene blue dye from stained spots of fucoidan-dye complex.

The role of induced mutation in conversion of photoperiod dependence in cotton.

Wild cotton germplasm resources are largely underutilized because of photoperiod-dependent flowering of "exotic" cottons. The objectives of this work were to explore the genome-wide effect of induced mutation in photoperiod-converted induced cotton mutants, estimating the genetic change between mutant and wild-type cottons using simple sequence repeats (SSRs) as well as understand the pattern of SSR mutation in induced mutagenesis. Three groups of photoperiod-converted radiomutants ((32)P) including their wild-type parental lines, A- and D-genome diploids, and typically grown cotton cultivars were screened with 250 cotton SSR primer pairs. Forty SSRs revealed the same SSR mutation profile in, at least, 2 independent mutant lines that were different from the original wild types. Induced mutagenesis both increased and decreased the allele sizes of SSRs in mutants with the higher mutation rate in SSRs containing dinucleotide motifs. Genetic distance obtained based on 141 informative SSR alleles ranged from 0.09 to 0.60 in all studied cotton genotypes. Genetic distance within all photoperiod-converted induced mutants was in a 0.09-0.25 range. The genetic distance among photoperiod-converted mutants and their originals ranged from 0.28 to 0.50, revealing significant modification of mutants from their original wild types. Typical Gossypium hirsutum cultivar, Namangan-77, revealed mutational pattern similar to induced radiomutants in 40 mutated SSR loci, implying possible pressure to these SSR loci not only in radiomutagenesis but also during common breeding process. Outcomes of the research should be useful in understanding the photoperiod-related mutations, and markers might help in mapping photoperiodic flowering genes in cotton.