Generation of two induced pluripotent stem cell lines from dilated cardiomyopathy patients carrying heterozygous FLNC mutations.

Dilated cardiomyopathy (DCM) is a heterogeneous cardiac disorder characterized by left ventricular dilatation and dysfunction. Mutations in dozens of cardiac genes have been connected to the development of DCM including the filamin C gene (FLNC). We generated two induced pluripotent stem cell (iPSCs) lines from DCM patients carrying single missense heterozygote FLNC mutations (c.6689G > A and c.3745G > A). Both lines expressed high levels of pluripotency markers, differentiated into derivatives of the three germ layers and possessed normal karyotypes. The derived iPSC lines can serve as powerful tools to model DCM in vitro and as a platform for therapeutic development.

A comparison of ImageJ and machine learning based image analysis methods to measure cassava bacterial blight disease severity.

BACKGROUND: Methods to accurately quantify disease severity are fundamental to plant pathogen interaction studies. Commonly used methods include visual scoring of disease symptoms, tracking pathogen growth in planta over time, and various assays that detect plant defense responses. Several image-based methods for phenotyping of plant disease symptoms have also been developed. Each of these methods has different advantages and limitations which should be carefully considered when choosing an approach and interpreting the results. RESULTS: In this paper, we developed two image analysis methods and tested their ability to quantify different aspects of disease lesions in the cassava-Xanthomonas pathosystem. The first method uses ImageJ, an open-source platform widely used in the biological sciences. The second method is a few-shot support vector machine learning tool that uses a classifier file trained with five representative infected leaf images for lesion recognition. Cassava leaves were syringe infiltrated with wildtype Xanthomonas, a Xanthomonas mutant with decreased virulence, and mock treatments. Digital images of infected leaves were captured overtime using a Raspberry Pi camera. The image analysis methods were analyzed and compared for the ability to segment the lesion from the background and accurately capture and measure differences between the treatment types. CONCLUSIONS: Both image analysis methods presented in this paper allow for accurate segmentation of disease lesions from the non-infected plant. Specifically, at 4-, 6-, and 9-days post inoculation (DPI), both methods provided quantitative differences in disease symptoms between different treatment types. Thus, either method could be applied to extract information about disease severity. Strengths and weaknesses of each approach are discussed.

Overexpression of Panax ginseng sesquiterpene synthase gene confers tolerance against Pseudomonas syringae pv. tomato in Arabidopsis thaliana.

Sesquiterpenes are an abundant group belonging to the terpenoid family, with a C15 structure comprise of three isoprene units. Many sesquiterpenes are volatile compounds and it act as chemical messenger in plant signalling, particularly in the defense mechanism against biotic and abiotic stresses. Panax ginseng Meyer is important medicinal herbs with various reported pharmacological efficacies in which its triterpenoid saponins, called ginsenosides, were mostly studied. However, there have been few studies on volatile sesquiterpenes compounds regulation on P. ginseng. As slow-growing perennial plant, P. ginseng received many kind of stresses during its cultivation. The pathogen attack is one of the most devastated perturbation for ginseng yield. Thus, we aimed to analyze P. ginseng STS gene (PgSTS) expressions in ginseng organs as well as mono-, sesquiterpenes contents from ginseng seedlings treated with elicitors. qRT-PCR and GC-MS analysis showed that two elicitors- salicylic acid (SA) and methyl jasmonate (MeJA) triggered PgSTS expression at different time points and significantly induced mono-, sesquiterpene yield. Overexpression of PgSTS in Arabidopsis also induced high terpene content and conferred tolerance against Pseudomonas syringae pv. tomato infection. These results suggested that PgSTS transcripts are involved in terpenoid biosynthesis in response to environmental stress mediated by MeJA and SA elicitors; thus, generate tolerance against pathogen attack.

Immunostimulatory effect of fermented red ginseng in the mouse model.

In this study, Woongjin fermented red ginseng extract (WFRG) was evaluated for its potential ability to act as an adjuvant for the immune response of mice. For the in vitro study, macrophages were treated with serial concentrations (1 µg/mL, 10 µg/mL, and 100 µg/mL) of WFRG. For in vivo studies, mice were administered different concentrations (10 mg/kg/day, 100 mg/kg/day, and 200 mg/kg/day) of WFRG orally for 21 days. In vitro, the production of nitric oxide and TNF-α by RAW 264.7 cells increased in a dose-dependent manner. In vivo, WFRG enhanced the proliferation of splenocytes induced by two mitogens (i.e., concanavalin A and lipopolysaccharide [LPS]) and increased LPS-induced production of TNF-α and IL-6, but not IL-1ß. In conclusion, WFRG has the potential to modulate immune function and should be further investigated as an immunostimulatory agent.

Screening and optimization of pectin lyase and polygalacturonase activity from ginseng pathogen Cylindrocarpon Destructans

Cylindrocarpon destructans isolated from ginseng field was found to produce pectinolytic enzymes. A Taguchi's orthogonal array experimental design was applied to optimize the preliminary production of polygalacturonase (PG) and pectin lyase (PL) using submerged culture condition. This method was applied to evaluate the significant parameters for the production of enzymes. The process variables were pH, pectin concentration, incubation time and temperature. Optimization of process parameters resulted in high levels of enzyme (PG and PL) production after ten days of incubation at a pH of 5.0 at 25°C in the presence of 1.5 percent pectin. Among different nitrogen sources, urea and peptone showed high production of PG and PL, respectively. The enzyme production and mycelial growth seems to have direct influence on the culture conditions; therefore, at stationary state high enzyme production and mycelial growth were obtained than agitation state. Along with this, optimization of enzyme activity was also determined using various physiological parameters like, temperature, incubation time and pH. Taguchi's data was also analyzed using one step ANOVA statistical method.

Lactobacillus kimchicus sp. nov., a ß-glucosidase-producing bacterium isolated from kimchi.

A Gram-reaction-positive, non-spore-forming, rod-shaped, non-motile, ß-glucosidase-producing bacterium, designated DCY51(T), was isolated from kimchi, a Korean fermented vegetable food. 16S rRNA gene sequence analysis revealed that strain DCY51(T) belonged to the genus Lactobacillus and exhibited highest 16S rRNA gene sequence similarity with Lactobacillus paracollinoides AB 74 (96.9 %), L. similis JCM 2765(T) (96.9 %), L. collinoides JCM 1123(T) (96.7 %) and L. hilgardii DSM 20176(T) (95.5 %). Subsequently, pheS sequence analysis confirmed that strain DCY51(T) formed a distinct lineage within the Lactobacillus plantarum group. The major polar lipid of strain DCY51(T) was phosphatidylethanolamine and minor amounts of phosphatidylglycerol and diphosphatidylglycerol were found. Phenotypic characteristics and DNA-DNA relatedness indicated that strain DCY51(T) was clearly distinguished from other type strains of species of the genus Lactobacillus. A novel species, Lactobacillus kimchicus sp. nov., is proposed, with type strain DCY51(T) ( = KCTC 12976(T)  = JCM 15530(T)).

Biotransformation of Ginsenoside Rb1 to Prosapogenins, Gypenoside XVII, Ginsenoside Rd, Ginsenoside F2, and Compound K by Leuconostoc mesenteroides DC102.

Ginsenoside Rb1is the main component in ginsenosides. It is a protopanaxadiol-type ginsenoside that has a dammarane-type triterpenoid as an aglycone. In this study, ginsenoside Rb1 was transformed into gypenoside XVII, ginsenoside Rd, ginsenoside F2 and compound K by glycosidase from Leuconostoc mesenteroides DC102. The optimum time for the conversion was about 72 h at a constant pH of 6.0 to 8.0 and the optimum temperature was about 30℃. Under optimal conditions, ginsenoside Rb1 was decomposed and converted into compound K by 72 h post-reaction (99%). The enzymatic reaction was analyzed by highperformance liquid chromatography, suggesting the transformation pathway: ginsenoside Rb1→ gypenoside XVII and ginsenoside Rd→ginsenoside F2→compound K.

Screening and optimization of pectin lyase and polygalacturonase activity from ginseng pathogen Cylindrocarpon Destructans.

Cylindrocarpon destructans isolated from ginseng field was found to produce pectinolytic enzymes. A Taguchi's orthogonal array experimental design was applied to optimize the preliminary production of polygalacturonase (PG) and pectin lyase (PL) using submerged culture condition. This method was applied to evaluate the significant parameters for the production of enzymes. The process variables were pH, pectin concentration, incubation time and temperature. Optimization of process parameters resulted in high levels of enzyme (PG and PL) production after ten days of incubation at a pH of 5.0 at 25°C in the presence of 1.5% pectin. Among different nitrogen sources, urea and peptone showed high production of PG and PL, respectively. The enzyme production and mycelial growth seems to have direct influence on the culture conditions; therefore, at stationary state high enzyme production and mycelial growth were obtained than agitation state. Along with this, optimization of enzyme activity was also determined using various physiological parameters like, temperature, incubation time and pH. Taguchi's data was also analyzed using one step ANOVA statistical method.

Rhodanobacter soli sp. nov., isolated from soil of a ginseng field.

Strain DCY45(T) was isolated from soil of a ginseng field in Pocheon Province, Korea. Strain DCY45(T) was Gram-negative, oxidase- and catalase-positive, motile and rod-shaped and produced yellow pigments on R2A agar. The organism grew optimally at 30 °C and at pH 7.0. The G+C content of the genomic DNA was 65.4 mol%. The predominant respiratory quinone was Q-8. The major fatty acids were iso-C(17 : 1)ω9c, iso-C(16 : 0) and iso-C(15 : 0). Phylogenetic analysis based on the 16S rRNA gene sequence was used to determine the taxonomic position of strain DCY45(T), which is most closely related to species of the genus Rhodanobacter, with similarity levels of 96.0-98.4 %; DNA-DNA relatedness with related strains was lower than 60 %. Strain DCY45(T) differed significantly from related type strains in phenotypic characteristics. On the basis of these phenotypic, genotypic and chemotaxonomic studies, strain DCY45(T) represents a novel species of the genus Rhodanobacter, for which the name Rhodanobacter soli sp. nov. is proposed. The type strain is DCY45(T) (=KCTC 22620(T) =JCM 16126(T)).

Microbacterium ginsengiterrae sp. nov., a beta-glucosidase-producing bacterium isolated from soil of a ginseng field.

Strain DCY37(T) was isolated from a soil sample of a ginseng field in the Republic of Korea and characterized in order to determine its taxonomic position. Cells were Gram-staining-positive, heterotrophic, strictly aerobic, non-motile short rods. 16S rRNA gene sequence analysis revealed that strain DCY37(T) belongs to the genus Microbacterium. According to 16S rRNA gene sequence analysis, it is closely related to Microbacterium aerolatum DSM 14217(T) (98.8 %), Microbacterium hydrocarbonoxydans DSM 16089(T) (98.5 %), Microbacterium natoriense JCM 12611(T) (98.5 %), Microbacterium foliorum (98.4 %) and Microbacterium phyllosphaerae (98.3 %). However, DNA-DNA hybridization studies showed reassociation values of less than 70 % between representative strains and DCY37(T). The DNA G+C content was 64.5 mol%. Strain DCY37(T) possessed chemotaxonomic markers that were consistent with classification in the genus Microbacterium, i.e. MK-12 and MK-13 as the major menaquinones and anteiso-C(15 : 0), anteiso-C(17 : 0) and iso-C(16 : 0) as the predominant cellular fatty acids. The major cell wall sugars were ribose, xylose and galactose. The diamino acid in cell-wall hydrolysates of strain DCY37(T) was ornithine and major cell-wall amino acids were alanine, glycine, d-glutamic acid and serine. The major polar lipids were glycolipid, phosphatidylglycerol, diphosphatidylglycerol and unknown aminolipids. Based on these data, DCY37(T) (=KCTC 19526(T) =JCM 15516(T)) should be classified as the type strain of a novel species of the genus Microbacterium, for which the name Microbacterium ginsengiterrae sp. nov. is proposed.

Solibius ginsengiterrae gen. nov., sp. nov., isolated from soil of a ginseng field, and emended description of the genus Sediminibacterium and of Sediminibacterium salmoneium.

A novel Gram-staining-negative bacterial strain, designated DCY13T, was isolated from soil of a ginseng field in Korea and characterized by using a polyphasic approach. The phylogenetic analysis based on 16S rRNA gene sequences revealed that strain DCY13T forms a cluster with members of the genera Sediminibacterium, Flavisolibacter, Niabella, Terrimonas, Niastella and Chitinophaga in the family 'Chitinophagaceae', phylum Bacteroidetes, and shares highest sequence similarity (95.2 %) with Sediminibacterium salmoneum NJ-44T. Sequence similarity with other members of the family is 87.6-91.4 %. Cells are non-spore-forming rods, catalase and oxidase-positive, motile by gliding that grow under strictly aerobic conditions. The predominant respiratory quinone is menaquinone MK-7, and the major fatty acids are anteiso-C15:0, iso C16:0, anteiso-C17:0, and iso-C15:0. The G+C content of the genomic DNA is 47.8 mol%. The major cell wall amino acids are D-aspartic acid, D-glycine, D-serine, L-lysine, D-glucosamine, and D-alanine. The major cell wall sugars are ribose, xylose, and galactose. The major polar lipids are phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. It is proposed that strain DCY13T represents a novel genus and species in the family 'Chitinophagaceae' for which the name Solibius ginsengiterrae gen. nov., sp. nov., is proposed. The type strain is DCY13T (= KCTC 12833T = JCM 15794T). Emended descriptions of the genus Sediminibacterium and of S. salmoneum NJ44T are also proposed.

Hydrogenophaga temperata sp. nov., a betaproteobacterium isolated from compost in Korea.

A Gram-negative, rod-shaped, non-spore-forming and motile bacterial strain TR7-01(T) was isolated from a compost soil in South Korea and subjected to a polyphasic taxonomic study. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain TR7-01(T) belonged to the genus Hydrogenophaga within the class Betaproteobacteria. Strain TR7-01(T) exhibited 16S rRNA gene sequence similarity values of 95.0-98.3% to members of the genus Hydrogenophaga: Hydrogenophaga bisanensis DSM12412(T) (98.3%), Hydrogenophaga flava DSM 619(T) (97.1%), Hydrogenophaga pseudoflava ATCC 33668(T) (96.8%), Hydrogenophaga intermedia S1(T) (96.4%), Hydrogenophaga atypica BSB 41.8(T) (95.8%), Hydrogenophaga defluvii BSB 9.5(T) (95.7%), Hydrogenophaga palleronii CCUG 20334(T) (95.6%), Hydrogenophaga caeni EMB71(T) (95.4%) and Hydrogenophaga taeniospiralis ATCC 49743(T) (95.0%). Chemotaxonomic data revealed that strain TR7-01(T) possesses ubiquinone Q-8, the G+C content was 69.9 mol%, and the predominant fatty acids were 16 : 1 ω7c/15 : 0 iso 2OH, 18 : 1 ω7c/ω9t/ω12t and C(16:0), all of which corroborated our assignment of the strain to the genus Hydrogenophaga. The results of DNA-DNA hybridization and physiological and biochemical tests clearly demonstrated that strain TR7-01(T) represents a distinct species. Based on these data, TR7-01(T) (= KCTC 12203(T) = DSM 18117(T)) should be classified as a novel Hydrogenophaga species, for which the name Hydrogenophaga temperata sp. nov. has been proposed.

Lysobacter soli sp. nov., isolated from soil of a ginseng field.

Strain DCY21(T), a Gram-negative, gliding and rod-shaped aerobic bacterium was isolated from soil of a ginseng field in the Republic of Korea and characterized using a polyphasic approach in order to determine its taxonomic position. Comparative 16S rRNA gene sequence analysis revealed that strain DCY21(T) clustered with the species of the genus Lysobacter. It was closely related to Lysobacter gummosus LMG 8763(T) (97.9 %), Lysobacter capsici YC5194(T) (97.6 %), Lysobacter antibioticus DSM 2044(T) (97.5 %), Lysobacter niastensis DSM 18481(T) (97.2 %) and Lysobacter enzymogenes DSM 2043(T) (96.9 %). The major cellular fatty acids of strain DCY21(T) were iso-C(15 : 0) (34.3 %), iso-C(17 : 1)omega9c (19.5 %) and iso-C(17 : 0) (17.2 %) and the major isoprenoid quinone was Q-8. The major polar lipids of strain DCY21(T) were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidyl-N-methylethanolamine. The G+C content of the total DNA was 65.4 mol%. The DNA-DNA relatedness values, and biochemical and physiological characteristics strongly supported the genotypic and phenotypic differentiation of strain DCY21(T) from species of the genus Lysobacter. Strain DCY21(T) therefore represents a novel species, for which the name Lysobacter soli sp. nov. is proposed. The type strain is DCY21(T) (=KCTC 22011(T) =LMG 24126(T)).

Stenotrophomonas ginsengisoli sp. nov., isolated from a ginseng field.

A Gram-negative, non-spore-forming, rod-shaped bacterium, designated strain DCY01(T), was isolated from soil from a ginseng field in South Korea and was characterized in order to determine its taxonomic position. 16S rRNA gene sequence analysis revealed that strain DCY01(T) belonged to the Gammaproteobacteria and was most closely related to Stenotrophomonas koreensis KCTC 12211(T) (98.4 % similarity), Stenotrophomonas humi R-32729(T) (97.2 %), Stenotrophomonas terrae R-32768 (97.1 %), Stenotrophomonas maltophilia DSM 50170(T) (96.9 %) and Stenotrophomonas nitritireducens DSM 12575(T) (96.8 %). Chemotaxonomic analyses revealed that strain DCY01(T) possessed a quinone system with Q-8 as the predominant compound, and iso-C(15 : 0) (28.2 %), C(16 : 0) 10-methyl (13.2 %), iso-C(15 : 1) F (10.8 %) and C(15 : 0) (7.5 %) as major fatty acids, corroborating assignment of strain DCY01(T) to the genus Stenotrophomonas. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The results of DNA-DNA hybridization and physiological and biochemical tests clearly demonstrated that strain DCY01(T) represents a species distinct from recognized Stenotrophomonas species. Based on these data, DCY01(T) (=KCTC 12539(T)=NBRC 101154(T)) should be classified as the type strain of a novel species of the genus Stenotrophomonas, for which the name Stenotrophomonas ginsengisoli sp. nov. is proposed.

Curtobacterium ginsengisoli sp. nov., isolated from soil of a ginseng field.

A Gram-positive, non-motile, pale-yellow, short rod-shaped bacterium, strain DCY26(T), was isolated from soil of a ginseng field in South Korea and was investigated to determine its taxonomic position. The organism grew optimally at 30-37 degrees C. The G+C content of its DNA was 65.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain DCY26(T) was related most closely to species of the genus Curtobacterium, in the family Microbacteriaceae. Strain DCY26(T) showed highest 16S rRNA gene sequence similarity to Curtobacterium pusillum DSM 20527(T) (96.3 %), Curtobacterium luteum DSM 20542(T) (96.2 %), Curtobacterium flaccumfaciens LMG 3645(T) (96.2 %), Curtobacterium citreum DSM 20528(T) (96.1 %), Curtobacterium albidum DSM 20512(T) (96.1 %) and Curtobacterium herbarum DSM 14013(T) (95.3 %). The predominant menaquinone of strain DCY26(T) was MK-9. Other chemotaxonomic data also supported the affiliation of strain DCY26(T) to the genus Curtobacterium. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain DCY26(T) is considered to represent a novel species of the genus Curtobacterium, for which the name Curtobacterium ginsengisoli sp. nov. is proposed. The type strain is DCY26(T) (=KCTC 13163(T) =JCM 14773(T)).

Microbacterium ginsengisoli sp. nov., a beta-glucosidase-producing bacterium isolated from soil of a ginseng field.

Strain Gsoil 259(T), a beta-glucosidase-producing bacterium, was isolated from a soil sample from a ginseng field in the Republic of Korea and characterized in order to determine its taxonomic position. Cells were Gram-positive, heterotrophic, strictly aerobic, non-motile short rods. 16S rRNA gene sequence analysis revealed that strain Gsoil 259(T) belonged to the genus Microbacterium and was closely related to Microbacterium arborescens IFO 3750(T) (98.5 %) and Microbacterium imperiale IFO 12610(T) (97.9 %). However, it has low values for DNA-DNA relatedness with the above strains (20.7 and 17.5 %, respectively). Strain Gsoil 259(T) possessed chemotaxonomic markers that were consistent with classification in the genus Microbacterium, i.e. MK-11 and MK-12 were the major menaquinones and anteiso-C(17 : 0), anteiso-C(15 : 0) and iso-C(16 : 0) were the predominant cellular fatty acids. The DNA G+C content was 69.4 mol%. The cell-wall sugar was rhamnose and the diamino acid in the cell-wall peptidoglycan was ornithine. On the basis of data from this polyphasic study, strain Gsoil 259(T) represents a novel species of the genus Microbacterium, for which the name Microbacterium ginsengisoli sp. nov. is proposed. The type strain is Gsoil 259(T) (=KCTC 19189(T) =DSM 18659(T)).

Paenibacillus soli sp. nov., a xylanolytic bacterium isolated from soil.

Two novel polysaccharide-degrading bacteria (strains DCY03(T) and DCY04) were isolated from a soil sample of a ginseng field in the Republic of Korea and were identified as representing members of the genus Paenibacillus on the basis of phenotypic characteristics and phylogenetic inference based on 16S rRNA gene sequences. Cells of the two isolates were Gram-positive, spore-forming, non-motile, straight rods. Based on DNA-DNA relatedness data, the strains were considered to belong to the same species. The DNA G+C content ranged from 56.6 to 57.0 mol%. The predominant cellular fatty acid was anteiso-C(15 : 0) (63.8-62.8 %). Levels of 16S rRNA gene sequence similarity between the two novel isolates and the type strains of recognized Paenibacillus species were 91.4-96.5 %. Strains DCY03(T) and DCY04 could clearly be distinguished from phylogenetically closely related Paenibacillus species on the basis of DNA-DNA relatedness data and phenotypic characteristics. Therefore, on the basis of these data, the two isolates are considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus soli sp. nov. is proposed. The type strain is DCY03(T) (=KCTC 13010(T)=LMG 23604(T)).

Burkholderia ginsengisoli sp. nov., a beta-glucosidase-producing bacterium isolated from soil of a ginseng field.

A bacterial strain (designated KMY03T) that possesses beta-glucosidase activity was isolated from soil from a ginseng field in South Korea and was characterized in order to determine its taxonomic position. The bacterium was found to comprise Gram-negative, rod-shaped, motile cells with unipolar polytrichous flagella. On the basis of 16S rRNA gene sequence similarity, strain KMY03T was shown to belong to the family Burkholderiaceae of the Betaproteobacteria, being most closely related to Burkholderia caledonica LMG 19076T (97.8%), Burkholderia terricola LMG 20594T (97.5%), Burkholderia xenovorans LMG 21463T (97.4%) and Burkholderia phytofirmans LMG 22146T (97.3%). Chemotaxonomic data (major ubiquinone, Q-8; major fatty acids, C17:0 cyclo, C16:0, C19:0 cyclo omega8c and summed feature 2) supported the affiliation of the novel strain with the genus Burkholderia. The results of DNA-DNA hybridizations and physiological and biochemical tests allowed the strain to be differentiated genotypically and phenotypically from Burkholderia species with validly published names. On the basis of these data, strain KMY03T represents a novel species of the genus Burkholderia, for which the name Burkholderia ginsengisoli sp. nov. is proposed. The type strain is KMY03T (=KCTC 12389T=NBRC 100965T).