Polygonum Tinctorium Lour Research Articles

Treatment with indigo plant (Polygonum tinctorium Lour) improves serum lipid profiles in Wistar rats fed a high-fat diet.

We investigated the effects of Polygonum tinctorium Lour (PTL), a plant commonly known as indigo, on biological parameters in an animal model of high-fat diet-induced obesity. Wistar rats fed a high-fat diet and treated with PTL showed lower serum levels of triglycerides and total cholesterol levels and a higher serum levels of HDL cholesterol than those in Wistar rats fed a high-fat diet without PTL treatment. The weight of mesenteric fat in PTL-treated rats was decreased compared to that in control rats not treated with PTL. In addition, energy metabolic rate in the dark period, but not in the light period, in PTL-treated rats was higher than that in control rats. Although a significant difference was not observed, body weight in PTL-treated rats tended to be decreased compared to that in control rats. The results show that PTL improves serum lipid profiles in Wistar rats with high-fat diet-induced obesity. J. Med. Invest. 67 : 158-162, February, 2020.

Constituents of flavonol O-glycosides and antioxidant activities of extracts from seeds, sprouts, and aerial parts of Polygonum tinctorium Lour.

Polygonum tinctorium Lour. (family Polygonaceae), known as indigo plant, has been useful as a medicinal or edible plant abundant in polyphenolic compounds. We have recently shown that flavonol O-glycosides with 3,5,4′-trihydroxy-6,7-methylenedioxyflavone (TMF) are predominant flavonoids in indigo leaves. However, no study has been performed regarding changes in the levels of flavonoid species during the germination and growth of indigo plant. Here, we attempted to determine the individual constituents of flavonol O-glycosides and the changes in their contents of the seeds, sprouts, and aerial parts. These results revealed that only the seeds predominantly contained flavonol O-(acetyl)-rhamnosides with quercetin or kaempferol as an aglycone. During the development of the sprouts and aerial parts, flavonol O-glycosides with TMF as an aglycone became mainly detectable and accounted for 79.4% and 74.9% of total flavonol O-glycosides from the extracts of aerial parts harvested in 2016 and 2017, respectively. Of the plant organs tested, the aerial parts exhibited the highest antioxidant activities concomitant with greatly increased levels of total polyphenols. Thus, we were able to conduct the identification and quantification of flavonol O-glycosides from the seeds, sprouts, and aerial parts of indigo plant and to evaluate antioxidant activities of their extracts. Taken together, our findings clearly provide the evidence that the aerial parts of indigo plant are a rich source of flavonol O-glycosides with TMF and exhibit much higher antioxidant activities, indicating the usefulness for the application to food and nutraceutical purposes.

Anti-neuroinflammatory effects of tryptanthrin from Polygonum tinctorium Lour. in lipopolysaccharide-stimulated BV2 microglial cells.

This study was conducted to isolate the anti-neuroinflammatory component(s) in the 80% EtOH extract of P. tinctoria, and to investigate underlying molecular mechanism of the anti-neuroinflammatory component(s) in LPS-induced BV2 microglial cells. To isolate the active component(s) in the extract, various chromatographic methods were employed, and the structures of the isolated secondary metabolites were determined mainly by analysis of spectroscopic data such as NMR and MS data. Tryptanthrin (1), isolated from P. tinctoria extract, significantly inhibited the protein expression of iNOS and COX-2, and reduced the levels of their products (NO and PGE2) in LPS-stimulated BV2 microglial cells. Tryptanthrin (1) also downregulated the production of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β. These anti-neuroinflammatory effects of tryptanthrin (1) was elucidated to be correlated with inactivating NF-κB pathway by interrupting the phosphorylation and degradation of the inhibitor of κB-α protein, and inhibiting the DNA binding activity of NF-κB. In addition, tryptanthrin (1) suppressed the activation of p38 MAPK pathway. Furthermore, tryptanthrin (1) inhibited the TLR4 and MyD88 protein expression in LPS-stimulated BV2 microglial cells. Taken together, it was suggested that tryptanthrin (1) have anti-neuroinflammatory effect by regulating TLR4-MyD88-mediated several inflammatory pathways including p38 and NF-κB pathways in LPS-induced BV2 microglial cells.

Bacillus fermenti sp. nov., an indigo-reducing obligate alkaliphile isolated from indigo fermentation liquor for dyeing

The indigo-reducing, facultatively anaerobic and obligately alkaliphilic strains Bf-1T, Bf-2 and Bf-4 were isolated from an indigo fermentation liquor used for dyeing, which uses sukumo [composted Polygonum indigo (Polygonum tinctorium Lour.) leaves] as a basic ingredient and was obtained from a craft centre in Date City, Hokkaido, Japan. The 16S rRNA gene sequence analyses indicated that the closest neighbours of strain Bf-1T are Bacillus maritimus DSM 100413T (98.3 % 16S rRNA gene sequence similarity), Bacillus persicus DSM 25386T (98.2 %) and Bacillus rigiliprofundi LMG 28275T (97.7 %). The 16S rRNA gene sequence of strain Bf-1T was almost identical to the sequences of strains Bf-2 and Bf-4 (99.9 %). Cells of strain Bf-1T stained Gram-positive and formed straight rods that achieved motility through a pair of subpolar flagella. Strain Bf-1T grew at temperatures of between 15 and 45 °C with optimum growth at 33‒40 °C. The strain grew in the pH range of pH 8‒12, with optimum growth at pH 10. The isoprenoid quinone detected was menaquinone-7 (MK-7), and the DNA G+C content was 41.7 %. The whole-cell fatty acid profile mainly (>10 %) consisted of iso-C15 : 0 and iso-C16 : 0. Phylogenetically related neighbours, although demonstrating high 16S rRNA gene sequence similarity (>97.6 %) with strain Bf-1T, exhibited less than 9 % relatedness in DNA-DNA hybridization experiments. Based on evidence from this polyphasic study, the isolates represent a novel species, for which the name Bacillus fermenti sp. nov. is proposed. The type strain of this species is Bf-1T (=JCM 31807T=NCIMB 15079T).

In Vitro Anti-inflammatory and Antioxidant Activities of 3,5,4'-Trihydroxy-6,7-Methylenedioxyflavone-O-Glycosides and Their Aglycone from Leaves of Polygonum tinctorium Lour.

Polygonum tinctorium Lour (indigo plant) has been regarded as a useful medicinal plant for traditional herbal medicine. The polyphenolic fraction of indigo leaves exhibited anti-inflammatory activities as determined by the suppressed synthesis of nitric oxide (NO) in cultured RAW264 macrophage cells. The acid hydrolysate of the fraction showed much more potent effect than the unhydrolyzed one. In sharp contrast, those fractions of indigo stems had almost no effect. 3,5,4'-Trihydroxy-6,7-methylenedioxyflavone (TMF)-O-glycosides and tryptanthrin were detected exclusively in the extracts of the leaves. The isolated flavonol species were furthermore tested for their anti-inflammatory activities against the synthesis of NO and prostaglandin E2 in the cultured macrophage cells. More potent anti-inflammatory effects were recognized with different aglycones of flavonols than their flavonol O-glycosides. Although the inhibitory effects of TMF were less effective than those of tryptanthrin, the levels of flavonol O-glycosides with TMF were much more abundant than those of tryptanthrin in the leaves. Oral administration of the fraction containing flavonol O-glycosides with TMF into mice revealed the detection of free TMF in the blood circulation, indicating that the aglycone moiety can be cleaved by digestive enzymes and absorbed in the gut. Alternatively, the assay of hydrophilic oxygen radical absorbance capacity revealed that the isolated species of flavonol O-glycosides with TMF and their aglycone had appreciable antioxidant activities. Taken together, our findings suggest that the predominant flavonol O-glycosides with TMF as an aglycone could be promising natural agents for the application to herbal medicine, nutraceuticals, and food additives.

植物色素の教材化 (2) : 藍

植物色素の教材化 (2) : 藍

Polygonibacillus indicireducens gen. nov., sp. nov., an indigo-reducing and obligate alkaliphile isolated from indigo fermentation liquor for dyeing

Obligately alkaliphilic and halophilic strains, designated In2-9T and D2-7, were isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample obtained from a craft centre in Date City, Hokkaido, Japan. The 16S rRNA gene sequence phylogeny suggested that strain In2-9T is a member of the genus Bacillus with the closest relatives being the alkaliphilic species of the genus Bacillus, Bacillus hemicellulosilyticusJCM 9152T (96.4 % 16S rRNA gene sequence similarity) and Bacillus alcalophilus DSM 485T (96.5 %). Cells of the isolate stained Gram-positive and were facultatively anaerobic straight rods that were motile by peritrichous flagella. Strain In2-9T grew between 13 and 45 °C with optimum growth at approximately 35-37 °C. The isolates grew in the pH range of 8-12 with optimum growth at pH 10. The isoprenoid quinone detected was menaquinone-6 (MK-6) and the DNA G+C content was 39.4 mol%. The whole-cell fatty acid profile mainly (>10 %) consisted of iso-C15 : 0, anteiso-C15 : 0 and C16 : 0. Spore shape and location and chemotaxonomic characteristics revealed that the isolates were distinctly different from phylogenetic neighbouring alkaliphilic species of the genus Bacillus. On the basis of phenotypic and chemotaxonomic characteristics and phylogenetic data, the isolates represent a novel species of a new genus, for which the name Polygonibacillusindicireducens gen. nov., sp. nov. is proposed. The type strain of the type species is In2-9T (=JCM 30831T=NCIMB 14982T), and strain D2-7 is an additional strain of the species.

Fermentibacillus polygoni gen. nov., sp. nov., an alkaliphile that reduces indigo dye.

Facultatively alkaliphilic strains, designated as strains IEB3T and IEB14, were isolated as indigo-reducing strains from a fermented Polygonum indigo (Polygonum tinctorium Lour) liquor sample prepared in our laboratory using a medium containing an indigo fermentation liquor as a sole substrate. The 16S rRNA gene sequence phylogeny and similarity suggested that strains IEB3T and IEB14 exhibit distinctive positions among the members of the genus Bacillus, and their closest neighbour was Bacillus nanhaiisediminis NH3T (similarity: 97.4 %) among the species with validly published names. The 16S rRNA sequence of strain IEB3Twas identical to that of strain IEB14. The cells of the isolates stained Gram-positive and were facultatively anaerobic, straight rods that were motile by a pair of subpolar flagella. Strains IEB3T and IEB14 grew at temperatures between 12 and 40 °C with optimum growth at 30‒33 °C and in the range of pH 7.5-12. Menaquinone-7 (MK-7) was detected as the major isoprenoid quinone. The DNA G+C contents of strains IEB3T and IEB14 were 39.0 and 39.1 mol%, respectively. The whole-cell fatty acid profile mainly (>10 %) consisted of iso-C14:0, iso-C15:0 and anteiso-C15:0. DNA-DNA hybridization revealed a low relatedness value between strain IEB3T and the phylogenetically most closely related species, Bacillus nanhaiisediminis JCM 16507T (<7 % ). On the basis of phenotypic and chemotaxonomic characteristics and phylogenetic data, the isolates represent a novel species within a novel genus, for which the name Fermentibacillus polygoni gen. nov., sp. nov. is proposed. The type strain is IEB3T (=JCM 30817T=NCIMB 14984T).

Identification of new flavonol O-glycosides from indigo (Polygonum tinctorium Lour) leaves and their inhibitory activity against 3-hydroxy-3-methylglutaryl-CoA reductase

Indigo plant (Polygonum tinctorium Lour) has been utilized as a medicinal plant with a variety of biological activities. We have recently detected higher levels of flavonoids in indigo leaves. This study was undertaken to conduct the simultaneous analysis of those flavonoids using total extracts from indigo leaves by ultra-performance liquid chromatography–electrospray ionization-time-of-flight/mass spectrometryE (UPLC–ESI-TOF/MSE). The analysis by UPLC–ESI-TOF/MSE allowed us to determine 11 peaks of flavonoid species. The chemical structures of these compounds were identified as flavonol O-glycosides with different types of aglycones by the combination of spectroscopic and chemical methods. The predominant compounds were flavonol O-glycosides with 3,5,4′-trihydroxy-6,7-methylenedioxyflavone as an aglycone. Of these, three compounds were elucidated as new compounds. All the isolated flavonol O-glycosides exhibited the inhibitory activity against 3-hydroxy-3-methylglutaryl-CoA reductase in a dose-dependent manner with different potencies. Taken together, our results suggest the potential usefulness of the major flavonol O-glycosides from indigo leaves in controlling cholesterol biosynthesis.

A natural dye, Niram improves atopic dermatitis through down-regulation of TSLP

Naju Jjok (Polygonum tinctorium Lour.) has been known to treat skin diseases in traditional Korean medicine. A natural textile dye, Niram made from Naju Jjok has traditionally been used to dye clothes. Thymic stromal lymphopoietin (TSLP) plays an important role in the development of atopic dermatitis (AD). Thus, we investigated that Niram might ameliorate AD through regulation of TSLP. Niram significantly inhibited the levels of TSLP through blockade of caspase-1/receptor-interacting protein 2 pathway in stimulated mast cells. Further, Niram ameliorated clinical symptoms in AD mouse. Niram significantly inhibited the infiltration of inflammatory cells in lesional skin. The levels of TSLP, caspase-1, IL-4, and IL-6 were inhibited in lesional skin applied topically with Niram. Niram significantly inhibited the serum levels of IgE and histamine in AD mouse. Finally, Niram significantly inhibited the levels of TSLP in polyriboinosinic polyribocytidylic acid-stimulated human keratinocyte HaCaT cells. These results establish Niram as a functional dye embracing the aspects of not only a traditional use but also a pharmacological effect.

Gracilibacillus alcaliphilus sp. nov., a facultative alkaliphile isolated from indigo fermentation liquor for dyeing.

A facultatively alkaliphilic, lactic-acid-producing and halophilic strain, designated SG103(T), was isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample for dyeing prepared in a laboratory. 16S rRNA gene sequence phylogeny suggested that SG103(T) is a member of the genus Gracilibacillus with the closest relatives being 'Gracilibacillus xinjiangensis' J2 (similarity: 97.06 %), Gracilibacillus thailandensis TP2-8(T) (97.06 %) and Gracilibacillus halotolerans NN(T) (96.87 %). Cells of the isolate stained Gram-positive and were facultatively anaerobic straight rods that were motile by peritrichous flagella. The strain grew at temperatures between 13 and 48 °C with the optimum at 39 °C. It grew in the range pH 7-10 with the optimum at pH 9. The isoprenoid quinone detected was menaquinone-7 (MK-7) and the DNA G+C content was 41.3 mol%. The whole-cell fatty acid profile mainly (>10 %) consisted of iso-C15 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. Unlike other reported species of the genus Gracilibacillus, the strain lacked diphosphatidylglycerol as a major polar lipid. DNA-DNA hybridization experiments with strains exhibiting greater than 96.87 % 16S rRNA gene sequence similarity, 'G. xinjiangensis' J2, G. thailandensis TP2-8(T) and G. halotolerans NN(T), revealed 2±4 %, 4±9 % and 3±2 % relatedness, respectively. On the basis of the differences in phenotypic and chemotaxonomic characteristics, and the results of phylogenetic analyses based on 16S rRNA gene sequences and DNA-DNA relatedness data from reported species of the genus Gracilibacillus, strain SG103(T) merits classification as a members of a novel species, for which the name Gracilibacillus alcaliphilus sp. nov. is proposed. The type strain is SG103(T) ( = JCM 17253(T) = NCIMB 14683(T)).

Anticancer and antioxidant effects of extracts from different parts of indigo plant

The water, methanol and ethanol extracts of flowers, leaves, stems, and seeds from indigo (Polygonum tinctorium Lour.) were tested in vitro to verify cytotoxic effects on human renal cell line (HEK 293) and inhibitory effects on the proliferation of colon cancer cell line (HCT-116), cervical carcinoma cells (HeLa), liver carcinoma cells (Hep3B), breast carcinoma cells (MCF-7), laryngeal cancer cell line (SNU-1066), and gastric cancer cell line (SNU-601), in order to further increase the use of the indigo plant. Significant differences were found in polyphenols and flavonoids in extracts of different parts of indigo. The highest amount of total polyphenols (22.94±2.18 and 22.81±2.04mgGAE/g DW) were found in methanol extracts of leaves and flowers and flavonoids (4.02±0.43, 3.70±0.34 and 3.28±0.26mgCE/gDW) – in methanol extracts of leaves, flowers and seeds. The highest antioxidant capacities by ABTS+ (99.12±9.18 and 96.35±8.75μMTE/gDW) and CUPRAC (78.37±7.86 and 86.22±8.43μMTE/gDW) were also found in methanol extracts of leaves and flowers. The correlation coefficient (R2) between antioxidant capacities and polyphenols of indigo parts in different extracts varied from 0.7142 to 0.9765, showing the highest correlation for leaves and flowers in methanol extracts. The survival rates at a concentration of 800mg/L for human renal cell line (HEK 293) in indigo plant water extracts from flowers and stems were 17.58%±9.29 and 61.78%±11.00 and in methanol extract of leaves and seeds estimated as 36.47%±18.70 and 28.7%±16.68, respectively. The extracts of stem prolonged the overall survival of cancer cells. The survival rates at a concentration of 800mg/L of colon cancer cell line (HCT-116) and cervical cancer cell line (HeLa) in the ethanol extract of indigo flowers were 5.10%±0.32 and 8.85%±0.11, respectively, and in seeds on HeLa cells was 16.93%±3.09. The survival rates of HeLa and HCT-116 in methanol extracts of indigo leaves at a concentration of 800mg/L were 6.89%±0.53 and 13.22%±8.51, respectively. Proliferative and antioxidant activities were correlated. The highest R2 between the antioxidant capacities and survival rates was in methanol extracts of leaves on HeLa cells. Therefore, investigation in vitro revealed that the extracts of the indigo plant were found to be effective in suppressing the proliferation of cancer cells, although the results varied depending on the different parts of P. tinctorium Lour., the concentrations of solvent and extracts, the antioxidant activities and the types of cancers.

Identification and quantitative analysis of polyphenolic compounds from the indigo plant (Polygonum tinctorium Lour)

The indigo plant (Polygonum tinctorium Lour) has been used traditionally as a medicinal plant with a variety of biological effects. Of these, polyphenolic ingredients are postulated to contribute to these activities. However, the identification and quantification of polyphenolic compounds in indigo plants have not been conducted comprehensively until now. This study was undertaken to identify the related ingredients by combined instrumental analyses using ultra-performance liquid chromatography electrospray-ionisation mass spectrometry and gas chromatography-mass spectrometry after the extracts of plant tissues were fractionated by absorption column chromatography. These analyses allowed the identification of kaempferol, quercetin-3-O-glucuronide, quercetin, kaempferol-3-O-glucopyranoside, caffeic acid, chlorogenic acid and tentative 3,5,4′-trihydroxy-6,7-methylenedioxyflavone. Furthermore, predominant polyphenolic compounds were quantified by reverse-phase high-performance liquid chromatography and capillary gas chromatography, revealing the higher proportions of kaempferol, quercetin-3-O-glucuronide and quercetin among them. The results indicate that the indigo plant is a promising source for flavonoids and the related compounds with beneficial medicinal effects.

Antioxidant and binding properties of methanol extracts from indigo plant leaves

AbstractThis study was conducted to clarify the effect of ultra-pressure treatment on the extraction of bioactive compounds from indigo plant leaves (Polygonum tinctorium Lour.) and on their properties. Leaves were harvested the in mid-August, 2013, from Naju City (Korea), and treated using two methods: ultra-pressure (550 MPa, 3 min) and hot-air (70°C, 24 h). Then, the leaves were ultrasonically extracted with methanol. The content of indirubin in leaves treated with ultra pressure and hot air was (535.55 ± 26.14) mg kg−1 and (52.63 ± 6.45) mg kg−1, respectively, and that of tryptanthrin was (165.55 ± 8.74) mg kg−1 and (153.00 ± 7.62) mg kg−1, respectively. Polyphenolic content in the leaves extract was (127.24 ± 13.67) mg kg−1 after the ultrapressure and (88.22 ± 5.33) mg kg−1 after the hot-air treatment. The content of flavonoids was (2298.67 ± 83.27) mg kg−1 after the ultra-pressure and (3224.00 ± 21.45) mg kg−1 after the hotair treatment. Di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH) radical scavenging activities of the indigo extract obtained by ultra-pressure and hot-air treatment methods at the concentration of 1 mg mL−1 were (80.25 ± 0.73) % and (66.54 ± 2.35) %, respectively, and 2,2′-azinobis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging activities were estimated as (90.14 ± 0.79) % and (64.45 ± 8.97) %, respectively. The methanol leaf extract after ultra-pressure treatment exhibited higher binding properties to human serum albumin in comparison with catechin and conventional treatments. Consequently, it was assumed that the ultra-pressure treatment is an effective method for the extraction of bioactive compounds from indigo leaves.

Evaluation of the effect of kaempferol in a murine allergic rhinitis model

Kaempferol (KP) is a major compound of Naju Jjok (Polygonum tinctorium Lour.). The effect of KP on allergic rhinitis (AR) has not been elucidated. Here, we report the effects and mechanisms of KP on new and predominant mediators of AR using an eosinophil cell line, Eol-1 and an ovalbumin (OVA)-induced AR mouse model. KP significantly inhibited the production of interleukin (IL)-32 and IL-8 and activation of caspase-1 in Eol-1 cells. Allergic symptoms and predominant mediators (IgE and histamine) in the KP-administered group were significantly lower than in the AR group. The levels of interferon-γ were enhanced while the levels of IL-4 were reduced in the KP group. KP significantly reduced the levels of IL-32 and thymic stromal lymphopoietin (TSLP) compared with the AR mice. KP reduced the levels of inflammation-related proteins. In the KP-administered groups, the infiltrations of eosinophils and mast cells increased by OVA were decreased. In addition, KP significantly reduced caspase-1 activity in nasal mucosa tissue of AR mice. Our findings indicate that KP has an anti-allergic effect through the regulation of the production of IL-32 and TSLP and caspase-1 activity in allergic diseases including AR.

Amphibacillus iburiensis sp. nov., an alkaliphile that reduces an indigo dye

An indigo-reducing alkaliphilic strain, designated strain N314(T), was isolated from a fermented polygonum indigo (Polygonum tinctorium Lour.) liquor sample, aged for 10 months, that was obtained from Date City, Iburi Branch, Hokkaido, Japan. The 16S rRNA gene sequence phylogeny suggested that strain N314(T) is a member of the genus Amphibacillus, with the closest relatives being Amphibacillus indicireducens (98.9 % similarity to the type strain) and Amphibacillus xylanus (98.0 % similarity to the type strain), the only species with 16S rRNA gene sequence similarities higher than 97 % to strain N314(T). The cells of the isolate stained Gram-positive and were facultatively anaerobic, straight rods that were motile by means of peritrichous flagella. The strain grew at 26-39 °C with optimum growth at 36 °C. It grew at pH 8.0-9.1, with optimum growth at pH 8.9-9.1. No isoprenoid quinone was detected, and the DNA G+C content was 38.4 mol%. The whole-cell fatty acid profile consisted mainly of iso-C15 : 0 and anteiso-C15 : 0. Analysis of DNA-DNA hybridization with the type strains of A. indicireducens and A. xylanus revealed 29±2 % and 10±2 % relatedness, respectively. Owing to differences in phenotypic characteristics from reported species of the genus A. and results of phylogenetic analyses based on 16S rRNA gene sequences and DNA-DNA relatedness data, the isolate merits classification within a novel species, for which the name Amphibacillus iburiensis sp. nov. is proposed. The type strain is N314(T) ( = JCM 18529(T) = NCIMB 14823(T)).

Oceanobacillus polygoni sp. nov., a facultatively alkaliphile isolated from indigo fermentation fluid

A facultatively alkaliphilic, lactic-acid-producing and halophilic strain, designated SA9(T), was isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample prepared in a laboratory. The 16S rRNA gene sequence phylogeny suggested that strain SA9(T) was a member of the genus Oceanobacillus with the closest relative being Oceanobacillus profundus KCCM 42318(T) (99.3% 16S rRNA gene sequence similarity). Cells of strain SA9(T) stained Gram-positive and were facultative anaerobic straight rods that were motile by peritrichous flagella. The strain grew between 5 and 48 °C (optimum, 35 °C) and at pH 7-12 (optimum, pH 9). The isoprenoid quinone detected was menaquinone-7 (MK-7) and the DNA G+C content was 40.6 ± 0.9 mol%. The whole-cell fatty acid profile mainly consisted of iso-C(15:0), anteiso-C(15:0), C(16:0) and anteiso-C(17:0). DNA-DNA hybridization with Oceanobacillus profundus DSM 18246(T) revealed a DNA-DNA relatedness value of 23 ± 2%. On the basis of the differences in phenotypic and chemotaxonomic characteristics, and the results of phylogenetic analyses based on 16S rRNA gene sequences and DNA-DNA relatedness data from recognized species of the genus Oceanobacillus, strain SA9(T) merits classification as a representative of a novel species of the genus Oceanobacillus, for which the name Oceanobacillus polygoni sp. nov. is proposed. The type strain is SA9(T) ( =JCM 17252(T) =NCIMB 14684(T)). An emended description of the genus Oceanobacillus is also provided.

Oceanobacillus indicireducens sp. nov., a facultative alkaliphile that reduces an indigo dye

An indigo-reducing facultatively alkaliphilic and halophilic strain, designated strain A21(T), was isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample aged for 4 days prepared in a laboratory. 16S rRNA gene sequence phylogeny suggested that strain A21(T) was a member of the genus Oceanobacillus with the closest relative being the type strain of Oceanobacillus chironomi (similarity: 96.0 %). The cells of the isolate stained Gram-positive and were facultatively anaerobic straight rods that were motile by peritrichous flagella. The strain grew between 18 and 48 °C with optimum growth at 39 °C. It grew in the pH range of 7-12. It hydrolysed casein, gelatin and Tween 20 but not Tweens 40, 60 and 80, starch or DNA. No isoprenoid quinone was detected and the DNA G+C content was 39.7 mol%. The whole-cell fatty acid profile mainly consisted of iso-C15 : 0, anteiso-C15 : 0 and C16 : 0. DNA-DNA hybridization experiments with O. chironomi revealed 13 % relatedness. Owing to the differences in phenotypic and chemotaxonomic characteristics, and phylogenetic analyses based on 16S rRNA gene sequences and DNA-DNA relatedness data from reported Oceanobacillus species, the isolate merits classification as a representative of a novel species, for which the name Oceanobacillus indicireducens sp. nov. is proposed. The type strain is A21(T) ( = JCM 17251(T) = NCIMB 14685(T)). The description of the genus Oceanobacillus is also emended.

Increased indigoid accumulation by plant defense activators in Polygonum tinctorium Lour.

Effect of plant defense activator on Polygonum tinctorium was evaluated to increase indigoid content in P. tinctorium. Three plant defense activators, 2,1,3-benzothiadiazole (BTH), 2,6-dichloroisonicotinic acid (INA), and acibenzolar-S methyl (ASM) were treated to two cultivars (Naju Local and Naram Blue) under the field condition at two different concentration regimens of 0.3 and 3 mM and harvested 7 days after treatment. The treatments increased indigo accumulation by 40 to 109% with statistical significance. Considering the treatment concentrations, BTH and INA at 0.3 mM were judged to be optimal with 2-fold increase in indigoid concentration. No statistical differences were found in the cultivars and maturity of leaves in the indigoid accumulation caused by plant activator treatment. This result is directly applicable to the P. tinctorium cultivation with high economic significance.

Amphibacillus indicireducens sp. nov., an alkaliphile that reduces an indigo dye

Two indigo-reducing alkaliphilic strains, designated strain C40(T) and strain N214, were isolated from a fermented Polygonum Indigo (Polygonum tinctorium Lour.) liquor sample aged for 10 months and obtained from Date City, Hokkaido, Japan. 16S rRNA gene sequence phylogeny suggested that strains C40(T) and N214 were members of the genus Amphibacillus with the closest relative being Amphibacillus xylanus JCM 7361(T) (97.5 % 16S rRNA gene sequence similarity with strain C40(T)), which is the only strain having a 16S rRNA gene sequence similarity higher than 97 % with strain C40(T). Cells of strain C40(T) were Gram-stain-positive, facultatively anaerobic, straight rods that were motile by means of peritrichous flagella. The strains grew between 17 and 39 °C (optimum, 35 °C) and in the pH range of 9.0-12.0. No isoprenoid quinone was detected and the DNA G+C content was 37.5-37.7 mol%. The whole-cell fatty acid profile mainly consisted of iso-C(15 : 0) and anteiso-C(15 : 0). DNA-DNA hybridization of strain C40(T) with Amphibacillus xylanus JCM 7361(T) revealed a DNA-DNA relatedness value of 10±3 %. Owing to the differences in phenotypic characteristics and phylogenetic analyses based on 16S rRNA gene sequences, as well as DNA-DNA relatedness data from reported species of the genus Amphibacillus, the isolates merit classification as a novel species in the genus Amphibacillus, for which the name Amphibacillus indicireducens sp. nov. is proposed. The type strain is C40(T) ( = JCM 17250(T) = NCIMB 14686(T)). An additional strain of the species is N214. An emended description of the genus Amphibacillus is provided.