Paenibacillus alkalitolerans sp. nov., a bacterium isolated from a salt lake of Turpan City in Xinjiang Province, north-west China.

A Gram stain-positive, rod-shaped, motile, aerobic and terminal endospore formation bacterium, designated YIM B00362T, was isolated from saline soil samples collected from a salt lake in Xinjiang Province, north-west China. Phylogenetic analysis based on the 16S rRNA gene sequences and whole genomes indicated that the isolate belongs to the genus Paenibacillus. However, the highest sequence similarity between strain YIM B00362T and the relatives was only 94.4%. Moreover, the DNA-DNA relatedness and ANI values between the novel isolate and the relative type strain, Paenibacillus antri SYSU K30003T was 13.6% and 70.3%, respectively. The major cellular fatty acids were anteiso-C15:0, C16:0 and the major quinone was MK-7. The isolate contained meso-diaminopimelic acid as the diagnostic diamino acid and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylglyceride, and two unidentified polar lipids. The genomic DNA G + C content was 50.9 mol%. The major whole-cell sugars contained glucose and galactose. On the basis of physiological, phenotypic, and chemotaxonomic data, strain YIM B00362T represents a novel species of genus Paenibacillus, for which the name Paenibacillus alkalitolerans sp. nov. is proposed. The type strain is YIM B00362T (= KCTC 43272 T = CGMCC 1.18801 T = NBRC 114667 T).

New antibacterial depsidones from an ant-derived fungus Spiromastix sp. MY-1.

Six new (1-6) and seven known depsidones (7-13) were isolated from the culture of an ant (Monomorium chinensis)-derived fungus Spiromastix sp. MY-1. Their structures were elucidated by extensive spectroscopic analysis including high resolution MS, 1D and 2D NMR data. The new bromide depsidones were obtained through supplementing potassium bromide in the fermentation medium of Spiromastix sp. MY-1. All isolated compounds showed various bioactivities against the tested phytopathogenic bacteria. Particularly, new bromide compound 4, named spiromastixone S, exhibited the strongest activity against Xanthomonas oryzae pv. oryzae with a MIC value of 5.2 µmol·-1.

Uniportal video-assisted thoracoscopic surgery for complex mediastinal mature teratoma: A case report.

BACKGROUND: Mediastinal mature teratoma is the most common histological type of primary extragonadal germ cell tumor. In this report, we describe a rare case of giant mature teratoma located primarily in the anterior mediastinum and causing partial atelectasis of the upper and middle lobes of the right lung, as well as extrinsic compression of the right atrium. CASE SUMMARY: A 31-year-old male with a giant mediastinal mature teratoma presented with progressive exertional dyspnea and chest pain for 1 mo. Computed tomography of the chest indicated the diagnosis of anterior mediastinal teratoma. The patient underwent right uniportal anterior approach video-assisted thoracoscopic surgery (VATS). En bloc resection of the giant teratoma, wedge resection of the upper and middle lobes of the right lung, resection of the thymus and partial excision of the pericardium were successfully performed. The pathological diagnosis revealed a mature cystic teratoma with foreign-body reaction that was closely related to the right lung, atrium dextrum, superior vena cava and ascending aorta. An atrophic thymic tissue was also discovered at the external teratoma surface. The patient was discharged on postoperative day 7. CONCLUSION: This is the first report of the use of uniportal VATS for complete resection of a teratoma in combination with wedge resection of the right upper and middle lung lobes and partial resection of the pericardium.

Alkalibacillus aidingensis sp. nov., an Bacterium Isolated from Aiding Lake in Xinjiang Province, North-West China.

A bacterial strain, Gram staining negative, aerobic, long rod, motile bacterium with flagellum, designated strain YIM 98829T, was isolated from the Aiding Lake in Xinjiang province, North-West China. The isolate produced oval subterminal endospores in swollen sporangia. The predominant menaquinone was MK-7. The cell wall peptidoglycan contained ornithine, serine, aspartic acid, glutamic acid, and alanine, while diaminopimelic acid could not be detected. The major whole-cell sugars contained xylose, glucose, galactose, and mannose. Diphosphatidylglycerol, phosphatidylglycerol, one unknown phospholipid, and two unidentified aminophospholipids were part of the polar lipid profile. Iso-C15:0 and anteiso-C15:0 were the major fatty acids. The DNA G + C content of the type strain was 38.0 mol%. Phylogenetic analysis indicated that the isolate belongs to the genus Alkalibacillus. However, it differed from its closest relatives, A. haloalkaliphilus DSM 5271T (97.04%), A. filiformis 4AGT (96.99%), and A. silvisoli BM2T (96.95%) in some physiological characteristics. DNA-DNA hybridization result indicated low levels of relatedness between strain YIM 98829T and A. haloalkaliphilus JCM 12303T (16.9%). On the basis of physiological, phenotypic, and chemotaxonomic data, strain YIM 98829T represents a novel species of genus Alkalibacillus, for which the name Alkalibacillus aidingensis sp. nov. is proposed. The type strain is YIM 98829T (= NBRC 114103T = CGMCC 1.17260T = DSM 112470T).

Oceanobacillus salinisoli sp. nov., a bacterium isolated from saline soil of Turpan city in Xinjiang province, north-west China.

YIM B00359T, a novel bacterial strain was isolated from the saline soil of Turpan city in Xinjiang province, north-west China. The strain was Gram-stain-positive, motile, aerobic, produced oval subterminal endospores in swollen sporangia. The whole-cell hydrolysates contain meso-diaminopimelic acid as the cell-wall diamino acid, with xylose, glucose, and ribose as the major whole-cell sugars. The phospholipids are diphosphatidylglycerol, phosphatidylglycerol, unidentified phospholipids, unidentified glycolipids, and one unidentified glycophospholipid. The predominant menaquinone is MK-7. The major fatty acids are anteiso-C15:0, iso-C14:0, iso-C15:0, and iso-C16:0. The DNA G + C content of the type strain is 37.5 mol%. Phylogenetic analysis indicated that the isolate belongs to the genus Oceanobacillus. However, it differed from its closest relatives, Oceanobacillus halophilus DSM 23996 T and Oceanobacillus senegalensis Marseille-P3587T in many physiological and chemotaxonomic characteristics. Based on comparative analysis of polyphasic taxonomic data, strain YIM B00359T represents a novel species of the genus Oceanobacillus, for which the name Oceanobacillus salinisoli sp. nov. is proposed. The type strain is YIM B00359T (= CGMCC 1.17509T = KCTC 43185T).

Haloechinothrix aidingensis sp. nov., an actinomycete isolated from salt lake in Xinjiang province, north-west China.

An actinomycete strain, designated YIM 98757T, was isolated from the hypersaline sediment of Aiding Lake in Xinjiang province, north-west China. The strain grew well on most media tested and no diffusible pigment was produced. The substrate mycelium was well developed and fragmented. No spores were formed. The whole-cell hydrolysates contained meso-diaminopimelic acid as the cell-wall diamino acid. Xylose, galactose, ribose were the major whole-cell sugars. The phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides and an unknown phospholipid. The predominant menaquinone was MK-8(H4). The major fatty acid was iso-C16:0. The DNA G + C content was 69.1 mol%. Phylogenetic analysis indicated that the isolate belongs to the genus Haloechinothrix. However, it differed from its closest relative, H. alba YIM 98757 T in many phenotypic and chemotaxonomic characteristics. Moreover, the DNA-DNA and ANI relatedness values between the novel isolate and H. alba YIM 93221 T were 53.3% and 92.5%, respectively. Based on comparative analysis of polyphasic taxonomic data, strain YIM 98757 T represents a novel species of the genus Haloechinothrix, for which the name Haloechinothrix aidingensis sp. nov. is proposed. The type strain is YIM 98757T (= CGMCC 4.7627T = CCTCC AA 2020012).

Alpinumisoflavone triggers GSDME-dependent pyroptosis in esophageal squamous cell carcinomas.

Esophageal squamous cell carcinoma (ESCC) presents a common human malignancy in the digestive system. We aimed to explore the critical effects of alpinumisoflavone (AIF) on ESCC in vitro and in vivo. The cell counting kit-8 assay was used to determine cell viability. Colony formation assay was employed to examine the effect of AIF on the long-term growth of ESCC cells. Cell apoptosis was determined by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay. Cell morphologies were observed by light microscopy. The enzyme-linked immunosorbent assay was performed to examine the lactate dehydrogenase release from AIF-treated cells. Immunofluorescent labeling was utilized to examine AIF-induced GSDME expression. Western blot was employed to determine the expression levels of the associated proteins. Immunohistochemistry was performed to determine the localization and expression of the associated proteins in mice tumor tissues. AIF inhibited ESCC cell viability and suppressed cell growth in a dose- and time-dependent fashion. Results showed that AIF promoted apoptosis in ESCC cells. Meanwhile, our results also showed that AIF triggered pyroptotic cell death in ESCC, which was mediated by gasdermin E (GSDME) cleavage. In addition, our experiments provided experimental evidence that AIF-induced GSDME cleavage was dependent on caspase-3 activation. Moreover, the inhibition of GSDSE by knockdown was able to switch the form of cell death from pyroptosis to apoptosis. Furthermore, the results from the xenograft animal model also supported our findings in vitro that AIF was able to promote GSDME-mediated pyroptotic cell death in ESCC. AIF inhibited ESCC growth in vitro and in vivo by triggering GSDME-mediated pyroptotic cell death, which is dependent on caspase-3 activation.

Paenibacillus turpanensis sp. nov., isolated from a salt lake of Turpan city in Xinjiang province, north-west China.

Strain YIM B00363T, a Gram-positive, aerobic, non-motile, rod-shaped, spore-forming bacterium, was isolated from saline soil samples collected from a salt lake in Xinjiang province, north-west China, and was characterized using a polyphasic approach. The optimum growth temperature was 37 °C and the optimum pH was 7.5-8.0. The major menaquinone was MK-7; anteiso-C15:0 (53.52%), iso-C15:0 (15.04%) and C16:0 (12.76%) were the predominant cellular fatty acids. The diagnostic diamino acid of the cell wall peptidoglycan was meso-diaminopimelic acid. The phospholipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, unidentified phospholipids, unidentified glycolipids and unknown lipids. The DNA G + C content of the type strain was 50.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain YIM B00363T belonged to a cluster comprising species of the genus Paenibacillus. The nearest relatives were P. residui MC-246T and P. senegalensis JC66T, with 93.2% and 92.8% gene sequence similarities, respectively. On the basis of its phenotypic characteristics and phylogenetic distinctivenes, strain YIM B00363T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus turpanensis sp. nov. is proposed. The type strain is YIM B00363T (= CGMCC 1.17507T = KCTC 43184T).

Oceanobacillus halotolerans sp. nov., a bacterium isolated from salt lake in Xinjiang province, north-west China.

A bacterial strain, designated YIM 98839T, was isolated from the hypersaline sediment of Aiding Lake in Xinjiang province, North-West China. The strain was Gram-stain-positive, motile, aerobic, produced oval subterminal or central endospores in swollen sporangia. The whole-cell hydrolysates contain meso-diaminopimelic acid as the diagnostic cell-wall diamino acid. Galactose, fucose and ribose are the major whole-cell sugars. The phospholipids are diphosphatidylglycerol, phosphatidylglycerol and one unknown phospholipid. The predominant menaquinone is MK-7. The major fatty acids are anteiso-C15:0, anteiso-C17:0 and iso-C15:0. The DNA G + C content of the type strain is 37.0 mol%. Phylogenetic analysis indicated that the isolate belongs to the genus Oceanobacillus. However, it differed from its closest relative, Oceanobacillus limi H9BT in many physiological characteristics. Moreover, the DNA-DNA relatedness values between the novel isolate and the relative type strain was 20.2%. Based on comparative analysis of polyphasic taxonomic data, strain YIM 98839T represents a novel species of the genus Oceanobacillus, for which the name Oceanobacillus halotolerans sp. nov. is proposed. The type strain is YIM 98839T (= CGMCC 1.17002T = KCTC 43140T).

Actinorugispora endophytica gen. nov., sp. nov., an actinomycete isolated from Daucus carota.

An actinomycete strain, designated YIM 690008T, was isolated from Daucus carota collected from South Korea and its taxonomic position was investigated by using a polyphasic approach. The strain grew well on most media tested and no diffusible pigment was produced. The aerial mycelium formed wrinkled single spores and short spore chains, some of which were branched. The whole-cell hydrolysates contained meso-diaminopimelic acid, glucose, mannose, ribose, galactose and rhamnose. The predominant menaquinones were MK-10(H4), MK-10(H6), MK-10(H8) and MK-10(H2). The polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannosides, some unknown phospholipids, glycolipids and polar lipids. The major fatty acids were i-C16 : 0, ai-C17 : 0 and C18 : 1ω9c. The DNA G+C content of the genomic DNA was 63.1 mol%. Phylogenetic analysis indicated that the isolate belongs to the family Nocardiopsaceae. However, based on phenotypic, chemotaxonomic and genotypic data, it was concluded that strain YIM 690008T represents a novel genus and novel species of the family Nocardiopsaceae, for which the name Actinorugispora endophytica gen. nov., sp. nov. (type strain YIM 690008T = DSM 46770T = JCM 30099T = KCTC 29480T) is proposed.

Drimane sesquiterpenoids from the Aspergillus oryzae QXPC-4.

Three new drimane sesquiterpenoids, astellolides C-E (1-3, resp.), four new drimane sesquiterpenoid p-hydroxybenzoates, astellolides F-I (4-7, resp.), together with two known compounds astellolides A and B (8 and 9, resp.), have been isolated from the liquid culture of Aspergillus oryzae (strain No. QXPC-4). Their structures were established by comprehensive analysis of spectroscopic data. The relative and absolute configurations were determined on the basis of NOESY and CD data, together with single-crystal X-ray diffraction analyses of compounds 1-3. The metabolites were evaluated for their cytotoxic activities, however, no compounds showed a significant cytotoxicity against the tested cell lines at a concentration of 20 µM.

Rothia endophytica sp. nov., an actinobacterium isolated from Dysophylla stellata (Lour.) Benth.

A novel endophytic actinobacterium, designated strain YIM 67072(T), was isolated from healthy roots of Dysophylla stellata (Lour.) Benth. Cells of this aerobic, cream-yellow-coloured strain occurred singly, in pairs or in tetrads, were Gram-stain-positive and ovoid- to spherical-shaped. Strain YIM 67072(T) grew at 4-45 °C, pH 5.0-10.0 and in the presence of 0-7 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 67072(T) belonged to the genus Rothia. The isolate contained MK-7 as the major component of the quinone system. The peptidoglycan type was A3α. The polar lipid profile consisted predominantly of diphosphatidylglycerol and glycolipids. The major fatty acids were anteiso-C15 : 0, iso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The DNA G+C content was 53.2 mol%. However, strain YIM 67072(T) differed from its closest relatives Rothia nasimurium CCUG 35957(T) (98.5 % 16S rRNA gene sequence similarity), Rothia amarae JCM 11375(T) (97.6 %) and Rothia terrae L-143(T) (97.3 %) in many phenotypic characteristics. Moreover, the levels of DNA-DNA relatedness between the novel isolate and the three above-mentioned type strains were 28.7±1.3 %, 36.5±1.2 %, 46.8±1.5 %, respectively. Based on comparative analysis of physiological and chemotaxonomic data, strain YIM 67072(T) represents a novel species of the genus Rothia, for which the name Rothia endophytica sp. nov. is proposed. The type strain is YIM 67072(T) ( = DSM 26247(T) = JCM 18541(T)).

Blastococcus endophyticus sp. nov., an actinobacterium isolated from Camptotheca acuminata.

A novel endophytic actinobacterium, designated strain YIM 68236(T), was isolated from healthy leaves of Camptotheca acuminata. and characterized by using a polyphasic approach. Cells of this strain occurred singly, in pairs or in tetrads. It grew at 10-45 °C, at pH 5.0-8.0 (optimum pH 7.0) and in the presence of 0-3% (w/v) NaCl. The DNA G+C content was 71.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 68236(T) belongs to the genus Blastococcus. However, it differed from its closest relatives, Blastococcus aggregatus DSM 4725(T), Blastococcus saxobsidens DSM 44509(T) and Blastococcus jejuensis DSM 19597(T) in many phenotypic characteristics. Moreover, the DNA-DNA relatedness values between the novel isolate and the three above-mentioned type strains were 49.0 ± 1.6%, 46.1 ± 3.2% and 39.8 ± 1.5%, respectively. Based on comparative analysis of physiological and chemotaxonomic data, strain YIM 68236(T) represents a novel species of the genus Blastococcus, for which the name Blastococcus endophyticus sp. nov. is proposed. The type strain is YIM 68236(T) ( =CCTCC AA 209045(T) =DSM 45413(T) =KCTC 19998(T)).

Streptomyces hundungensis sp. nov., a novel actinomycete with antifungal activity and plant growth promoting traits.

A novel actinobacterium MBRL 251(T), isolated from a sample collected from a limestone quarry at Hundung, Manipur, India, was characterized using a polyphasic approach. The 16S ribosomal RNA gene sequence of strain MBRL 251(T) showed closest similarities with Streptomyces xanthochromogenes NRRL B-5410(T) (99.6%) and Streptomyces michiganensis NBRC 12797(T) (99.6%). The DNA relatedness between MBRL 251(T) and S. xanthochromogenes NBRC 12828(T), and S. michiganensis NBRC 12797(T) was 46.6% and 40.7%, respectively. Strain MBRL 251(T) contained LL-diaminopimelic acid as the diagnostic diamino acid, with glucose and xylose as the main cell wall sugars, whereas small amounts of galactose, mannose, rhamnose and ribose were also detected in the whole-cell wall hydrolysates. The major fatty acids identified were anteiso-C15:0 (35.1%), iso-C16:0 (21.1%) and anteiso-C17:1 (13.2%). The predominant menaquinones detected were MK-9(H6) and MK-9(H8), whereas the polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositolmannosides. The G+C content of the genomic DNA was 72.3%. The phenotypic and genotypic data showed that strain MBRL 251(T) merits the recognition as a representative of a novel species of the genus Streptomyces. It is proposed that the isolate should be classified in the genus Streptomyces as a novel species, Streptomyces hundungensis sp. nov. The type strain is MBRL 251(T) (=JCM 17577(T)=KCTC 29124(T)).

Rhodococcus canchipurensis sp. nov., an actinomycete isolated from a limestone deposit site.

A novel actinobacterial strain, MBRL 353(T), was isolated from a sample collected from a limestone quarry at Hundung, Manipur, India. Comparison of 16S rRNA gene sequences of strain MBRL 353(T) and other members of the genus Rhodococcus showed sequence similarities ranging from 95.5 to 98.2 %, with strain MBRL 353(T) showing closest sequence similarity to Rhodococcus triatomae IMMIB RIV-085(T) (98.2 %) and Rhodococcus equi DSM 20307(T) (97.2 %). DNA-DNA hybridization results, however, revealed that DNA-DNA relatedness values between strain MBRL 353(T) and R. triatomae DSM 44892(T) (43.4 %) and R. equi DSM 20307(T) (33.4 %) were well below the 70 % limit for species identification. Strain MBRL 353(T) contained meso-diaminopimelic acid as the diagnostic diamino acid and galactose and arabinose in the cell wall. Mycolic acids were present. The major fatty acids were C(16 : 0) (45.7 %), C(18 : 1)ω9c (18.2 %) and 10-methyl C(18 : 0) (11.3 %). The only menaquinone detected was MK-8(H(2)), while the major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and one unknown phospholipid. The G+C content of the genomic DNA was 69.2 mol%. The phenotypic and genotypic data showed that strain MBRL 353(T) merits recognition as a representative of a novel species of the genus Rhodococcus for which the name Rhodococcus canchipurensis sp. nov. is proposed; the type strain is MBRL 353(T) (= KCTC 19851(T) = JCM 17578(T)).

Streptomyces endophyticus sp. nov., an endophytic actinomycete isolated from Artemisia annua L.

Three filamentous actinomycetes, strains YIM 65594(T), YIM 65638 and YIM 65642, were isolated from the surface-sterilized roots of Artemisia annua L. collected from Yunnan province, south-west China. These strains were found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces. The organisms formed an extensively branched substrate mycelium, with abundant aerial hyphae that differentiated into spores. The cell wall of the isolates contained ll-diaminopimelic acid and the menaquinones were MK-9(H(8)) and MK-9(H(6)). The major fatty acids were anteiso-C(15 : 0), anteiso-C(17 : 0) and iso-C(16 : 0). Phylogenetic analysis of the 16S rRNA gene sequences of these strains revealed that the strains clustered together and were most closely related to Streptomyces kunmingensis NBRC 14463(T), with 98.5-98.6 % 16S rRNA gene sequence similarity. The results of DNA-DNA hybridization and physiological tests allowed the genotypic and phenotypic differentiation of strains YIM 65594(T), YIM 65638 and YIM 65642 from related species. However, the high level of DNA-DNA relatedness between them showed that these three strains belong to the same species. Strain YIM 65594(T) (= DSM 41984(T) = CCTCC AA 209036(T)) was selected as the type strain to represent this novel species, for which the name Streptomyces endophyticus sp. nov. is proposed.

An endophytic Pseudonocardia species induces the production of artemisinin in Artemisia annua.

Endophytic actinobacteria colonize internal tissues of their host plants and are considered as a rich and reliable source of diverse species and functional microorganisms. In this study, endophytic actinobacterial strain YIM 63111 was isolated from surface-sterilized tissue of the medicinal plant Artemisia annua. We identified strain YIM 63111 as a member of the genus Pseudonocardia. A. annua seedlings grown under both sterile and greenhouse conditions were inoculated with strain YIM 63111. The growth of A. annua seedlings was strongly reduced when YIM 63111 was inoculated at higher concentrations under sterile conditions. However, no growth inhibition was observed when A. annua was grown under greenhouse conditions. Using an enhanced green fluorescent protein (EGFP) expressing YIM 63111 strain, we also observed the endophytic colonization of A. annua seedling using confocal laser-scanning microscopy. The transcription levels of the key genes involved in artemisinin biosynthesis were investigated using real time RT-PCR, revealing that cytochrome P450 monooxygenase (CYP71AV1) and cytochrome P450 oxidoreductase (CPR) expression were up-regulated in A. annua upon inoculation with strain YIM 63111 under certain conditions. The up-regulation of these genes was associated with the increased accumulation of artemisinin. These results suggest that endophytic actinobacteria effectively stimulate certain plant defense responses. Our data also demonstrate the use of Pseudonocardia sp. strain YIM 63111 as a promising means to enhance artemisinin production in plants.

Streptomyces manipurensis sp. nov., a novel actinomycete isolated from a limestone deposit site in Manipur, India.

A novel actinobacterium, designated MBRL 201(T), was isolated from a sample collected from a limestone quarry at Hundung, Manipur, India. The strain was characterized using polyphasic taxonomy. Comparison of the 16S rRNA gene sequence of strain MBRL 201(T) and other Streptomyces species showed sequence similarities ranging from 93.0 to 99.6 % and strain MBRL 201(T) showed closest similarities to Streptomyces virginiae NBRC 12827(T) (99.6 %) and Streptomyces cinnamonensis NBRC 15873(T) (99.6 %). The DNA relatedness between MBRL 201(T) and the type strains of S. virginiae NBRC 12827(T) and S. cinnamonensis NBRC 15873(T) were 44.5 and 35.6 % respectively. Strain MBRL 201(T) contained LL: -diaminopimelic acid (A(2)pm) as the diagnostic diamino acid, with glucose as the main sugar, while small amounts of galactose, glucose, mannose, rhamnose, ribose and xylose were also present in cell-wall hydrolysates. The major fatty acids identified were anteiso-C(15:0) (38.9 %), iso-C(15:0) (19.9 %) and anteiso-C(17:1) (14.7 %). The predominant menaquinones detected were MK-9(H(6)) and MK-9(H(8)), while the polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositolmannosides, with other unknown phospholipids and lipids. The G+C content of the genomic DNA was 72.9 %. The phenotypic and genotypic data showed that strain MBRL 201(T) merits recognition as a representative of a novel species of the genus Streptomyces. It is proposed that the isolate should be classified in the genus Streptomyces as a novel species, Streptomyces manipurensis sp. nov. The type strain is MBRL 201(T) (=DSM 42029(T) = JCM 17351(T)).

Abundant and diverse endophytic actinobacteria associated with medicinal plant Maytenus austroyunnanensis in Xishuangbanna tropical rainforest revealed by culture-dependent and culture-independent methods.

Endophytes are now considered as an important component of biodiversity. However, the diversity of endophytic actinobacteria associated with tropical rainforest native medicinal plants is essentially unknown. In this study, the diversity of endophytic actinobacteria residing in root, stem and leaf tissues of medicinal plant Maytenus austroyunnanensis collected from tropical rainforest in Xishuangbanna, China was investigated with a combination of cultivation and culture-independent analysis on the basis of 16S rRNA gene sequencing. By using different selective isolation media and methods, a total of 312 actinobacteria were obtained, and they were affiliated with the order Actinomycetales (distributed into 21 genera). Based on a protocol for endophytes enrichment, three 16S rRNA gene clone libraries were constructed and 84 distinct operational taxonomic units were identified and they distributed among the orders Actinomycetales and Acidimicrobiales, including eight suborders and at least 38 genera with a number of rare actinobacteria genera. Phylogenetic analysis showed that 32% of the clones in the libraries had lower than 97% similarities with related type strains. Interestingly, six genera from the order Actinomycetales and uncultured clones from Acidimicrobiales have not, to our knowledge, been previously reported as endophytes. Our study confirms abundant endophytic actinobacterial consortium in tropical rainforest native plant and suggests that this special habitat represents an underexplored reservoir of diverse and novel actinobacteria of potential interest for bioactive compounds discovery.

Rhodococcus artemisiae sp. nov., an endophytic actinobacterium isolated from the pharmaceutical plant Artemisia annua L.

A Gram-positive, non-motile actinobacterium, designated YIM 65754T, was isolated from the stem of Artemisia annua L., collected from Yunnan province, south-west China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YIM 65754T comprised an evolutionary lineage within the genus Rhodococcus. The isolate clustered with Rhodococcus pyridinivorans PDB9T, Rhodococcus gordoniae W 4937T and Rhodococcus rhodochrous DSM 43241T, with which it shared 98.4, 97.9 and 97.8 % 16S rRNA gene sequence similarities, respectively. However, DNA-DNA relatedness demonstrated that strain YIM 65754T was distinct from its closest phylogenetic neighbours. The cell-wall peptidoglycan contained meso-diaminopimelic acid, arabinose, galactose, mannose and glucose (cell-wall chemotype IV). The major menaquinone was MK-8(H2) and the predominant fatty acids were C16:0 (27.83 %), iso-C15:0 2-OH and/or C16:1ω7c (20.21 %) and 10-methyl C18:0 (17.50 %). The DNA G+C content was 66.2 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic evidence, the isolate represents a novel species of the genus Rhodococcus, for which the name Rhodococcus artemisiae sp. nov. is proposed; the type strain is YIM 65754T (=CCTCC AA 209042T=DSM 45380T).