Peimenine unleashes therapeutic promise in urothelial bladder cancer: inhibition of proliferation, induction of cell death and modulation of key pathways.

Peimenine (PEI) is a steroid alkaloid substance isolated from Fritillaria thunbergii bulbs. It has various pharmacological activities, such as relief from coughs and asthma, expectorant properties, antibacterial effects, sedative qualities, and anti-inflammatory properties. Notably, PEI can effectively inhibit the proliferation and tumor formation of liver cancer and osteosarcoma cells by inducing autophagic cell death. However, the precise effect and mechanisms of PEI on urothelial bladder cancer (UBC) cells remain uncertain. Thus, this study aims to investigate the impact of PEI on UBC cells both in vivo and in vitro. The IC50 values of BIU-87 and EJ-1 cells after 48 h were 710.3 and 651.1 µg/mL, respectively. Additionally, PEI blocked the cell cycle in BIU-87 and EJ-1 cells during the G1 phase. Furthermore, it hindered the migration of BIU-87 and EJ-1 cells substantially. PEI significantly inhibited the tumor development of EJ-1 cells within the xenograft tumor model in vivo. Mechanically, PEI augmented the protein and mRNA expression of BIM, BAK1, and Cytochrome C (CYCS) in UBC cells. Taken together, PEI suppressed the proliferation of UBC cells both in vitro and in vivo by inducing cell death and cell cycle arrest, suggesting that PEI could be applied in the treatment of UBC.

Three-dimensional chromatin landscapes in hepatocellular carcinoma associated with hepatitis B virus.

BACKGROUND: Three-dimensional (3D) chromatin architecture frequently altered in cancer. However, its changes during the pathogenesis of hepatocellular carcinoma (HCC) remained elusive. METHODS: Hi-C and RNA-seq were applied to study the 3D chromatin landscapes and gene expression of HCC and ANHT. Hi-C Pro was used to generate genome-wide raw interaction matrices, which were normalized via iterative correction (ICE). Moreover, the chromosomes were divided into different compartments according to the first principal component (E1). Furthermore, topologically associated domains (TADs) were visualized via WashU Epigenome Browser. Furthermore, differential expression analysis of ANHT and HCC was performed using the DESeq2 R package. Additionally, dysregulated genes associated with 3D genome architecture altered were confirmed using TCGA, qRT-PCR, immunohistochemistry (IHC), etc. RESULTS: First, the intrachromosomal interactions of chr1, chr2, chr5, and chr11 were significantly different, and the interchromosomal interactions of chr4-chr10, chr13-chr21, chr15-chr22, and chr16-chr19 are remarkably different between ANHT and HCC, which resulted in the up-regulation of TP53I3 and ZNF738 and the down-regulation of APOC3 and APOA5 in HCC. Second, 49 compartment regions on 18 chromosomes have significantly switched (A-B or B-A) during HCC tumorigenesis, contributing to up-regulation of RAP2A. Finally, a tumor-specific TAD boundary located on chr5: 6271000-6478000 and enhancer hijacking were identified in HCC tissues, potentially associated with the elevated expression of MED10, whose expression were associated with poor prognosis of HCC patients. CONCLUSION: This study demonstrates the crucial role of chromosomal structure variation in HCC oncogenesis and potential novel biomarkers of HCC, laying a foundation for cancer precision medicine development.

Micromonospora profundi TRM 95458 converts glycerol to a new osmotic compound.

Plant growth and agricultural productivity was greatly limited by soil salinity and alkalization. The application of salt-tolerant rhizobacteria could effectively improve plant tolerance to saline-alkali stress. Micromonospora profundi TRM 95458 was obtained from the rhizosphere of chickpea (Cicer arietinum L.) as a moderate salt-tolerant rhizobacteria. A new osmotic compound (ABAGG) was isolated from the fermentation broth of M. profundi TRM 95458. The chemical structure of the new compound was elucidated by analyzing nuclear magnetic resonance (NMR) and high-resolution mass (HRMS) data. M. profundi TRM 95458 could convert glycerol into ABAGG. The accumulation of ABAGG varied depending on the amount of glycerol and glycine added to the fermentation medium. In addition, the concentration of NaCl affected the ABAGG content obviously. The highest yield of ABAGG was observed when the salt content of the fermentation medium was 10 g/L. The study indicated that salt stress led to the accumulation of ABAGG using glycerol and glycine as substrates, suggesting ABAGG might aid in the survival and adaptation of the strain in saline-alkaline environments as a new osmotic compound.

Specific subsets of urothelial bladder carcinoma infiltrating T cells associated with poor prognosis.

Comprehensive investigation of tumor-infiltrating lymphocytes in cancer is crucial to explore the effective immunotherapies, but the composition of infiltrating T cells in urothelial bladder carcinoma (UBC) remains elusive. Here, single-cell RNA sequencing (scRNA-seq) were performed on total 30,905 T cells derived from peripheral blood, adjacent normal and tumor tissues from two UBC patients. We identified 18 distinct T cell subsets based on molecular profiles and functional properties. Specifically, exhausted T (TEx) cells, exhausted NKT (NKTEx) cells, Ki67+ T cells and B cell-like T (B-T) cells were exclusively enriched in UBC. Additionally, the gene signatures of TEx, NKTEx, Ki67+ T and B-T cells were significantly associated with poor survival in patients with BC and various tumor types. Finally, IKZF3 and TRGC2 are the potential biomarkers of TEx cells. Overall, our study demonstrated an exhausted context of T cells in UBC, which layed a theoretical foundation for the development of effective tumor immunotherapies.

Three novel Actinoplanes species isolated by using polyaspartic acid as a water-retaining agent for the enrichment in situ.

Three novel actinomycete strains, designated TRM66264-DLMT, TRM88002T and TRM88003T, were isolated by using polyaspartic acid as a water-retaining agent for the enrichment in situ. The 16S rRNA gene sequence and phylogenetic analyses of three strains indicated that they belonged to the genus Actinoplanes. The phylogenetically closest strains of TRM66264-DLMT, TRM88002T and TRM88003T were Actinoplanes bogorensis LIPI11-2-Ac043T (98.4 %), Actinoplanes abujensis A4029T (98.0 %) and Actinoplanes ferrugineus IFO15555T (98.1 %), respectively. The major polar lipids of strains TRM66264-DLMT and TRM88002T were phosphatidylethanolamine and disphosphatidylglycerol, while strain TRM88003T only had phosphatidylethanolamine. The predominant menaquinones of strain TRM66264-DLMT were identified as MK-9(H4) and MK-9 (H6). Strains TRM88002T and TRM88003T had MK-9(H4). The cell-wall peptidoglycan of three strains contained meso-diaminopimelic acid. The whole-cell sugars of strain TRM66264-DLMT were identified as arabinose, glucose, galactose and xylose. Strains TRM88002T and TRM88003T mainly had arabinose and glucose. The DNA G+C content of strains TRM66264-DLMT, TRM88002T and TRM88003T were 70.48, 70.46 and 70.64 mol%, respectively. Genotypic and phenotypic analysis confirmed that all three strains sre new members of the genus Acinoplanes. Therefore, it is proposed that strains TRM66264-DLMT, TRM88002T and TRM88003T represent three novel species of the genus Actinoplanes, for which the names Actinoplanes polyasparticus sp. nov. (type strain TRM66264-DLMT=CCTCC AA 2021015T=LMG 32389T), Actinoplanes hotanensis sp. nov. (type strain TRM88002T=CCTCC AA 2021036T=LMG 32621T) and Actinoplanes aksuensis sp. nov. (type strain TRM88003T=CCTCC AA 2021037 T=LMG 32622T) are proposed.

A secreted effector with a dual role as a toxin and as a transcriptional factor.

Bacteria have evolved multiple secretion systems for delivering effector proteins into the cytosol of neighboring cells, but the roles of many of these effectors remain unknown. Here, we show that Yersinia pseudotuberculosis secretes an effector, CccR, that can act both as a toxin and as a transcriptional factor. The effector is secreted by a type VI secretion system (T6SS) and can enter nearby cells of the same species and other species (such as Escherichia coli) via cell-cell contact and in a contact-independent manner. CccR contains an N-terminal FIC domain and a C-terminal DNA-binding domain. In Y. pseudotuberculosis cells, CccR inhibits its own expression by binding through its DNA-binding domain to the cccR promoter, and affects the expression of other genes through unclear mechanisms. In E. coli cells, the FIC domain of CccR AMPylates the cell division protein FtsZ, inducing cell filamentation and growth arrest. Thus, our results indicate that CccR has a dual role, modulating gene expression in neighboring cells of the same species, and inhibiting the growth of competitors.

Effect of polyaspartic acid on soil water storage, soil microbial diversity, cotton yield and fiber quality.

Water-saving is one of the most important problems in agricultural development, especially in arid and semi-arid areas. The effects of polyaspartic acid (PASP) on soil water storage, soil microbial community, soil physiochemical properties, cotton yield and fiber quality were studied to find water-saving material utilized in cotton field. The experiment was divided into two parts, the first part concerned the direct application of three different amounts of PASP under field conditions. In the second part, PASP was mixed with soil in different proportions and the mixtures were put into bottles, which were then buried in the cotton field. The application of PASP improved the water-holding capacity and thus increased water content available to the cotton root system in the cotton field for a long time, and significantly (p < 0.05) increased the content of soil organic matter, available P and ammonium-N. Relative abundances of Methylophaga, Sphingomonas, Cupriavidus, Pseudeurotium, Fusarium and Nectria were significantly affected by applying PASP. Compared to the control group, 15, 75 and 150 kg ha-1 of PASP increased seed cotton yield by 3.94, 8.31 and 7.71%, respectively. The application of PASP also increased the reflectance degree, Micronaire and short fiber index of cotton. These results suggested that 75 kg ha-1 of PASP can be appropriate to alleviate drought stress in arid and semi-arid areas.

Rhizobium alarense sp. nov. and Rhizobium halophilum sp. nov. isolated from the nodule and rhizosphere of Lotus japonicus.

Two strains (TRM95111T and TRM95001T) of Gram-stain-negative, aerobic, rod-shaped microbes were isolated from the nodule and rhizosphere of Lotus japonicus grown in the campus of Tarim University in Alar, Xinjiang, China. Strain TRM95111T and strain TRM95001T shared 93.1% 16S rRNA gene sequences similarity with each other and had 98.2 and 97.9% 16S rRNA gene sequence similarity to the closest species Rhizobium subbaraois JC85T and R. halotolerans AB21T by EzBioCloud blast, respectively. Phylogenetic analyses based on 16S rRNA gene sequences, housekeeping gene sequences and core-proteome average amino acid identity (cpAAI) showed that two strains belonged to the genus Rhizobium. The value of digital DNA-DNA hybridization (dDDH) between strain TRM95111T and the closest strain R. subbaraonis JC85T was 21.8%, respectively. The dDDH value between strain TRM95001T and the closest strains R. tarimense PL-41T was 27.1%. Whole-genome average nucleotide identity (ANI) values of the strain TRM95111T were 75.6-79.3% and strain TRM95001T were 79.2-83.8%, compared to their closely related strains. The G + C content values of strain TRM95111T and TRM95001T were 65.1 and 60.7 mol%, respectively. Two isolates contained predominant quinone of Q-10 and the major fatty acids was C18:1ω7c and they were sensitive to 1 µg of amikacin and kanamycin. The polar lipids of strain TRM95111T included unidentified aminophospho lipids (APL1-3), unidentified phospholipids (PL1-2), phosphatidylcholine (PC), unidentified lipids (L1-5) and phospholipids of unknown structure containing glucosamine (NPG), compared to the polar lipids of strain TRM95001T including unidentified aminophospho lipids (APL1-3), unidentified phospholipids (PL1-2), phosphatidylcholine (PC), unidentified lipids (L2-5), hydroxy phosphatidyl ethanolamine (OH-PE) and phospholipids of unknown structure containing glucosamine (NPG). Nodulation tests showed that two strains could induce nodules formation in L. japonicus. Based on the genomic, phenotypic and phylogenetic analyses, strain TRM95111T and strain TRM95001T are suggested to represent two new species of the genus Rhizobium, whose names are proposed as Rhizobium alarense sp. nov. and Rhizobium halophilum sp. nov. The type strains are TRM95111T (=CCTCC AB 2021116T =JCM34826T) and TRM950011T (=CCTCC AB 2021095T =JCM34967T), respectively.

Streptomyces pimonensis sp. nov., isolated from the Taklimakan desert in Xinjiang, China.

A novel Streptomyces strain, designated TRM 75549T, was isolated from a sample of sand in Pimo reclamation area, Taklimakan desert, Xinjiang, North-West China. Phylogenetic analyses of the 16S rRNA gene sequences placed strain TRM75549T within the genus Streptomyces with the highest similarities to Streptomyces pilosus NBRC 12772T (98.7%). Nonetheless, average nucleotide identity (ANI) value and the digital DNA-DNA hybridization (dDDH) value between strain TRM75549T and S. pilosus NBRC 12772T were, respectively, 88.2% and 44.1%, and well below 95-96% and 70% cutoff point recommended for recognizing genomospecies, respectively. A multi-locus sequence analysis of five house-keeping genes (atpD, gyrB, recA, rpoB and trpB) and phylogenomic analysis also illustrated that strain TRM75549T should also be assigned to the genus Streptomyces. Strain TRM75549T contained MK-9 (H6) and MK-9 (H8) as predominant menaquinones. The diagnostic diamino acid of cell walls were identified as LL-diaminopimelic acid and meso-diaminopimelic. The whole-cell sugar pattern of strain TRM 75549T consisted of mannose and glucose. The major fatty acids (> 5%) were iso-C14:0, iso-C15:0, anteiso-C15:0, iso-C16:1H, iso-C16:0. The polar lipids were diphosphatidylglycerol, lysophosphatidylglycerol, phosphatidylethanolamine, phospholipids, phosphatidylglycerol, phosphatidylinositol, phosphatiylinositol mannosides and unidentified phospholipids. Strain TRM75549T could be differentiated from S. pilosus NBRC 12772T, based on physiological and biochemical characteristics. Thus, strain TRM75549T is representative of a novel species of the genus Streptomyces, for which the name Streptomyces pimonensis sp. nov. is proposed. The type strain is TRM75549T (= CCTCC AA 2020054T = LMG 32221T).

Screening of Microbial Fermentation Products for Anti-M. tuberculosis Activity.

Tuberculosis (TB), caused by M. tuberculosis (M.tb), is the leading infectious cause of mortality worldwide. The emergence of drug-resistant M.tb has made the control of TB more difficult. In our study, we investigated the ability of microorganism fermentation products from the soil to inhibit M.tb. We successfully identified four fermentation products (Micromonospora chokoriensis, Micromonospora purpureochromogenes, Micromonospora profundi, Streptomyces flavofungini) that inhibited the growth of M.tb in vitro and in intracellular bacteria at 25 µg/mL MIC. Importantly, the fermentation products decreased some essential gene expression levels for M.tb growth. Our data provide the possibility that microbial fermentation products have potential development value for anti-M.tb drugs.

Halomonas jincaotanensis sp. nov., isolated from the Pamir Plateau degrading polycyclic aromatic hydrocarbon.

A Gram-strain-negative, rod-shaped, aerobic bacterium, designated strain TRM 85114T, was isolated from the Jincaotan wetland in the Pamir Plateau of China. This strain grew optimally at 30 °C and pH 6.0 in the presence of 3% (w/v) NaCl. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain TRM 85114T was affiliated with the genus Halomonas, and shared high sequence similarity with Halomonas korlensis XK1T (97.3%) and Halomonas tibetensis pyc13T (96.4%). Strain TRM 85114T contained C16:0 and C19:0 cyclo ω8c as primary cellular fatty acids, Q-9 as predominate respiratory quinone, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phospholipids of unknown structure containing glucosamine, unidentified aminophospholipids, unidentified lipids and three unidentified phospholipids as the major polar lipids. The complete genome of TRM 85114T comprised 3,902 putative genes with a total of 4,126,476 bp and a G + C content of 61.6%. The average nucleotide identity and digital DNA-DNA hybridization values between strain TRM 85114T and related type Halomonas strains of H. korlensis XK1T, H. tibetensis pyc13T, Chromohalobacter salexigens DSM 6768T, and Halomonas urumqiensis BZ-SZ-XJ27T were 75.4-88.9% and 22.9-39.2%, respectively. Based on phenotypic, chemotaxonomic, and molecular features, strain TRM 85114T represents a novel species of the genus Halomonas, for which the name is proposed as Halomonas jincaotanensis sp. nov.. The type strain is TRM 85114T (CCTCC AB 2021006T = LMG 32311T). The amount of 1-naphthylamine degradation by strain TRM 85114T reached up to 32.0 mg/L in 14 days.

Streptomyces griseicoloratus sp. nov., isolated from soil in cotton fields in Xinjiang, China.

A novel bacterium of the genus Streptomyces, designated TRM S81-3T, was isolated from soil in cotton fields of Xinjiang, China. Comparative 16S rRNA gene sequence analysis indicated that strain TRM S81-3T is most closely related to Streptomyces viridiviolaceus NBRC 13359T (98.9% sequence similarity); however, the average nucleotide identity (ANI) between strains TRM S81-3T and S. viridiviolaceus NBRC 13359T is relatively low (91.6%). Strain TRM S81-3T possesses LL-diaminopimelic acid as the diagnostic cell-wall diamino acid, MK-9(H4), MK-9(H6), and MK-9(H10) as the major menaquinones, and polar lipids including diphosphatidylglycerol (DPG), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylmethyl ethanolamine (PME), phosphotidylinositolone (PI), phospholipid of unknown structure containing glucosamine (NPG), and two unidentified phospholipids (PLs).The major fatty acids are iso-C16:0, anteiso-C15:0, anteiso-C17:1 ω9c, anteiso-C17:0, iso-C15:0, and C14:0. The genomic DNA G + C content is 72.1%. Based on the evidence from this polyphasic study, strain TRM S81-3T represents a novel species of Streptomyces, for which the name Streptomyces grisecoloratus is proposed. The type strain is TRM S81-3T (= CCTCC AA 2020002T = LMG 31942T).

Anti-inflammatory effect of a polysaccharide fraction from Craterellus cornucopioides in LPS-stimulated macrophages.

Immunocytes-involved inflammation is considered to modulate the damage in various diseases. Oxidative stress is initiated by oxidative agents such as LPS and ROS, which are strongly involved in chronic inflammation. Our previous study found that a polysaccharide fraction from Craterellus cornucopioides (CCPP-1) showed good antioxidant activity. However, the anti-inflammatory effect of CCPP-1 was still elusive. The objective of this study was to evaluate the anti-inflammatory activity of CCPP-1 and its potential mechanism in LPS-stimulated RAW264.7 macrophages. The results showed that CCPP-1 could inhibit LPS-induced ROS and NO accumulation. Additionally, CCPP-1 could decrease pro-inflammatory cytokines production (TNF-α, IL-1ß, and IL-18) and inflammatory mediator (iNOS) expression, which might be associated with its capacity to inhibit NF-κB signaling pathway and NLRP3 inflammasome activation. Therefore, this study suggested that CCPP-1 had an ameliorative effect on the inflammation response and was potential to develop into functional food for treating chronic inflammation. PRACTICAL APPLICATIONS: Craterellus cornucopioides is an edible fungus widely distributed in Southwestern China. It was reported that C. cornucopioides polysaccharide (CCPP-1), as important active ingredient, showed good antioxidant activity. However, the anti-inflammatory effect was still elusive. This study showed that CCPP-1 possessed anti-inflammatory activity. The molecular mechanism might be associated with its capacity to inhibit NF-κB signaling pathway and NLRP3 inflammasome activation. Therefore, polysaccharides from C. cornucopioides have potential to develop into functional food to combat inflammatory condition and thus indirectly halt the progression of various inflammatory response-related chronic diseases.

Streptomyces polyasparticus sp. nov. isolated from cotton field soil by a medium applied with polyaspartic acid.

A novel Streptomyces strain (TRM66268-LWLT) was isolated from cotton field soil by a medium supplied with polyaspartic acid (PASP) at Alar, Xinjiang, Northwest PR China, and characterized using a polyphasic taxonomic approach. The strain was found to degrade PASP, and grow well on the medium to take PASP as the sole carbon source. The TRM66268-LWLT fermentation broth was applied to the surface of PASP, and there were pores on the surface of PASP after a period of time. The strain was observed to be Gram-stain-positive and to form greyish-white aerial mycelia that differentiated into straight spore chains with round spores. The whole-cell sugar pattern of TRM 66268-LWLT consisted of ribose, mannose and arabinose, and the principal phospholipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannoside, phosphatidylinositol and two undetermined polar lipids. The predominant menaquinones were MK-7, MK-7(H4), MK-9(H8), MK-10(H6). The diagnostic cell wall amino acid was identified as LL-diaminopimelic acid. The G+C content of strain TRM66268-LWLT was 70.11 mol%. The average nucleotide identity value between strain TRM66268-LWLT and the phylogenetically related strain Streptomyces indicus IH32-1T was calculated to be 85.49%. The digital DNA-DNA hybridization value between them was 30.40%. A multilocus sequence analysis of five house-keeping genes (atpD, gyrB, rpoB, recA and trpB) also illustrated that strain TRM66268-LWLT should be assigned to the genus Streptomyces. On the basis of evidence from polyphasic study, strain TRM66268-LWLT is designated as representing a novel species of the genus Streptomyces, for which the name Streptomyces polyasparticus sp. nov. is proposed. The type strain is TRM66268-LWLT (CCTCC AA 2020003T = LMG32106T).

A new chlorine-containing iridoid glycoside from Plantago maxima.

A phytochemical investigation on the whole plant of Plantago maxima Juss. ex Jacq led to the isolation of a new and rare chlorinated iridoid glycoside named plantomoside (1), along with three known compounds, geniposidic acid (2), 10-deoxygeniposidic acid (3), and viteoid II (4). The structure of 1 was determined through 1 D and 2 D NMR spectroscopic data analysis, HR-ESI-MS, and acid hydrolysis.

Amicoumacins from a desert bacterium: quorum sensing inhibitor against Chromobacterium violaceum.

In our study, the anti-quorum sensing (QS) activity of fermentation broth from TRM B-02, a bacterium isolated from Taklimakan desert, was investigated using the biosensor bioassay on Chromobacterium violaceum ATCC12472. TRM B-02 was 100% similar to Bacillus subtilis subsp. Inaquosorum KCTC 13429(T) by genotypic and phenotypic analyses. Based on anti-QS activity tracking, six known amicoumacins, named as AI-77-H (1), AI-77-F (2), amicoumacin B (3), amicoumacin C (4), AI-77-C (5) and bacilosarcins D (6), were isolated and identified. Among them, compounds 1-3 exhibited a better inhibitory effect on C. violaceum ATCC12472. Further research suggested that compounds 1-3 could significantly down-regulate the expressions of violacein operon A (vioA), vioB, vioD and vioE and up-regulate vioC. Docking experiments indicated that compounds 1-3 may act as an inhibitor of violacein biosynthetic pathway competitively inhibiting the binding of flavin adenine dinucleotide (FAD) with the vioD enzyme.[Figure: see text].

Streptomyces lycii sp. nov., an endogenous actinomycete isolated from Lycium ruthenicum.

A novel endogenous actinobacteria strain, designated TRM 66187T, was isolated from Lycium ruthenicum sampled at Alar, Xinjiang, Northwest PR China, and characterized using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain TRM 66187T with the genus Streptomyces. The whole-cell sugar pattern of TRM 66187T consisted of galactose, glucose and ribose. The predominant menaquinones were MK-9(H4) and MK-9(H6). Major cellular fatty acids were iso-C14:0, iso-C15:0, anteiso-C15:0 and anteiso-C16:0. The detected polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and two unidentified polar lipids. The G+C content of strain TRM 66187T was 71.8 mol%. Results of phylogenetic analysis showed that strain TRM 66187T had 98.48% 16S rRNA gene sequence similarity to the closest described species Streptomyces qinglanensis DSM 42035T. The average nucleotide identity value between strain TRM 66187T and the closest related strain Streptomyces qinglanensis DSM 42035T was calculated to be 77.2%. The digital DNA-DNA hybridization value between them was 22.4%. Multilocus sequence analyses based on five housekeeping genes (atpD, gyrB, recA, rpoB and trpB) also indicated that strain TRM 66187T should be assigned to the genus Streptomyces. On the basis of evidence from this polyphasic study, strain TRM 66187T should be designated as representing a novel species of the genus Streptomyces, for which the name Streptomyces lycii sp. nov. is proposed. The type strain is TRM 66187T (=LMG 31493T=CCTCC AA 2018094T).

Streptomyces apocyni sp. nov., an endogenous actinomycete isolated from Apocynum venetum.

A novel actinomycete, designated strain TRM 66233T, was isolated from Apocynum venetum L. collected from the Xinjiang Uygur Autonomous Region of China and characterized using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain TRM 66233T with the genus Streptomyces. Strain TRM 66233T showed a high similarity value to Streptomyces bikiniensis NRRL B-1049T (98.07 %) based on the 16S rRNA gene phylogenetic tree. The whole-cell sugar pattern of TRM 66233T consisted of glucose, galactose, mannose and ribose. The predominant menaquinones were MK-9(H2), MK-9(H6), MK-9(H8) and MK-9(H10). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and four unidentified lipids. The major fatty acids were iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0, C16 : 0 and iso-C17 : 0. The G+C content of the DNA was 70.35 mol%. The DNA-DNA relatedness and average nucleotide identity values as well as evolutionary distances based on multilocus (atpD, gyrB, recA, rpoB and trpB) sequences between strain TRM 66233T and closely related type strains were significantly lower than the recommended threshold values. The whole-genome average nucleotide identity and digital DNA-DNA hybridization values between strain TRM 66233T and S. bikiniensis NRRL B-1049T were 78.86 and 23.2 %, respectively. On the basis of evidence from this polyphasic study, strain TRM 66233T should represent a novel species of the genus Streptomyces, for which the name Streptomyces apocyni sp. nov. is proposed. The type strain is TRM 66233T (=CCTCC AA 2019056T=LMG 31559T).

Colonization on Cotton Plants with a GFP Labeled Strain of Bacillus axarquiensis.

Verticillium dahliae was one of the most important diseases caused Verticillium wilt of cotton. In our previous study, Bacillus axarquiensis TUBP1 was screened and found to be an antagonistic strain against V. dahliae with 43% biocontrol effect in the cotton field. In order to uncover the functional mechanism of B. axarquiensis against Verticillium wilt in cotton, the colonization of B. axarquiensis labeled with a green fluorescent protein (GFP) was investigated in cotton plants and the rhizosphere soil. Firstly, a plasmid (pHT-315) containing gfp gene was successfully transformed into wild B. axarquiensis TUBP1 and labeled a green fluorescence by electroporation, which didn't change the bioactivity in vitro. In gnotobiotic conditions, cotton seeds were then inoculated with the gfp-labeled strain and grown in green house. Observation with a confocal laser scanning microscope and a scanning electron microscope showed that GFP-labeled B. axarquiensis TUBP1 infected cotton roots and widely distributed in epidermis, cortical parenchyma, intercellular spaces, the xylem vessels, and pith cells as well as root hair cells through cracks formed at the lateral root junctions, followed by a slow migration from roots to stems and leaves. Quantitative fluorescence and flow cytometry (FACS) approaches showed a gradual decrease in the number of TUBP1-315gfp with increasing inoculation time. However, TUBP1-315gfp levels were detectable till 45 days after planting. In contrast, no fluorescence signal was detected in the non-inoculated groups. Therefore, GFP-labeled B. axarquiensis TUBP1 exhibited colonization in different parts of cotton plants from the rhizosphere soil.

Streptomyces albicerus sp. nov., a novel actinomycete isolated from the sediments of the Tailan River in Xinjiang, China.

A novel actinomycete, designated TRM68295T, was isolated from the sediments of the Tailan River, Xinjiang, northwest China. The study of the polyphasic approach showed that the characteristics of the strain were consistent with the genus Streptomyces. Phylogenetic analysis indicated that the strain had a high sequence similarity with Streptomyces phaeochromogenes ATCC 3338T (97.9%). The diagnostic diamino acid of cell walls was identified as LL-diaminopimelic acid. The whole cell sugars were identified as ribose, xylose, glucose and galactose. The major fatty acids were iso-C14:0, iso-C15:0, anteiso-C15:0, iso-C16:0, C16:1ω9c, C16:0 and anteiso-C17:0. The major menaquinones were MK-9 (H10), MK-9 (H6) and MK-9 (H2). The polar lipids were composed of diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylcholine (PC), phosphatidylinositol (PI) and phosphatidylinositol mannoside (PIM). The G+C content in the draft genome sequence of the strain was identified as 70.0 mol%. The average nucleotide identity (ANI) between strains TRM68295T and S. phaeochromogenes ATCC 3338T was 88.1%. Digital DNA-DNA hybridization (dDDH) value between strain TRM68295T and S. phaeochromogenes ATCC 3338T was 40.7%. The multilocus sequence analysis of five house-keeping genes (atpD, gyrB, recA, rpoB, and trpB) indicated that the MLSA distances between the strain and similar species were greater than the threshold of 0.007. Based on the above studies, strain TRM68295T is considered as a novel species of the genus Streptomyces, for which the name Streptomyces albicerus sp. nov. is proposed. The type strain is TRM68295T (= CCTCC AA 2018085T = KCTC 49272T).