Phaeotremella camelliae sp. nov. (Phaeotremellaceae, Tremellales), A Novel Yeasts Isolated from Tea-Oil Fruits in Jiangxi Province, China.

Sixty-two isolates among the 65 yeast strains isolated from Jiangxi province, China, were identified into 15 known species based on the sequence analysis of the D1/D2 domains of the LSU rRNA and ITS region. The other three strains, GaoanZ14, GaoanC57, and GaoanC191, isolated from tea-oil fruits, were identified as two undescribed species of Phaeotremella based on the multiple gene sequence analysis, physiological, and biochemical comparisons. Strains GaoanC57 and GaoanC191 had one substitution difference both in the D1/D2 domains of the LSU rRNA and ITS region. They formed a separate branch from the other Phaeotremella species in the D1/D2 and multiple genes trees, and differed from the known species by at least 10 nucleotide substitutions in the D1/D2 domains and more than 6% mismatches in the ITS region. The phylogenetic analysis indicated that those two strains represent a novel species of Phaeotremella, for which the names Phaeotremella camelliae sp. nov. (Holotype CGMCC 2.6141, Mycobank MB832699) is proposed. Only one strain, GaoanZ14, represents the other undescribed species of Phaeotremella, so it will be described in latter when more strains are found.

[Percutaneous transforaminal endoscopic discectomy via lateral position assisted with local anesthesia for lumbar disc herniation in elder with systemic diseases].

OBJECTIVE: To explore the clinical effect of percutaneous transforaminal endoscopic discectomy via lateral position assisted with local anesthesia for lumbar disc herniation in elder with systemic diseases. METHODS: The clinical data of 44 elder patients with lumbar disc herniation and systemic diseases treated from June 2013 to June 2017 were retrospectively analyzed. Including 26 males and 18 females, aged 70 to 86 years old with an average of(77.5±3.5) years, course of disease was 3 weeks to 6 months. Percutaneous transforaminal endoscopic discectomy via lateral position assisted with local anesthesia was performed in the patients. Postoperative straight leg-raising degree change and Nakai criteria were used to evaluate the operative effect. Visual analogue scale(VAS) and JOA scoring system were used to compare the symptom improvement between preoperative and postoperative. RESULTS: No serious complications occurred during and after the operation, and the coexisting medical diseases were not aggravated. All patients safely survived the perioperative period. All the 44 cases were followed up from 6 to 30 months with an average of 20 months. Postoperative straight leg raising angle was obviously improved. At final follow-up, according to Nakai standard, 41 cases got excellent results, 3 good. VAS scores were significantly decreased and JOA scores were significantly increased in final follow-up(P<0.01). CONCLUSIONS: Percutaneous transforaminal endoscopic discectomy via lateral position assisted with local anesthesia is a safe, effective, less invasive technique for the treatment of lumbar disc herniation in elder with systemic diseases. Multidisciplinary collaboration and perioperative well controlled disease are essential for early rehabilitation of such patients.

Selenylation of Polysaccharide from the Sweet Potato and Evaluation of Antioxidant, Antitumor, and Antidiabetic Activities.

Interest in sweet potato as a functional food is growing. A polysaccharide (SWP) was isolated from the sweet potato tuber and elucidation of its structure as composed of rhamnose, glucose, and galactose undertaken. To improve its activity, selenylation of this novel polysaccharide (Se-SWP) was undertaken by using microwave synthesis. In vitro evaluation showed that the Se-SWP has excellent antioxidant activity on scavenging free radicals and reducing capacity. In vivo antitumor evaluation showed selenylation polysaccharide could effectively inhibit tumor growth (>50%) and adjust immune factor levels in the mice (IL-2, TNF-α, and VEGF). The antidiabetic potential of Se-SWP was tested in STZ-induced diabetic rats. The results indicated that the Se-SWP treatment significantly reduced the levels of malondialdehyde and other disadvantageous factors that were increased by the STZ treatment. Meanwhile, the Se-SWP treatment caused a significant increase in the activities of enzymatic antioxidants and the levels of nonenzymatic antioxidants in the organs of diabetic rats. All of the activity evaluations indicated that the selenylation method could improve the activity of sweet potato polysaccharide and its efficacy as a potential therapeutic, which will be the focus of further study.

Cloning and Characterization of a Squalene Synthase Gene from the Chaga Medicinal Mushroom, Inonotus obliquus (Agaricomycetes).

Squalene synthase catalyzes the condensation of 2 molecules of farnesyl diphosphate to produce squalene, the first committed precursor for sterol, brassinosteroid, and triterpene biosynthesis. A squalene synthase gene, designated IoSQS, was isolated from Inonotus obliquus, a medicinal mushroom that produces a plethora of bioactive triterpenes. IoSQS complementary DNA was found to contain an open reading frame of 1476 bp, encoding a protein of 491 amino acids with a calculated molecular mass of 55.85 kDa. The IoSQS genomic DNA sequence consisted of 1813 bp and contained 4 exons and 3 introns. The restriction fragment polymorphisms revealed by Southern blot analysis suggested that IoSQS was a single-copy gene. Promoter analysis indicated that the 5' upstream region of IoSQS possessed various potential elements associated with physiological and environmental factors. The expression pattern of IoSQS in different stages and under methyl jasmonate treatment correlated with the accumulation of total triterpenoids and was consistent with the predicted results of the IoSQS promoter region. The N-terminal 466 residues of the hydrophilic sequence were expressed as a His-tagged protein in Escherichia coli, and the resultant bacterial crude extract was incubated with farnesyl diphosphate and NADPH. Squalene was detected in vitro in reaction mixture by high-performance liquid chromatography analysis. These results suggest that the IoSQS enzyme is involved in squalene production in I. obliquus.

Complete genome sequence of Kibdelosporangium phytohabitans KLBMP 1111(T), a plant growth promoting endophytic actinomycete isolated from oil-seed plant Jatropha curcas L.

Kibdelosporangium phytohabitans KLBMP 1111(T) is a plant growth promoting endophytic actinomycete isolated from the oil-seed plant Jatropha curcas L. collected from dry-hot valley, in Sichuan, China. The complete genome sequence of this actinomycete consists of one chromosome (11,759,770bp) with no plasmid. From the genome, we identified gene clusters responsible for polyketide and nonribosomal peptide synthesis of natural products, and genes related to the plant growth promoting, such as zeatin, 1-aminocyclopropane-1-carboxylate deaminase (ACCD) and siderophore. The complete genome information may be useful to understand the beneficial interactions between K. phytohabitans KLBMP 1111(T) and host plants.

Nocardia rhizosphaerae sp. nov., a novel actinomycete isolated from the coastal rhizosphere of Artemisia Linn., China.

A novel actinomycete, designated strain KLBMP S0043(T), was isolated from the rhizosphere soil of Artemisia Linn. collected from the coastal region of Lianyungang, Jiangsu Province, in east China and was studied in detail for its taxonomic position. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain KLBMP S0043(T) is a member of the genus Nocardia. The 16S rRNA gene sequence similarity indicated that strain KLBMP S0043(T) is closely related to Nocardia asteroides NBRC 15531(T) (97.61 %) and Nocardia neocaledoniensis SBHR OA6(T) (97.38 %); similarity to other type strains of the genus Nocardia was found to be less than 97.2 %. The organism has chemical and morphological features consistent with its classification in the genus Nocardia such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan and arabinose and galactose as the diagnostic sugars. The predominant menaquinone was identified as MK-8(H4ω-cycl). Mycolic acids were detected. The diagnostic phospholipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides. The predominant cellular fatty acids were identified as C16:0, C18:0, C18:1ω9c, 10-methyl C18:0 [tuberculostearic acid (TBSA)] and summed feature 3 (C16:1ω7c/C16:1ω6c). The G+C content of the genomic DNA was determined to be 71.4 mol%. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of the strain from its most closely related strains. Based on morphological, chemotaxonomic and phylogenetic data, strain KLBMP S0043(T) is considered to represent a novel species of the genus Nocardia, for which the name Nocardia rhizosphaerae sp. nov. is proposed. The type strain is KLBMP S0043(T) (=CGMCC 4.7204 (T) = KCTC 29678(T)).

Nocardioides panzhihuaensis sp. nov., a novel endophytic actinomycete isolated from medicinal plant Jatropha curcas L.

A novel Gram-positive, aerobic, rod-shaped and mycelia-producing bacterial strain, designated KLBMP 1050(T), was isolated from the stem of the oil-seed plant Jatropha curcas L. collected from Sichuan Province, south-west China. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the isolate KLBMP 1050(T) belonged to the genus Nocardioides, with the highest sequence similarity to Nocardioides albus KCTC 9186(T) (99.38 %) and Nocardioides luteus KCTC 9575(T) (99.03 %). However, the DNA-DNA relatedness of isolate KLBMP 1050(T) to these two type strains were 37.5 ± 3.5 and 33 ± 2.3 %, respectively. Strain KLBMP 1050(T) grew at the pH range 6-11, temperature range 10-32 °C and with 0-12 % NaCl. The physiological properties of strain KLBMP 1050(T) differ from those of N. albus KCTC 9186(T) and N. luteus KCTC 9575(T). The cell-wall peptidoglycan contained LL: -diaminopimelic acid and MK-8(H(4)) was the major respiratory quinone. The predominant cellular fatty acid of strain KLBMP 1050(T) was iso-C(16:0) (23.3 %). The total DNA G+C content was 70.1 mol%. On the basis of the phenotypic, chemotaxonomic and phylogenetic data, strain KLBMP 1050(T) represents a novel species of the genus Nocardioides, for which the name Nocardioides panzhihuaensis sp. nov. is proposed. The type strain is KLBMP 1050(T) (= KCTC 19888(T) = NBRC 108680(T)).

Streptomyces phytohabitans sp. nov., a novel endophytic actinomycete isolated from medicinal plant Curcuma phaeocaulis.

A novel actinomycete, designated strain KLBMP 4601(T), was isolated from the root of the medicinal plant Curcuma phaeocaulis collected from Sichuan Province, south-west China. The strain produced extensively branched substrate and aerial hyphae that carried straight to flexuous spore chains. Chemotaxonomic properties of this strain were consistent with those of members of the genus Streptomyces. The cell wall of strain KLBMP 4601(T) contained LL-diaminopimelic acid as the characteristic diamino acid. The major menaquinone was MK-9(H(4)), with minor amounts of MK-9(H(6)), MK-9(H(8)) and MK-10(H(2)). The major fatty acids were C(16:0), iso-C(16:0), C(18:1)ω9c and C(16:1), iso G. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain KLBMP 4601(T) belongs to the genus Streptomyces and is most closely related to Streptomyces armeniacus JCM 3070(T) (97.9 %), Streptomyces pharmamarensis PM267(T) (97.6 %) and Streptomyces artemisiae YIM 63135(T) (97.5 %). The 16S rRNA gene sequence similarity between strain KLBMP 4601(T) and other members of this genus were lower than 97.5 %. DNA-DNA hybridization studies of strain KLBMP 4601(T) with the three closest species showed relatedness values of 36.3 ± 4.2 %, 27.3 ± 0.6 %, and 30.9 ± 2.5 %, respectively. On the basis of chemotaxonomic, phenotypic and genotypic characteristics, it is evident that strain KLBMP 4601(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces phytohabitans sp. nov. is proposed. The type strain is KLBMP 4601(T) (=KCTC 19892(T) = NBRC 108772(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.

Promicromonospora xylanilytica sp. nov., an endophytic actinomycete isolated from surface-sterilized leaves of the medicinal plant Maytenus austroyunnanensis.

A novel xylan-degrading actinomycete, strain YIM 61515(T), was isolated from surface-sterilized leaves of the medicinal plant Maytenus austroyunnanensis. Cells were Gram-positive and non-spore-forming, produced primary branches and formed white to yellowish white colonies on the media tested. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 61515(T) was most similar to Promicromonospora aerolata V54A(T) and Promicromonospora vindobonensis V45(T) (99.4 and 99.1% sequence similarity, respectively). The isolate formed a separate lineage in a cluster containing P. aerolata V54A(T). 16S rRNA gene sequence similarity between the isolate and other members of the genus Promicromonospora ranged from 96.3 to 98.4%. Chemotaxonomic data, including major menaquinones, fatty acid compositions and polar lipid profiles, supported the placement of strain YIM 61515(T) in the genus Promicromonospora. DNA-DNA relatedness, physiological and biochemical data showed that strain YIM 61515(T) could be distinguished from members of all known species of the genus Promicromonospora and therefore represented a novel species. The name Promicromonospora xylanilytica sp. nov. is proposed, with YIM 61515(T) (=DSM 21603(T)=CCTCC AA 208046(T)) as type strain.

In vitro antitumor activity and structure characterization of ethanol extracts from wild and cultivated Chaga medicinal mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyllophoromycetideae).

Inonotus obliquus (Pers.:Fr.) Pilát has been traditionally used as a folk remedy for treatment of cancers, cardiovascular disease and diabetes in Russia, Poland, and most of the Baltic countries, but natural reserves of this fungus have nearly been exhausted. This study was designed to investigate the artificial cultivation of I. obliquus and the antitumor activity of its tissues. The ethanol extract of cultivated sclerotium had the highest cell growth inhibitory rate (74.6%) as determined by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. 78% of the bags produced sclerotia and only 6.17 g/bag of sclerotium was obtained. Extracts of the cultivated fruiting body showed 44.2% inhibitory activity against tumor cells. However, the yield was as high as 18.24 g/bag, and 98% of the bags produced fruiting body. The results of gas chromatography-mass spectroscopy (GC-MS) showed that similar compounds were extracted from the wild and cultivated samples. The principal compounds observed were lanosterol, inotodiol, and ergosterol. Their percentages of the mass fraction were 86.1, 59.9, and 71.8% of the total, for the wild sclerotium, cultivated sclerotium, and cultivated fruiting body, respectively. Ergosterol was found to be much higher (27.32%) in cultivated fruiting body. We conclude that cultivated fruiting body of I. obliquus obtained by inoculation of the substrate with spawn mycelium of the fifth generation could serve as an ideal substitute for the wild I. obliquus.

A new cytotoxic casbane diterpene from Euphorbia pekinensis.

A new cytotoxic casbane diterpene, named pekinenal, was isolated from the roots of Euphorbia pekinensis. Its structure was elucidated as 5alpha-hydroxy-1betaH,2alphaH-casba-3Z,7E,11E-triene-18-al by a combination of 1D- and 2D-NMR techniques and confirmed by X-ray crystallography. Pekinenal showed cytotoxic activity against all four human cancer cell lines tested.

Endophytic fungal metabolite fumigaclavine C causes relaxation of isolated rat aortic rings.

Two indole alkaloids were isolated from the culture of Aspergillus fumigatus (strain No. CY018), an endophytic fungus harboring inside the elder leaf of Cynodon dactylon. These two chemicals were identified as fumigaclavine C and fumitremorgin C. In screening the bioactivity of these two indole alkaloids, their vasorelaxant effects on isolated rat thoracic aortic rings were observed. The results showed that fumigaclavine C exhibited potent concentration-dependent vasorelaxant actions in isolated rat aortic rings pre-contracted by high K+ or phenylephrine (with EC50 values of 5.62 micromol/L and 1.58 micromol/L, respectively) whereas fumitremorgin C displayed a weaker vasorelaxation. A detailed investigation was therefore performed with fumigaclavine C. The vasorelaxing action of fumigaclavine C is independent of the presence of endothelium, suggesting its effect of vasorelaxation was not related to endothelial mediators. Blockage of L-type voltage-dependent calcium channels, activation of ATP-sensitive potassium channels and inhibition of Ca2+ release from intracellular Ca2+ stores may be involved in fumigaclavine C induced relaxation of rat isolated aortic rings. These results demonstrate that fumigaclavine C from the endophytic fungus has a potential capacity in vascular protection and thus may have therapeutic use in protection against cardiovascular disease.