First Report of Anthracnose on Hymenocallis littoralis Caused by Colletotrichum siamense in China.

Spider lily (Hymenocallis littoralis (Jacq.) Salisb.) is a widely cultivated horticultural plant worldwide and has ornamental and medicinal value. Spider lily plants were seriously affected by a leaf spot disease in the campus of Guangdong Ocean University and gardens in Zhanjiang city in February 2018 with an incidence of 30 to 100%. Affected leaves usually developed small circular purple spots, which enlarged to oval spots and large irregular spots. The spots were brown at the center, deep purple at the border and surrounded by a yellow halo. Diseased cultivars were collected in Zhanjiang city, Gangzhou city in Guangdong province and and Zhangping city in Fujian province. Symptomatic leaf samples were disinfested with 1% NaOCl, and cultured on sucrose agar (PSA) at 28 °C for one week. Ten single-spore isolates were recovered from PSA medium. Colonies developing on PSA were grayish white with a regular border. Conidia were straight, hyaline with rounded ends, 4.3 to 6.1×12.8 to 32.1µm (n = 50 conidia of each isolate). Fungal mycelia were hyaline, septate, and branched. Conidia were born on a long conidiogenous cell, appressoria were oval, 6.7 to 10.7 × 5.2 to 6.2 µm (n=50). The isolates were morphologically identified as Colletotrichum sp. (Weir et al. 2012). Tests of pathogenicity were performed according to Koch's postulates using three isolates. Fresh wounds were made with a sterile needle on the healthy surface of leaves of H. littoralis at the 4- to 6-leaf stage and each leaf was covered with a piece of cotton drenched with 200 µL of conidial suspension (106 conidia/ml) from each isolate. Control seedlings were inoculated identically except sterile water was used to drench the cotton. Inoculated plants were placed in a moisturizing light incubator at 25℃ and 80% humidity under a 12-h light/dark cycle for 20 days and examined daily to monitor disease symptom development. Small round brown spots were observed at the inoculation sites 3 days after the inoculation. The brown spots developed into large brown lesions 5 days after inoculation. There were no symptoms observed in the water-inoculated plants. A Colletotrichum spp. strain based on morphology was consistently reisolated from leaves lesions fulfilling Koch's postulates. For molecular identification, the internal transcribed spacer (ITS) region of ribosomal DNA, calmodulin (CAL), Tublin (Tub) and Apmat loci of three isolates were amplified using primer pairs of ITS4/ITS5, CL1C/CL2C, T1/T2 and AM-F/AM-R (Sharma et al. 2015). A phylogenetic tree derived from a neighbor-joining analysis of a concatenated alignment of ITS, CAL, Tub and ApMAT sequences was created. The accession numbers of three isolates GZHLCG, ZJHLCG and FJHLCG used in this study were MW553083, MN540457, MN540458 for ITS, MW553087- MW553089 for CL, MW553090-MW553092 for Tub and MW553084-MW553086 for ApMAT. The sequences of the three isolates were aligned with related species of Colletotrichum (Sharma et al. 2015). Analyses based on concatenated data sets of four genes showed that the sequences had high levels of identity to those of the C. siamense strains. According to both morphological and sequence analyses, the H. littoralis pathogen was identified as C. siamense. There is a report of foliar diseases on H. littoralis caused by Colletotrichum sp. (Tan et al., 2009). To our knowledge, this is the first report of anthracnose on H. littoralis caused by C. siamense in China. Identification of the pathogen provide valuable information for diagnosis and controlling this disease in H. littoralis.

Geodermatophilus ruber sp. nov., isolated from rhizosphere soil of a medicinal plant.

A novel actinobacterial strain, designated CPCC 201356(T), was isolated from a rhizosphere soil sample of the medicinal plant Astragalus membranaceus and subjected to a polyphasic taxonomic analysis. Good growth occurred at 20-32 °C, at pH 7.0-7.5 and with 0-1 % (w/v) NaCl. Colonies on R2A and ISP 2 agar were light red to red, round and lacked aerial mycelium; cells adhered to the agar. The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinones were MK-9(H(4)) and MK-9. Polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine and two unknown phospholipids. The major cellular fatty acids were iso-C(16 : 0), iso-C(15 : 0) and C(17 : 1)ω8c. The G+C content of the genomic DNA was 72.8 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain CPCC 201356(T) belonged to the family Geodermatophilaceae and consistently formed a distinct sub-branch with Geodermatophilus obscurus DSM 43160(T). The organism showed 16S rRNA gene sequence similarity of 97.7 % with G. obscurus DSM 43160(T). DNA-DNA hybridization between strain CPCC 201356(T) and G. obscurus DSM 43160(T) was 17.4 %. On the basis of evidence from this polyphasic taxonomic study, a novel species, Geodermatophilus ruber sp. nov., is proposed; the type strain is CPCC 201356(T) (=DSM 45317(T) =CCM 7619(T)).

Alloactinosynnema album gen. nov., sp. nov., a member of the family Actinosynnemataceae isolated from soil.

The taxonomic position of a Gram-stain-positive, aerobic strain, designated 03-9939(T), isolated from a soil sample collected from Xinjiang Province, China, was established using a polyphasic approach. Whole-cell hydrolysates of strain 03-9939(T) contained galactose and ribose as diagnostic sugars and meso-diaminopimelic acid as the diamino acid. The predominant menaquinone was MK-9(H4). The phospholipids consisted of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylcholine. The major fatty acids were iso-C(16 : 0) (61.5 %) and iso-C(16 : 1) H (11.6 %). The genomic DNA G+C content was 68.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 03-9939(T) should be placed within the family Actinosynnemataceae, in which the strain formed a distinct lineage. Signature nucleotides in the 16S rRNA gene sequence showed that the strain contained a genus-specific diagnostic nucleotide signature pattern. The combination of phylogenetic analysis, phenotypic characteristics and chemotaxonomic data supported the conclusion that strain 03-9939(T) represents a novel species in a new genus of the family Actinosynnemataceae, for which the name Alloactinosynnema album gen. nov., sp. nov. is proposed. Strain 03-9939(T) (=DSM 45114(T) =KCTC 19294(T) =CCM 7461(T)) is the type strain of Alloactinosynnema album.

Actinoplanes sichuanensis sp. nov. and Actinoplanes xinjiangensis sp. nov.

Two motile actinomycetes, designated strains 03-723(T) and 03-8772(T), which had potent inhibitory activity against Enterococcus faecium peptide deformylase and several clinical Gram-positive, antibiotic-resistant strains, were isolated from two soil samples collected from Sichuan Province and Xinjiang Uyghur Autonomous Region in China, respectively. The taxonomic status of these two organisms was established by using a polyphasic approach. The taxonomic data were consistent with the assignment of the strains to the genus Actinoplanes. The neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that the two isolates formed a branch with the type strains of Actinoplanes lobatus, Actinoplanes auranticolor, Actinoplanes capillaceus, Actinoplanes campanulatus and Actinoplanes philippinensis in the clade of Actinoplanes species. This branching pattern was also supported by the tree constructed with the maximum-parsimony method. Levels of 16S rRNA gene sequence similarity between strains 03-723(T) and 03-8772(T) and their phylogenetic neighbours ranged from 98.0 to 98.8 % and 97.4 to 98.1 %, respectively. However, the two strains shared low levels of DNA-DNA relatedness with the type strains of closely related Actinoplanes species and were readily distinguished by using a combination of phenotypic properties. Therefore, it is proposed that strains 03-723(T) and 03-8772(T) represent two novel species of the genus Actinoplanes, for which the names Actinoplanes sichuanensis sp. nov. (type strain 03-723(T)=KCTC 19460(T)=CCM 7526(T)) and Actinoplanes xinjiangensis sp. nov. (type strain 03-8772(T)=KCTC 19461(T)=CCM 7527(T)) are proposed.

Roseomonas vinacea sp. nov., a Gram-negative coccobacillus isolated from a soil sample.

Strain CPCC 100056(T), which was isolated from a soil sample collected from the Qinghai-Tibet plateau, China, was subjected to a polyphasic taxonomic study. The organism was coccobacillus-shaped, non-motile and formed vinaceous colonies on ISP2 agar medium. The respiratory quinone was ubiquinone-10. The major fatty acids were C(18:1)omega7c and C(16:1)omega7c and/or C(16:1)omega6c. The G+C content of the genomic DNA was 67.3 mol%. A comparison of sequences in GenBank revealed that strain CPCC 100056(T) exhibited highest 16S rRNA gene sequence similarity (84.5-95.5%) with Roseomonas species. Strain CPCC 100056(T) could be distinguished from all Roseomonas species with validly published names by differences in phenotypic and genotypic properties. In view of the combined phenotypic, chemotaxonomic and phylogenetic data, strain CPCC 100056(T) should be classified as a representative of a novel species in the genus Roseomonas, Roseomonas vinacea sp. nov.; the type strain is CPCC 100056(T) (=KCTC 22045(T) =CCM 7468(T)).

Nesterenkonia halophila sp. nov., a moderately halophilic, alkalitolerant actinobacterium isolated from a saline soil.

A Gram-positive, non-motile, moderately halophilic, alkalitolerant actinobacterium, designated strain YIM 70179(T), was isolated from a saline soil sample collected from Xinjiang Province, north-west China, and was subjected to a polyphasic taxonomic study. The cell-wall peptidoglycan type of strain YIM 70179(T) was A4alpha, l-Lys-Gly-l-Glu. Cells of the isolate contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and an unknown glycolipid, MK-8 as major menaquinone and anteiso-C(15 : 0), anteiso-C(17 : 0) and iso-C(16 : 0) as major fatty acids. The DNA G+C content was 68.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 70179(T) fell within the radiation of species of the genus Nesterenkonia. Levels of 16S rRNA gene sequence similarity between strain YIM 70179(T) and the type strains of recognized Nesterenkonia species were below 97 %, except to Nesterenkonia halobia DSM 20541(T) (99.6 %), but these two strains exhibited a low level of DNA-DNA relatedness (18.4 %). Based on genetic and phenotypic evidence, it is proposed that strain YIM 70179(T) represents a novel species of the genus Nesterenkonia, for which the name Nesterenkonia halophila sp. nov. is proposed. The type strain is YIM 70179(T) (=DSM 16378(T) =KCTC 19048(T)).

Streptomyces emeiensis sp. nov., a novel streptomycete from soil in China.

An actinomycete, strain 4776(T), was isolated from soil collected from Emei Mountain in Sichuan Province, China. The taxonomic status of this strain was established using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of streptomycetes. Phylogenetic analysis based on the almost complete 16S rRNA gene sequence indicated that the novel isolate belongs to the genus Streptomyces and consistently falls into a clade together with Streptomyces prasinopilosus DSM 40098(T), Streptomyces prasinus JCM 4603(T), Streptomyces bambergiensis DSM 40590(T), Streptomyces hirsutus DSM 40095(T) and Streptomyces cyanoalbus DSM 40198(T). However, DNA-DNA relatedness and phenotypic data distinguished strain 4776(T) from these phylogenetically related type strains. It is therefore concluded that strain 4776(T) (=CGMCC 4.3504(T)=DSM 41884(T)) represents a novel species of the genus Streptomyces, for which the name Streptomyces emeienseis sp. nov. is proposed.

Deinococcus yunweiensis sp. nov., a gamma- and UV-radiation-resistant bacterium from China.

A Gram-negative, non-spore-forming, non-motile, rod-shaped, red-pigmented strain, designated YIM 007T, was found as a contaminant on an agar plate in the laboratory of Yunnan Institute of Microbiology, China. The optimum growth pH and temperature for the isolate were 7.0-7.5 and 30 degrees C, respectively. The predominant respiratory quinone was MK-8. The polar lipid profile consisted mainly of various unknown phosphoglycolipids and glycolipids. The major cellular fatty acids were C(16 : 1)omega7c, C(16 : 0), C(17 : 0) and C(17 : 1)omega8c. l-Ornithine was detected in its peptidoglycan. The DNA G+C content was 64.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YIM 007(T) showed 16S rRNA gene sequence similarity levels of 86.8-92.1 % to the other described Deinococcus species. Based on the high 16S rRNA gene sequence divergence and phenotypic differences, it is proposed that the unknown strain should be classified as a novel species in the genus Deinococcus with the name Deinococcus yunweiensis sp. nov. The type strain is YIM 007T (=KCTC 3962T=DSM 17005T).

A case of chylothorax treated curatively with Sapylin, a streptococcus preparation.

Chylothorax is an uncommon disease where fatty fluid accumulates within the chest cavity. Conservative management, including repeated thoracentesis or pleurodesis, seems to be suitable to most cases. Herein, we present a case of efficacious pleurodesis by intrapleural injection of Sapylin, a streptococcus preparation, for the treatment of chylothorax. A 52-year-old non-smoking female farmer was diagnosed as idiopathic chylothorax after we ruled out possible causes including chest trauma, lymphoma, lung cancer, filariasis, tuberculosis, and etc. Two-time intra-thoracic injection of 3 Klinische Einheit (KE) Sapylin achieved rapid and effective control of chylothorax with no severe side effects. Sapylin may facilitate pleurodesis by producing a strong inflammatory response.

Sinococcus qinghaiensis gen. nov., sp. nov., a novel member of the order Bacillales from a saline soil in China.

A Gram-positive, non-spore-forming isolate, designated YIM 70212(T), was isolated from a hypersaline soil sample collected from Qinghai, north-west China. Cells of the isolate were orange-pigmented, motile cocci with multiple flagella. A polyphasic taxonomic investigation was carried out on the isolate. The organism grew at 10-45 degrees C and pH 7.5-11.0, with optimum growth at 28 degrees C and pH 8.0-9.5. Strain YIM 70212(T) grew optimally in the presence of 10 % NaCl, KCl or MgCl(2).6H(2)O and growth was observed in 1-25 % NaCl, KCl or MgCl(2).6H(2)O. The peptidoglycan type was A1gamma. Ribose and minor amounts of galactose were detected as the whole-cell sugars. MK-5 was the only menaquinone. The major cellular fatty acids were ai-C(15 : 0) (52.4 %) and ai-C(17 : 0) (26.5 %). The DNA G+C content was 47.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YIM 70212(T) formed a distinct lineage within the order Bacillales and was most closely related to members of the genus Marinococcus, showing 16S rRNA gene sequence similarity levels of 91.0-91.4 %. Based on the high 16S rRNA gene sequence divergence and differences in phenotypic characteristics, it is proposed that the unknown strain be classified in a novel genus and species with the name Sinococcus qinghaiensis gen. nov., sp. nov.; the type strain of Sinococcus qinghaiensis is YIM 70212(T) (=KCTC 3943(T)=DSM 17008(T)).