Nocardia cavernae sp. nov., a novel actinobacterium isolated from a karst cave sample.

A novel actinobacterial strain, designated YIM A1135T, was isolated from a soil sample collected from a karst cave in Xingyi county, Guizhou province, south-western China. The taxonomic position of the strain was investigated using a polyphasic approach. Cells of the strain were aerobic, Gram-stain-positive and partially acid-alcohol-fast. Strain YIM A1135T shared 98.3 % 16S rRNA gene sequence similarity with Nocardia jejuensis NBRC 103114T and 97.6 % with Nocardia alba YIM 30243T. DNA-DNA hybridization values between strain YIM A1135T and related type strains of the genus Nocardia were less than 70 %. In addition, meso-diaminopimelic acid was the diagnostic diamino acid in cell-wall peptidoglycan. The whole-cell sugars were fructose, mannose, galactose and glucose. The major isoprenoid quinone was MK-8(H4,ω-cycl), while the major fatty acids (>10 %) were C16 : 0, C18 : 1ω9c, C18 : 0 10-methyl and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). The polar lipids contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. Mycolic acids were present. The genomic DNA G+C content of strain YIM A1135T was 66.7 mol%. Based on the results of the molecular studies supported by its morphological, physiological, chemotaxnomic and other differential phenotypic characteristics, strain YIM A1135T is considered to represent a novel species within the genus Nocardia, for which the name Nocardia cavernae sp. nov. is proposed. The type strain is YIM A1135T (=KCTC 39595T=CCTCC AA 2017030T).

Lentzea cavernae sp. nov., an actinobacterium isolated from a karst cave sample, and emended description of the genus Lentzea.

A novel actinobacterial strain, designated SYSU K10001T, was isolated from a limestone sample collected from a karst cave in Xingyi county, Guizhou province, south-western China. The taxonomic position of the strain was investigated using a polyphasic approach. Cells of the strain were aerobic and Gram-stain-positive. On the basis of 16S rRNA gene sequence analysis, strain SYSU K10001T was most closely related to the type strains of the genus Lentzea, Lentzea albida NBRC 16102T (98.8 % similarity) and Lentzea waywayandensis NRRL B-16159T (98.6 %), and is therefore considered to represent a member of the genus Lentzea. DNA-DNA hybridization values between strain SYSU K10001T and related type strains of the genus Lentzea were less than 70 %. In addition, meso-diaminopimelic acid was the diagnostic diamino acid in the cell-wall peptidoglycan. The whole-cell sugars were arabinose, fructose, mannose and xylose. The major isoprenoid quinone was MK-9(H4), while the major fatty acids (>10 %) were iso-C16 : 0 and C14 : 0. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, hydroxy-phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside, one unidentified phospholipid and one unidentified lipid. The genomic DNA G+C content of strain SYSU K10001T was 69.4 mol%. On the basis of phenotypic, genotypic and phylogenetic data, strain SYSU K10001T represents a novel species of the genus Lentzea, for which the name Lentzea cavernae sp. nov. is proposed. The type strain is SYSU K10001T (=KCTC 39804T=CGMCC 4.7367T=NBRC 112394T).

Nocardioides cavernae sp. nov., an actinobacterium isolated from a karst cave.

A novel actinobacterial strain, designated YIM A1136T, was isolated from a soil sample collected from a karst cave in Xingyi county, Guizhou province, south-western China. The taxonomic position of the strain was investigated using a polyphasic approach. Cells of the strain were aerobic, Gram-staining-positive and rod-shaped. Colonies of the strain were circular, convex, opaque and yellowish-white in colour. On the basis of 16S rRNA gene sequence analysis, strain YIM A1136T was most closely related to the type strains Nocardioides ganghwensis JC2055T (98.3 % sequence similarity), Nocardioides exalbidus RC825T (98.2 %), Nocardioides alpinus Cr7-14T (98.2 %), Nocardioides hwasunensis HFW-21T (98.0 %), Nocardioides oleivorans DSM 16090T (97.9 %) and Nocardioides furvisabuli SBS-26T (97.8 %) and is therefore considered to represent a member of the genus Nocardioides. DNA-DNA hybridization values between strain YIM A1136T and related type strains of the genus Nocardioides were less than 70 %. ll-Diaminopimelic acid was the diagnostic diamino acid in the cell-wall peptidoglycan. The whole-cell sugars were galactose, glucose, mannose, rhamnose and ribose. The major isoprenoid quinone was MK-8(H4), while the major fatty acids (>10 %) were C16 : 0, summed feature 3 and summed feature 8. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and two unidentified phospholipids. The genomic DNA G+C content was 71.4 mol%. On the basis of phenotypic, genotypic and phylogenetic data, strain YIM A1136T merits representation of a novel species of the genus Nocardioides, for which the name Nocardioides cavernae sp. nov. is proposed. The type strain is YIM A1136T (=KCTC 39551T=DSM 29950T).

Alcaligenes endophyticus sp. nov., isolated from roots of Ammodendron bifolium.

A Gram-stain-negative, rod-shaped, motile bacterium, designated AER10T, was isolated from the roots of Ammodendron bifolium collected from Takeermohuer desert in Xinjiang Uygur Autonomous Region, northwestern China. Growth was found to occur from 10 to 45 °C, at pH 5.0-9.0, and could tolerate up to 10 % (w/v) NaCl. 16S rRNA gene sequence result indicated that the strain AER10T belongs to the genus Alcaligenes and was closely related to Alcaligenes aquatilis (98.4 %), Alcaligenes faecalissubsp. parafaecalis (98.4 %), Alcaligenes faecalissubsp. faecalis (98.1 %) and Alcaligenes faecalissubsp. phenolicus (97.9 %). However, the DNA-DNA hybridization values between the strain AER10T and the above strains were less than the threshold value (below 70 %) for the delineation of genomic species. The DNA G+C content was 53.3 mol%. Ubiquinone-8 (Q-8) was the only quinone system present. The major fatty acids were summed feature 8 (C18 : 1ω7c, 25 %), C16 : 0 (24.2 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c, 19.3 %) and cyclo-C17 : 0 (10.5 %). The polar lipid profile of the strain AER10T consists of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, two unidentified aminolipids and five unknown polar lipids. On the basis of the evidence presented in this study, strain AER10T is a representative of a novel species in the genus Alcaligenes, for which the name Alcaligenes endophyticus sp. nov. is proposed. The type strain is AER10T (=DSM 100498T=KCTC 42688T).

Actinomadura amylolytica sp. nov. and Actinomadura cellulosilytica sp. nov., isolated from geothermally heated soil.

Two aerobic, Gram-positive actinomycetes, designated YIM 77502(T) and YIM 77510(T), were isolated from geothermally heated soil of Tengchong county, Yunnan province, south-west China. The taxonomic position of strains YIM 77502(T) and YIM 77510(T) were investigated by a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that strains YIM 77502(T) and YIM 77510(T) belong to the genus Actinomadura. Both strains form extensively-branched substrate and aerial mycelia which differentiated into short spore chains. The cell wall of the two strains contained meso-diaminopimelic acid, while the whole-cell sugars detected were glucose, madurose, mannose and rhamnose. The polar lipid profile of strain YIM 77502(T) was found to consist of diphosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylinositol, two unidentified phospholipids and an unidentified polar lipid, while strain YIM 77510(T) consisted of diphosphatidylglycerol, phosphatidylinositol mannoside and phosphatidylinositol. The respiratory quinones of strains YIM 77502(T) and YIM 77510(T) were MK-9(H6) and MK-9(H8). The major fatty acids (>10 %) of strain YIM 77502(T) were C17:0, iso-C16:0, C17:010-methyl and iso-C18:0, and those of strain YIM 77510(T) were iso-C16:0, C17:010-methyl and iso-C18:0. The G+C contents of strains YIM 77502(T) and YIM 77510(T) were determined to be 71.3 and 70.2 mol%, respectively. The DNA-DNA hybridization values of strains YIM 77502(T), YIM 77510(T) and their closest phylogenetic neighbours Actinomadura echinospora BCRC 12547(T) and Actinomadura umbrina KCTC 9343(T) were less than 70 %. Based on the morphological and physiological properties, and phylogenetic analyses, strains YIM 77502(T) and YIM 77510(T) are considered to represent two novel species of the genus Actinomadura, for which the names Actinomadura amylolytica sp. nov. (type strain YIM 77502(T) = DSM 45822(T) = CCTCC AA 2012024(T)) and Actinomadura cellulosilytica sp. nov. (type strain YIM 77510(T) = DSM 45823(T) = CCTCC AA 2012023(T)) are proposed.

Pseudonocardia xishanensis sp. nov., an endophytic actinomycete isolated from the roots of Artemisia annua L.

A Gram-positive, non-motile, endophytic actinomycete, designated strain YIM 63638(T), was isolated from the surface-sterilized roots of Artemisia annua L. The isolate grew optimally with 1-3 % (w/v) NaCl, at pH 6.0-7.0 and at 20-37 °C. Comparative analysis of the 16S rRNA gene sequence showed that the isolate belonged to the genus Pseudonocardia and showed highest sequence similarity with Pseudonocardia oroxyli D10(T) (98.9 %), Pseudonocardia ailaonensis YIM 45505(T) (98.3 %) and Pseudonocardia halophobica IMSNU 21327(T) (98.0 %). Phylogenetic distance from other type strains of species with validly published names within the genus Pseudonocardia was greater than 2.3 %. Strain YIM 63638(T) had a genomic DNA G+C content of 72.1 mol% and MK-8(H(4)) as the predominant respiratory quinone. The major fatty acids were iso-C(16 : 0) (36.32 %) and 10-methyl-C(16 : 0) (19.78 %). On the basis of phenotypic properties and phylogenetic distinctiveness, strain YIM 63638(T) represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia xishanensis sp. nov. is proposed. The type strain is YIM 63638(T) ( = JCM 17906(T)  = KCTC 29005(T)).

Pseudonocardia yuanmoensis sp. nov., a novel actinobacterium isolated from soil in Yunnan, south-west China.

A novel Gram-stain positive, aerobic, non-motile, spore-forming actinobacterium, designated YIM 75926(T), was isolated from a soil sample collected at soil forest in Yuanmo county of Yunnan province, south-west China. Its taxonomic position was investigated by a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that the novel strain YIM 75926(T) belongs to the genus Pseudonocardia and was closely related to Pseudonocardia halophobica DSM 43089(T) (98.1% similarity). Strain YIM 75926(T) had MK-8 (H(4)) as the predominant menaquinone. The whole organism hydrolysates mainly consisted of meso-diaminopimelic acid, mannose, glucose, galactose and arabinose. The major cellular fatty acids were iso-C(16:0) (37.16%) and C(16:0) (12.43%). The DNA G+C content of strain YIM 75926(T) was 70.6 mol%. The resultant phylogenetic trees further showed that strain YIM 75926(T) belong to Pseudonocardia and had a distinct subclade within the evolutionary radiation of the genus Pseudonocardia. On the basis of its comparative analysis of phenotypic and genotypic characteristics, it is proposed that strain YIM 75926(T) represent a novel species of the genus Pseudonocardia, named Pseudonocardia yuanmoensis sp. nov. The type strain is YIM 75926(T) (=CCTCC AA 2011017(T) = JCM 18055(T)).

Paenibacillus thermophilus sp. nov., a novel bacterium isolated from a sediment of hot spring in Fujian province, China.

A Gram-positive, thermophilic, strictly aerobic bacterium, designated WP-1(T), was isolated from a sediment sample from a hot spring in Fujian province of China and subjected to a polyphasic taxonomic study. Cells of strain WP-1(T) were rods (~0.6-0.8 × 2.5-3.5 µm) and motile by means of peritrichous flagella. Endospores were ellipsoidal in terminal or subterminal positions. Strain WP-1(T) grew at 37-60 °C (optimum 42-45 °C), 0-3 % NaCl (optimum 1 %, w/v) and pH 3.0-9.0 (optimum pH 6.5-7.0). The predominant menaquinone was MK-7. The major fatty acids were anteiso-C(15:0), iso-C(16:0), C(16:0) and anteiso-C(17:0). The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, two glycolipids, two unidentified phospholipids and two unknown polar lipids. The cell-wall peptidoglycan contained meso-diaminopimelic acid (meso-DAP). The G + C content of the genomic DNA was 52.5 %. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain WP-1(T) is a member of the genus Paenibacillus and exhibited sequence similarity of 99.3 % to Paenibacillus macerans DSM 24(T) and both strains represented a separate lineage from all other Paenibacillus species. However, the level of DNA-DNA relatedness between strain WP-1(T) and P. macerans DSM 24(T) was 34.0 ± 4.7 %. On the basis of phylogenetic, physiological and chemotaxonomic analysis data, strain WP-1(T) is considered to represent as a novel species of the genus Paenibacillus, for which the name Paenibacillus thermophilus sp. nov., is proposed, with the type strain WP-1(T) (=DSM 24746(T) = JCM 17693(T) = CCTCC AB 2011115(T)).

Lysobacter thermophilus sp. nov., isolated from a geothermal soil sample in Tengchong, south-west China.

A Gram-negative and aerobic bacterium, designated YIM 77875(T), was isolated from a geothermal soil sample collected at Rehai National Park, Tengchong, Yunnan Province, south-west China. Bacterial growth occurred from 37 to 65 °C (optimum 50 °C), pH 6.0-8.0 (optimum pH 7.0) and 0-1 % NaCl (w/v). Cells were rod-shaped and colonies were convex, circular, smooth, yellow and non-transparent. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM 77875(T) belongs to the genus Lysobacter. The 16S rRNA gene sequence similarity values between strain YIM 77875(T) and other species of the genus Lysobacter were all below 94.7 %. The polar lipids of strain YIM 77875(T) were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and five unknown phospholipids. The predominant respiratory quinone was Q-8 and the G+C content was 68.8 mol%. Major fatty acids were iso-C(16:0), iso-C(15:0) and iso-C(11:0). On the basis of the morphological and chemotaxonomic characteristics, as well as genotypic data, strain YIM 77875(T) represents a novel species, Lysobacter thermophilus sp. nov., in the genus Lysobacter. The type strain is YIM 77875(T) (CCTCC AB 2012064(T) = KCTC 32020(T)).

Lihuaxuella thermophila gen. nov., sp. nov., isolated from a geothermal soil sample in Tengchong, Yunnan, south-west China.

A novel filamentous bacterium, designated YIM 77831(T), was isolated from a geothermal soil sample collected at Rehai National Park, Tengchong, Yunnan province, south-west China. Growth occurred from 28 to 65 °C (optimum 50 °C), pH 6.0-8.0 (optimum pH 7.0). The strain formed branched substrate mycelia, endospores were produced on the substrate mycelium and aerial mycelium was not produced on any of the growth media tested. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YIM 77831(T) was affiliated with the family Thermoactinomycetaceae. The stain YIM 77831(T) contained meso-diaminopimelic acid in the cell wall. Whole-cell hydrolysates contained glucose, galactose, mannose, ribose and rhamnose. The polar lipids were phosphatidylmethylethanolamine, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, an unidentified aminophospholipid and four unknown phospholipids. The only menaquinone was MK-7. Major fatty acids were iso-C(15:0), anteiso-C(15:0) and anteiso-C(17:0). The G+C content was 55.6 mol%. On the basis of the morphological and chemotaxonomic characteristics as well as genotypic data, strain YIM 77831(T) represents a novel genus and species, Lihuaxuella thermophila gen. nov., sp. nov., in the family Thermoactinomycetaceae. The type strain is YIM 77831(T) (CCTCC AA 2011024(T) = JCM 18059(T)).

Insights on the Effects of Heat Pretreatment, pH, and Calcium Salts on Isolation of Rare Actinobacteria from Karstic Caves.

The phylum Actinobacteria is one of the most ubiquitously present bacterial lineages on Earth. In the present study, we try to explore the diversity of cultivable rare Actinobacteria in Sigangli Cave, Yunnan, China by utilizing a combination of different sample pretreatments and under different culture conditions. Pretreating the samples under different conditions of heat, setting the isolation condition at different pHs, and supplementation of media with different calcium salts were found to be effective for isolation of diverse rare Actinobacteria. During our study, a total of 204 isolates affiliated to 30 genera of phylum Actinobacteria were cultured. Besides the dominant Streptomyces, rare Actinobacteria of the genera Actinocorallia, Actinomadura, Agromyces, Alloactinosynnema, Amycolatopsis, Beutenbergia, Cellulosimicrobium, Gordonia, Isoptericola, Jiangella, Knoellia, Kocuria, Krasilnikoviella, Kribbella, Microbacterium, Micromonospora, Mumia, Mycobacterium, Nocardia, Nocardioides, Nocardiopsis, Nonomuraea, Oerskovia, Pseudokineococcus, Pseudonocardia, Rhodococcus, Saccharothrix, Streptosporangium, and Tsukamurella were isolated from these cave samples.

Pseudonocardia antimicrobica sp. nov., a novel endophytic actinomycete associated with Artemisia annua L. (sweet wormwood).

A Gram-reaction-positive, non-motile, endophytic actinomycete, designated strain YIM 63235(T), was isolated from the surface-sterilized stems of Artemisia annua L., and characterized to determine its taxonomic position. The strain YIM 63235(T) formed well-differentiated aerial and substrate mycelia on media tested. The phylogenetic tree based on 16S rRNA gene sequences showed that the new isolate formed a distinct lineage within the genus Pseudonocardia, and the strain YIM 63235(T) was closely related to Pseudonocardia parietis 04-St-002(T) (99.1%). However, DNA-DNA relatedness demonstrated that strain YIM 63235(T) was distinct from the closest phylogenetic neighbor. The chemotaxonomic properties of strain YIM 63235(T) were consistent with those of the genus Pseudonocardia: the diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid and MK-8(H(4)) was the predominant menaquinone. The major fatty acids were iso-C(16:0) and iso-C(16:1) H. The DNA G+C content of strain YIM 63235(T) was 71.0 mol%. On the basis of the phenotypic and phylogenetic distinctiveness, the novel isolate was identified as representing a novel species of the genus Pseudonocardia, for which the name Pseudonocardia antimicrobica sp. nov. (type strain YIM 63235(T) =CCTCC AA 208080(T)=DSM 45303(T)) is proposed.