A primary assessment of the endophytic bacterial community in a xerophilous moss (Grimmia montana) using molecular method and cultivated isolates.

Investigating the endophytic bacterial community in special moss species is fundamental to understanding the microbial-plant interactions and discovering the bacteria with stresses tolerance. Thus, the community structure of endophytic bacteria in the xerophilous moss Grimmia montana were estimated using a 16S rDNA library and traditional cultivation methods. In total, 212 sequences derived from the 16S rDNA library were used to assess the bacterial diversity. Sequence alignment showed that the endophytes were assigned to 54 genera in 4 phyla (Proteobacteria, Firmicutes, Actinobacteria and Cytophaga/Flexibacter/Bacteroids). Of them, the dominant phyla were Proteobacteria (45.9%) and Firmicutes (27.6%), the most abundant genera included Acinetobacter, Aeromonas, Enterobacter, Leclercia, Microvirga, Pseudomonas, Rhizobium, Planococcus, Paenisporosarcina and Planomicrobium. In addition, a total of 14 species belonging to 8 genera in 3 phyla (Proteobacteria, Firmicutes, Actinobacteria) were isolated, Curtobacterium, Massilia, Pseudomonas and Sphingomonas were the dominant genera. Although some of the genera isolated were inconsistent with those detected by molecular method, both of two methods proved that many different endophytic bacteria coexist in G. montana. According to the potential functional analyses of these bacteria, some species are known to have possible beneficial effects on hosts, but whether this is the case in G. montana needs to be confirmed.

Mucilaginibacter polytrichastri sp. nov., isolated from a moss (Polytrichastrum formosum), and emended description of the genus Mucilaginibacter.

A Gram-staining-negative, rod-shaped, non-spore-forming bacterium, designated strain RG4-7(T), was isolated from the moss Polytrichastrum formosum collected from Gawalong glacier in Tibet, China, and characterized by using a polyphasic taxonomic approach. The predominant fatty acids of strain RG4-7(T) were summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), iso-C15 : 0 and iso-C17 : 0 3-OH. The major polar lipids were phosphatidylethanolamine, one unidentified aminophospholipid, one unidentified phospholipid, two unidentified aminolipids and one unidentified lipid. Strain RG4-7(T) contained MK-7 as the dominant menaquinone and the G+C content of its genomic DNA was 39.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain RG4-7(T) was affiliated to species of the genus Mucilaginibacter, and its closest relative was Mucilaginibacter jinjuensis YC7004(T) (97.0 % sequence similarity). However, the DNA-DNA relatedness between this strain and strain RG4-7(T) was only 49.1±3.7 %. Based on phylogenetic inference and phenotypic data, strain RG4-7(T) is considered to represent a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter polytrichastri sp. nov. is proposed. The type strain is RG4-7(T) ( = CGMCC 1.12493(T) = DSM 26907(T)). An emended description of the genus Mucilaginibacter is also proposed.

Cnuella takakiae gen. nov., sp. nov., a member of the phylum Bacteroidetes isolated from Takakia lepidozioides.

A Gram-staining-negative, rod-shaped and non-spore-forming bacterium, designated strain RG1-1(T), was isolated from Takakia lepidozioides collected from Gawalong glacier in Tibet, China, and characterized by using a polyphasic taxonomic approach. The predominant fatty acids of strain RG1-1(T) were iso-C(15 : 0) (19.8%), summed feature 3 (C(16 : 1)ω7c and/or C(16 : 1)ω6c, 17.0%), C(16 : 0 (9.9)%) and iso-C(17 : 0) 3-OH (9.4%); its major polar lipids were phosphatidylethanolamine, four unidentified aminolipids, one unidentified phospholipid, one unidentified aminoglycolipid, one unidentified glycolipid, and three unidentified lipids. Strain RG1-1(T) contained MK-7 as the dominant menaquinone, and the G+C content of its genomic DNA was 49.1 mol%. Strain RG1-1(T) exhibited the highest 16S rRNA gene sequence similarity (91.8%) with Flavisolibacter ginsengiterrae Gsoil 492(T) and Flavisolibacter ginsengisoli Gsoil 643(T). Phylogenetic analysis showed that strain RG1-1(T) was a member of the family Chitinophagaceae, phylum Bacteroidetes. On the basis of 16S rRNA gene sequence analysis, and phenotypic and chemotaxonomic data, strain RG1-1(T) is considered to represent a novel species of a novel genus, for which the name Cnuella takakiae gen. nov., sp. nov. is proposed. The type strain is RG1-1(T) ( = CGMCC 1.12492(T) = DSM 26897(T)).

A primary assessment of the endophytic bacterial community in a xerophilous moss (Grimmia montana) using molecular method and cultivated isolates

Investigating the endophytic bacterial community in special moss species is fundamental to understanding the microbial-plant interactions and discovering the bacteria with stresses tolerance. Thus, the community structure of endophytic bacteria in the xerophilous moss Grimmia montana were estimated using a 16S rDNA library and traditional cultivation methods. In total, 212 sequences derived from the 16S rDNA library were used to assess the bacterial diversity. Sequence alignment showed that the endophytes were assigned to 54 genera in 4 phyla (Proteobacteria, Firmicutes, Actinobacteria and Cytophaga/Flexibacter/Bacteroids). Of them, the dominant phyla were Proteobacteria (45.9%) and Firmicutes (27.6%), the most abundant genera included Acinetobacter, Aeromonas, Enterobacter, Leclercia, Microvirga, Pseudomonas, Rhizobium, Planococcus, Paenisporosarcina and Planomicrobium. In addition, a total of 14 species belonging to 8 genera in 3 phyla (Proteobacteria, Firmicutes, Actinobacteria) were isolated, Curtobacterium, Massilia, Pseudomonas and Sphingomonas were the dominant genera. Although some of the genera isolated were inconsistent with those detected by molecular method, both of two methods proved that many different endophytic bacteria coexist in G. montana. According to the potential functional analyses of these bacteria, some species are known to have possible beneficial effects on hosts, but whether this is the case in G. montana needs to be confirmed.

Paenibacillus typhae sp. nov., isolated from roots of Typha angustifolia L.

A Gram-stain-positive, facultatively anaerobic and rod-shaped bacterium, designated strain xj7(T), was isolated from roots of Typha angustifolia L. growing in Beijing Cuihu Wetland, China. The isolate was identified as a member of the genus Paenibacillus based on phenotypic characteristics and phylogenetic inference. The novel strain was spore-forming, motile, catalase-positive and oxidase-negative. Optimal growth of strain xj7(T) occurred at 28-30 °C and pH 7.0-7.5. Diphosphatidylglycerol was the most abundant polar lipid and occurred along with phosphatidylglycerol, phosphatidylethanolamine, one unknown phospholipid and three unknown aminophospholipids. The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. The predominant isoprenoid quinone was MK-7. The major fatty acid components were anteiso-C15 : 0 (56.1 %), iso-C16 : 0 (9.1 %), C16 : 0 (8.0 %), iso-C14 : 0 (6.3 %) and iso-C15 : 0 (5.1 %). The G+C content of genomic DNA was 47.9 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain xj7(T) fell within the evolutionary radiation encompassed by the genus Paenibacillus, its closest neighbours were Paenibacillus borealis KK19(T) (97.5 %) and Paenibacillus durus DSM 1735(T) (97.1 %). However, the DNA-DNA relatedness values between strain xj7(T) and P. borealis KK19(T) and between strain xj7(T) and P. durus DSM 1735(T), were both 35 %. Based on phenotypic, chemotaxonomic and phylogenetic properties, strain xj7(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus typhae sp. nov. is proposed. The type strain is xj7(T) ( = CGMCC 1.11012(T) = DSM 25190(T)).

Massilia namucuonensis sp. nov., isolated from a soil sample.

A Gram-staining-negative, rod-shaped and non-spore-forming bacterium, designated strain 333-1-0411(T), was isolated from a soil sample collected from Namucuo, Tibet Autonomous Region, China and characterized in a taxonomic study using a polyphasic approach. The major fatty acid components of strain 333-1-0411(T) were summed feature 3 (C(16 : 1)ω7c and/or C(16 : 1)ω6c) and C(16 : 0); its major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. Q-8 was the dominant ubiquinone, and the G+C content of the genomic DNA was 66.7 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that strain 333-1-0411(T) fell within the evolutionary radiation encompassed by the genus Massilia. The 16S rRNA gene sequence similarities between strain 333-1-0411(T) and recognized species of the genus Massilia ranged from 95.4 % to 97.2 %, and the most closely related strains were Massilia flava Y9(T) (97.2 %) and Massilia albidiflava 45(T) (97.0 %). However, the DNA-DNA relatedness values between strain 333-1-0411(T) and M. flava Y9(T) and M. albidiflava 45(T) were 20.2 % and 27.2 %, respectively. Based on the phenotypic, chemotaxonomic and phylogenetic properties, strain 333-1-0411(T) is considered to represent a novel species of the genus Massilia, for which the name Massilia namucuonensis sp. nov. is proposed. The type strain is 333-1-0411(T) (= CGMCC 1.11014(T) = DSM 25159(T)).

Burkholderia grimmiae sp. nov., isolated from a xerophilous moss (Grimmia montana).

A Gram-staining-negative, rod-shaped, non-spore-forming bacterium, designated strain R27(T), was isolated from the moss Grimmia montana, collected from Beijing Songshan National Nature Reserve, China, and characterized by using a polyphasic taxonomic approach. The predominant fatty acids of strain R27(T) were C18:1ω7c (33.6%), C16:0 (16.3%), summed feature 3 (C16:1ω7c and/or C16:1ω6c; 15.8%) and C17:0 cyclo (8.7%) and its major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, three uncharacterized aminolipids and an unknown phospholipid. Strain R27(T) contained Q-8 as the dominant isoprenoid quinone and the G+C content of its genomic DNA was 64.6 mol%. On the basis of 16S rRNA gene sequence comparison, strain R27(T) showed 99.1% similarity to the closest related type strain, Burkholderia zhejiangensis OP-1(T), and 97.6% similarity to Burkholderia glathei ATCC 29195(T). However, the DNA-DNA relatedness between strain R27(T) and B. zhejiangensis CCTCC AB 2010354(T) and B. glathei ATCC 29195(T) was 10.2 and 14.9%, respectively. Based on 16S rRNA and rpoB gene sequence similarities and phenotypic and chemotaxonomic data, strain R27(T) is considered to represent a novel species of the genus Burkholderia, for which the name Burkholderia grimmiae sp. nov. is proposed. The type strain is R27(T) (=CGMCC 1.11013(T) =DSM 25160(T)).