Williamsia phyllosphaerae sp. nov., isolated from the surface of Trifolium repens leaves

Abstract

A Gram-positive-staining, non-endospore-forming actinobacterium, designated C7T , was isolated from the leaf surface of Trifolium repens. On the basis of 16S rRNA gene sequence analysis, strain C7 T was shown to belong to the genus Williamsia and was most closely related toWilliamsia maris SJS0289/JS1 T (98.0 % 16S rRNA gene sequence similarity), Williamsia deligens IMMIB RIV-956 T (96.4 %) and Williamsia serinedens IMMIB SR-4 T (95.7 %). The quinone system consisted predominantly of the menaquinones MK-9(H 2 ), MK-8(H 2 ) and MK-7(H 2 ). The major components in the polar lipid profile were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol. Mycolic acids were present. These chemotaxonomic traits and the major fatty acids, which were C 16 : 1 v7c, Strain C7 T was isolated from the leaf surface of Trifolium repens, which was collected from a meadow in the HainichDün region of Germany (Fischer et al., 2010). The strain was isolated after growth on M125 medium (http://www. dsmz.de/) at 25 u C for 14 days and maintained on nutrient agar (Oxoid) at 30 u C for 48 h. On M125 medium, the colony type exhibited by the isolate was present only occasionally, while other colony morphologies dominated.The strain was preserved in nutrient broth (Oxoid) with 20 % (v/v) glycerol at 280 u C or by lyophilization.Cell morphology was determined by phase-contrast microscopy as described by Kämpfer & Kroppenstedt (2004). Gram-staining was performed as described by Gerhardt et al. (1994).The 16S rRNA gene sequence of strain C7 T (1372 bp) was determined and analysed as described previously (Kämpfer & Kroppenstedt, 2004) , 2007). Genetic distances were calculated on the basis of pairwise comparisons and cluster analyses were performed with neighbour joining as well as maximum likelihood with fastDNAml (Olsen et al., 1994) using ARB. Tree reconstruction using the maximum-likelihood method was performed with a 30 % conservation filter (only alignment columns in which the frequency of the most abundant nucleotide is ¢30 % were included) as well as without filters. A maximum-parsimony tree was calculated with MEGA4. No significant differences could be observed between these trees. A neighbour-joining tree (without filters) is shown in Fig. 1. All of the nodes were also recovered in the maximum-likelihood and maximum-parsimony trees. Sequence similarity calculations on the basis of pairwise comparison indicated that the closest relatives of strain C7 T Abbreviations: NA, nitroanilide; NP, nitrophenyl.The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain C7T is FR691321. For analysis of mycolic acids, quinones and polar lipids, cells were grown in nutrient broth for 48 h at 28 u C in shake flasks at 180 r.p.m. The occurrence of mycolic acids was determined by TLC as described by Minnikin et al. (1975). Menaquinones were extracted and analysed as described by Collins et al. (1979) and Groth et al. (1996). Polar lipids were extracted by the method of Minnikin et al. (1979)...

Role of Williamsia and Segniliparus in human infections with the approach taxonomy, cultivation, and identification methods

“…At the time of writing, the specific media have not been described for the isolation of Williamsia from human clinical samples. In literature, various media have been used for Williamsia isolation from various sources; however, those associated with good growth or appropriate for morphological examination are columbia agar supplemented with 5% sheep blood agar and brain heart infusion (BHI) agar [ 40 , 43 ], M3 agar supplemented with cycloheximide and nystatin [ 44 ], glucose/yeast extract agar (GYEA) plates [ 44 , 45 ], raffinose–histidine agar plate supplemented with cycloheximide and nystatin [ 45 ], tryptic soy agar (trypticase soy agar/tryptone soy agar) [ 2 , 46 – 48 ], starch-casein agar supplemented with cycloheximide [ 47 ], nystatin and rifampicin and ISP media 2–7 [ 47 ], modified Bennett’s agar [ 47 ], glucose-yeast extract malt extract agar [ 47 ], nutrient agar [ 47 , 49 , 50 ], Gauze’s medium with cycloheximide, nalidixic acid, novobiocin, and nystatin [ 51 ], M125 medium [ 49 ], tap water agar and ISP medium 2 [ 48 ], Reasoner's 2A agar (R2A) [ 2 ], GC agar [ 52 ], serum broth [ 52 ], and M1 agar plate [ 53 ].…”

“…Phenotypic characterizations are the first step for these bacteria identification at the genus and species levels. In the Williamsia there is 12 species names validly published includes Williamsia aurantiacus [ 53 ], W. deligens [ 40 ], Williamsia faeni [ 51 ], Williamsia herbipolensis [ 50 ], Williamsia limnetica [ 47 ], Williamsia maris [ 44 ], Williamsia marianensis [ 45 ], Williamsia muralis [ 2 ], Williamsia phyllosphaerae [ 49 ], Williamsia serinedens [ 43 ], Williamsia spongiae [ 46 ], and Williamsia sterculiae [ 48 ]. The species of this genus are distributed in different environments; however, they have also been isolated from clinical specimens [ 40 ].…”

“…The temperature range for the growth for Williamsia spp. including W. limnetica , W. sterculiae , W. maris , W. muralis is 10 to 37 °C [ 2 , 44 , 47 , 48 ], for W. aurantiacus and W. spongiae is 10 to 45 °C [ 46 , 53 ], for W. phyllosphaerae and W. herbipolensis is 25 to 30 °C [ 49 , 50 ], for W. deligens is 37 °C [ 40 ], for W. serinedens is 22 to 30 °C [ 43 ], for W. marianensis is 4 to 30 °C [ 45 ], and for W. faeni is 10 to 30 °C [ 51 ]. Also, the pH range for the growth is between 4.0 and 10.0 for W. aurantiacus and W. spongiae [ 46 , 53 ] and 5.0–8.0 for W. sterculiae [ 48 ].…”

Draft genome of the Arabidopsis thaliana phyllosphere bacterium, Williamsia sp. ARP1

“…At the time of writing, only one other draft genome of Williamsia sp. D3 was publicly available [ 4 ] and nine species of this taxon were recognized with valid scientific names: Williamsia deligens [ 5 ], Williamsia faeni [ 6 ], Williamsia limnetica [ 7 ], Williamsia marianensis [ 8 ], Williamsia maris [ 9 ], Williamsia muralis [ 1 ], Williamsia phyllosphaerae [ 10 ], Williamsia serinedens [ 11 ] and Williamsia sterculiae [ 12 ]. Further this genus has been linked with the degradation of hexahydro-1,3,5-trinitro-1,3,5-triazine in soils as a sole nitrogen source [ 13 ], the degradation of carbonyl sulfide in soils [ 14 ] and polychlorinated biphenyls in tree habitats [ 15 ].…”