Klugiella xanthotipulae gen. nov., sp. nov., a novel member of the family Microbacteriaceae

Abstract

Strain 44C3T was isolated from the hindgut of Tipula abdominalis larvae as described by Cook et al. (2007). T. abdominalis is an aquatic crane fly, larvae of which are primary shredders of leaf litter in small, riparian streams. The hindgut of T. abdominalis larvae hosts a dense and diverse bacterial community (Klug & Kotarski, 1980), which is suggested to facilitate digestion of their lignocellulosic diet (Lawson & Klug, 1989). Strain 44C3T was maintained as 40 % (w/v) glycerol suspensions at 220 u C. Culture for biochemical and molecular studies was obtained by cultivation on trypticase soy agar (TSA; Difco) or in trypticase soy broth (TSB; Difco) at 28 u C for 48 h. Cultures were incubated at 4, 10, 22, 28, 30, 37 and 45 u C to determine the range and optimum temperature for growth. At 28 u C, growth was tested at pH 6-12 and in the presence of NaCl concentrations of 0.5-9 % to determine the pH and NaCl optima and range. Colony morphology was observed on TSA after 48 h growth at 28 u C. Gram staining was performed and standard physiological tests were performed with API NE, API Staph, API Strep and API Coryne test kits (bioMérieux). For phase-contrast microscopy observation, cells were viewed at 6100 magnification with a Leica SP2 upright microscope (Leica Microsystems Inc.) and images were captured with a Zeiss AxioCam (Carl Zeiss MicroImaging, Inc.) at the Center for Advanced Ultrastructure Research at the University of Georgia. A pixel to micrometre ratio was calculated via imaging software and this ratio was used to determine cell size. Strain 44C3T stained Gram-variable, but was Gram-type positive. It was aerobic, grew optimally at 28 u C and was able to grow at 4-30 u C. Although limited growth did occur at 4 u C, this was not observed until 672 h (4 weeks) of incubation. Irregular rod-shaped cells (0.6-3.460.4-0.8 mm) were observed, but spores were not found. Small, smooth, yellow colonies formed on TSA. Detailed biochemical and physiological characteristics of the strain are given in the genus and species descriptions below.Analyses of cell-wall sugars, menaquinones, amino acids and acyl type were performed by the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ) under the direction of Dr Peter Schumann according to recognized methods (Groth et al., 1996;Schleifer, 1985;Schleifer & Kandler, 1972;Staneck & Roberts, 1974;Uchida et al., 1999). Total cellular fatty acids were analysed by using the MIDI-FAME procedure essentially as described by Haack et al. (1994); gas chromatographs were comparedThe GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain 44C3T is AY372075.A minimum-evolution phylogenetic dendrogram based on 16S rRNA gene sequence similarity is available as supplementary material with the online version of this paper.

Glaciihabitans tibetensis gen. nov., sp. nov., a psychrotolerant bacterium of the family Microbacteriaceae, isolated from glacier ice water

“…Amended descriptions of the family were subsequently presented by Stackebrandt et al (1997) and Zhi et al (2009). At the time of writing, the family comprises 39 recognized genera such as Alpinimonas (Schumann et al , 2012), Amnibacterium (Kim & Lee, 2011), Chryseoglobus (Baik et al , 2010), Naasia (Weon et al , 2013), Compostimonas (Kim et al , 2012b), Marisediminicola (Li et al , 2010), Glaciibacter (Katayama et al , 2009), Klugiella (Cook et al , 2008) and Labedella (Lee, 2007). A common feature of the genera within the family Microbacteriaceae is the B-type cell-wall peptidoglycan.…”

“…Members of the family have been isolated from various habitats, including mushroom compost, water, plant material, soil, dusts, the gut of larvae, dried seaweed and cheese (Kim et al , 2012a, b; Kim & Lee, 2011; Dorofeeva et al , 2002; Zhang et al , 2008; Reddy et al , 2010; Kämpfer et al , 2000; Cook et al , 2008; Lee, 2007; Tsukamoto et al , 2001). The first psychrophilic member of the family Microbacteriaceae to be described was Cryobacterium with Cryobacterium psychrophilum as the type species (Suzuki et al , 1997).…”

Pontimonas salivibrio gen. nov., sp. nov., a new member of the family Microbacteriaceae isolated from a seawater reservoir of a solar saltern

“…Polar lipids were extracted using the procedures described by Minnikin et al (1984), separated by two-dimensional TLC and identified by spraying with appropriate detection reagents (Komagata & Suzuki, 1987). The quinone system was extracted according to Minnikin et al (1984) and analysed by HPLC as described by Collins (1985) using Yonghaparkia alkaliphila KCTC 19126 T (Yoon et al , 2006) and Klugiella xanthotipulae KACC 21121 T (Cook et al , 2008) as reference strains. The presence or absence of diaminopimelic acid in the peptidoglycan was determined according to the method described by Komagata & Suzuki (1987).…”

Compostimonas suwonensis gen. nov., sp. nov., isolated from spent mushroom compost

“…At the time of writing, the family includes 34 genera that are differentiated on the basis of chemotaxonomic characteristics, such as the diagnostic diamino acids of the cell-wall peptidoglycan (2,4-diaminobutyric acid, lysine and ornithine), the major respiratory quinones (menaquinones 7-14), the cellular fatty acids (anteiso-C 15 : 0 , anteiso-C 17 : 0 , iso-C 14 : 0 , iso-C 15 : 0 and iso-C 16 : 0 ) and the DNA G+C content (59-75 mol%). Members of the family have been isolated from different habitats, such as distilled water, freshwater, groundwater, ice, insect gut, lichen, plant, sea sediment, seawater, seaweed, sewage and soil (An et al, 2008;Cook et al, 2008;Evtushenko et al, 2000;Han et al, 2003;Katayama et al, 2009;Lee, 2007;Lee et al, 2008;Li et al, 2010;Männistö et al, 2000;Sheridan et al, 2003;Tiago et al, 2005Tiago et al, , 2006Vaz-Moreira et al, 2008;Yoon et al, 2006).…”

“…Table 1. Differential characteristics of strain SMC46 T and closely related genera of the family Microbacteriaceae Genera: 1, strain SMC46 T ; 2, Amnibacterium (Kim & Lee, 2011); 3, Clavibacter (Bendinger et al, 1992;Collins & Bradbury, 1992;Groth et al, 1996;Evtushenko et al, 2000); 4, Cryobacterium (Suzuki et al, 1997;Zhang et al, 2007a;Dastager et al, 2008a;Reddy et al, 2010);5, Frigoribacterium (Kämpfer et al, 2000;Dastager et al, 2008b); 6, Frondihabitans (Zhang et al, 2007b;Greene et al, 2009;Lee, 2010); 7, Klugiella (Cook et al, 2008); 8, Labedella (Lee, 2007); 9, Marisediminicola (Li et al, 2010); 10, Mycetocola (Tsukamoto et al, 2001;Bora et al, 2008); 11, Rathayibacter (Zgurskaya et al, 1993;Sasaki et al, 1998;Dorofeeva et al, 2002);12, Subtercola (Männistö et al, 2000). DAB, Diaminobutryic acid; DMA, dimethyl acetal; DPG, diphosphatidylglycerol; GL, unknown glycolipid; L, unknown lipid; PG, phosphatidylglycerol; PL, unknown phospholipid; +, positive; V, variable; 2, negative; ND, no data available.…”