Downloaded from www.microbiologyresearch.org by IP: 54.70.40.11 On: Thu, 06 Dec 2018 23:45:38 Brevundimonas canariensis sp. nov., isolated from roots of Triticum aestivum Esther Men endez, 1,2 Juan P erez-Yepes, 3 Lorena Carro, 4 Mercedes Fern andez-Pascual, 5 Martha-Helena Ramírez-Bahena, 2,6 Hans-Peter Klenk, 4 Milagros León Barrios, 3 Alvaro Peix 2,6 and Encarna Vel azquez 1,2, * Abstract A bacterial strain designated GTAE24 T was isolated from a root of wheat growing in soil from the Canary Islands, Spain. Phylogenetic analyses based on 16S rRNA gene sequences placed the isolate in the genus Brevundimonas with Brevundimonas abyssalis TAR-001 T as its closest relative at 99.4 % similarity. DNA–DNA hybridization studies showed an average of 38 % relatedness between strain GTAE24 T and the type strain of B. abyssalis. Cells were Gram-stain-negative and motile by polar flagella. The strain was positive for oxidase and weakly positive for catalase. Gelatin, starch and casein were not hydrolysed. Growth was supported by many carbohydrates and organic acids as carbon source. Ubiquinone Q-10 was the predominant isoprenoid quinone and C 18 : 1 !7c/C 18 : 1 !6c (summed feature 8) and C 16 : 0 were the major fatty acids. The major polar lipids were phosphatidylglycerol, 1,2-di-O-acyl-3-O-[D-glucopyranosyl-(1,4)-a-D-glucopyranuronosyl] glycerol, 1,2- diacyl-3-O-[6¢-phosphatidyl-a-D-glucopyranosyl] glycerol, 1,2-di-O-acyl-3-O-a-D-glucopyranosyl glycerol, and 1,2-di-O-acyl-3- O-a-D-glucopyranuronosyl glycerol. The DNA G+C content was 63.9 mol%. Phylogenetic, chemotaxonomic and phenotypic analyses showed that strain GTAE24 T should be considered as representing a novel species of the genus Brevundimonas, for which the name Brevundimonas canariensis sp. nov. is proposed. The type strain is GTAE24 T (=LMG 29500 T =CECT 9126 T ). The genus Brevundimonas comprises Gram-negative rod- shaped, vibrioid or subvibrioid cells. Members of this genus are strictly aerobic, chemo-organotrophic and oligotrophic [1]. This genus currently contains 25 validly described spe- cies (http://www.bacterio.net/brevundimonas.html), some of them isolated from soil, but to date no novel Brevundimo- nas species have been isolated from plant root tissues. In this work we characterize a strain, named GTAE24 T , iso- lated from roots of wheat and based on its genotypic, chemotaxonomic and phenotypic characteristics we propose its classification into a novel species of the genus Brevundimonas. Strain GTAE24 T was isolated from a root of Triticum aesti- vum (wheat) growing in soil from Lanzarote (Canary Islands, Spain) during a study of root endophytes of this plant. The roots were surface-disinfected with 70 % ethanol for 1 min, 2 % NaClO for 3 min and 70 % ethanol for 30 s. Then, roots were rinsed five times with sterile distilled water and crushed into a tube containing 10 ml sterile distilled water. Samples were incubated at room temperature and 160 r.p.m. for 1 h. Aliquots of the homogenized root tissue were inoculated on tripticase soy agar (TSA; Difco, Becton Dickinson) and the plates were incubated at 28  C for 4 days. In parallel, some of the disinfected entire root frag- ments were incubated in the same medium to ensure their complete external disinfection and no growth was observed around these root fragments. Colonies of strain GTAE24 T were white to cream-coloured, round, smooth and convex with approximate diameters of 1–3 mm. The strain was grown on nutrient agar (NA; Sigma) for 48 h at 22  C to check for motility by phase- contrast microscopy using the hanging drop method. Gram staining was carried out by the procedure described by Doetsch [2] after 24 h of incubation at 28  C. The flagella- tion type was determined by electron microscopy after 48 h of incubation of strain GTAE24 T on NA at 22  C. The cells Author affiliations: 1 Departamento de Microbiología y Gen etica, Universidad de Salamanca, Salamanca, Spain; 2 Unidad Asociada Grupo de Interacción Planta-Microorganismo, Universidad de Salamanca-IRNASA-CSIC, Salamanca, Spain; 3 Departamento de Bioquímica, Microbiología, Biología Celular y Gen etica, Universidad de La Laguna, Tenerife, Spain; 4 School of Biology, Newcastle University, Newcastle-upon-Tyne, UK; 5 Instituto de Ciencias Agrarias, Consejo Superior de Investigaciones Científicas (ICA-CSIC), Madrid, Spain; 6 Instituto de Recursos Naturales y Agrobiología de Salamanca, Consejo Superior de Investigaciones Científicas (IRNASA-CSIC), Salamanca, Spain. *Correspondence: Encarna Vel azquez, evp@usal.es Keywords: Brevundimonas; endophytic bacteria; Triticum aestivum; roots; Canary Islands. The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain GTAE24 T is KX898252. Two supplementary figures are availlable with the online Supplementary Material. NOTE Men endez et al., Int J Syst Evol Microbiol 2017;67:969–973 DOI 10.1099/ijsem.0.001725 001725 ã 2017 IUMS 969