Methylopila carotae sp. nov., a facultative methylotroph, isolated from a root of Daucus carota L

Antonie Van Leeuwenhoek. 2019 Sep;112(9):1307-1316. doi: 10.1007/s10482-019-01263-x. Epub 2019 Apr 10.

Abstract

An aerobic facultatively methylotrophic bacterium, designated strain Das4.1T, was isolated from a root of Daucus carota L. The cells of this strain were observed to be Gram-stain negative, asporogenous, non-motile short rods multiplying by binary fission. Strain Das4.1T can utilise methanol, methylamine and a variety of polycarbon compounds as carbon and energy sources. C1-compounds were found to be assimilated via the isocitrate lyase-negative variant of the serine pathway. On medium with 0.5% methanol, growth of strain Das4.1T was observed at pH 5.5-9.0 (optimum, pH 6.0-7.0) and 18-37 °C (optimum, 24-29 °C) and in the presence of 0-2% (w/v) NaCl (optimum, 0.05%). Cells are catalase and oxidase positive and synthesise indole from L-tryptophan. The major fatty acids of methanol-grown cells were identified as C18:1ω7c, C18:0 and 11-methyl-C18:1ω7c. The predominant phospholipids were found to be phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylmonomethylethanolamine. The major respiratory quinone was identified as Q-10. The DNA G + C content of strain Das4.1T was determined to be 67.3 mol% (Tm). Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain Das4.1T belongs to the genus Methylopila and shows high sequence similarity to Methylopila oligotropha 2395AT (98.4%) and Methylopila capsulata IM1T (98.0%). However, the DNA-DNA relatedness of strain Das4.1T with M. oligotropha 2395AT was only 22 ± 3%. Based on genotypic, chemotaxonomic and physiological characterisation, the isolate can be classified as a novel species of the genus Methylopila, for which the name Methylopila carotae sp. nov. is proposed. The type strain is Das4.1T (= VKM B-3244T = CCUG 72399T).

Keywords: Methylopila carotae sp.nov.; Phytosymbiont; Taxonomy.

MeSH terms

  • Bacterial Typing Techniques
  • Base Composition
  • Cluster Analysis
  • Cytosol / chemistry
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Ribosomal / chemistry
  • DNA, Ribosomal / genetics
  • Daucus carota / microbiology*
  • Fatty Acids / analysis
  • Methylocystaceae / classification*
  • Methylocystaceae / genetics
  • Methylocystaceae / isolation & purification*
  • Methylocystaceae / physiology
  • Nucleic Acid Hybridization
  • Phospholipids / analysis
  • Phylogeny
  • Plant Roots / microbiology
  • Quinones / analysis
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA

Substances

  • DNA, Bacterial
  • DNA, Ribosomal
  • Fatty Acids
  • Phospholipids
  • Quinones
  • RNA, Ribosomal, 16S