A Mesorhizobium loti mutant with reduced glucan content shows defective invasion of its host plant Lotus japonicus, by Y. Kawaharada, S. Eda, K. Minamisawa and H. Mitsui

Microbiology vol. 153, part 12, pp. 3983 - 3993

Fig. S1. Determination of sugar composition of carbohydrates in the major peak from the gel filtration of ML001 (wild-type) cell extract (Fig. 2a of main paper). The sample was hydrolysed in 2 M trifluoroacetic acid at 96 °C for 6 h. The liberated sugars were trimethylsilylated with silylation reagent (TMSH-C; GL Sciences), and separated on a capillary dimethylpolysiloxane column (OV-1, 25 m×0.25 mm×0.3 μm; GL Sciences) at a flow rate of 1.22 ml min^-1 of carrier gas (He), which was mounted on a G-3500 gas chromatograph (Hitachi High-Technologies). The column temperature was programmed to 1 min at 140 °C followed by a 2 °C min^-1 increase to 220 °C, and the injector and detector temperatures were both 220 °C. A chromatogram of the sample is shown together with that of mixture of glucose and galactose used as standards. Triplets derived from glucose and galactose are marked by asterisks and plus signs, respectively, in the chromatograms. The relative amount of glucose was calculated to be 76% from peak areas as the ratio of glucose to all the materials detected. The other peaks were not characterized to determine whether they are derived from sugar components or different organic compounds. [PDF] (320 kb)

Fig. S2. Thin-layer chromatography profiles of cell-associated glucans from Mesorhizobium loti strains. A sample containing a fixed amount (15 μg) in glucose equivalents, which was derived from the major peak on the gel-filtration chromatography of ethanol extract of cells, was spotted onto Silica gel 60 TLC plates (Merck). The plates were developed with 1-butanol/ethanol/water (5:5:4), sprayed with 5% (v/v) sulfuric acid in ethanol, and heated at 120 °C for 30 min as described by Breedveld et al. (1995). Lane 1, ML001 (wild-type); lane 2, MYa179 (cep::TnphoA mutant); lane 3, MYa179 complemented with pYK33 (cep⁺); lane 4, YML1011 (cep::aadA mutant). Positions of anionic glucans and neutral glucans are indicated. [PDF] (420 kb)

REFERENCE
Breedveld, M. W., Hadley, J. A. & Miller, K. J. (1995). A novel cyclic β-1,2-glucan mutant of Rhizobium meliloti. J Bacteriol 177, 6346-6351.

This Article

	Abstract
	Full Text

Services

	Email this article to a friend
	Alert me to new issues of the journal


Citing Articles

	Citing Articles via CrossRef

INT J SYST EVOL MICROBIOL	MICROBIOLOGY	J GEN VIROL
J MED MICROBIOL	ALL SGM JOURNALS