A Rhizobium leguminosarum AcpXL Mutant Produces Lipopolysaccharide Lacking 27-Hydroxyoctacosanoic Acid

The structure of the lipid A from Rhizobium etli and Rhizobium leguminosarum lipopolysaccharides (LPSs) lacks phosphate and contains a galacturonosyl residue at its 4′ position, an acylated 2-aminogluconate in place of the proximal glucosamine, and a very long chain ω-1 hydroxy fatty acid, 27-hydroxyoctacosanoic acid (27OHC28:0). The 27OHC28:0 moiety is common in lipid A's among members of the Rhizobiaceae and also among a number of the facultative intracellular pathogens that form chronic infections, e.g., Brucella abortus, Bartonella henselae, and Legionella pneumophila. In this paper, a mutant of R. leguminosarum was created by placing a kanamycin resistance cassette within acpXL, the gene which encodes the acyl carrier protein for 27OHC28:0. The result was an LPS containing a tetraacylated lipid A lacking 27OHC28:0. A small amount of the mutant lipid A may contain an added palmitic acid residue. The mutant is sensitive to changes in osmolarity and an increase in acidity, growth conditions that likely occur in the nodule microenvironment. In spite of the probably hostile microenvironment of the nodule, the acpXL mutant is still able to form nitrogen-fixing root nodules even though the appearance and development of nodules are delayed. Therefore, it is possible that the acpXL mutant has a host-inducible mechanism which enables it to adapt to these physiological changes

Cell Wall Carbohydrate Compositions of Strains from the Bacillus cereus Group of Species Correlate with Phylogenetic Relatedness▿

Members of the Bacillus cereus group contain cell wall carbohydrates that vary in their glycosyl compositions. Recent multilocus sequence typing (MLST) refined the relatedness of B. cereus group members by separating them into clades and lineages. Based on MLST, we selected several B. anthracis, B. cereus, and B. thuringiensis strains and compared their cell wall carbohydrates. The cell walls of different B. anthracis strains (clade 1/Anthracis) were composed of glucose (Glc), galactose (Gal), N-acetyl mannosamine (ManNAc), and N-acetylglucosamine (GlcNAc). In contrast, the cell walls from clade 2 strains (B. cereus type strain ATCC 14579 and B. thuringiensis strains) lacked Gal and contained N-acetylgalactosamine (GalNAc). The B. cereus clade 1 strains had cell walls that were similar in composition to B. anthracis in that they all contained Gal. However, the cell walls from some clade 1 strains also contained GalNAc, which was not present in B. anthracis cell walls. Three recently identified clade 1 strains of B. cereus that caused severe pneumonia, i.e., strains 03BB102, 03BB87, and G9241, had cell wall compositions that closely resembled those of the B. anthracis strains. It was also observed that B. anthracis strains cell wall glycosyl compositions differed from one another in a plasmid-dependent manner. When plasmid pXO2 was absent, the ManNAc/Gal ratio decreased, while the Glc/Gal ratio increased. Also, deletion of atxA, a global regulatory gene, from a pXO2− strain resulted in cell walls with an even greater level of Glc

Analysis of Business Organising in Ornamental Plant Cooperatives

Namen diplomskega dela je bil raziskati pripravljenost pridelovalcev okrasnih rastlin za povezovanje v poslovno organiziranje zadružništvo. Za pridobitev informacij je bilo v maju in juniju 2017 anketiranih 17 pridelovalcev okrasnih rastlin iz severovzhodne Slovenije. Rezultati ankete so pokazali, da so pridelovalci v glavnem seznanjeni z obliko poslovnega organiziranja zadružništvo. Večina pridelovalcev (94 %) je mnenja, da bi bilo potrebno na območju SV Slovenije ustanoviti zadrugo pridelovalcev okrasnih rastlin. Ugotovili smo, da bi v primeru ustanovitve zadruge bila več kot polovica pridelovalcev pripravljena postati član zadruge, iz česar sledi, da so pripravljeni na povezovanje v zadrugo. V primeru ustanovitve zadruge pa pridelovalci ne bi bili pripravljeni zaposliti osebo, ki bi skrbela za nabavo repromateriala, trženje, promocijo in drugo. Raziskava je pokazala, da večina (69 %) pridelovalcev ni zadovoljna s sodelovanjem slovenskih vrtnarjev med seboj.The purpose of the degree was to investigate the willingness of producers of ornamental plants to join business organising cooperatives. To acquire information on the topic, a survey was carried out among 17 growers of ornamental plants from north-eastern Slovenia, and it was carried out in May and June 2017. The results of the survey showed that the growers are mostly familiar with the business organising cooperatives form. Most Growers’ (94%) opinion was that they would be beneficial for the north-eastern area of Slovenia and that Ornamental Plant Cooperatives should be established. We found that, in the event of the establishment of a cooperative, more than half of the growers would be willing to become members of the cooperative and are thus ready to join the cooperative. In case a cooperative should be established, growers would not be willing to employ people who would be responsible for the procurement of raw materials, marketing, promotion and more. The survey showed that a high percentage (69%) of growers are not satisfied with the cooperation of Slovenian gardens with each other

Reactivity of monoclonal antiserum EAII-6G6-2-3, specific for <i>Ba</i> neutral cell wall polysaccharide [19], with HF-SCWPs from <i>Bc</i> group strains.

<p>Meso Scale Discovery (MSD) multi-array high bind 96 well plates were coated with a fixed concentration of HF-SCWP antigen (2 μg/ml) from different <i>Bacillus</i> species and probed with the monoclonal antibody in serial two fold dilutions. Bound antibody was detected by using 2.5 μg/ml of sulfo-tagged goat anti-mouse IgM detection antibody. Data points are the average of three independent experiments. Error bars represent one standard error. Reactivity reported as effective concentration (EC₅₀) titer in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0183115#pone.0183115.t004" target="_blank">Table 4</a></b>.</p

Percent and type of structural substituent at the 3-position of backbone ManNAc residues in HF-SCWPs from examined infection-associated <i>B</i>. <i>cereus</i> strains.

<p>Percent and type of structural substituent at the 3-position of backbone ManNAc residues in HF-SCWPs from examined infection-associated <i>B</i>. <i>cereus</i> strains.</p

Expanded region of proton NMR spectra identifying the locations and area ratios of the Gal substituents on HF-SCWPs from <i>B</i>. <i>cereus</i> strains.

<p><b>(A)</b> human isolate <i>B</i>. <i>cereus</i> G9241; <b>(B)</b> great ape isolate <i>B</i>. <i>cereus</i> CA (Cameroon); <b>(C)</b> great ape isolate <i>Bc</i> CI (Côte d’Ivoire). The α-anomeric signal originates from the free reducing end of all these polysaccharides, and co-migrates with a new residue (<b>K</b>) in the <i>Bc</i> CA and <i>Bc</i> CI SCWPs. This reducing end (α-GlcNAc residue) was removed by borodeuteride reduction for subsequent experiments, to facilitate integration and characterization of the residue <b>K</b> system. The percentage of Gal (<b>G</b>) and Gal-Gal disaccharide (<b>J</b>+<b>K</b>) substitution at ManNAc residues (<b>B</b>+<b>B′</b>+<b>B″</b>) is estimated by examination of the signal areas for these residues. The residue <b>G</b> spin system undergoes a shift at several positions, and is designated residue <b>J</b>, when it is substituted by residue <b>K</b> (refer to <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0183115#pone.0183115.t002" target="_blank">Table 2</a></b>).</p

<i>Bacillus anthracis</i> and <i>B</i>. <i>cereus</i> strains used in this study.

<p><i>Bacillus anthracis</i> and <i>B</i>. <i>cereus</i> strains used in this study.</p

Immunochemical analysis of HF-SCWP with monoclonal and polyclonal antisera.

<p>Immunochemical analysis of HF-SCWP with monoclonal and polyclonal antisera.</p