7-methylpterin in methanogenic bacteria

Abstract A blue fluorescent compound was extracted and purified from cells of Methanobacterium thermoautotrophicum . The compound was identified as 7-methylpterin on the basis of its (physico-) chemical properties and by comparison with 7-methylpterin prepared by organic synthesis. The compound is present in all methanogenic bacteria studied so far and it provides methanogenic bacteria the characteristic blue fluorescence observed upon fluorescence microscopy.     

Nitrogen retranslocation in plants of maize,lupin and cocklebur

Summary In a split root experiment translocation of N from shoot to root was studied using¹⁵NO ₃ ^– . The three plant species selected for this experiment differed significantly with respect to root NRA. For lupin, maize and cocklebur about 80, 50 and 6% of all absorbed NO ₃ ^– was assmilated in the roots, respectively.Although NO ₃ ^– was reduced in the roots of lupin and maize plants to a greater extent than required for the roots' demand for organic N, a significant phloem flow of N from shoot to roots was found in these plants. Unexpectedly, for cocklebur, the plant with the very low root NRA, the fraction of total N present in the root that has been imported from the shoot was only half that as found for lupin and maize.     

ATP synthesis from 2,3-diphosphoglycerate by cell-free extract of Methanobacterium thermoautotrophicum (strain ΔH)

Cell-free extracts of Methanobacterium thermoautotrophicum were found to catalyze ATP synthesis from an endogeneous substrate. Synthesis was stimulated under hydrogen atmosphere and inhibited by KCL (K _i =150 mM). Comparison of the properties of a number of cell constituents showed the endogeneous substrate to be 2,3-diphosphoglycerate. The compound is converted into 3-phosphoglycerate, and via 2-phosphoglycerate and phosphoenolpyruvate into pyruvate, at which the latter reaction is linked with ATP synthesis.Abbreviations HS-CoM Coenzyme M, 2-mercaptoethanesulfonate - CH₃S-CoM methylcoenzyme m, 2-(methylthio)ethanesulfonate - HS-HTP 7-mercaptoheptanoyl-l-threonine phosphate - CoM-SS-HTP the heterodisulfide of HS-CoM and HS-HTP - BCFE bolled cell-free extract - TES N-tris(hydroxymethyl)methyl-2-aminoethanesulfonate - HEPES N-2-hydroxyethylpiperazine-N-ethanesulfonic acid - PEP phosphoenolpyruvate - 2,3-DPG 2,3-diphosphoglycerate - cDPG cyclic 2,3-diphosphoglycerate - 3-PG 3-phosphoglycerate - 2-PG 2-phosphoglycerate     

Identification of pseudomurein cell wall binding domains

Methanothermobacter thermautotrophicus is a methanogenic Gram-positive microorganism with a cell wall consisting of pseudomurein. Currently, no information is available on extracellular pseudomurein biology and so far only two prophage pseudomurein autolysins, PeiW and PeiP, have been reported. In this paper we show that PeiW and PeiP contain two different N-terminal pseudomurein cell wall binding domains. This finding was used to identify a novel domain, PB007923, on the M. thermautotrophicus genome present in 10 predicted open reading frames. Three homologues were identified in the Methanosphaera stadtmanae genome. Binding studies of fusion constructs of three separate PB007923 domains to green fluorescent protein revealed that it also constituted a cell wall binding domain. Both prophage domains and the PB007923 domain bound to the cell walls of Methanothermobacter species and fluorescence microscopy showed a preference for the septal region. Domain specificities were revealed by binding studies with other pseudomurein-containing archaea. Localized binding was observed for M. stadtmanae and Methanobrevibacter species, while others stained evenly. The identification of the first pseudomurein cell wall binding domains reveals the dynamics of the pseudomurein cell wall and provides marker proteins to study the extracellular pseudomurein biology of M. thermautotrophicus and of other pseudomurein-containing archaea.     

Evaluating the contribution of magnesium deficiency in the aluminium toxicity syndrome in twelve sorghum genotypes

The contribution of Mg deficiency to Al stress in twelve different sorghum (Sorghum bicolor (L.) Moench) genotypes was investigated in nutrient solution culture under conditions of low Mg supply (between 50 and 1000 M) at two pH values. At pH 4.2, 30 M Al strongly inhibited Mg uptake. When dry matter yield was plotted as a function of the plant Mg concentration, similar response curves were obtained in the absence and the presence of Al with three genotypes. With many other genotypes dry matter yields of the control (without Al treatment) and Al-stressed plants were remarkably different at similar internal Mg concentrations, suggesting that growth had been suppressed not by Mg deficiency but by another factor, i.e. Al-induced root damage. At pH 4.8, 30 M Al hardly induced root damage but reduced Mg uptake and Al-induced Mg deficiency could almost completely account for the growth reaction of all genotypes. Therefore, at this pH the efficiency of uptake or use of Mg in different genotypes was the basis of their respective susceptibility to Al toxicity. When specific root length surpassed a certain critical range below 80–100 m per g dry root, growth control by Al-induced Mg deficiency was nearly abolished. The pH and Al concentration where this range was reached depended on the Al sensitivity of the genotypes.     

Methanobacterium thermoautotrophicum (strain ΔH) contains a membrane-bound cyclic 2,3-diphosphoglycerate hydrolase

Cyclic 2,3-diphosphoglycerate (cDPG) hydrolase activity was demonstrated in cofactor-free extract of Methanobacterium thermoautotrophicum (strain H), but not in crude extract. Only after ultrafiltration or dialysis of crude extract cDPG hydrolase activity could be shown. cCPG hydrolysis was optimal at pH 6.0 and 60°C. Hydrolysis of cDPG occurred under nitrogen or hydrogen atmosphere and was completely inhibited by oxygen. Phosphate and potassium chloride were also strong inhibitors: 50% inhibition occurred at 0.6–0.7 mM phosphate or 0.2 M KCl. The enzyme was localized in the membrane fraction and could be solubilized for approximately 60% by treatment with 25 mM of the detergent CHAPS. The K _m and the V _max for cDPG were determined at 60°C and were 59 mM and 216 mU/mg, respectively. Furthermore, cDPG hydrolase was dependent on the presence of Co²⁺. The role of cDPG and cDPG hydrolase is discussed.Abbreviations cDPG cyclic 2,3-diphosphoglycerate - 2,3-DPG 2,3-diphosphoglycerate - 2-PG 2-phosphoglycerate - 3-PG 3-phosphoglycerate - PG phosphoglycerate - PEP phosphoenolpyruvate - TES N-tris(hydroxymethyl)methyl-2-aminoethanesulfonate - TRIS tris(hydroxymethyl)-aminomethane - DTT dithiothreitol - CHAPS 3-([3-cholamidopropyl]-dimethylammonio)-1-propanesulfonate - MOPS 3-(N-morpholino) propanesulfonic acid     

Acid soil damage in sorghum genotypes: Role of magnesium deficiency and root impairment

The effects of liming and Mg fertilization on growth, specific root length (root length per unit of root dry weight; SRL) and nutrient uptake of twelve sorghum genotypes (Sorghum bicolor (L.) Moench) were studied in two pot experiments. Liming increased the pH of the sandy loam from pH 4.3 (unlimed) to 4.7 (with 0.5 g Ca(OH)₂ kg^-1 soil) and to 6.1 (with 2.5 g Ca(OH)₂ kg^-1 soil). Liming increased the dry matter yield of the genotypes by factors of 1.2 to 6.0 (between pH 4.3 and 4.7) and by 1.1 to 2.4 (between pH 4.7 and 6.1). In absence of Mg at soil pH of 4.3 and 4.7, all genotypes suffered from Mg deficiency, as indicated by low Mg concentrations in the shoots (26–94 mmol Mg kg^-1 DM) and visible Mg deficiency symptoms. At pH 4.7 several of the genotypes responded to Mg application and produced significantly more dry matter. At pH 4.3, however, none of the genotypes responded to Mg, even though the internal Mg concentrations were increased by applied Mg. The relative increase in dry matter yield between pH 4.3 and 4.7 was closely correlated to the relative change in specific root length in the same soil pH interval, especially when the soil was fertilized with Mg (r²=0.91^**). The group of genotypes where SRL and dry matter yield were reduced by soil acidity was not the same as the group that responded positively to Mg application at pH 4.7.It is concluded that the growth of sorghum genotypes on acid soils is determined by two independent characteristics: the sensitivity of root development to soil acidity and the efficiency of the uptake and utilization of Mg. The first characteristic is predminant at high soil acidity whilst the latter is dominant at moderate soil acidity.