ABSTRACT
This study describes habitual physical activity (HPA) of Bolivian boys living at different altitudes and from different socioeconomic status. The boys were living at high altitude (HA) in La Paz (4000 m) and at low altitude (LA) in Santa Cruz (400 m). At both altitudes samples of 10- to 12-year-old boys were chosen from a relatively low socioeconomic status (LSES) and a relatively high socioeconomic status (HSES). At HA 19 boys from LSES and 10 boys from HSES were measured and at LA 14 boys from LSES and 13 boys from HSES. HPA was measured by 24-h heart rate (HR) monitoring. Also an interview was completed to recall the HPA. By comparing the registered HR data with the time they were asleep the mean HR during sleep was calculated (HRsleep). The maximal HR (HRmax) was measured from a maximal exercise test on a bicycle ergometer. Heart rate reserve (HRR = HRmax-HRsleep) was used to measure the mean level of physical activity of the subjects. The results show that HRsleep (= HRrest) in HA boys with 70 (+/- 6) beats/min was significantly lower (p < 0.05) than in LA boys with 77 (+/- 10) beats/min. HRmax was also significantly lower (p < 0.05) in HA boys (187 +/- 12 beats/min) compared to LA boys (195 +/- 8 beats/min). Because HA influences HRsleep and HRmax in the same way, HRR is not significantly different between boys of HA and LA. The mean heart rate over 24 h (HRmean) in HA boys (87 +/- 7 beats/min) was significantly lower than in LA boys (93 +/- 8 beats/min).(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Altitude , Motor Activity/physiology , Social Class , Body Constitution , Bolivia , Child , Heart Rate , Humans , Male , Monitoring, PhysiologicABSTRACT
To investigate the role of dicarboxylate transport in nitrogen-fixing symbioses between Rhizobium and tropical legumes, we made a molecular genetic analysis of the bacterial transport system in Rhizobium sp. NGR234. This braod host range strain fixes nitrogen in association with evolutionarily divergent legumes. Two dicarboxylate transport systems were cloned from Rhizobium NGR234. One locus was chromosomally located, whereas the other was carried on the symbiotic plasmid (pSym) and contained a dctA carrier protein gene, which was analyzed in detail. Although the DNA and derived amino acid sequences of the structural gene were substantially homologous to that of R. meliloti, its promoter sequences was quite distinct, and the upstream sequence also exhibited no homology to dctB, which is found at this position in R. meliloti. A site-directed internal deletion mutant in dctA of NGR234 exhibited a (unique) exclusively symbiotic phenotype that could grow on dicarboxylates ex planta, but could not fix nitrogen in planta. This phenotype was found for tested host plants of NGR234 with either determinate- or indeterminate-type nodules, confirming for the first time that symbiosis-specific uptake of dicarboxylates is a prerequisite for nitrogen fixation in tropical legume symbioses.
Subject(s)
Bacterial Proteins/genetics , Carrier Proteins/genetics , Dicarboxylic Acid Transporters , Dicarboxylic Acids/metabolism , Fabaceae/microbiology , Nitrogen Fixation/genetics , Plants, Medicinal , Rhizobium/metabolism , Amino Acid Sequence , Base Sequence , Biological Transport/genetics , Cloning, Molecular , DNA, Bacterial , Genes, Bacterial , Membrane Transport Proteins/genetics , Molecular Sequence Data , Mutagenesis, Site-Directed , Phenotype , Restriction Mapping , Rhizobium/genetics , SymbiosisABSTRACT
Rhizobium sp. NGR234 is a broad-host range strain. The rpoN gene of this organism encodes a sigma factor which is a primary co-regulator of endosymbiosis. We characterized the locus upstream of rpoN, and identified a contiguous open reading frame, here termed ORF1. DNA sequence analysis of this ORF showed that it encoded a polypeptide highly conserved with a corresponding ORF of Rhizobium meliloti. The gene product contained two ATP/GTP binding pockets. Codon usage in the ORF and the nitrogenase operon nifKDH of NGR234 was similar. Although we used a non-transposable cassette flanked by appropriate sized DNA fragments, we were unable to isolate site-directed mutants in the ORF, whose ATP/GTP binding protein product is thus probably of essential biological function. ORF1 and rpoN exhibited conserved linkage among diverse rhizobia, and in Azotobacter vinelandii. Intragenomic and interspecific homology studies confirmed directly that ORF1 (NGR234) belonged to a large family of ATP-binding protein genes.
Subject(s)
Bacterial Proteins/genetics , Open Reading Frames/genetics , Rhizobium/genetics , Adenosine Triphosphate/metabolism , Amino Acid Sequence , Base Sequence , Binding Sites/genetics , Blotting, Southern , Codon/genetics , Genetic Linkage/genetics , Gram-Negative Bacteria/genetics , Guanosine Triphosphate/metabolism , Molecular Sequence Data , Multigene Family/genetics , Mutagenesis, Site-Directed/genetics , Nitrogenase/genetics , Sequence Homology, Nucleic Acid , Sigma Factor/geneticsABSTRACT
Nodule senescence is one aspect of nitrogen fixation that is important to study from the perspective of improving the host-bacteroid interaction. In winged bean nodules, a 21-kilodalton protein is specifically expressed when senescence begins. Using subcellular fractionation, we observed that this plant protein interacts with the bacteroids. Microsequencing of the protein allowed us to obtain a specific oligonucleotide that was used to isolate the corresponding nodule cDNA. Sequence analysis of this cDNA revealed that the 21-kilodalton protein has all of the features of a legume Kunitz protease inhibitor. Subsequent analysis confirmed that this nodulin is indeed a protease inhibitor. Immunocytochemical study showed that the protease inhibitor is exclusively localized in infected senescent cells of the nodule, particularly in disorganized bacteroids, the peribacteroid membrane, vacuole membranes, and in the vacuole fluid. The specific expression of a protease inhibitor at senescence may be of particular interest if the targeted proteolytic activity is important for the symbiotic relationship. This point is discussed in relation to the known nodule proteases.
Subject(s)
Fabaceae/enzymology , Membrane Proteins , Plant Proteins/genetics , Plants, Medicinal , Protease Inhibitors , Aging , Amino Acid Sequence , Base Sequence , DNA/genetics , Fabaceae/genetics , Fabaceae/microbiology , Molecular Sequence Data , Nitrogen Fixation , Plant Proteins/biosynthesis , Protease Inhibitors/metabolism , Rhizobium/physiology , Sequence Alignment , Trypsin Inhibitor, Kunitz Soybean/geneticsABSTRACT
We report the nucleotide sequence of the rpoN gene from broad-host-range Rhizobium sp. strain NGR234 and analyze the encoded RPON protein, a sigma factor. Comparative analysis of the deduced amino acid sequence of RPON from NGR234 with sequences from other gram-negative bacteria identified a perfectly conserved RPON box unique to RPON sigma factors. Symbiotic regulatory phenotypes were defined for a site-directed internal deletion within the coding sequence of the rpoN gene of Rhizobium strain NGR234: they included quantitative nodulation kinetics on Vigna unguiculata and microscopic analysis of the Fix- determinate nodules of V. unguiculata and Macroptilium atropurpureum. RPON was a primary coregulator of nodulation and was implicated in establishment or maintenance of the plant-synthesized peribacteroid membrane. Phenotypes of rpoN in Rhizobium strain NGR234 could be grouped as symbiosis related, rather than simply pleiotropically physiological as in free-living bacteria such as Klebsiella pneumoniae and Pseudomonas putida.
