Nitrification, a key process in the global nitrogen cycle that generates nitrate through microbial activity, may enhance losses of fertilizer nitrogen by leaching and denitrification. Certain plants can suppress soil-nitrification by releasing inhibitors from roots, a phenomenon termed biological nitrification inhibition (BNI). Here, we report the discovery of an effective nitrification inhibitor in the root-exudates of the tropical forage grass Brachiaria humidicola (Rendle) Schweick. Named ''brachialactone,'' this inhibitor is a recently discovered cyclic diterpene with a unique 5-8-5-membered ring system and a ␥-lactone ring. It contributed 60 -90% of the inhibitory activity released from the roots of this tropical grass. Unlike nitrapyrin (a synthetic nitrification inhibitor), which affects only the ammonia monooxygenase (AMO) pathway, brachialactone appears to block both AMO and hydroxylamine oxidoreductase enzymatic pathways in Nitrosomonas. global warming ͉ nitrogen pollution ͉ nitrous oxide emissions ͉ root exudation ͉ climate change M ost modern agricultural systems are based on large inputs of inorganic nitrogen (N), with ammonium (NH 4 ϩ ) being the primary N source (1, 2). Also, current crop management practices result in the development of highly nitrifying soil environments (3, 4). Nitrification results in the transformation of the relatively immobile NH 4 ϩ to highly mobile nitrate (NO 3 Ϫ ), making inorganic N susceptible to losses through leaching of NO 3 Ϫ and/or gaseous N emissions, potentially initiating a cascade of environmental and health problems (1, 2, 5, 6). Nitrous oxide (N 2 O) is one of the three major biogenic greenhouse gases contributing to global warming, produced primarily from denitrification processes in agricultural systems (5, 7). Also, assimilation of NO 3 Ϫ by plants can result in further N 2 O emissions directly from plant canopies (8). The low agronomic N-use efficiency (NUE) found in many agricultural systems is largely the result of N losses associated with nitrification (i.e., N losses from NO 3 Ϫ leaching and denitrification) (9-11). Most plants have the ability to assimilate both NH 4 ϩ and NO 3 Ϫ (12); therefore, nitrification does not need to be a dominant process in the N cycle for efficient N use.Nitrification is low in some forest and grassland soils (13-17). Since the early 1960s, some tropical grasses have been suspected of having the capacity to inhibit nitrification (18-21). However, this concept remained controversial due to the lack of direct evidence showing such inhibitory effects or the identification of specific inhibitors (22).We adopted a very sensitive bioassay using a recombinant luminescent Nitrosomonas europaea to detect biological nitrification inhibition (BNI) in plant-soil systems with the inhibitory activity of roots expressed in allylthiourea units (ATU) (23). Using this methodology, we were able to show that certain plants release nitrification inhibitors from their roots (23-26). Such BNI capacity appears to be relatively widespread among...

Lectin-like, oxidized low-density lipoprotein (LDL) receptor 1, LOX-1, is the major receptor for oxidized LDL (OxLDL) in endothelial cells. We have determined the crystal structure of the ligand binding domain of LOX-1, with a short stalk region connecting the domain to the membrane-spanning region, as a homodimer linked by an interchain disulfide bond. In vivo assays with LOX-1 mutants revealed that the "basic spine," consisting of linearly aligned arginine residues spanning over the dimer surface, is responsible for ligand binding. Single amino acid substitution in the dimer interface caused a severe reduction in LOX-1 binding activity, suggesting that the correct dimer arrangement is crucial for binding to OxLDL. Based on the LDL model structure, possible binding modes of LOX-1 to OxLDL are proposed.

We show that selection of drug-resistant bacterial mutants allows the discovery of antibacterial compounds. Mutant strains of a soil-isolated Streptomyces species that does not produce antibacterials synthesize a previously unknown class of antibacterial, which we name piperidamycin. Overall, 6% of non-Streptomyces actinomycetes species and 43% of Streptomyces species that do not produce antibacterials are activated to produce them. The antibacterial-producing mutants all carried mutations in RNA polymerase and/or the ribosomal protein S12.

Plant cell walls constitute the bulk of the earth renewable source of energy and are a component in the diet of humans and herbivores. l-Arabinofuranosyl (Araf) residues are a quantifiably important constituent of these walls. Plants use uridine diphosphate (UDP)-l-arabinofuranose (UDP-Araf) to donate Araf residues in the biosynthesis of Araf-containing polysaccharides, proteoglycans, and glycoproteins. However, little is known about the formation of UDP-Araf. We now describe the purification and partial characterization of a rice UDP-arabinopyranose mutase (UAM) that catalyzes the formation of UDP-Araf from UDP-arabinopyranose (UDP-Arap). The reaction is reversible and at thermodynamic equilibrium the pyranose form is favored over the furanose form (90 : 10). Three related proteins that are encoded by rice gene loci Os03g40270, Os04g56520, and Os07g41360 were identified from partial amino acid sequences of UAM. These proteins have >80% sequence identity with polypeptides that are reversibly glycosylated in the presence of UDP-sugars. The rice mutase and two functionally active recombinant mutases were shown to be reversibly glycosylated in the presence of UDP-Glc. The cofactor, flavin-adenine-dinucleotide (FAD), is required for the catalytic activity of UDP-galactose mutases of prokaryotes, fungi, and protozoa. The plant mutases, which do not require a cofactor, must therefore have a different catalytic mechanism. Putative UAM-encoding genes are present in the green algae Chlamydomonas reinhardtii, the moss Physcomitrella patens, the gymnosperm Pinus taeda (loblolly pine), and in numerous dicots and monocots, indicating that UAMs are widespread in green plants.

We found that a polycistronic operon (ywfBCDEFG) and a monocistronic gene (ywfH) are required for the biosynthesis of bacilysin in Bacillus subtilis. The disruption of these genes by plasmid integration caused loss of the ability to produce bacilysin, accompanied by a lack of bacilysin synthetase activity in the crude extract. We investigated the regulatory mechanism for bacilysin biosynthesis using the transcriptional lacZ fusion system. The transcription of these genes was found to be induced at the transition from exponential to stationary phase. Induction of transcription was accelerated by depleting a required amino acid, which was done by transferring the wild-type (rel ؉ ) cells to an amino acidlimited medium. In contrast, no enhancement of the gene expression was detected in relA mutant cells. In wild-type (rel ؉ ) cells, a forced reduction of intracellular GTP, brought about by addition of decoyinine, which is a GMP synthetase inhibitor, enhanced the expression of both the ywfBCDEFG operon and the ywfH gene, resulting in a 2.5-fold increase in bacilysin production. Disruption of the codY gene, which regulates stationary phase genes by detecting the level of GTP, also induced transcription of these genes. In contrast, the expression of ywfBCDEFG in relA cells was not activated either by decoyinine addition or codY disruption, although the expression of ywfH was induced. Moreover, the codY disruption resulted in an increase of bacilysin production only in rel ؉ cells. These results indicate that guanosine 5-diphosphate 3-diphosphate (ppGpp) plays a crucial role in transcription of the ywfBCDEFG operon and that the transcription of these genes are dependent upon the level of intracellular GTP which is transmitted as a signal via the CodY-mediated repression system. We propose that, unlike antibiotic production in Streptomyces spp., bacilysin production in B. subtilis is controlled by a dual regulation system composed of the guanine nucleotides ppGpp and GTP.The stringent response is one of the most important adaptations, by which bacteria have to survive in a nutrient limited environment. This response leads to the repression of stable RNA synthesis (rRNA and tRNA) and gene expression for various translational factors and ribosomal proteins. The stringent response also activates the expression of certain genes, including the amino acids biosynthesis genes. Numerous studies have indicated that the stringent response depends on a transient increase of the hyperphosphorelated guanosine nucleotides, guanosine 5Ј-diphosphate 3Ј-diphosphate (ppGpp), 1 in response to the binding of uncharged tRNA to the ribosomal A site (1). Mutant cells that are unable to repress stable RNA synthesis under depleted amino acid conditions have been termed "relaxed." In many cases, these mutations are found in the relA gene, which encodes the ppGpp synthetase, or the relC (ϭ rplK) gene, which codes for the ribosomal protein L11. These relaxed (rel) mutants are unable to initiate ribosome-mediated synthesis of ppGpp (2-5). Therefore...