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In this review, we consider the exchange of nutrients between the host plant and the bacterial microsymbiont in nitrogen-fixing legume root nodules. During nodule formation, the host tissues and the bacterial microsymbiont develop in response to each other to form a specialized tissue that maintains an environment where nitrogen fixation can occur (Brewin, 2004;Mergaert et al., 2006;Prell and Poole, 2006). This complex development will not be considered here but, at the end of the process, specialized, nitrogen-fixing forms of the bacteria, known as bacteroids, reside in the plant cytosol, enclosed within plant-derived membranes. These organelle-like structures are known as symbiosomes; the plant-derived membrane that surrounds the bacteroid is the symbiosome (or peribacteroid) membrane and the space between the two is the symbiosome (or peribacteroid) space. An infected plant cell may be packed with thousands of symbiosomes. The exchange of nutrients that is fundamental to N 2 fixation therefore involves metabolism in the plant to provide carbon and nitrogen compounds to the bacteroids and to assimilate the metabolites that bacteroids release. Nutrients transferred between the symbionts must traverse both the symbiosome and the bacteroid membranes. It is clear that there is more than one pattern whereby successful nutrient exchange can take place: There are two basic types of legume nodules, determinate and indeterminate, and there are fundamental differences between the two in how they develop and in their carbon and nitrogen metabolism. This review focuses on the metabolism of carbon and nitrogen compounds in the symbionts and on the exchange of nutrients across the bacteroid and symbiosome membranes. Particular attention is paid to the movement of primary carbon and nitrogen sources and how they are utilized by both bacteroids and the plant.

One of the largest contributions to biologically available nitrogen comes from the reduction of N 2 to ammonia by rhizobia in symbiosis with legumes. Plants supply dicarboxylic acids as a carbon source to bacteroids, and in return they receive ammonia. However, metabolic exchange must be more complex, because effective N 2 fixation by Rhizobium leguminosarum bv viciae bacteroids requires either one of two broad-specificity amino acid ABC transporters (Aap and Bra). It was proposed that amino acids cycle between plant and bacteroids, but the model was unconstrained because of the broad solute specificity of Aap and Bra. Here, we constrain the specificity of Bra and ectopically express heterologous transporters to demonstrate that branched-chain amino acid (LIV) transport is essential for effective N 2 fixation. This dependence of bacteroids on the plant for LIV is not due to their known down-regulation of glutamate synthesis, because ectopic expression of glutamate dehydrogenase did not rescue effective N 2 fixation. Instead, the effect is specific to LIV and is accompanied by a major reduction in transcription and activity of LIV biosynthetic enzymes. Bacteroids become symbiotic auxotrophs for LIV and depend on the plant for their supply. Bacteroids with aap bra null mutations are reduced in number, smaller, and have a lower DNA content than wild type. Plants control LIV supply to bacteroids, regulating their development and persistence. This makes it a critical control point for regulation of symbiosis.mutualism ͉ nitrogen fixation ͉ peas ͉ symbiosis T he largest input of available nitrogen in the biosphere comes from biological reduction of atmospheric N 2 to ammonium (1). Most of this comes from legume-Rhizobium symbioses, arising from infection of host plants and resulting in root structures called nodules (2). These symbioses are initiated by plant-released flavonoids and related compounds, which elicit synthesis of lipochitooligosaccharide Nod factors by rhizobia. Bacteria are trapped by curling root hairs that they enter via infection threads. These grow into the root cortex, into a zone of newly induced meristematic cells forming the origin of the nodule. Bacteria are released from infection threads by endocytosis and are surrounded by a plant-derived symbiosome membrane. In nodules of galegoid legumes (a clade in the subfamily Papilionoideae, such as Medicago, Pisum, or Vicia), bacteria undergo dramatic increases in size, shape, and DNA content (3) before they start to reduce N 2 . Plants provide differentiated bacteria (bacteroids) with dicarboxylic acids, which energize N 2 reduction to ammonium for secretion back to the plant (4).A simple exchange of dicarboxylates and ammonium is the classical model of nutrient exchange in nodules, but amino acid transport by bacteroids has also been shown to be essential (5). Rhizobium leguminosarum mutated in 2 broad-specificity amino acid ABC transporters (Aap and Bra) formed N 2 -fixing pea bacteroids that appeared morphologically normal in electron micrographs,...

Rhizobium leguminosarum bv. viciae forms nitrogen-fixing nodules on several legumes, including pea (Pisum sativum) and vetch (Vicia cracca), and has been widely used as a model to study nodule biochemistry. To understand the complex biochemical and developmental changes undergone by R. leguminosarum bv. viciae during bacteroid development, microarray experiments were first performed with cultured bacteria grown on a variety of carbon substrates (glucose, pyruvate, succinate, inositol, acetate, and acetoacetate) and then compared to bacteroids. Bacteroid metabolism is essentially that of dicarboxylate-grown cells (i.e., induction of dicarboxylate transport, gluconeogenesis and alanine synthesis, and repression of sugar utilization). The decarboxylating arm of the tricarboxylic acid cycle is highly induced, as is ␥-aminobutyrate metabolism, particularly in bacteroids from early (7-day) nodules. To investigate bacteroid development, gene expression in bacteroids was analyzed at 7, 15, and 21 days postinoculation of peas. This revealed that bacterial rRNA isolated from pea, but not vetch, is extensively processed in mature bacteroids. In early development (7 days), there were large changes in the expression of regulators, exported and cell surface molecules, multidrug exporters, and heat and cold shock proteins. fix genes were induced early but continued to increase in mature bacteroids, while nif genes were induced strongly in older bacteroids. Mutation of 37 genes that were strongly upregulated in mature bacteroids revealed that none were essential for nitrogen fixation. However, screening of 3,072 mini-Tn5 mutants on peas revealed previously uncharacterized genes essential for nitrogen fixation. These encoded a potential magnesium transporter, an AAA domain protein, and proteins involved in cytochrome synthesis.

Abstract. The potential impact of climate change on agriculture is uncertain. In addition, agriculture could influence above-and below-ground carbon storage. Development of models that represent agriculture is necessary to address these impacts. We have developed an approach to integrate agriculture representations for three crop types -maize, soybean, and spring wheat -into the coupled carbon-nitrogen version of the Community Land Model (CLM), to help address these questions. Here we present the new model, CLMCrop, validated against observations from two AmeriFlux sites in the United States, planted with maize and soybean. Seasonal carbon fluxes compared well with field measurements for soybean, but not as well for maize. CLM-Crop yields were comparable with observations in countries such as the United States, Argentina, and China, although the generality of the crop model and its lack of technology and irrigation made direct comparison difficult. CLM-Crop was compared against the standard CLM3.5, which simulates crops as grass. The comparison showed improvement in gross primary productivity in regions where crops are the dominant vegetation cover. Crop yields and productivity were negatively correlated with temperature and positively correlated with precipitation, in agreement with other modeling studies. In case studies with the new crop model looking at impacts of residue management and planting date on crop yield, we found that increased residue returned to the litter pool increased crop yield, while reduced residue returns resulted in yield decreases. Using climate controls to signal planting date caused different responses in different crops. Maize and soybean had opposite reactions: when low temperature threshold resulted in early planting, maize responded with a loss of yield, but soybean yields increased.Our improvements in CLM demonstrate a new capability in the model -simulating agriculture in a realistic way, complete with fertilizer and residue management practices. Results are encouraging, with improved representation of human influences on the land surface and the potentially resulting climate impacts.