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PubMed2005

The stringent response is required for amino acid and nitrate utilization, nod factor regulation, nodulation, and nitrogen fixation in Rhizobium etli.

A Rhizobium etli Tn5 insertion mutant, LM01, was selected for its inability to use glutamine as the sole carbon and nitrogen source. The Tn5 insertion in LM01 was localized to the rsh gene, which encodes a member of the RelA/SpoT family of proteins. The LM01 mutant was affected in the ability to use amino acids and nitrate as nitrogen sources and was unable to accumulate (p)ppGpp when grown under carbon and nitrogen starvation, as opposed to the wild-type strain, which accumulated (p)ppGpp under these conditions. The R. etli rsh gene was found to restore (p)ppGpp accumulation to a DeltarelA DeltaspoT mutant of Escherichia coli. The R. etli Rsh protein consists of 744 amino acids, and the Tn5 insertion in LM01 results in the synthesis of a truncated protein of 329 amino acids; complementation experiments indicate that this truncated protein is still capable of (p)ppGpp hydrolysis. A second rsh mutant of R. etli, strain AC1, was constructed by inserting an Omega element at the beginning of the rsh gene, resulting in a null allele. Both AC1 and LM01 were affected in Nod factor production, which was constitutive in both strains, and in nodulation; nodules produced by the rsh mutants in Phaseolus vulgaris were smaller than those produced by the wild-type strain and did not fix nitrogen. In addition, electron microscopy revealed that the mutant bacteroids lacked poly-beta-hydroxybutyrate granules. These results indicate a central role for the stringent response in symbiosis.

Amino Acid Sequence↗
PubMed2006

The effect of cutting, mulching and applications of farmyard manure on nitrogen fixation in a red clover/grass sward.

In organic farming, maximising the amount of nitrogen (N) which is fixed and retained within the soil is of paramount importance for the yield of the following crop. The aim of this study was to establish the extent to which increased soil fertility, farmyard manure (FYM) applications and/or mulching, could adversely affect fixation. At two sites, situated in the South West (SW) and North East (NE) of England, N(2) fixation was estimated in 'organically' managed red clover/grass plots, both with and without green manure (i.e. surface mulched) and/or the addition of FYM. The FYM was incorporated into the seedbeds at both sites in autumn 2002 at the rate of 170 kg total Nha(-1), as either well-composted (SW site), or not actively-composted (NE site) manures. The same FYM application rate was repeated as top-dressings to both sites in autumn 2003. The plots were cut three or four times each year over two growing seasons. In the first harvest year (2003), incorporation of FYM had beneficial effects of increasing dry matter and N yields significantly at the first cut, but there were no significant differences in subsequent cuts. The same pattern was found in the second harvest year (2004) after the top dressings of FYM, suggesting that most of the N in both types of FYM was in recalcitrant forms. Over the two growing seasons, mulching did not affect red clover/grass dry matter or N yields, but did reduce the proportion of N(2) fixed, by up to 60 kg Nha(-1) when compared with plots from which the clover/grass herbage was cut and removed. Thus, the gain in N from FYM or green manure tended to be offset by a similar reduction in N(2) fixation. These results demonstrate the close association between the availability of soil N and the feed-back system which operates on N(2) fixation by red clover.

Agriculture↗