Rhizobium Meliloti Symbiotic Nitrogen Fixation: Identification of NIF and FIX Genes; The NIF FIX Cascade Regulatory Pathway

Author(s):  
Pierre Boistard
Genome ◽  
1989 ◽  
Vol 31 (1) ◽  
pp. 354-360 ◽  
Author(s):  
San Chiun Shen ◽  
Shui Ping Wang ◽  
Guan Qiao Yu ◽  
Jia Bi Zhu

Genes that specify nodulation (nod genes) are only active in the free-living rhizobia or in the nodule initiation state of rhizobia. As soon as the repression of nod genes occurs in the bacteroids of the nodule, nifA is induced, while ntrC is inactivated and thus the nifA-mediated nif/fix genes are turned on. Limitation of available oxygen brings about the induction of nifA, which reflects the actual status of nif/fix gene activities in symbiotic state of rhizobia. Oxygen thus appears to be a major symbiotic signal to the expression of bacteroid nif/fix genes. Mutation of nifA or shortage of nifA product in wild-type rhizobia caused by the inhibition of multicopy nifH/fixA promoters leads to an abnormal development of nodules and premature degradation of bacteroids in nodules.Key words: nitrogen fixation, nodulation, nif/fix regulation, nifA mutant.


1992 ◽  
Vol 38 (6) ◽  
pp. 555-562 ◽  
Author(s):  
Vipin Rastogi ◽  
Monika Labes ◽  
Turlough Finan ◽  
Robert Watson

Symbiotic nitrogen fixation may be limited by the transport of C4 dicarboxylates into bacteroids in the nodule for use as a carbon and energy source. In an attempt to increase dicarboxylate transport, a plasmid was constructed in which the Rhizobium meliloti structural transport gene dctA was fused to a tryptophan operon promoter from Salmonella typhimurium, trpPO. This resulted in a functional dctA gene that was no longer under the control of the dctBD regulatory genes, but the recombinant plasmid was found to be unstable in R. meliloti. To stably integrate the trpPO-dctA fusion, it was recloned into pBR325 and recombined into the R. meliloti exo megaplasmid in the dctABD region. The resultant strain showed constitutive dctA-specific mRNA synthesis which was about 5-fold higher than that found in fully induced wild-type cells. Uptake assays showed that [14C]succinate transport by the trpPO-dctA fusion strain was constitutive, and the transport rate was the same as that of induced control cells. Acetylene reduction assays indicated a significantly higher rate of nitrogen fixation in plants inoculated with the trpPO-dctA fusion strain compared with the control. Despite this apparent increase, the plants had the same top dry weights as those inoculated with control cells. Key words: acetylene reduction, genetic engineering, nodule, plasmid stability, promoter.


1980 ◽  
Vol 60 (1) ◽  
pp. 209-212 ◽  
Author(s):  
H. ANTOUN ◽  
L. M. BORDELEAU ◽  
D. PRÉVOST ◽  
R. A. LACHANCE

Specific activities of the assimilatory and "regulatory" types of nitrate reductase were studied in 41 strains of Rhizobium meliloti having different symbiotic nitrogen fixation activities. Both nitrate reductase enzymes were present in very effective and ineffective strains and no significant correlation was found between the specific activities of the two enzymes and the dry matter yields of alfalfa obtained with the 41 strains. Measurements of the specific activities of the two nitrate reductase enzymes in the vegetative bacteria cannot be used as a rapid physiological test for the selection of very effective strains of R. meliloti.


1979 ◽  
Vol 25 (3) ◽  
pp. 298-301 ◽  
Author(s):  
Ilona Barabás ◽  
Tibor Sik

In two out of three pleiotropic mutants of Rhizobium meliloti, defective in nitrate reductase induced by amino acid utilization in vegetative bacteria and in symbiotic nitrogen fixation, nitrogenase activity could be restored completely by purines and partially by the amino acids L-glutamate, L-aspartate, L-glutamine, and L-asparagine. The compounds restoring effectiveness in nitrogen fixation did not restore nitrate reductase activity in vegetative bacteria. The restoration of effectiveness supports our earlier conclusion that the mutation is not in the structural gene for a suggested common subunit of nitrogenase and nitrate reductase.


1977 ◽  
Vol 57 (2) ◽  
pp. 433-439 ◽  
Author(s):  
L. M. BORDELEAU ◽  
H. ANTOUN ◽  
R. A. LACHANCE

Symbiotic nitrogen fixation with 49 isolates of Rhizobium meliloti was studied under controlled environment with alfalfa cv. Saranac. It was shown that plant yield in dry weight can be used as an indirect measurement of nitrogen fixation, and as a criterion for selecting efficient strains of R. meliloti. Statistical study on yields of three cuttings has established that the second cutting gives the most necessary information to correctly evaluate the symbiotic efficiency of the isolates. Six very efficient strains were selected.


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