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  • Rhizobium  (7)
  • Springer  (7)
  • Munksgaard International Publishers
  • Wiley-Blackwell
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 12 (1989), S. 181-188 
    ISSN: 1573-5028
    Keywords: nodulation ; Rhizobium ; lac-fusions ; promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nodulation regulon of Rhizobium meliloti AK631 includes several operons (nodABC, hsnABC, hsnD, efn locus) which have in common a consensus promoter sequence called the nod box. A synthetic nod box probe was used to identify two additional nod boxes, n4 and n5, which were subcloned for study. By constructing lac fusions, we show that n4 and n5 sponsor induction of downstream regions as previously shown for n1-nodABC and n2-hsnABC. Using site-directed Tn5 mutagenesis, we find that the n5 locus plays a significant role in nodulation of alfalfa and sweetclover, whereas the n4 locus is important for alfalfa, but not for sweetclover. Hybridization data suggest that the n5 locus is conserved among Rhizobium species. In contrast, the n4 locus seems to be unique to Rhizobium meliloti strains, in agreement with the host-specific phenotype of n4 locus mutants. Thus, the use of a promoter probe allows us to identify nodulation genes which may be overlooked by standard methods such as random Tn5 mutagenesis.
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  • 2
    ISSN: 1573-5028
    Keywords: host recognition ; hsnD (nodH) ; nod genes ; Rhizobium ; root hair deformation ; symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The role of the hsnD (nodH) gene in the determination of the host-specific nodulation ability of Rhizobium meliloti was studied by expressing the common nodulation genes (nodABC) with or without the hsnD gene in Escherichia coli and testing for biological activity on various leguminous plants. In this way, four categories of plants were established. Upon infection with E. coli carrying the nodABC construct, root hair deformation (Had) was detected on clovers while the hsnD gene was additionally needed for the elicitation of the same response on alfalfa and sweet clover. A weak root hair deformation was seen on siratro by inoculation with E. coli harbouring the nodABC genes and was highly increased when hsnD was also introduced. Cowpea and Desmodium did not respond to any of the E. coli strains constructed. Exudates or cytosolicfractions of the respective E. coli derivatives elicited the same root hair deformation as the intact bacteria. These data indicate that not only the nodABC gene products but also the hsnD product are involved in the synthesis of Had factors. Subclones expressing only the nodA, nodB, or nodC genes or the same genes in pairs (nodAB, nodBC, nodAC) did not provide a compound with activity comparable to the NodABC factor, suggesting that all three genes are required for the production of the Had factor which is active on clover. Coinoculation of alfalfa plants with two strains of E. coli, one carrying the nodABC genes and the other expressing only hsnD, or combining exudates or cytosolic fractions from these strains did not result in root hair deformation on alfalfa. These data indicate that the HsnD protein itself or its product is not an additional alfalfa-specific extracellular signal but more likely is enzymatically involved in the modification of the basic compound determined by the nodABC genes.
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  • 3
    ISSN: 1432-2048
    Keywords: Key words: Cytosolic free calcium ; Lipochitooligo saccharide ; Nodulation ; Rhizobium ; Signal transduction ; Tip growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Using Ca2+-selective microelectrodes, the concentration of free calcium ([Ca2+]) in the cytosol has been measured in root hair cells of Medicago sativa L. in the presence of nodulation (Nod) factors. Growing root hairs of M. sativa displayed a steep apical [Ca2+] gradient, i.e. 604–967 nM in the tip compared with 95–235 nM in the basal region. When tested within the first 5 to 10 μm of the tip, addition of NodRm-IV(C16:2,S) decreased the cytosolic [Ca2+], whereas an increase was observed when tested behind the tip. Overall, this led to a partial dissipation of the [Ca2+] gradient. The Ca2+ response was specific: it was equally well observed in the presence of NodRm-IV(Ac,C16:2,S), reduced with NodRm-IV(C16:0,S), but not with chitotetraose, the nonactive glucosamine backbone. In contrast to growing root hairs, non-growing root hairs without a tip-to-base cytosolic [Ca2+] gradient responded to NodRm-IV(C16:2,S) with an increase in cytosolic [Ca2+] at the tip as well as at the root hair base. We suggest that the response to Nod factors depends on the stage of development of the root hairs, and that changes in cytosolic [Ca2+] may play different roles in Nod-factor signaling: changes of cytosolic [Ca2+] in the apical part of the root hair may be related to root hair deformation, while the increase in [Ca2+] behind the tip may be essential for the amplification of the Nod signal, for its propagation and transduction to trigger downstream events.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 226 (1991), S. 337-340 
    ISSN: 1617-4623
    Keywords: Rhizobium ; Nodulation ; Transcriptional regulation ; Nuclear receptor ; Steroid hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The signal specificity and structure of sensor-activator proteins from different species (NodD of Rhizobium bacteria and vertebrate nuclear receptors) were compared. Several compounds (including flavonoids, coumestrol and estradiol) that bind to mammalian receptors also interact with NodD proteins. NodD-dependent synergism of the signal compounds luteolin and catechin was observed suggesting that these compounds bind directly to NodD. Two regions comprising 63 and 37 amino acids in NodD showed 45% and 36% homology, respectively, with the estrogen receptor. These regions, designated as modules M1 and M2, coincide with conserved parts of the ligand-binding domains of the nuclear receptors. A part of NodD overlapping with the M1 module was predicted to be membrane associated and was 46% homologous to a membrane-spanning sensory segment of the Agrobacterium VirA protein. We suggest that the homologous polypeptide modules detected in NodD and the nuclear receptors originate from a common ancestor protein and may be directly involved in ligand binding.
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  • 5
    ISSN: 1617-4623
    Keywords: Nitrogen regulation (ntr) ; nodABC ; nodD3 ; Nodulation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The presence of combined nitrogen in the soil suppresses the formation of nitrogen-fixing root nodules by Rhizobium. We demonstrate that bacterial genes determining early nodulation functions (nodABC) as well as the regulatory gene nodD3 are under nitrogen (NH 4 + ) control. Our results suggest that the gene product of nodD3 has a role in mediating the ammonia regulation of early nod genes. The general nitrogen regulatory (ntr) system as well as a chromosomal locus mutated in Rhizobium meliloti were also found to be involved in the regulation of nod gene expression. A R. meliloti mutant with altered sensitivity to ammonia regulation was isolated, capable of more efficient nodulation of alfalfa than the wild-type strain in the presence of 2 mM ammonium sulfate.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 240 (1993), S. 435-444 
    ISSN: 1617-4623
    Keywords: Nitrogen regulation ; nodD3 ; syrM ; Common nod genes ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of the nodulation genes nodABC of Rhizobium meliloti, which determine early response functions to plant host signals, is regulated by the level of ammonia, the primary product of symbiotic nitrogen fixation. We show that the pathway that links the ammonia-induced signal to the transcriptional control of the nodABC genes involves at least two regulatory levels. The fluctuating nitrogen level is sensed and the signal is mediated by the members of the general nitrogen regulatory (ntr) system, then transmitted to the syrM-nodD3 genes representing the nod-specific level of ammonia regulation. At low ammonia concentration, the activator protein NtrC exerts its effect via nodD3. In conditions of nitrogen excess ntrR, involved in the repression of nod genes, may function in coordination with the syrM gene. Finally, the NodD3 protein may relay the nitrogen status signal to the transcriptional control of the nodABC genes.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 8 (1987), S. 61-75 
    ISSN: 1573-5028
    Keywords: host-specific nodulation genes ; nod gene homology ; Rhizobium ; symbiotic genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The internal structural portion of genes nodC and nodD (representatives of the two transcription units coding for common nodulation functions) and of hsnB and hsnD (genes from the two transcription units determining host-specificity of nodulation) have been cloned from Rhizobium meliloti into M13 vectors and used as probes against genomic DNAs from different Rhizobium strains and species. nodC and nodD were found in all species with one exception, indicating that they are common and widely spread genes, though the nodD gene hybridized only very weakly with slow-growing rhizobia. Interestingly, reiteration of nodD sequences was observed in almost all fast-growing strains (with the exception of R. leguminosarum). hsnB and, more so, hsnD are present only in a few species tested, supporting their specific involvement in R. meliloti-Medicago sativa symbiosis. In several cases the hybridizing bands from total Rhizobium DNA were compared to those found in recombinant plasmids carrying functional nodulation regions, and these analyses supported the notion that the bands indicate the presence of functional genes.
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