Structures and expression patterns of two cDNA clones encoding S-adenosyl-L-methionine synthetase from the root nodule of Elaeagnus umbellata

2001 ◽  
Vol 28 (9) ◽  
pp. 951
Author(s):  
Sang Ho Lee ◽  
Ho Bang Kim ◽  
Chung Sun An
Keyword(s):  
Amino Acid ◽  
Expression Pattern ◽  
Root Nodule ◽  
Vascular System ◽  
Expression Patterns ◽  
Amino Acid Level ◽  
Nodule Development ◽  
Pcr Analysis ◽  
Cdna Clones ◽  
Elaeagnus Umbellata

This paper originates from an address at the 8th International Symposium on Nitrogen Fixation with Non-Legumes, Sydney, NSW, December 2000 Two cDNA clones encoding S-adenosyl-L-methionine synthetase (SAMS) were isolated from the root nodule cDNA library of Elaeagnus umbellata Thunberg and analysed on the basis of deduced amino acid sequence and expression pattern. Two EuSAMS clones shared 75–84% identity at the nucleotide level, and 85–95% identity at the amino acid level, with the other plant SAMS genes. Genomic Southern hybridization revealed the presence of more than two copies of SAMS genes in the genome of E. umbellata. Reverse transcriptase-mediated polymerase chain reaction (RT–PCR) analysis showed EuSAMS1 transcripts were more abundant than those of EuSAMS2. Similar to the expression pattern of other plant SAMS genes, both genes were expressed at higher levels in root than in leaf. During nodule development, expression of both genes was increased, with the highest level at 6–8 week after inoculation, and decreased rapidly thereafter. In situ hybridization analysis also showed both SAMS transcripts in the meristem zone, the infected cells of the fixation zone and in the central vascular system of root nodules. However, EuSAMS2 transcripts were strongly detected in the prefixation zone, whereas EuSAMS1 transcripts were hardly detected. These results suggest different regulatory mechanisms for the two genes in the root nodule. The expression pattern of SAMS genes in the root nodule may correlate mostly with cell wall synthesis, polyamine biosynthesis and other methylation-mediated functions.

scholarly journals Differential Expression Patterns of an Acidic Chitinase and a Basic Chitinase in the Root Nodule of Elaeagnus umbellata

2002 ◽  
Vol 15 (3) ◽  
pp. 209-215 ◽  
Author(s):  
Ho Bang Kim ◽  
Chung Sun An
Keyword(s):  
Root Nodule ◽  
Catalytic Domain ◽  
Vascular System ◽  
Root Nodules ◽  
Expression Patterns ◽  
Protein Structures ◽  
Cdna Clones ◽  
Outer Cortex ◽  
Elaeagnus Umbellata ◽  
Defensive Role

Two cDNA clones encoding chitinase were isolated from a root nodule cDNA library of Elaeagnus umbellata by the hybridization-competition method. The two clones, EuNOD-CHT1 and EuNOD-CHT2, encode for 335 and 317 amino acid residues with the molecular mass of mature proteins being 33.3 and 31.1 kDa, respectively. The two chitinases showed similar protein structures consisting of four domains: hydrophobic signal peptide domain, cysteine-rich chitin-binding domain, hinge domain, and catalytic domain. The EuNOD-CHT1 gene showed similar expression levels in root nodules and leaves, with no detection of transcripts in the roots. The EuNOD-CHT2 gene was expressed at similarly high levels in the roots and root nodules, but at a very low level in the leaves. In situ hybridization showed that EuNOD-CHT1 transcripts were strongly detected in the meristem zone, but weakly detected in the outer cortex layer of the root nodule and in the uninfected cells of the fixation zone. On the other hand, EuNOD-CHT2 transcripts were strongly detected in the infected cells of the fixation zone and central vascular system, but weakly detected in the senescence zone. Our results suggest that the two chitinases may play different biological roles in the root nodule. EuNOD-CHT2 may be involved in a defense response against internal symbionts, external pathogens, or both, while EuNOD-CHT1 may be involved in normal plant development as well as in a defensive role against external pathogens.

Sequence analysis and expression patterns of two nifA genes from Frankia EuIK1

2001 ◽  
Vol 28 (9) ◽  
pp. 939
Author(s):  
Hyoungseok Lee ◽  
Si Bum Sung ◽  
Ho Bang Kim ◽  
Chung Sun An
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Nitrogenase Activity ◽  
Sequence Similarity ◽  
Expression Patterns ◽  
Amino Acid Sequences ◽  
Start Codon ◽  
Nodule Development ◽  
Intergenic Sequence ◽  
Elaeagnus Umbellata

This paper originates from an address at the 8th International Symposium on Nitrogen Fixation with Non-Legumes, Sydney, NSW, December 2000 Two nifA genes were cloned from Frankia EuIK1 strain, a symbiont of Elaeagnus umbellata Thunb. and analysed on the basis of their deduced amino acid sequences and expression patterns. The complete nucleotide sequence of 1926 bp of nifA1 and 1524 bp of nifA2 was determined, respectively. A putative NifA-binding site was found –95 to about –80 bp upstream of start codon for nifA1 ORF as TGT-N10-ACA, but the clpB ORF was followed by nifA2 ORF with 15 bp of intergenic sequence. Deduced amino acid sequence showed that two nifA genes encode typical NifA, having three major domains and two linkers, and their central domains of NifA1 and NifA2 showed sequence similarity of 70–75% with those from other NifA proteins. However, entire NifA2 ORF is more similar to alternative NifA (60%) than to typical NifA (53%). Conserved amino acid sequence in helix-turn-helix motif of typical NifA was also found in NifA1, but it was not conserved in NifA2, which is also common in alternative NifA proteins. Moreover, the expression of nifA1 during nodule development was similar to that of Rhizobium meliloti in that it was expressed at low level constitutively, while that of nifA2 was similar to the pattern of nifH, structural gene for nitrogenase reductase, in that its transcripts level was changed in accordance with nitrogenase activity. These results indicate that nifA1 and nifA2 might be classified into typical nifA and alternative nifA, respectively. This is the first report on the presence of two nifA genes in Frankia.

Isolation and characterization of a cDNA clone encoding polyubiquitin from the root nodule ofElaeagnus umbellata

1999 ◽  
Vol 77 (9) ◽  
pp. 1270-1278
Author(s):  
Ho Bang Kim ◽  
Chung Sun An
Keyword(s):  
Amino Acids ◽  
In Situ Hybridization ◽  
Amino Acid ◽  
Cdna Clone ◽  
Root Nodule ◽  
Vascular System ◽  
Northern Hybridization ◽  
Elaeagnus Umbellata ◽  
Isolation And Characterization

A cDNA clone encoding polyubiquitin was isolated from a root nodule cDNA library of Elaeagnus umbellata Thunb. by differential hybridization, and its molecular aspects were characterized. The polyubiquitin clone pEuNOD-PUB1 has an insert size of 1642 bp and has the capacity to code for a 458 amino acid residue polyubiquitin protein. The derived amino acid sequence indicates that pEuNOD-PUB1 encodes a polyprotein consisting of six repeats of ubiquitin monomer, except for the last repeat, which has two additional amino acids (Asp-Phe) to be removed in the course of polyubiquitin processing into monomer. The molecular mass of ubiquitin monomer, consisting of 76 amino acids, was predicted to be 8524 Da, and the pI value was predicted to be 7.57. The nucleotide sequence of ubiquitin monomers from pEuNOD-PUB1 showed 73.1-86.0% sequence similarity with those from other organisms. The polyubiquitin mRNA content was four to six times higher in the root nodules than in the leaves and roots. In situ hybridization results showed polyubiquitin transcripts were strongly detected in the meristem zone, in infected cells of the fixation zone, and in the central vascular system. Genomic Southern hybridization revealed that polyubiquitin genes are present as a small multigene family in the genome of E. umbellata.Key words: Elaeagnus umbellata, root nodule, cDNA, polyubiquitin, Northern hybridization, in situ hybridization.

Cloning of rabbit prolactin cDNA and prolactin gene expression in the rabbit mammary gland

1996 ◽  
Vol 16 (1) ◽  
pp. 27-37 ◽  
Author(s):  
L Gabou ◽  
M Boisnard ◽  
I Gourdou ◽  
H Jammes ◽  
J-P Dulor ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mammary Gland ◽  
Amino Acid ◽  
Untranslated Regions ◽  
Pcr Analysis ◽  
Cdna Clones ◽  
Rt Pcr ◽  
Prolactin Gene ◽  
New Zealand White Rabbits ◽  
Rna Probe

ABSTRACT cDNA clones coding for rabbit prolactin were isolated from a pituitary library using a rat prolactin RNA probe. One cDNA contained 873 bases including the entire coding sequence of rabbit prolactin, its signal peptide and the 5′ and 3′ untranslated regions of 44 and 145 nucleotides respectively. The deduced amino acid sequence of the cloned prolactin cDNA presented a 93–78% identity with mink, porcine and human prolactins. The prolactin gene transcription was investigated by RT-PCR analysis in several organs of midlactating New Zealand White rabbits. The ectopic transcription of the prolactin gene was examined in more detail in the mammary gland. A strong PCR signal was detected in the mammary gland of virgin does and was also observed during pregnancy and at the beginning of lactation. This PCR signal was very weak in mid-lactating and absent in post-weaning mammary gland.

scholarly journals Comparison of Nodule Induction in Legume and Actinorhizal Symbioses: The Induction of Actinorhizal Nodules Does Not Involve ENOD40

2003 ◽  
Vol 16 (9) ◽  
pp. 808-816 ◽  
Author(s):  
Carole Santi ◽  
Uritza von Groll ◽  
Ana Ribeiro ◽  
Maurizio Chiurazzi ◽  
Florence Auguy ◽  
...  
Keyword(s):  
Expression Pattern ◽  
Host Plants ◽  
Root Nodule ◽  
Higher Plants ◽  
Expression Patterns ◽  
Lotus Japonicus ◽  
Casuarina Glauca ◽  
Actinorhizal Plant ◽  
Allocasuarina Verticillata ◽  
Actinorhizal Nodules

Two types of root nodule symbioses are known for higher plants, legume and actinorhizal symbioses. In legume symbioses, bacterial signal factors induce the expression of ENOD40 genes. We isolated an ENOD40 promoter from an actinorhizal plant, Casuarina glauca, and compared its expression pattern in a legume (Lotus japonicus) and an actinorhizal plant (Allocasuarina verticillata) with that of an ENOD40 promoter from the legume soybean (GmENOD402). In the actinorhizal Allocasuarina sp., CgENOD40-GUS and GmENOD40-2-GUS showed similar expression patterns in both vegetative and symbiotic development, and neither promoter was active during nodule induction. The nonsymbiotic expression pattern of CgENOD40-GUS in the legume genus Lotus resembled the nonsymbiotic expression patterns of legume ENOD40 genes however, in contrast to GmENOD40-2-GUS, CgENOD40-GUS was not active during nodule induction. The fact that only legume, not actinorhizal, ENOD40 genes are induced during legume nodule induction can be linked to the phloem unloading mechanisms established in the zones of nodule induction in the roots of both types of host plants.

scholarly journals Induction and Spatial Organization of Polyamine Biosynthesis During Nodule Development in Lotus japonicus

2004 ◽  
Vol 17 (12) ◽  
pp. 1283-1293 ◽  
Author(s):  
Emmanouil Flemetakis ◽  
Rodica C. Efrose ◽  
Guilhem Desbrosses ◽  
Maria Dimou ◽  
Costas Delis ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Expression Patterns ◽  
Lotus Japonicus ◽  
Cyclin D3 ◽  
Nodule Development ◽  
Cdna Clones ◽  
Long Distance ◽  
Polyamine Biosynthesis ◽  
Transcript Levels

Putrescine and other polyamines are produced by two alternative pathways in plants. One pathway starts with the enzyme arginine decarboxylase (ADC; EC 4.1.1.19), the other with ornithine decarboxylase (ODC; EC 4.1.1.17). Metabolite profiling of nitrogen-fixing Lotus japonicus nodules, using gas chromatography coupled to mass spectrometry, revealed a two- to sixfold increase in putrescine levels in mature nodules compared with other organs. Genes involved in polyamine biosynthesis in L. japonicus nodules were identified by isolating cDNA clones encoding ADC (LjADC1) and ODC (LjODC) from a nodule library. Searches of the public expressed sequence tag databases revealed the presence of a second gene encoding ADC (LjADC2). Real-time reverse-transcription-polymerase chain reaction analysis showed that LjADC1 and LjADC2 were expressed throughout the plant, while LjODC transcripts were detected only in nodules and roots. Induction of LjODC and LjADC gene expression during nodule development preceded symbiotic nitrogen fixation. Transcripts accumulation was maximal at 10 days postinfection, when a 6.5-fold increase in the transcript levels of LjODC was observed in comparison with the uninfected roots, while a twofold increase in the transcript levels of LjADC1 and LjADC2 was detected. At later stages of nodule development, transcripts for ADC drastically declined, while in the case of ODC, transcript accumulation was higher than that in roots until after 21 days postinfection. The expression profile of genes involved in putrescine biosynthesis correlated well with the expression patterns of genes involved in cell division and expansion, including a L. japonicus Cyclin D3 and an α-expansin gene. Spatial localization of LjODC and LjADC1 gene transcripts in developing nodules revealed that both transcripts were expressed in nodule inner cortical cells and in the central tissue. High levels of LjADC1 transcripts were also observed in both nodule and connecting root vascular tissue, suggesting that putrescine and other polyamines may be subject to long-distance transport. Our results indicate that polyamines are primarily involved in physiological and cellular processes involved in nodule development, rather than in processes that support directly symbiotic nitrogen fixation and assimilation.

scholarly journals Evolution of the Heme Peroxidases of Culicidae (Diptera)

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Austin L. Hughes
Keyword(s):  
Amino Acid ◽  
Gene Duplication ◽  
Common Ancestor ◽  
Expression Patterns ◽  
Amino Acid Level ◽  
Recent Common Ancestor ◽  
Amino Acid Sequence Level ◽  
Most Recent Common Ancestor ◽  
High Level ◽  
Heme Peroxidases

Phylogenetic analysis of heme peroxidases (HPXs) of Culicidae and other insects revealed six highly conserved ancient HPX lineages, each of which originated by gene duplication prior to the most recent common ancestor (MRCA) of Hemimetabola and Holmetabola. In addition, culicid HPX7 and HPX12 arose by gene duplication after the MRCA of Culicidae and Drosophilidae, while HPX2 orthologs were not found in any other order analyzed except Diptera. Within Diptera, HPX2, HPX7, and HPX12 were relatively poorly conserved at the amino acid level in comparison to the six ancient lineages. The genome ofAnopheles gambiaeincluded genes ecoding five proteins (HPX10, HPX11, HPX13, HXP14, and HPX15) without ortholgs in other genomes analyzed. Overall, gene expression patterns did not seem to reflect phylogenetic relationships, but genes that evolved rapidly at the amino acid sequence level tended to have divergent expression patterns as well. The uniquely high level of duplication of HPXs inA. gambiaemay have played a role in coevolution with malaria parasites.

scholarly journals Genome-wide survey of soybean papain-like cysteine proteases and their expression analysis in root nodule symbiosis

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Songli Yuan ◽  
Danxia Ke ◽  
Rong Li ◽  
Xiangyong Li ◽  
Lei Wang ◽  
...  
Keyword(s):  
Root Nodule ◽  
Expression Patterns ◽  
Cysteine Proteases ◽  
Nodule Development ◽  
Whole Genome ◽  
Root Nodule Symbiosis ◽  
Genome Wide ◽  
A Genome ◽  
Genome Wide Survey ◽  
Nodule Symbiosis

Abstract Background Plant papain-like cysteine proteases (PLCPs) are a large class of proteolytic enzymes and play important roles in root nodule symbiosis (RNS), while the whole-genome studies of PLCP family genes in legume are quite limited, and the roles of Glycine max PLCPs (GmPLCPs) in nodulation, nodule development and senescence are not fully understood. Results In the present study, we identified 97 GmPLCPs and performed a genome-wide survey to explore the expansion of soybean PLCP family genes and their relationships to RNS. Nineteen paralogous pairs of genomic segments, consisting of 77 GmPLCPs, formed by whole-genome duplication (WGD) events were identified, showing a high degree of complexity in duplication. Phylogenetic analysis among different species showed that the lineage differentiation of GmPLCPs occurred after family expansion, and large tandem repeat segment were specifically in soybean. The expression patterns of GmPLCPs in symbiosis-related tissues and nodules identified RNS-related GmPLCPs and provided insights into their putative symbiotic functions in soybean. The symbiotic function analyses showed that a RNS-related GmPLCP gene (Glyma.04G190700) really participate in nodulation and nodule development. Conclusions Our findings improved our understanding of the functional diversity of legume PLCP family genes, and provided insights into the putative roles of the legume PLCPs in nodulation, nodule development and senescence.

scholarly journals Identification and tissue distribution of mRNAs encoding salmon-type calcitonins-IV and -V in the rainbow trout

2004 ◽  
Vol 32 (3) ◽  
pp. 963-974 ◽  
Author(s):  
Y Hidaka ◽  
M Suzuki
Keyword(s):  
Rainbow Trout ◽  
Amino Acid ◽  
Tissue Distribution ◽  
Restriction Enzymes ◽  
Tissue Expression ◽  
Pcr Analysis ◽  
Cdna Clones ◽  
Rt Pcr ◽  
Parenchymal Cells ◽  
Ultimobranchial Gland

Four types of calcitonin are produced in salmonid fish, although their functional diversity is almost unknown. To explore the significance of these isoforms, we have characterized salmon-type calcitonin (sCT) mRNAs in the rainbow trout (Oncorhynchus mykiss), and examined their tissue distribution. In addition to the previously isolated sCT-I cDNAs, two new forms of sCT cDNA were cloned from the ultimobranchial gland, and one of them (sCT-IV cDNA) was predicted to encode an N-terminal peptide of 80 amino acid residues, a putative cleavage site Lys-Arg, sCT-IV, a cleavage and amidation sequence Gly-Lys-Lys-Arg, and a C-terminal peptide of 18 amino acids. The sCT-IV precursor was 78% identical with the rainbow trout sCT-I precursors. The other cloned cDNA encoded a precursor for a novel CT, sCT-V. The sCT-V peptide was different from sCT-IV by only one amino acid residue: Val at position 8 in the latter was replaced by Met. The sCT-V precursor had 80 and 90% identity with the sCT-I and -IV precursors respectively. No cDNA clones were obtained for sCTs-II or -III.Tissue distribution of sCT-I, -IV and -V mRNAs was examined by RT-PCR and specific cleavage with restriction enzymes. An amplified fragment from sCT-I mRNA was detected not only in the ultimobranchial gland, but also in the gills, testis and ovary. RT-PCR analysis coupled to restriction digestion further revealed that sCT-IV mRNA was expressed in both the testis and the ultimobranchial gland. The expression sites of sCT-IV mRNA were localized to the Leydig cells of the testis and to the parenchymal cells of the ultimobranchial gland, by in situ hybridization histochemistry. Although the amino acid sequence of sCT-V peptide was nearly the same as that of sCT-IV, the sCT-V gene showed a much wider pattern of expression: the band amplified by RT-PCR was detected in all the tissues examined except the kidney, gills and blood cells. The sCT-V mRNA was shown to be localized in the parenchymal cells of the ultimobranchial gland, but not in other tissues at the cellular level, suggesting very low expression of sCT-V mRNA in those tissues. Our results show different patterns of tissue expression of three types of sCT genes in the rainbow trout, suggesting that sCTs-I, -IV and -V might differ in their local actions.

scholarly journals Disparate Oxygen Responsiveness of Two Regulatory Cascades That Control Expression of Symbiotic Genes in Bradyrhizobium japonicum

2003 ◽  
Vol 185 (18) ◽  
pp. 5639-5642 ◽  
Author(s):  
Michel-Angelo Sciotti ◽  
Astrid Chanfon ◽  
Hauke Hennecke ◽  
Hans-Martin Fischer
Keyword(s):  
Bradyrhizobium Japonicum ◽  
Target Genes ◽  
Root Nodule ◽  
Expression Patterns ◽  
Nodule Development ◽  
Symbiotic Genes ◽  
Free Living ◽  
Regulatory Systems ◽  
Regulatory Cascades ◽  
Oxygen Conditions

ABSTRACT Two oxygen-responsive regulatory systems controlling numerous symbiotic genes in Bradyrhizobium japonicum were assayed in free-living cultures for their capacity to activate target genes under different oxygen conditions. NifA- and FixLJ-controlled target genes showed disparate relative expression patterns. Induction of NifA-dependent genes was observed only at oxygen concentrations below 2% in the gas phase, whereas that of FixLJ-controlled targets progressively increased when the oxygen concentration was lowered from 21 to 5, 2, or 0.5%. We propose that this reflects a response to a gradient of increasing oxygen deprivation as bacteria invade their host during root nodule development.

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