| nucleotide sequence of rhizobium meliloti rcr2011 genes involved in host specificity of nodulation. | a 6 kb dna segment of the r. meliloti 2011 psym megaplasmid, which contains genes controlling host specificity of root hair infection and of nodulation, was cloned and sequenced. the dna sequence analysis, in conjunction with previous genetic data, allowed identification of four nod genes designated as e, f, g and h. nodh is divergently transcribed with respect to nodfe and nodg. a conserved nucleotide sequence was found around 200 bp upstream of the translation start of nodf, nodh and noda. thi ... | 1986 | 3020515 |
| the vird operon of agrobacterium tumefaciens encodes a site-specific endonuclease. | tumor formation by agrobacterium tumefaciens involves the transfer and integration of a defined segment (t-dna) of tumor-inducing (ti) plasmid dna into the plant nuclear genome. a set of plasmid genes outside the t-dna, the vir genes, are thought to mediate the transfer process. we report here that the vird operon encodes a site-specific endonuclease that cleaves at a unique site within each of the 24 bp direct repeats that flank the t-dna. the endonuclease function was further localized to the ... | 1986 | 3021341 |
| organization of the adenyl cyclase (cya) locus of rhizobium meliloti. | a cya-like gene encoding adenyl cyclase from rhizobium meliloti was localized to a 0.8-kb psti-ecori fragment by subcloning experiments. experiments in escherichia coli 'maxicells' identified a r. meliloti cya gene product of 28 kda, which is significantly smaller than the corresponding protein from enteric bacteria. a control region for the expression of the cya gene in e. coli was found on an adjacent 2.6-kb bglii-bamhi sequence by insertional mutagenesis with tn5 and phage mudi (apr lac). the ... | 1986 | 3021593 |
| a gene essential for agrobacterium virulence is homologous to a family of positive regulatory loci. | the vir region of agrobacterium tumefaciens spans at least six transcriptional loci required for crown gall tumorigenesis. the transcriptional induction of two of these vir loci in response to cocultivation with tobacco suspension cells was measured by using bacteria containing mutations in each of the six vir loci located on the ti plasmid. induction of these vir genes occurred only in bacteria that had functional copies of vira and virg. the nucleic acid sequence of a 1.25-kilobase clone encom ... | 1986 | 3022288 |
| cloning and regulation of erwinia herbicola pigment genes. | the genes coding for yellow pigment production in erwinia herbicola eho10 (atcc 39368) were cloned and localized to a 12.4-kilobase (kb) chromosomal fragment. a 2.3-kb avai deletion in the cloned fragment resulted in the production of a pink-yellow pigment, a possible precursor of the yellow pigment. production of yellow pigment in both e. herbicola eho10 and pigmented escherichia coli clones was inhibited by glucose. when the pigment genes were transformed into a cya (adenylate cyclase) e. coli ... | 1986 | 3023282 |
| assignment of symbiotic developmental phenotypes to common and specific nodulation (nod) genetic loci of rhizobium meliloti. | rhizobium meliloti nodulation (nod) genes required for specific infection and nodulation of alfalfa have been cloned. transposon tn5 mutagenesis defined three nod regions spanning 16 kilobases of the psym megaplasmid. genetic and cytological studies of 62 nodulation-defective mutants allowed the assignment of symbiotic developmental phenotypes to common and specific nod loci. root hair curling was determined by both common (region i) and specific (region iii) nod transcription units; locus iiib ... | 1986 | 3023297 |
| t-dna and opine synthetic loci in tumors incited by agrobacterium tumefaciens a281 on soybean and alfalfa plants. | we report here the molecular characterization of transferred dna (t-dna) in leguminous tumors incited by agrobacterium tumefaciens a281 harboring the tumor-inducing plasmid ptibo542. the t-dna is composed of two regions named tl (left portion)-dna and tr (right portion)-dna, in accordance with the nomenclature for the octopine strains. tl-dna is defined by several internal hindiii restriction fragments totaling 10.8 kilobase pairs (kbp) in uncloned soybean and alfalfa tumors. alfalfa tumor dna m ... | 1986 | 3023301 |
| upstream sequences required for efficient expression of a soybean heat shock gene. | a soybean gene (gmhsp17.5-e) encoding a small heat shock protein was introduced into primary sunflower tumors via t-dna-mediated transformation. rna blot hybridizations and s1-nuclease hybrid protection studies indicated that the heat shock gene containing 3.25 kilobases of 5'-flanking sequences was strongly transcribed in a thermoinducible (40 degrees c) manner. transcriptional induction also occurred to a lesser extent upon treatment of whole tumors with sodium arsenite and cdcl2. basal (26 de ... | 1986 | 3023855 |
| biosynthesis and degradation of nodule-specific rhizobium loti compounds in lotus nodules. | two nodule-specific rhizobium loti compounds were identified in lotus tenuis and lotus pedunculatus nodules induced by strain nzp2037. one, a silver nitrate-positive cation called rhizolotine, has been characterized as the riboside of a novel alpha-hydroxyimino acid containing a 1,4,5,6-tetrahydropyrimidine ring (g. j. shaw, r. d. wilson, g. a. lane, l. d. kennedy, d. b. scott, and g. j. gainsford, j. chem. soc. chem. commun., p. 180-181, 1986), and the other, yellow-1, stains yellow with ninhyd ... | 1987 | 3025173 |
| characterization of nonattaching mutants of agrobacterium tumefaciens. | the first step in tumor formation by agrobacterium tumefaciens is the site-specific binding of the bacteria to plant host cells. transposon mutants of the bacteria which fail to attach to carrot suspension culture cells were isolated. these mutants showed no significant attachment to carrot cells with either microscopic or viable cell count assays of bacterial binding. the nonattaching mutants were all avirulent. when revertants of the mutants were obtained by enriching for bacteria which do bin ... | 1987 | 3025176 |
| isolation of competition-defective mutants of rhizobium fredii. | we coupled tn5 mutagenesis with a competition assay to isolate mutants of rhizobium fredii usda 257 that are defective in competition for nodulation of soybeans. two mutants with single tn5 inserts in the chromosome showed reduced competitiveness in vermiculite but were identical to the wild-type strain in symbiotic properties when inoculated alone. recombination of tn5 and flanking genomic regions cloned from the mutants into the parent strain showed that tn5 was responsible for the mutant phen ... | 1987 | 3025184 |
| nitrogen fixation ability of exopolysaccharide synthesis mutants of rhizobium sp. strain ngr234 and rhizobium trifolii is restored by the addition of homologous exopolysaccharides. | several transposon tn5-induced mutants of the broad-host-range rhizobium sp. strain ngr234 produce little or no detectable acidic exopolysaccharide (eps) and are unable to induce nitrogen-fixing nodules on leucaena leucocephala var. peru or siratro plants. the ability of these exo- mutants to induce functioning nodules on leucaena plants was restored by coinoculation with a sym plasmid-cured (nod- exo+) derivative of parent strain ngr234, purified eps from the parent strain, or the oligosacchari ... | 1987 | 3025187 |
| [the use of transposons for the mutagenesis of r. phaseoli and r. japonicum]. | plasmid psup2011 has been used to transfer transposon tn5 into the cells of r. japonicum 110 and r. phaseoli 693. transposition of tn5 into the chromosomes of r. japonicum and r. phaseoli has been demonstrated,resulting in isolation of auxotrophic and symbiotic mutants of both strains. the frequencies of selected auxotrophic mutations have reached 4% in r. japonicum 110 and 0.6% in r. phaseoli 693. streptomycin resistance gene locating on tn5 has been found to be phenotypically expressed in r. j ... | 1985 | 3025720 |
| [characteristics of derivatives of the plasmid rp4 in a broad range of hosts with altered properties of maintenance and inheritance]. | hydroxylamine-induced mutants of the plasmid ppd6 (8.4 kb) were isolated which are resistant to high doses of tetracycline. one of the plasmids studied--ppd21 is a multicopy mutant, another one, ppd12 is a dimeric form of the ppd6 plasmid. the ppd12 plasmid is very unstable, its derivative, ppd13 spontaneous mutant acquiring stability but not the ability to resolve dna multimeric forms into monomeric forms. multicopy bireplicon ppd619 plasmid was constructed by joining in vitro ppd6 and puc19 pl ... | 1986 | 3026896 |
| integration of the delta-endotoxin gene of bacillus thuringiensis into the chromosome of root-colonizing strains of pseudomonads using tn5. | the delta-endotoxin gene (tox) from bacillus thuringiensis subsp. kurstaki hd-1 was cloned into tn5 and the resulting tn5-tox element transposed from a vector plasmid into the chromosome of six corn-root-colonizing strains of pseudomonas fluorescens and agrobacterium radiobacter. chromosomal integration of the tox gene maximized stability and minimized the potential for horizontal transfer of the tox gene to other bacterial species. expression of the tox gene was demonstrated by western blot ana ... | 1986 | 3026918 |
| nucleotide sequence of the virulence gene virg of the agrobacterium tumefaciens octopine ti plasmid: significant homology between virg and the regulatory genes ompr, phob and dye of e. coli. | the entire nucleotide sequence of the virg locus, from the octopine ti plasmid of agrobacterium tumefaciens strain 15955, has been determined. the virg gene is 801 nucleotides in length and has one open reading frame which encodes a protein of mr 29,995. the virg gene is involved in the transcriptional activation of the ti plasmid vir-loci, which occurs after induction by specific compounds present in plant exudate. sequence analysis of the agrobacterium virg protein showed significant homology ... | 1986 | 3027669 |
| a spontaneous insertion in the agrocin sensitivity region of the ti-plasmid of agrobacterium tumefaciens c58. | a spontaneous agrocin-resistant mutant of agrobacterium tumefaciens strain c58 was shown to have an insertion of 1.2 kb in the agrocin-sensitivity region of ptic58. the insertion was cloned from the ti-plasmid, and a subclone containing only dna internal to the insertion was used to probe the ti-plasmid and chromosomal dna of the wild-type strain c58. the probe showed homology to chromosomal sequences but showed no homology to wild-type ptic58. homology was also detected with chromosomal sequenc ... | 1986 | 3027729 |
| studies of the structure and functional organization of foreign dna integrated into the genome of nicotiana tabacum. | in transgenic plants obtained either by agrobacterium tumefaciens-mediated transformation or by direct dna transfer, the structure of integrated chimeric donor dna remains stable during vegetative proliferation, during sexual transmission, and under various selection conditions. we correlate the level of expression of the introduced gene in independent transformants and their offspring with the particular arrangement and modification of their integrated dnas. genetic analysis of transgenic plant ... | 1986 | 3028737 |
| processing of the t-dna of agrobacterium tumefaciens generates border nicks and linear, single-stranded t-dna. | transfer and integration of a defined region (t-dna) of the tumor-inducing (ti) plasmid of agrobacterium tumefaciens is essential for tumor formation. we used a physical assay to study structural changes induced in agrobacterium t-dna by cocultivation with plant cells. we show that nicks are introduced at unique, identical locations in each of the 24-base-pair imperfect direct repeats which flank the t-dna and present evidence that a linear, single-stranded molecule is generated. we propose that ... | 1987 | 3029014 |
| conservation of structure and location of rhizobium meliloti and klebsiella pneumoniae nifb genes. | using transposon tn5-mediated mutagenesis, an essential rhizobium meliloti nitrogen fixation (nif) gene was identified and located directly downstream of the regulatory gene nifa. maxicell and dna sequence analysis demonstrated that the new gene is transcribed in the same direction as nifa and codes for a 54-kilodalton protein. in klebsiella pneumoniae, the nifbq operon is located directly downstream of a gene which is structurally and functionally homologous to the r. meliloti nifa gene. the dn ... | 1987 | 3029020 |
| genetic and structural analysis of the rhizobium meliloti fixa, fixb, fixc, and fixx genes. | the fixa, fixb, fixc, and fixx genes of rhizobium meliloti 1021 constitute an operon and are required for nitrogen fixation in alfalfa nodules. dna homologous to the r. meliloti fixabc genes is present in all other rhizobium and bradyrhizobium species examined, but fixabc-homologous sequences were found in only one free-living diazotroph, azotobacter vinelandii. to determine whether the fixabcx genes share sequence homology with any of the 17 klebsiella pneumoniae nif genes, we determined the en ... | 1987 | 3029021 |
| alteration of surface properties in a tn5 mutant strain of rhizobium trifolii 0403. | a symbiotically defective mutant strain of rhizobium trifolii, ur251, was obtained by transposon tn5 mutagenesis of r. trifolii 0403 rif and recognized by its partially ineffective (fix +/-) phenotype on white clover plants. ur251 had a single tn5 insertion in plasmid dna, a wild-type plasmid pattern, and no detectable mu dna sequences originally present in the vector used for tn5 mutagenesis. agglutination by the clover lectin trifoliin a and attachment to clover root hairs was higher with ur25 ... | 1987 | 3029022 |
| organization and partial sequence of a dna region of the rhizobium leguminosarum symbiotic plasmid prl6ji containing the genes fixabc, nifa, nifb and a novel open reading frame. | by hybridization and heteroduplex studies the fixabc and nifa genes of the rhizobium leguminosarum symbiotic plasmid prl6ji have been identified. dna sequencing of the region containing nifa showed an open reading frame of 1557 bp encoding a protein of 56, 178 d. based on sequence homology, this orf was confirmed to correspond to the nifa gene. comparison of three nifa proteins (klebsiella pneumoniae, rhizobium meliloti, rhizobium leguminosarum) revealed only a weak relationship in their n-termi ... | 1987 | 3029674 |
| construction of a tn5 derivative determining resistance to gentamicin and spectinomycin using a fragment cloned from r1033. | a physical and genetic map of the incp plasmid r1033 was constructed: restriction fragments were subcloned and antibiotic resistance genes were located. the map is consistent with previous reports that r1033 is a derivative of rp4 carrying a 16-kb transposon tn1696 which contains the antibiotic-resistance determinants present on r1033 but not on rp4. a bamhi fragment from r1033, determining resistance to gentamicin, spectinomycin and streptomycin, was cloned into tn5, replacing the central bg/ii ... | 1986 | 3030896 |
| rhizobium meliloti insertion element isrm2 and its use for identification of the fixx gene. | two of the three plasmids of the wild-type rhizobium meliloti 41 (prme41a and prme41c) carry a copy of isrm2, a 2.7-kilobase-long transposable element. isrm2 is terminated by 22-base-pair (bp) inverted repeat sequences, exhibiting some homology to the inverted repeats of elements generating 9-bp target sequence duplication. transposition of isrm2 results in a duplication of 8 bp in length, rather rare among transposable elements. dna sequences homologous to an internal fragment of isrm2 were fou ... | 1987 | 3031010 |
| isolation and characterization of symbiotic mutants of bradyrhizobium sp. (arachis) strain nc92: mutants with host-specific defects in nodulation and nitrogen fixation. | random transposon tn5 mutagenesis of bradyrhizobium sp. (arachis) strain nc92, a member of the cowpea cross-inoculation group, was carried out, and kanamycin-resistant transconjugants were tested for their symbiotic phenotype on three host plants: groundnut, siratro, and pigeonpea. two nodulation (nod- phenotype) mutants were isolated. one is unable to nodulate all three hosts and appears to contain an insertion in one of the common nodulation genes (nodabcd); the other is a host-specific nodula ... | 1987 | 3032910 |
| transcription of rhizobium meliloti nodulation genes. identification of a nodd transcription initiation site in vitro and in vivo. | nodulation genes in rhizobium are required for invasion of the host plant. the nodabc operon is induced by plant activator molecules; this activation requires the gene product of the constitutively expressed nodd locus, which is transcribed divergently from nodabc. we are employing in vitro transcription to elucidate the molecular mechanism of nod gene activation. we used a micropurification technique to obtain rna polymerase from rhizobium meliloti, and here demonstrate that it initiated and te ... | 1987 | 3032978 |
| physical characterization of rhizobium meliloti megaplasmids. | intact megaplasmids of rhizobium meliloti 2011 have been isolated and visualized by electron microscopy. the contour lengths of 64 megaplasmid molecules were determined. one definite class of molecules of 400 micron length and a range of larger molecules with lengths of up to 560 micron was observed. the contour lengths of the megaplasmids prme2011a and prme2011b were measured after isolation from plasmid-free agrobacterium strains into which they had been individually transferred. plasmid prme2 ... | 1987 | 3033717 |
| rhizobium meliloti ntra (rpon) gene is required for diverse metabolic functions. | we report the identification and cloning of an ntra-like (glnf rpon) gene of rhizobium meliloti and show that the r. meliloti ntra product (ntra) is required for c4-dicarboxylate transport as well as for nitrate assimilation and symbiotic nitrogen fixation. dna sequence analysis showed that r. meliloti ntra is 38% homologous with klebsiella pneumoniae ntra. subcloning and complementation analysis suggested that the r. meliloti ntra promoter lies within 125 base pairs of the initiation codon and ... | 1987 | 3034856 |
| transposon-induced symbiotic mutants of bradyrhizobium japonicum: isolation of two gene regions essential for nodulation. | two strains of the soybean endosymbiont bradyrhizobium japonicum, usda 110 and 61 a101 c, were mutagenized with transposon tn5. after plant infection tests of a total of 6,926 kanamycin and streptomycin resistant transconjugants, 25 mutants were identified that are defective in nodule formation (nod-) or nitrogen fixation (fix-). seven nod- mutants were isolated from strain usda110 and from strain 61 a101 c, 4 nod- mutants and 14 fix- mutants were identified. subsequent auxotrophic tests on thes ... | 1987 | 3037278 |
| interposon mutagenesis of soil and water bacteria: a family of dna fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. | we have constructed a series of derivatives of the omega interposon [prentki and krisch, gene 29 (1984) 303-313] that can be used for in vitro insertional mutagenesis. each of these dna fragments carries a different antibiotic or hg2+ resistance gene (apr, cmr, tcr, kmr or hgr) which is flanked, in inverted orientation, by transcription and translation termination signals and by synthetic polylinkers. the dna of these interposons can be easily purified and then inserted, by in vitro ligation, in ... | 1987 | 3038679 |
| deletion analysis of the mannopine synthase gene promoter in sunflower crown gall tumors and agrobacterium tumefaciens. | we have used deletion mutagenesis to analyze a tr-dna promoter from the octopine-type ti plasmid ptib6806. the promoter for the gene encoding mannopine synthase (mas) was cloned upstream of the bacterial kanamycin-resistance gene neomycin phosphotransferase ii (npt ii). bal31 deletion mutagenesis was used to generate deletion derivatives of the mas/nptii gene beginning 1353 bp upstream of the initiation of transcription and extending to 120 bp downstream from the mrna start site. deletions that ... | 1987 | 3039293 |
| the noc region of ti plasmid c58 codes for arginase and ornithine cyclodeaminase. | plant tumors induced by agrobacterium tumefaciens synthesize a group of substances (opines) which can serve as sole source of carbon and nitrogen for the bacteria. we investigate ti-plasmid-coded genes and enzymes involved in catabolism of the opine n2-(1,3-dicarboxypropyl)-l-arginine (nopaline) with a novel approach: expression and mapping of protein-coding regions in escherichia coli minicells, followed by identification of enzyme functions in the heterologous e. coli background. the results s ... | 1987 | 3040404 |
| transcription of a rhizobium leguminosarum biovar phaseoli gene needed for melanin synthesis is activated by nifa of rhizobium and klebsiella pneumoniae. | the rhizobium leguminosarum biovar phaseoli symbiotic plasmid prp2ji carries a gene, mela, specifying the enzyme tyrosinase, which is responsible for the production of the pigment melanin in these bacteria. transcription of mela is activated by the nifa gene of rhizobium and, when the cloned mela gene is transferred to escherichia coli, mela is expressed if the recipients contain nifa gene of klebsiella pneumoniae. this nifa-dependent activation was temperature sensitive and required the ntra ge ... | 1988 | 3041240 |
| the ndva gene product of rhizobium meliloti is required for beta-(1----2)glucan production and has homology to the atp-binding export protein hlyb. | the ndva locus of rhizobium meliloti is homologous to and can substitute for the chva locus of agrobacterium tumefaciens. we have previously shown that an ndva mutant exhibited reduced motility and formed small, white, empty nodules on alfalfa roots. here we show that this ndva mutant is defective in the production of the cyclic extracellular polysaccharide beta-(1----2)glucan, even though a 235,000-dalton protein intermediate, known to be involved in the synthesis of this molecule, is present a ... | 1988 | 3042754 |
| transfer of the ti plasmid from agrobacterium tumefaciens into escherichia coli cells. | we have screened strains of agrobacterium tumefaciens for spontaneous mutants showing constitutive transfer of the nopaline ti plasmid ptic58 during conjugation. the ti plasmid derivatives obtained could be transferred not only to a. tumefaciens but also to e. coli cells. the ti plasmid cannot survive as a freely replicating plasmid in e. coli, but it can occasionally integrate into the e. coli chromosome. however, insertion in tandem of plasmids carrying fd replication origins (pfd plasmids) in ... | 1988 | 3049936 |
| flavonoids in the environment: structure-activity relationships. | | 1988 | 3051025 |
| conformational changes in the membrane-bound hydrogenase of bradyrhizobium japonicum. evidence that the redox state of the enzyme affects its accessibility to protease and membrane-impermeant reagents. | the sensitivity of the membrane-bound hydrogenase of bradyrhizobium japonicum to inactivation by proteases and membrane-impermeant protein modification reagents was compared under hydrogen versus oxygen. in membrane vesicles, the half-life of enzyme inactivation by trypsin of the h2-reduced enzyme was approximately 10 min, whereas o2-oxidized enzyme was much less sensitive to trypsin inactivation (half-life of over 90 min). diazobenzene sulfonate (dabs) affected the enzyme activity in a manner s ... | 1988 | 3053719 |
| further analysis of nitrogen fixation (nif) genes in azotobacter chroococcum: identification and expression in klebsiella pneumoniae of nifs, nifv, nifm, and nifb genes and localization of nife/n-, nifu-, nifa- and fixabc-like genes. | the results presented extend previous investigations on the genetics of nitrogen fixation in azotobacter chroococcum and indicate that nif- and fix-like dna is located in at least five different regions of the genome. region i contains functional copies of nifs,v and m, as well as nifh, d and k, all of which complemented mutants of klebsiella pneumoniae. in addition, nife- and/or nifn-like and nifu-like dna is located in this region. the organization of the nif cluster in region i closely resemb ... | 1988 | 3053983 |
| rhizobium meliloti host range nodh gene determines production of an alfalfa-specific extracellular signal. | the rhizobium meliloti nodh gene is involved in determining host range specificity. by comparison with the wild-type strain, nodh mutants exhibit a change in host specificity. that is, although nodh mutants lose the ability to elicit root hair curling (hac-), infection threads (inf-), and nodule meristem formation (nod-) on the homologous host alfalfa, they gain the ability to be hac+ inf+ nod+ on a nonhomologous host such as common vetch. using root hair deformation (had) bioassays on alfalfa a ... | 1988 | 3056902 |
| high-frequency transformation of rhizobium meliloti. | transformation of r factor rp4 and its derivative prk290 from escherichia coli to rhizobium meliloti is reported. the efficiency of transformation was in the range of 10(-5) per viable cell. in addition, chromosomal dna prepared from one r. meliloti strain resistant to streptomycin was transferred to the isoleucine-valine-requiring mutant susceptible to streptomycin. | 1988 | 3056925 |
| bidirectional transfer from a 24 bp border repeat of agrobacterium tumefaciens. | t-region transfer from wild-type agrobacterium strains is thought to be an orientated process, starting at the right border repeat and terminating at the left border repeat of the t-region. here we demonstrate that a right border repeat in the inverted orientation relative to the onc-genes can also mediate transfer of the t-region to the plant cell, although with lower efficiency as a border repeat in the native orientation. transfer mediated by an inverted right border repeat is stimulated by t ... | 1988 | 3057440 |
| effect of agrobacterium tumefaciens lipopolysaccharide on hemolytic complement system. | | 1988 | 3058583 |
| factors affecting the survival and growth of bacteria introduced into lake water. | the populations of pseudomonas sp. b4, escherichia coli, klebsiella pneumoniae, micrococcus flavus, and rhizobium leguminosarum biovar phaseoli declined rapidly in lake water. the initially rapid decline of the two pseudomonads and r. phaseoli was followed by a period of slow loss of viability, but viable cells of the other species were not found after 10 days. the rapid initial phase of decline was not a result of bdellovibrio spp., bacteriophages, or toxins in the water since bdellovibrio spp. ... | 1988 | 3060035 |
| cosmids. | | 1988 | 3061512 |
| the dna-binding domain of the transcriptional activator protein nifa resides in its carboxy terminus, recognises the upstream activator sequences of nif promoters and can be separated from the positive control function of nifa. | the positive control protein nifa activates transcription of nitrogen fixation promoters in klebsiella pneumoniae. nifa is believed to bind to specific sites, the upstream activator sequences (uas's), of the nif promoters which it activates. we have now shown by mutation of the carboxy terminus of nifa that this is the dna-binding domain and that the dna-binding and positive activator functions of nifa can be separated. mutational analysis of the nifh uas and in vivo methylation protection analy ... | 1988 | 3062575 |
| detection of ligand-protein binding by direct electrophoresis of the complex. | | 1988 | 3069855 |
| basic processes underlying agrobacterium-mediated dna transfer to plant cells. | | 1988 | 3071244 |
| foreign genes in plants: transfer, structure, expression, and applications. | | 1988 | 3071255 |
| nickel as a micronutrient element for plants. | the detrimental effects of excessive ni on plant growth have been well known for many years. more recent evidence indicates that ni is required in small amounts for normal plant growth and development. ni is an essential component of urease in plants and microorganisms. a deficiency of ni in plants is reported to result in necrotic lesions in leaves in response to toxic accumulations of urea. urease plays an essential role in mobilization of nitrogenous compounds in plants, a process that is esp ... | 1988 | 3076427 |
| flavones and isoflavones as inducing substances of legume nodulation. | rhizobia are soil bacteria that can form symbiotic associations with leguminous plants leading to the fixation of atmospheric nitrogen to ammonia which the plant can use. this is an interaction which involves the exchange of many signals between the plant and the bacterium. to start this interaction, rhizobia have adapted to use flavonoid compounds, released by the plant root, as part of a regulatory system to initiate the transcription of their infection (nodulation, nod) genes. the development ... | 1988 | 3076431 |
| development and differentiation of the root nodule. involvement of plant and bacterial genes. | | 1988 | 3077981 |
| [nitrogenase, hydrogenase and nitrate reductase activities, oxygen consumption, and atp content in nodules formed by strains of rhizobium leguminosarum 128c53 and 300 in symbiosis with pea plants]. | the nitrogenase activity, nitrate reductase activity and oxygen uptake as well as the hydrogen incorporation and atp content were examined in the root nodules and bacteroids, respectively, formed by rhizobium leguminosarum strains 128c53 (hydrogenase positive) and 300 (hydrogenase negative) in symbiosis with pisum sativum plants grown in the presence of 2 mm kno3. the strain 128c53 showed the greatest values for all parameters analyzed, except for the nitrate reductase activity, which was higher ... | 1986 | 3078142 |
| use of agrobacterium radiobacter in agricultural ecosystems. | agrobacterium tumefaciens is a soil inhabitant that infects many dicotyledonous plants, causing crown gall disease. the success of agrobacterium radiobacter k84 to control this disease in agroecosystems throughout the world has been impressive. this review describes the attributes of k84, its mechanism of control, its failures, and other alternative biocontrol agents. | 1988 | 3079224 |
| kinetics of monoclonal antibody production in low serum growth medium. | factors affecting growth and monoclonal antibody production in vitro by a mouse-mouse hybridoma cell line have been investigated in a series of studies. the goal was to maximize antibody yields and demonstrate that antibodies can be produced efficiently on a large-scale in fermentors. this initial report describes (i) development of a radial immunodiffusion assay for accurate determination of antibody levels in culture, (ii) a culture medium formulation that allowed for reduction in the amount o ... | 1986 | 3080528 |
| role of galactosyltransferase activity in phage sensitivity and nodulation competitiveness of rhizobium meliloti. | a stock culture of rhizobium meliloti 102f51 contains colonies of two distinct phenotypes (handelsman et al., j. bacteriol. 157:703-707, 1984); one colony type is agglutinated by high dilutions of the alfalfa agglutinin, is sensitive to phage f20, and is resistant to phage 16b, and the other is agglutinated only by low dilutions of the alfalfa agglutinin, is resistant to phage f20, and is sensitive to phage 16b. cells of the latter phenotype have an inner-membrane-bound galactosyltransferase act ... | 1986 | 3082851 |
| plant phenolic compounds induce expression of the agrobacterium tumefaciens loci needed for virulence. | the virulence loci of agrobacterium tumefaciens are a set of linked transcriptional units that play an essential role in the early stages of plant tumorigenesis. these loci are induced upon cocultivation of the bacteria with plant cells. seven phenolic compounds that are widely distributed among the angiosperm plants--catechol, gallic acid, pyrogallic acid, p-hydroxybenzoic acid, protocatechuic acid, beta-resorcylic acid, and vanillin--are able to induce the expression of the virulence loci. the ... | 1986 | 3085219 |
| activation of the bradyrhizobium japonicum nifh and nifdk operons is dependent on promoter-upstream dna sequences. | previous analysis of b. japonicum nifh'- and nifd'-'lacz translational fusions showed that these promoters could be activated by the k. pneumoniae nifa plus the e. coli ntra gene products. to study the functions of the dna 5' to these promoters, plasmids carrying deletions in this region were constructed and analyzed in vivo in a heterologous system consisting of an e. coli (ntra+) background with a plasmid that constitutively expresses the k. pneumoniae nifa gene. activation of the b. japonicum ... | 1986 | 3086837 |
| agrobacterium ti and ri plasmids specify enzymic lactonization of mannopine to agropine. | a novel enzymic activity, responsible for the conversion of mannopine to agropine by lactonization, has been identified in agrobacterium strains. this activity is encoded by octopine-type and agropine-type ti or ri plasmids, and is inducible by mannopine and agropine. in crude extracts it is stable for long periods and can be used for preparative synthesis of agropine from mannopine. the physiological role of this activity is not understood. however, it is probably involved in degradation of opi ... | 1986 | 3098912 |
| klebsiella pneumoniae nif-lac fusions are expressed in agrobacterium tumefaciens c58. | plasmids containing hybrid genes, in which different klebsiella pneumoniae nif (nitrogen-fixation) promoters were fused with the structural part of the escherichia coli lac operon, were introduced into a double auxotrophic derivative of agrobacterium tumefaciens c58. a study of their expression in the new host was made simple by the inherent inability of a. tumefaciens c58 to produce beta-galactosidase unless provided with the wild-type lac operon of e. coli. as shown by quantitative measurement ... | 1987 | 3108628 |
| visual assays of transformation in plant cells. | we discuss the utility of visual assays for the expression of genes introduced into plant cells. such assays are valuable for both transient and stable gene expression studies. we review the properties of three visual assays that are already in use or under development phase for maize and other cereal crops. these assays depend on the expression of beta-galactosidase, luciferase, or structural genes required for anthocyanin pigment biosynthesis. | 1987 | 3109371 |
| expression vectors based on the agrobacterium rhizogenes ri plasmid transformation system. | this article describes several new expression vectors that capitalize on the ability of agrobacterium rhizogenes to transfer dna from its ri plasmid to the plant nuclear genome. the intermediate vectors described include an expression cassette based on one of the three following promoters: the nopaline synthase promoter, or the cauliflower mosaic virus (camv) promoters responsible for transcription of either the 19s or 35s camv rna. the termination and polyadenylation signals are either from the ... | 1987 | 3111548 |
| genetic analysis of mannityl opine catabolism in octopine-type agrobacterium tumefaciens strain 15955. | the genetic organization of functions responsible for mannityl opine catabolism of the ti plasmid of agrobacterium tumefaciens strain 1,5955 was investigated. a partial hindiii digest of pti1,5955 was cloned into a broad host range cosmid and the clones obtained were tested for ability to confer mannityl opine degradation upon agrobacterium. inserts containing genes for catabolism of mannopinic acid, mannopine, agropine, and agropinic acid were obtained, spanning a segment of 43 kb on the ti pla ... | 1987 | 3112522 |
| microbes in or near field-testing. | | 1987 | 3114880 |
| analysis of lectin binding by bradyrhizobium japonicum strains grown on nitrocellulose filters using peroxidase-labeled lectin. | a procedure was developed to assess the ability of wild-type and mutant strains of bradyrhizobium japonicum to bind soybean lectin. the lectin-binding ability of bacteria grown on nitrocellulose filters was determined using peroxidase-labeled soybean lectin. the assay produced clear differences between strains known to be unable to bind soybean lectin and those which can. the assay gave results identical to those of the fluorescein isothiocyanate-soybean lectin-binding assay of t. v. bhuvaneswar ... | 1987 | 3118739 |
| characterization of the rhizobium leguminosarum genes nodlmn involved in efficient host-specific nodulation. | three nodulation genes, nodl, nodm and nodn, were isolated from rhizobium leguminosarum and their dna sequences were determined. the three genes are in the same orientation as the previously described nodfe genes and the predicted molecular weights of their products are 20,105 (nodl), 65,795 (nodm) and 18,031 (nodn). analysis of gene regulation using operon fusions showed that nodl, nodm and nodn are induced in response to flavanone molecules and that this induction is nodd-dependent. in additio ... | 1988 | 3132583 |
| hydrogen metabolism in rhizobium: energetics, regulation, enzymology and genetics. | | 1988 | 3132815 |
| escherichia coli lacz gene as a biochemical and histochemical marker in plant cells. | several lacz chimeric genes were constructed by fusing the truncated lacz sequence of escherichia coli to n-terminal sequences of few other genes. promoters used to direct expression of the chimeric genes were the promoter for 35s rna of cauliflower mosaic virus (p35s) as well as those of the small subunit gene of ribulose bisphosphate carboxylase and the octopine synthase gene. these constructs were introduced into tobacco cells using a ti plasmid of agrobacterium tumefaciens, and beta-galactos ... | 1988 | 3138164 |
| differentiation of some gram-negative glucose nonfermenting bacteria using miniaturized carbon sources assimilation tests. | in water and soil the gram-negative nonfermenting bacteria play an important role in the biological mineralization process. to improve the methods for species differentiation of these heterogenous bacterial group, a total of 481 reference strains of gram-negative glucose nonfermenting bacteria belonging to the genera pseudomonas, alcaligenes, bordetella, agrobacterium, moraxella, acinetobacter, flavobacterium and some cdc groups have been investigated for their ability to utilize 42 different ca ... | 1988 | 3142164 |
| role of sublethal injury in decline of bacterial populations in lake water. | following their addition to lake water, the populations of escherichia coli and of antibiotic-resistant strains of pseudomonas fluorescens, agrobacterium tumefaciens, micrococcus flavus, rhizobium meliloti, and klebsiella pneumoniae declined rapidly, as determined by counting on media containing antibacterial compounds. the estimates of population sizes were occasionally higher if procedures were used that permitted possible resuscitation of injured cells. no resuscitation procedure yielded cons ... | 1988 | 3145714 |
| cloning and characterization of a novel beta-galactosidase-coding gene from rhizobium meliloti. | the rhizobium meliloti (rm) lacz gene provides a convenient model to investigate patterns of gene regulation in these agronomically important bacteria. a gene encoding beta-galactosidase (beta gal) activity was cloned from r. meliloti by complementing a lactose-negative escherichia coli mutant. a series of sau3a subclones was generated in pbr322, and the coding region for the beta gal-coding gene was localized to a 2.4-kb core fragment. in e. coli 'maxicells', these lacz subclones produced a 79- ... | 1988 | 3145908 |
| evidence for a role of a vicinal dithiol in the transport of gamma-butyrobetaine in agrobacterium sp. | an agrobacterium sp. isolated from soil is able to use gamma-butyrobetaine as its sole source of carbon and nitrogen. the involvement of thiol groups for active transport of gamma-butyrobetaine was investigated by use of the thiol alkylating reagent n-ethylmaleimide (nem) and the dithiol specific reagent phenylarsine oxide (pao). both reagents strongly inhibited gamma-butyrobetaine uptake, but also induced the release of the accumulated substrate, suggesting that the transport system either cont ... | 1988 | 3148329 |
| agrobacterium rhizogenes as a vector for transforming higher plants. | agrobacterium rhizogenes is a natural vector for genetically transforming higher plants. a protocol is given for using this system to insert foreign genes into host plants. improvements currently being developed in several laboratories are also discussed. | 1987 | 3153165 |
| characterization of adenosine triphosphatase from rhizobium japonicum. | | 1985 | 3161817 |
| identification of cytoplasmic nodule-associated forms of malate dehydrogenase involved in the symbiosis between rhizobium leguminosarum and pisum sativum. | the malate dehydrogenase activity (ec 1.1.1.37), present in the cytoplasm of pisum sativum root nodules, can be separated by ion-exchange chromatography into four different fractions. malate dehydrogenase activity present in the cytoplasm of roots elutes mainly as a single peak. during nodule development an increase in malate dehydrogenase activity per gram of material was observed. this increase occurred concomitantly with the increase in nitrogenase activity. the kinetic properties of the sepa ... | 1988 | 3162212 |
| relative strengths of the 35s cauliflower mosaic virus, 1', 2', and nopaline synthase promoters in transformed tobacco sugarbeet and oilseed rape callus tissue. | the 35s promoter of cauliflower mosaic virus and promoters from the nopaline synthase, 1' and 2' genes of agrobacterium tumefaciens t-dna were fused to the bacterial octopine synthase and chitinase gene coding regions. these chimaeric gene constructions were introduced into tobacco, sugarbeet and oilseed rape cells and their relative levels of expression measured by primer extension analysis of rna isolated from pooled populations of stably transformed calli. in tobacco callus, the 35s promoter ... | 1988 | 3163765 |
| expression of tandem gene fusions in transgenic tobacco plants. | we have studied the expression of four sets of tandem gene fusions in transgenic tobacco plants. this was to determine if the problem of between-transformant variability in expression of introduced genes could be overcome by using a linked reference gene as a co-ordinately expressed control. tandem gene fusions containing identical 5' flanking regions (ssu301-ocs with either ssu301-cat or ssu301-ssu911) were not co-ordinately expressed in the transgenic tobacco plants whereas the tandem gene fus ... | 1988 | 3166132 |
| rola locus of the ri plasmid directs developmental abnormalities in transgenic tobacco plants. | plants containing the left t-dna (tl) of agrobacterium rhizogenes show a variety of developmental abnormalities that include severely wrinkled leaves, loss of apical dominance, reduced geotropism of roots, reduced internode distances, and floral hyperstyly. the tl-dna also affects the morphology of tumor tissue at the site of inoculation on kalanchoe diagremontiana leaves. single mutations at four loci of the tl-dna (rola, rolb, rolc, and rold) are known to affect tumor morphology on k. diagremo ... | 1988 | 3166443 |
| phosphoglycerol substituents present on the cyclic beta-1,2-glucans of rhizobium meliloti 1021 are derived from phosphatidylglycerol. | the synthesis of periplasmic cyclic beta-1,2-glucans is a property unique to species of the family rhizobiaceae. for this reason, it is generally believed that these molecules may play an important role in the plant infection process. in the present study, we determined that the cyclic beta-1,2-glucans produced by rhizobium meliloti 1021 were predominantly anionic in character and contained both phosphoglycerol and succinic acid substituents. in addition, we demonstrated that phosphatidylglycero ... | 1988 | 3170478 |
| flavonoids in the rhizobium-legume symbiosis. | | 1988 | 3174708 |
| isolation and characterization of root nodule proteins from lupin. | a group of root nodule-specific plant proteins (nodulins) has been isolated from yellow lupin (lupinus luteus) by immunoaffinity chromatography. the cytoplasmic nodule protein extract was initially enriched in nodulins on a column with immobilized igg fraction. it was then purified by chromatography on sepharose 4b - bound igg against uninfected root proteins and finally on sepharose 4b - bound igg against rhizobium lupini proteins. rocket immunoelectrophoresis showed that the nodulin preparatio ... | 1988 | 3176786 |
| deoxyribonuclease i sensitivity of the t-dna ipt gene is associated with gene expression. | we have analyzed the chromatin structure of the t-dna isopentenyl transferase gene, ipt, in four nicotiana tabacum crown gall tumor lines. these four transformed lines contain identical t-dna inserts and are derivatives of a single clone that did not exhibit any tumorous properties and contained a highly methylated, nonexpressed copy of t-dna. one of the derivatives also does not exhibit tumorous properties, and the t-dna of this line is not expressed. the other three lines have reverted to tumo ... | 1988 | 3179274 |
| role of iron in the enhancement by agrobacterium tumefaciens infection in mice. | microorganisms require iron for their growth and usually compete with their host for available iron from the system. iron supplementation to host causes an increase of available iron both to host and to potential microbial invaders and favours the latter more than the former as the bacteria release siderophores which are responsible for iron transport mechanism. in view of this observation a study was done to deal with the distribution of storage and injected iron given as an overload within a p ... | 1988 | 3181831 |
| purification and properties of the gamma-butyrobetaine-binding protein from an agrobacterium sp. | a binding protein for gamma-butyrobetaine was purified from osmotic shock fluid of an agrobacterium sp. it was a monomeric protein with an apparent molecular weight of 52,000 or 53,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, respectively. the isoelectric point was 4.3, as determined by isoelectric focusing. amino acid analysis of the protein showed that asx and glx were predominant components and that the protein contained no cysteine. the d ... | 1988 | 3182728 |
| isolation and characterization of azospirillum brasilense loci that correct rhizobium meliloti exob and exoc mutations. | the occurrence in azospirillum brasilense of genes that code for exopolysaccharide (eps) synthesis was investigated through complementation studies of rhizobium meliloti exo- mutants. these mutants are deficient in the synthesis of the major acidic eps of rhizobium species and form empty, non-nitrogen-fixing root nodules on alfalfa (j. a. leigh, e. r. signer, and g. c. walker, proc. natl. acad. sci. usa 82:6231-6235, 1985). we demonstrated that the exoc mutation of r. meliloti could be corrected ... | 1988 | 3182731 |
| symbiotic phenotypes of auxotrophic mutants of rhizobium meliloti 104a14. | auxotrophic mutants of rhizobium meliloti 104a14 were isolated using nitrous acid mutagenesis followed by penicillin enrichment. mutants in ornithine transcarbamylase, argininosuccinate synthetase or serine-glycine biosynthesis formed nitrogen-fixing (fix-nodules on the roost of alfalfa (medicago sativa). mutants with defects in ornithine, pyrimidine, purine, asparagine, leucine, methionine or tyrosine biosynthesis, in one-carbon metabolism or in carbamoylphosphate synthetase formed nodules but ... | 1988 | 3183624 |
| organization and characterization of the vircd genes from agrobacterium rhizogenes. | we have precisely localized virulent (vir) genes of the hairy root-inducing plasmid pria4b on the basis of sequence similarity with the tumor-inducing plasmid ptia6nc, and shown that the overall organizations of vir genes in both plasmids are fairly analogous, although sizes and spacer lengths in some genes differ from each other. among the vir genes thus mapped, the virc and vird loci were characterized in detail. transposon insertions in vird led to loss of tumorigenicity on kalanchoe stems an ... | 1988 | 3185501 |
| conservation of nif sequences in frankia. | southern blots of frankia total dnas were hybridized with nifhdk probes from rhizobium meliloti, klebsiella pneumoniae and frankia strain arl3. differences between strains were noted in the size of the hybridizing restriction fragments. these differences were more pronounced among elaeagnus-compatible strains than among alnus- or casuarina-compatible strains. gene banks constructed for frankia strains eun1f, hrn18a, ced and acon24d were used to isolate nif-hybridizing restriction fragments for s ... | 1988 | 3185502 |
| storage of competent cells for agrobacterium transformation. | | 1988 | 3186459 |
| role of the overdrive sequence in t-dna border cleavage in agrobacterium. | the t-dna of the ti plasmid of agrobacterium is flanked by 25-base-pair imperfect direct repeats that are required in cis for transfer to the genome of the plant host. another sequence, designated overdrive, is located adjacent to the right-border repeats and functions in cis to enhance tumor formation. we have examined the effect of the overdrive sequence on the early steps in t-dna processing. we report here that overdrive greatly enhances cleavage by the site-specific endonuclease in agrobact ... | 1988 | 3186745 |
| selective quantitative determination of tobramycin from fermentation broth. | a derivative of rhizobium meliloti 41 was constructed (strain gy654) which was resistant to apramycin and kanamycin but remained sensitive to tobramycin. strain gy654 selectively measures tobramycin and 6"-o-carbamoyltobramycin in the presence of apramycin, kanamycin and 6"-o-carbamoylkanamycin b, therefore it is suitable for the rapid quantitation of 6"-o-carbamoyltobramycin and tobramycin in fermentation broths of streptomyces tenebrarius and solvents containing antibiotic mixtures. | 1988 | 3188834 |
| molybdate transport by bradyrhizobium japonicum bacteroids. | bacteroid suspensions of bradyrhizobium japonicum usda 136 isolated from soybeans grown in mo-deficient conditions were able to transport molybdate at a nearly constant rate for up to 1 min. the apparent km for molybdate was 0.1 microm, and the vmax was about 5 pmol/min per mg (dry weight) of bacteroid. supplementation of bacteroid suspensions with oxidizable carbon sources did not markedly increase molybdate uptake rates. anaerobically isolated bacteroids accumulated twice as much mo in 1 h as ... | 1988 | 3192511 |
| glycine betaine allows enhanced induction of the agrobacterium tumefaciens vir genes by acetosyringone at low ph. | we established growth conditions for efficient induction of the vir genes of agrobacterium tumefaciens by acetosyringone. optimal induction was attained at a ph below 5.2 in an ab minimal medium-derived high-osmotic-strength medium containing glycine betaine. this natural osmoprotectant accelerated the adaptation of the bacteria to these conditions. we established the kinetics of induction for virb, vird, vire, and virg by using lacz fusions, and we found that the virb mutant strain could not ad ... | 1988 | 3192516 |
| vir-induced recombination in agrobacterium. physical characterization of precise and imprecise t-circle formation. | induction of ti plasmid virulence (vir) gene expression during the early stages of plant cell transformation by agrobacterium tumefaciens initiates the generation of several t-dna-associated molecular events: (1) site-specific nicks at t-dna border sequences (border nicks); (2) free, unipolar, linear, single-stranded t-dna copies (t-strands); and (3) double-stranded, circular t-dna molecules (t-circles). the first two t-dna products have been detected in a. tumefaciens, while t-circles have only ... | 1988 | 3199438 |
| effects of aldicarb on the biochemical composition of rhizobium meliloti. | | 1988 | 3207910 |
| comparison of the chemotactic behaviour of rhizobium leguminosarum with and without the nodulation plasmid. | the chemotactic behaviour of a strain of rhizobium leguminosarum biovar viciae was investigated. the flavanoids apigenin and naringenin, inducers of transcription of the nodulation (nod) genes, were both potent attractants but hesperitin, another flavone nod gene inducer, was not. the response of strains containing the sym plasmid prl1jl to apigenin and naringenin was significantly greater than the response of a strain cured of the plasmid, although both strains gave a positive response. additio ... | 1988 | 3210967 |
| genetic diversity and relationships among isolates of rhizobium leguminosarum biovar phaseoli. | fifty-one isolates of rhizobium leguminosarum biovar phaseoli from various geographic and ecological sources, largely in mexico, were characterized by the electrophoretic mobilities of 15 metabolic enzymes, and 46 distinctive multilocus genotypes (electrophoretic types [ets]) were distinguished on the basis of allele profiles at the enzyme loci. mean genetic diversity per enzyme locus among the 46 ets was 0.691, the highest value yet recorded for any species of bacterium. the occurrence of stron ... | 1988 | 3214160 |
| characterization of the fixabc region of azorhizobium caulinodans ors571 and identification of a new nitrogen fixation gene. | the fast growing strain, azorhizobium caulinodans ors571, isolated from stem nodules of the tropical legume sesbania rostrata, can grow in the free-living state at the expense of molecular nitrogen. five point mutants impaired in nitrogen fixation in the free-living state have been complemented by a plasmid containing the cloned fix-abc region of strain ors571. genetic analysis of the mutants showed that one was impaired in fixc, one in fixa and the three others in a new gene, located upstream f ... | 1988 | 3216855 |
| respiration supported nitrogenase activity of isolated rhizobium meliloti bacteroids. | bacteroids having a high level of respiration-supported nitrogenase activity were isolated from nitrogen-fixing alfalfa root nodules. gentle maceration under anaerobic conditions in the presence of sodium succinate and a fatty acid scavenging agent were employed in this method. a large proportion of isolated bacteroids retained a triple membrane structure as shown by transmission electron microscopy. dicarboxylic acids of the tca cycle (malate, fumarate, succinate), but not glutamate or aspartat ... | 1988 | 3220879 |
| growth dependent enzymatic profiles of some gram-negative nonfermentative bacteria of clinical significance. | a total of 734 strains of gram-negative nonfermentative bacteria (46 species and biochemically defined groups) of the genera pseudomonas, alcaligenes, bordetella, moraxella, acinetobacter, agrobacterium, and flavobacterium were investigated for their ability to hydrolyze 25 different chromogenic substrates. all tests were carried out in growth-stimulating media. results were read photometrically and evaluated automatically following a 24-h incubation. many of the 46 different species and biochem ... | 1988 | 3223128 |