Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Jul 1997; 179(14): 4464–4472.
PMCID: PMC179280

Characterization of the alginate biosynthetic gene cluster in Pseudomonas syringae pv. syringae.


Alginate, a copolymer of D-mannuronic acid and L-guluronic acid, is produced by a variety of pseudomonads, including Pseudomonas syringae. Alginate biosynthesis has been most extensively studied in P. aeruginosa, and a number of structural and regulatory genes from this species have been cloned and characterized. In the present study, an alginate-defective (Alg-) mutant of P. syringae pv. syringae FF5 was shown to contain a Tn5 insertion in algL, a gene encoding alginate lyase. A cosmid clone designated pSK2 restored alginate production to the algL mutant and was shown to contain homologs of algD, alg8, alg44, algG, algX (alg60), algL, algF, and algA. The order and arrangement of the structural gene cluster were virtually identical to those previously described for P. aeruginosa. Complementation analyses, however, indicated that the structural gene clusters in P. aeruginosa and P. syringae were not functionally interchangeable when expressed from their native promoters. A region upstream of the algD gene in P. syringae pv. syringae was shown to activate the transcription of a promoterless glucuronidase (uidA) gene and indicated that transcription initiated upstream of algD as described for P. aeruginosa. Transcription of the algD promoter from P. syringae FF5 was significantly higher at 32 degrees C than at 18 or 26 degrees C and was stimulated when copper sulfate or sodium chloride was added to the medium. Alginate gene expression was also stimulated by the addition of the nonionic solute sorbitol, indicating that osmolarity is a signal for algD expression in P. syringae FF5.

Full Text

The Full Text of this article is available as a PDF (186K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barta TM, Kinscherf TG, Willis DK. Regulation of tabtoxin production by the lemA gene in Pseudomonas syringae. J Bacteriol. 1992 May;174(9):3021–3029. [PMC free article] [PubMed]
  • Bender CL, Cooksey DA. Indigenous plasmids in Pseudomonas syringae pv. tomato: conjugative transfer and role in copper resistance. J Bacteriol. 1986 Feb;165(2):534–541. [PMC free article] [PubMed]
  • Bender CL, Young SA, Mitchell RE. Conservation of Plasmid DNA Sequences in Coronatine-Producing Pathovars of Pseudomonas syringae. Appl Environ Microbiol. 1991 Apr;57(4):993–999. [PMC free article] [PubMed]
  • Berry A, DeVault JD, Chakrabarty AM. High osmolarity is a signal for enhanced algD transcription in mucoid and nonmucoid Pseudomonas aeruginosa strains. J Bacteriol. 1989 May;171(5):2312–2317. [PMC free article] [PubMed]
  • Boyd A, Ghosh M, May TB, Shinabarger D, Keogh R, Chakrabarty AM. Sequence of the algL gene of Pseudomonas aeruginosa and purification of its alginate lyase product. Gene. 1993 Sep 6;131(1):1–8. [PubMed]
  • Campos M, Martínez-Salazar JM, Lloret L, Moreno S, Núez C, Espín G, Soberón-Chávez G. Characterization of the gene coding for GDP-mannose dehydrogenase (algD) from Azotobacter vinelandii. J Bacteriol. 1996 Apr;178(7):1793–1799. [PMC free article] [PubMed]
  • Chitnis CE, Ohman DE. Cloning of Pseudomonas aeruginosa algG, which controls alginate structure. J Bacteriol. 1990 Jun;172(6):2894–2900. [PMC free article] [PubMed]
  • Chitnis CE, Ohman DE. Genetic analysis of the alginate biosynthetic gene cluster of Pseudomonas aeruginosa shows evidence of an operonic structure. Mol Microbiol. 1993 May;8(3):583–593. [PubMed]
  • Chu L, May TB, Chakrabarty AM, Misra TK. Nucleotide sequence and expression of the algE gene involved in alginate biosynthesis by Pseudomonas aeruginosa. Gene. 1991 Oct 30;107(1):1–10. [PubMed]
  • Darzins A, Chakrabarty AM. Cloning of genes controlling alginate biosynthesis from a mucoid cystic fibrosis isolate of Pseudomonas aeruginosa. J Bacteriol. 1984 Jul;159(1):9–18. [PMC free article] [PubMed]
  • Darzins A, Frantz B, Vanags RI, Chakrabarty AM. Nucleotide sequence analysis of the phosphomannose isomerase gene (pmi) of Pseudomonas aeruginosa and comparison with the corresponding Escherichia coli gene manA. Gene. 1986;42(3):293–302. [PubMed]
  • Darzins A, Wang SK, Vanags RI, Chakrabarty AM. Clustering of mutations affecting alginic acid biosynthesis in mucoid Pseudomonas aeruginosa. J Bacteriol. 1985 Nov;164(2):516–524. [PMC free article] [PubMed]
  • Deretic V, Gill JF, Chakrabarty AM. Pseudomonas aeruginosa infection in cystic fibrosis: nucleotide sequence and transcriptional regulation of the algD gene. Nucleic Acids Res. 1987 Jun 11;15(11):4567–4581. [PMC free article] [PubMed]
  • Ertesvåg H, Doseth B, Larsen B, Skjåk-Braek G, Valla S. Cloning and expression of an Azotobacter vinelandii mannuronan C-5-epimerase gene. J Bacteriol. 1994 May;176(10):2846–2853. [PMC free article] [PubMed]
  • Ertesvåg H, Høidal HK, Hals IK, Rian A, Doseth B, Valla S. A family of modular type mannuronan C-5-epimerase genes controls alginate structure in Azotobacter vinelandii. Mol Microbiol. 1995 May;16(4):719–731. [PubMed]
  • Fett WF, Osman SF, Dunn MF. Characterization of exopolysaccharides produced by plant-associated fluorescent pseudomonads. Appl Environ Microbiol. 1989 Mar;55(3):579–583. [PMC free article] [PubMed]
  • Fett WF, Osman SF, Fishman ML, Siebles TS. Alginate production by plant-pathogenic pseudomonads. Appl Environ Microbiol. 1986 Sep;52(3):466–473. [PMC free article] [PubMed]
  • Fett WF, Wijey C, Lifson ER. Occurrence of alginate gene sequences among members of the pseudomonad rRNA homology groups I-IV. FEMS Microbiol Lett. 1992 Dec 1;78(2-3):151–157. [PubMed]
  • Fialho AM, Zielinski NA, Fett WF, Chakrabarty AM, Berry A. Distribution of alginate gene sequences in the Pseudomonas rRNA homology group I-Azomonas-Azotobacter lineage of superfamily B procaryotes. Appl Environ Microbiol. 1990 Feb;56(2):436–443. [PMC free article] [PubMed]
  • Figurski DH, Helinski DR. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. [PMC free article] [PubMed]
  • Franklin MJ, Chitnis CE, Gacesa P, Sonesson A, White DC, Ohman DE. Pseudomonas aeruginosa AlgG is a polymer level alginate C5-mannuronan epimerase. J Bacteriol. 1994 Apr;176(7):1821–1830. [PMC free article] [PubMed]
  • Franklin MJ, Ohman DE. Identification of algF in the alginate biosynthetic gene cluster of Pseudomonas aeruginosa which is required for alginate acetylation. J Bacteriol. 1993 Aug;175(16):5057–5065. [PMC free article] [PubMed]
  • Franklin MJ, Ohman DE. Identification of algI and algJ in the Pseudomonas aeruginosa alginate biosynthetic gene cluster which are required for alginate O acetylation. J Bacteriol. 1996 Apr;178(8):2186–2195. [PMC free article] [PubMed]
  • Goldberg JB, Gorman WL, Flynn JL, Ohman DE. A mutation in algN permits trans activation of alginate production by algT in Pseudomonas species. J Bacteriol. 1993 Mar;175(5):1303–1308. [PMC free article] [PubMed]
  • Hershberger CD, Ye RW, Parsek MR, Xie ZD, Chakrabarty AM. The algT (algU) gene of Pseudomonas aeruginosa, a key regulator involved in alginate biosynthesis, encodes an alternative sigma factor (sigma E). Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7941–7945. [PMC free article] [PubMed]
  • Jones JD, Gutterson N. An efficient mobilizable cosmid vector, pRK7813, and its use in a rapid method for marker exchange in Pseudomonas fluorescens strain HV37a. Gene. 1987;61(3):299–306. [PubMed]
  • Keen NT, Tamaki S, Kobayashi D, Trollinger D. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988 Oct 15;70(1):191–197. [PubMed]
  • Kidambi SP, Sundin GW, Palmer DA, Chakrabarty AM, Bender CL. Copper as a signal for alginate synthesis in Pseudomonas syringae pv. syringae. Appl Environ Microbiol. 1995 Jun;61(6):2172–2179. [PMC free article] [PubMed]
  • KING EO, WARD MK, RANEY DE. Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med. 1954 Aug;44(2):301–307. [PubMed]
  • Konyecsni WM, Deretic V. Broad-host-range plasmid and M13 bacteriophage-derived vectors for promoter analysis in Escherichia coli and Pseudomonas aeruginosa. Gene. 1988 Dec 30;74(2):375–386. [PubMed]
  • Leitão JH, Fialho AM, Sá-Correia I. Effects of growth temperature on alginate synthesis and enzymes in Pseudomonas aeruginosa variants. J Gen Microbiol. 1992 Mar;138(3):605–610. [PubMed]
  • Lloret L, Barreto R, León R, Moreno S, Martínez-Salazar J, Espín G, Soberón-Chávez G. Genetic analysis of the transcriptional arrangement of Azotobacter vinelandii alginate biosynthetic genes: identification of two independent promoters. Mol Microbiol. 1996 Aug;21(3):449–457. [PubMed]
  • Maharaj R, May TB, Wang SK, Chakrabarty AM. Sequence of the alg8 and alg44 genes involved in the synthesis of alginate by Pseudomonas aeruginosa. Gene. 1993 Dec 22;136(1-2):267–269. [PubMed]
  • Martin DW, Schurr MJ, Yu H, Deretic V. Analysis of promoters controlled by the putative sigma factor AlgU regulating conversion to mucoidy in Pseudomonas aeruginosa: relationship to sigma E and stress response. J Bacteriol. 1994 Nov;176(21):6688–6696. [PMC free article] [PubMed]
  • May TB, Chakrabarty AM. Isolation and assay of Pseudomonas aeruginosa alginate. Methods Enzymol. 1994;235:295–304. [PubMed]
  • May TB, Chakrabarty AM. Pseudomonas aeruginosa: genes and enzymes of alginate synthesis. Trends Microbiol. 1994 May;2(5):151–157. [PubMed]
  • Monday SR, Schiller NL. Alginate synthesis in Pseudomonas aeruginosa: the role of AlgL (alginate lyase) and AlgX. J Bacteriol. 1996 Feb;178(3):625–632. [PMC free article] [PubMed]
  • Rehm BH. The Azotobacter vinelandii gene algJ encodes an outer-membrane protein presumably involved in export of alginate. Microbiology. 1996 Apr;142(Pt 4):873–880. [PubMed]
  • Rehm BH, Ertesvåg H, Valla S. A new Azotobacter vinelandii mannuronan C-5-epimerase gene (algG) is part of an alg gene cluster physically organized in a manner similar to that in Pseudomonas aeruginosa. J Bacteriol. 1996 Oct;178(20):5884–5889. [PMC free article] [PubMed]
  • Rich JJ, Willis DK. A single oligonucleotide can be used to rapidly isolate DNA sequences flanking a transposon Tn5 insertion by the polymerase chain reaction. Nucleic Acids Res. 1990 Nov 25;18(22):6673–6676. [PMC free article] [PubMed]
  • Schiller NL, Monday SR, Boyd CM, Keen NT, Ohman DE. Characterization of the Pseudomonas aeruginosa alginate lyase gene (algL): cloning, sequencing, and expression in Escherichia coli. J Bacteriol. 1993 Aug;175(15):4780–4789. [PMC free article] [PubMed]
  • Shankar S, Ye RW, Schlictman D, Chakrabarty AM. Exopolysaccharide alginate synthesis in Pseudomonas aeruginosa: enzymology and regulation of gene expression. Adv Enzymol Relat Areas Mol Biol. 1995;70:221–255. [PubMed]
  • Shinabarger D, Berry A, May TB, Rothmel R, Fialho A, Chakrabarty AM. Purification and characterization of phosphomannose isomerase-guanosine diphospho-D-mannose pyrophosphorylase. A bifunctional enzyme in the alginate biosynthetic pathway of Pseudomonas aeruginosa. J Biol Chem. 1991 Feb 5;266(4):2080–2088. [PubMed]
  • Shinabarger D, May TB, Boyd A, Ghosh M, Chakrabarty AM. Nucleotide sequence and expression of the Pseudomonas aeruginosa algF gene controlling acetylation of alginate. Mol Microbiol. 1993 Sep;9(5):1027–1035. [PubMed]
  • Staskawicz BJ, Dahlbeck D, Keen NT. Cloned avirulence gene of Pseudomonas syringae pv. glycinea determines race-specific incompatibility on Glycine max (L.) Merr. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6024–6028. [PMC free article] [PubMed]
  • Sundin GW, Bender CL. Ecological and genetic analysis of copper and streptomycin resistance in Pseudomonas syringae pv. syringae. Appl Environ Microbiol. 1993 Apr;59(4):1018–1024. [PMC free article] [PubMed]
  • Van den Eede G, Deblaere R, Goethals K, Van Montagu M, Holsters M. Broad host range and promoter selection vectors for bacteria that interact with plants. Mol Plant Microbe Interact. 1992 May-Jun;5(3):228–234. [PubMed]
  • Wang SK, Sa'-Correia I, Darzins A, Chakrabarty AM. Characterization of the Pseudomonas aeruginosa alginate (alg) gene region II. J Gen Microbiol. 1987 Aug;133(8):2303–2314. [PubMed]
  • Xiao Y, Lu Y, Heu S, Hutcheson SW. Organization and environmental regulation of the Pseudomonas syringae pv. syringae 61 hrp cluster. J Bacteriol. 1992 Mar;174(6):1734–1741. [PMC free article] [PubMed]
  • Zielinski NA, Chakrabarty AM, Berry A. Characterization and regulation of the Pseudomonas aeruginosa algC gene encoding phosphomannomutase. J Biol Chem. 1991 May 25;266(15):9754–9763. [PubMed]
  • Zukowski MM, Gaffney DF, Speck D, Kauffmann M, Findeli A, Wisecup A, Lecocq JP. Chromogenic identification of genetic regulatory signals in Bacillus subtilis based on expression of a cloned Pseudomonas gene. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1101–1105. [PMC free article] [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...