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J Bacteriol. Sep 1997; 179(17): 5372–5379.
PMCID: PMC179406

Attachment of Agrobacterium tumefaciens to carrot cells and Arabidopsis wound sites is correlated with the presence of a cell-associated, acidic polysaccharide.

Abstract

An early step in crown gall tumor formation involves the attachment of Agrobacterium tumefaciens to host plant cells. A. tumefaciens C58::A205 (C58 attR) is a Tn3HoHo1 insertion mutant that was found to be avirulent on Bryophyllum daigremontiana and unable to attach to carrot suspension cells. The mutation mapped to an open reading frame encoding a putative protein of 247 amino acids which has significant homology to transacetylases from many bacteria. Biochemical analysis of polysaccharide extracts from wild-type strain C58 and the C58::A205 mutant showed that the latter was deficient in the production of a cell-associated polysaccharide. Anion-exchange chromatography followed by 1H nuclear magnetic resonance and gas chromatography-mass spectrometry analyses showed that the polysaccharide produced by strain C58 was an acetylated, acidic polysaccharide and that the polysaccharide preparation contained three sugars: glucose, glucosamine, and an unidentified deoxy-sugar. Application of the polysaccharide preparation from strain C58 to carrot suspension cells prior to inoculation with the bacteria effectively inhibited attachment of the bacteria to the carrot cells, whereas an identical preparation from strain C58::A205 had no inhibitory effect and did not contain the acidic polysaccharide. Similarly, preincubation of Arabidopsis thaliana root segments with the polysaccharide prevented attachment of strain C58 to that plant. This indicates that the acidic polysaccharide may play a role in the attachment of A. tumefaciens to host soma plant cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. [PubMed]
  • Baev N, Kondorosi A. Nucleotide sequence of the Rhizobium meliloti nodL gene located in locus n5 of the nod regulon. Plant Mol Biol. 1992 Feb;18(4):843–846. [PubMed]
  • Blumenkrantz N, Asboe-Hansen G. New method for quantitative determination of uronic acids. Anal Biochem. 1973 Aug;54(2):484–489. [PubMed]
  • Cangelosi GA, Hung L, Puvanesarajah V, Stacey G, Ozga DA, Leigh JA, Nester EW. Common loci for Agrobacterium tumefaciens and Rhizobium meliloti exopolysaccharide synthesis and their roles in plant interactions. J Bacteriol. 1987 May;169(5):2086–2091. [PMC free article] [PubMed]
  • Carlson RW, Sanders RE, Napoli C, Albersheim P. Host-Symbiont Interactions: III. Purification and Partial Characterization of Rhizobium Lipopolysaccharides. Plant Physiol. 1978 Dec;62(6):912–917. [PMC free article] [PubMed]
  • Chilton MD, Drummond MH, Merio DJ, Sciaky D, Montoya AL, Gordon MP, Nester EW. Stable incorporation of plasmid DNA into higher plant cells: the molecular basis of crown gall tumorigenesis. Cell. 1977 Jun;11(2):263–271. [PubMed]
  • Douglas CJ, Halperin W, Nester EW. Agrobacterium tumefaciens mutants affected in attachment to plant cells. J Bacteriol. 1982 Dec;152(3):1265–1275. [PMC free article] [PubMed]
  • Forsberg LS, Reuhs BL. Structural characterization of the K antigens from Rhizobium fredii USDA257: evidence for a common structural motif, with strain-specific variation, in the capsular polysaccharides of Rhizobium spp. J Bacteriol. 1997 Sep;179(17):5366–5371. [PMC free article] [PubMed]
  • Gish W, States DJ. Identification of protein coding regions by database similarity search. Nat Genet. 1993 Mar;3(3):266–272. [PubMed]
  • Gurlitz RH, Lamb PW, Matthysse AG. Involvement of Carrot Cell Surface Proteins in Attachment of Agrobacterium tumefaciens. Plant Physiol. 1987 Mar;83(3):564–568. [PMC free article] [PubMed]
  • Henikoff S, Henikoff JG. Automated assembly of protein blocks for database searching. Nucleic Acids Res. 1991 Dec 11;19(23):6565–6572. [PMC free article] [PubMed]
  • Nam J, Matthysse AG, Gelvin SB. Differences in susceptibility of Arabidopsis ecotypes to crown gall disease may result from a deficiency in T-DNA integration. Plant Cell. 1997 Mar;9(3):317–333. [PMC free article] [PubMed]
  • Jann K, Jann B. Biochemistry and expression of bacterial capsules. Biochem Soc Trans. 1991 Aug;19(3):623–628. [PubMed]
  • Kaneko T, Sato S, Kotani H, Tanaka A, Asamizu E, Nakamura Y, Miyajima N, Hirosawa M, Sugiura M, Sasamoto S, et al. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res. 1996 Jun 30;3(3):109–136. [PubMed]
  • Kiss E, Reuhs BL, Kim JS, Kereszt A, Petrovics G, Putnoky P, Dusha I, Carlson RW, Kondorosi A. The rkpGHI and -J genes are involved in capsular polysaccharide production by Rhizobium meliloti. J Bacteriol. 1997 Apr;179(7):2132–2140. [PMC free article] [PubMed]
  • Klein P, Kanehisa M, DeLisi C. The detection and classification of membrane-spanning proteins. Biochim Biophys Acta. 1985 May 28;815(3):468–476. [PubMed]
  • Matthysse AG. Role of bacterial cellulose fibrils in Agrobacterium tumefaciens infection. J Bacteriol. 1983 May;154(2):906–915. [PMC free article] [PubMed]
  • Matthysse AG. Initial interactions of Agrobacterium tumefaciens with plant host cells. Crit Rev Microbiol. 1986;13(3):281–307. [PubMed]
  • Matthysse AG. Characterization of nonattaching mutants of Agrobacterium tumefaciens. J Bacteriol. 1987 Jan;169(1):313–323. [PMC free article] [PubMed]
  • Matthysse AG, Holmes KV, Gurlitz RH. Elaboration of cellulose fibrils by Agrobacterium tumefaciens during attachment to carrot cells. J Bacteriol. 1981 Jan;145(1):583–595. [PMC free article] [PubMed]
  • Matthysse AG, Yarnall HA, Young N. Requirement for genes with homology to ABC transport systems for attachment and virulence of Agrobacterium tumefaciens. J Bacteriol. 1996 Sep;178(17):5302–5308. [PMC free article] [PubMed]
  • Parent R, Roy PH. The chloramphenicol acetyltransferase gene of Tn2424: a new breed of cat. J Bacteriol. 1992 May;174(9):2891–2897. [PMC free article] [PubMed]
  • Petrovics G, Putnoky P, Reuhs B, Kim J, Thorp TA, Noel KD, Carlson RW, Kondorosi A. The presence of a novel type of surface polysaccharide in Rhizobium meliloti requires a new fatty acid synthase-like gene cluster involved in symbiotic nodule development. Mol Microbiol. 1993 Jun;8(6):1083–1094. [PubMed]
  • Rende-Fournier R, Leclercq R, Galimand M, Duval J, Courvalin P. Identification of the satA gene encoding a streptogramin A acetyltransferase in Enterococcus faecium BM4145. Antimicrob Agents Chemother. 1993 Oct;37(10):2119–2125. [PMC free article] [PubMed]
  • Reuhs BL, Kim JS, Badgett A, Carlson RW. Production of cell-associated polysaccharides of Rhizobium fredii USDA205 is modulated by apigenin and host root extract. Mol Plant Microbe Interact. 1994 Mar-Apr;7(2):240–247. [PubMed]
  • Reuhs BL, Carlson RW, Kim JS. Rhizobium fredii and Rhizobium meliloti produce 3-deoxy-D-manno-2-octulosonic acid-containing polysaccharides that are structurally analogous to group II K antigens (capsular polysaccharides) found in Escherichia coli. J Bacteriol. 1993 Jun;175(11):3570–3580. [PMC free article] [PubMed]
  • Reuhs BL, Williams MN, Kim JS, Carlson RW, Côté F. Suppression of the Fix- phenotype of Rhizobium meliloti exoB mutants by lpsZ is correlated to a modified expression of the K polysaccharide. J Bacteriol. 1995 Aug;177(15):4289–4296. [PMC free article] [PubMed]
  • Robertson JL, Holliday T, Matthysse AG. Mapping of Agrobacterium tumefaciens chromosomal genes affecting cellulose synthesis and bacterial attachment to host cells. J Bacteriol. 1988 Mar;170(3):1408–1411. [PMC free article] [PubMed]
  • Stachel SE, An G, Flores C, Nester EW. A Tn3 lacZ transposon for the random generation of beta-galactosidase gene fusions: application to the analysis of gene expression in Agrobacterium. EMBO J. 1985 Apr;4(4):891–898. [PMC free article] [PubMed]
  • Swart S, Lugtenberg BJ, Smit G, Kijne JW. Rhicadhesin-mediated attachment and virulence of an Agrobacterium tumefaciens chvB mutant can be restored by growth in a highly osmotic medium. J Bacteriol. 1994 Jun;176(12):3816–3819. [PMC free article] [PubMed]
  • Swart S, Smit G, Lugtenberg BJ, Kijne JW. Restoration of attachment, virulence and nodulation of Agrobacterium tumefaciens chvB mutants by rhicadhesin. Mol Microbiol. 1993 Nov;10(3):597–605. [PubMed]
  • Tennigkeit J, Matzura H. Nucleotide sequence analysis of a chloramphenicol-resistance determinant from Agrobacterium tumefaciens and identification of its gene product. Gene. 1991 Feb 1;98(1):113–116. [PubMed]
  • Thomashow MF, Karlinsey JE, Marks JR, Hurlbert RE. Identification of a new virulence locus in Agrobacterium tumefaciens that affects polysaccharide composition and plant cell attachment. J Bacteriol. 1987 Jul;169(7):3209–3216. [PMC free article] [PubMed]
  • Tsai CM, Frasch CE. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. [PubMed]
  • Wagner VT, Matthysse AG. Involvement of a vitronectin-like protein in attachment of Agrobacterium tumefaciens to carrot suspension culture cells. J Bacteriol. 1992 Sep;174(18):5999–6003. [PMC free article] [PubMed]
  • WEISSBACH A, HURWITZ J. The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. J Biol Chem. 1959 Apr;234(4):705–709. [PubMed]
  • Whatley MH, Bodwin JS, Lippincott BB, Lippincott JA. Role of Agrobacterium cell envelope lipopolysaccharide in infection site attachment. Infect Immun. 1976 Apr;13(4):1080–1083. [PMC free article] [PubMed]
  • Whatley MH, Spiess LD. Role of bacterial lipopolysaccharide in attachment of agrobacterium to moss. Plant Physiol. 1977 Nov;60(5):765–766. [PMC free article] [PubMed]

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