Skip to main content
Springer Nature Link
Log in

Expression of heterologous xyloglucan xylosyltransferases in Arabidopsis to investigate their role in determining xyloglucan xylosylation substitution patterns

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Main conclusion

Putative XyG xylosyltransferases from Tropaeolum majus (nasturtium) and Solanum lycopersicum (tomato) homologous to characterized Arabidopsis genes were identified and shown to functionally complement Arabidopsis mutants lacking xyloglucan demonstrating they represent xyloglucan xylosyltransferases.

Xyloglucan is a major hemicellulose in the plant cell wall and is important for the structural organization of the wall. The fine structure of xyloglucan can vary dependent on plant species and tissue type. Most vascular seed-bearing plants including Arabidopsis thaliana and nasturtium (Tropaeolum majus) have a xyloglucan structure, in which three out of four backbone glucosyl-residues are substituted with xylosyl-residues. In contrast, the xyloglucan found in plants of the Solanaceae family, which includes tomato (Solanum lycopersicum), is typically less xylosylated with only two of the four backbone glucosyl-residues substituted with xylosyl-residues. To investigate the genetics of xyloglucan xylosylation, candidate xyloglucan xylosyltransferase genes (XXTs) homologous to known A. thaliana XXTs were cloned from nasturtium and tomato. These candidate XXTs were expressed in the A. thaliana xxt1/2 double and xxt1/2/5 triple mutant, whose walls lack detectable xyloglucan. Expression of the orthologs of XXT5 resulted in no detectable xyloglucan in the transgenic A. thaliana plants, consistent with a lack of xyloglucan in the A. thaliana xxt1/2 double mutant. However, transformation of both the tomato and nasturtium orthologs of AtXXT1 and AtXXT2 resulted in the production of xyloglucan with a xylosylation pattern similar to wild type A. thaliana indicating that both SlXXT2 and TmXXT2 likely have xylosyltransferase activity. As the expression of the SlXXT2 did not result in xyloglucan with a decreased xylosylation frequency found in tomato, this gene is not responsible for the unique xylosylation pattern found in the solanaceous plants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Ade CP, Bemm F, Dickson JM, Walter C, Harris PJ (2014) Family 34 glycosyltransferase (GT34) genes and proteins in Pinus radiata (radiata pine) and Pinus taeda (loblolly pine). Plant J 78(2):305–318

    Article  CAS  PubMed  Google Scholar 

  • Andrew IG, Little JWL (1997) A xyloglucan in etiolated seedlings of Pinus radiata. Phytochemistry 46:203–207

    Article  CAS  Google Scholar 

  • Bombarely A, Menda N, Tecle IY, Buels RM, Strickler S, Fischer-York T, Pujar A, Leto J, Gosselin J, Mueller LA (2011) The Sol Genomics Network (solgenomics.net): growing tomatoes using Perl. Nucleic Acids Res 39:D1149–D1155

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Buckeridge MS, Crombie HJ, Mendes CJ, Reid JS, Gidley MJ, Vieira CC (1997) A new family of oligosaccharides from the xyloglucan of Hymenaea courbaril L. (Leguminosae) cotyledons. Carbohydr Res 303:233–237

    Article  CAS  PubMed  Google Scholar 

  • Cavalier DM, Keegstra K (2006) Two xyloglucan xylosyltransferases catalyze the addition of multiple xylosyl residues to cellohexaose. J Biol Chem 281:34197–34207

    Article  CAS  PubMed  Google Scholar 

  • Cavalier DM, Lerouxel O, Neumetzler L, Yamauchi K, Reinecke A, Freshour G, Zabotina OA, Hahn MG, Burgert I, Pauly M, Raikhel NV, Keegstra K (2008) Disrupting two Arabidopsis thaliana xylosyltransferase genes results in plants deficient in xyloglucan, a major primary cell wall component. Plant Cell 20:1519–1537

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Chou YH, Pogorelko G, Zabotina OA (2012) Xyloglucan xylosyltransferases XXT1, XXT2, and XXT5 and the glucan synthase CSLC4 form Golgi-localized multiprotein complexes. Plant Physiol 159:1355–1366

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16(6):735–743

    Article  CAS  PubMed  Google Scholar 

  • Cocuron J-C, Lerouxel O, Drakakaki G, Alonso AP, Liepman AH, Keegstra K, Raikhel N, Wilkerson CG (2007) A gene from the cellulose synthase-like C family encodes a b-1,4 glucan synthase. Proc Natl Acad Sci USA 104:8550–8555

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Del Bem LE, Vincentz MG (2010) Evolution of xyloglucan-related genes in green plants. BMC Evol Biol 10:341

    Article  PubMed Central  PubMed  Google Scholar 

  • Edwards ME, Dickson CA, Chengappa S, Sidebottom C, Gidley MJ, Reid JS (1999) Molecular characterisation of a membrane-bound galactosyltransferase of plant cell wall matrix polysaccharide biosynthesis. Plant J 19(6):691–697

    Article  CAS  PubMed  Google Scholar 

  • Edwards ME, Choo TS, Dickson CA, Scott C, Gidley MJ, Reid JS (2004) The seeds of Lotus japonicus lines transformed with sense, antisense, and sense/antisense galactomannan galactosyltransferase constructs have structurally altered galactomannans in their endosperm cell walls. Plant Physiol 134:1153–1162

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Faik A, Price NJ, Raikhel NV, Keegstra K (2002) An Arabidopsis gene encoding an alpha-xylosyltransferase involved in xyloglucan biosynthesis. Proc Natl Acad Sci USA 99:7797–7802

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Fanutti C, Gidley MJ, Reid JS (1991) A xyloglucan-oligosaccharide-specific alpha-d-xylosidase or exo-oligoxyloglucan-alpha-xylohydrolase from germinated nasturtium (Tropaeolum majus L.) seeds : purification, properties and its interaction with a xyloglucan-specific endo-(1→4)-beta-d-glucanase and other hydrolases during storage-xyloglucan mobilisation. Planta 184:137–147

    Article  CAS  PubMed  Google Scholar 

  • Fry SC, Smith RC, Renwick KF, Martin DJ, Hodge SK, Matthews KJ (1992) Xyloglucan endotransglycosylase, a new wall-loosening enzyme activity from plants. Biochem J 282(Pt 3):821–828

    PubMed Central  CAS  PubMed  Google Scholar 

  • Fry SC, York WS, Albersheim P, Darvill A, Hayashi T, Joseleau JP, Kato Y, Lorences EP, Maclachlan GA, Mcneil M, Mort AJ, Reid JSG, Seitz HU, Selvendran RR, Voragen AGJ, White AR (1993) An unambiguous nomenclature for xyloglucan-derived oligosaccharides. Physiol Plantarum 89:1–3

    Article  CAS  Google Scholar 

  • Gibeaut DM, Pauly M, Bacic A, Fincher GB (2005) Changes in cell wall polysaccharides in developing barley (Hordeum vulgare) coleoptiles. Planta 221:729–738

    Article  CAS  PubMed  Google Scholar 

  • Gouy M, Guindon S, Gascuel O (2010) SeaView version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Mol Biol Evol 27:221–224

    Article  CAS  PubMed  Google Scholar 

  • Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 59:307–321

    Article  CAS  PubMed  Google Scholar 

  • Gunl M, Pauly M (2011) AXY3 encodes a alpha-xylosidase that impacts the structure and accessibility of the hemicellulose xyloglucan in Arabidopsis plant cell walls. Planta 233:707–719

    Article  PubMed Central  PubMed  Google Scholar 

  • Hantus S, Pauly M, Darvill AG, Albersheim P, York WS (1997) Structural characterization of novel l-galactose-containing oligosaccharide subunits of jojoba seed xyloglucans. Carbohydr Res 304:11–20

    Article  CAS  PubMed  Google Scholar 

  • Hanus J, Mazeau K (2006) The xyloglucan-cellulose assembly at the atomic scale. Biopolymers 82:59–73

    Article  CAS  PubMed  Google Scholar 

  • Hayashi T (1989) Xyloglucans in the primary-cell wall. Annu Rev Plant Phys 40:139–168

    Article  CAS  Google Scholar 

  • Hayashi T, Kaida R (2011) Functions of xyloglucan in plant cells. Mol Plant 4:17–24

    Article  CAS  PubMed  Google Scholar 

  • Hoffman M, Jia Z, Pena MJ, Cash M, Harper A, Blackburn AR 2nd, Darvill A, York WS (2005) Structural analysis of xyloglucans in the primary cell walls of plants in the subclass Asteridae. Carbohydr Res 340:1826–1840

    Article  CAS  PubMed  Google Scholar 

  • Hsieh YS, Harris PJ (2009) Xyloglucans of monocotyledons have diverse structures. Mol Plant 2:943–965

    Article  CAS  PubMed  Google Scholar 

  • Hsieh YSY, Harris PJ (2012) Structures of xyloglucans in primary cell walls of gymnosperms, monilophytes (ferns sensu lato) and lycophytes. Phytochemistry 79:87–101

    Article  CAS  PubMed  Google Scholar 

  • Jensen JK, Schultink A, Keegstra K, Wilkerson CG, Pauly M (2012) RNA-Seq analysis of developing nasturtium seeds (Tropaeolum majus): identification and characterization of an additional galactosyltransferase involved in xyloglucan biosynthesis. Mol Plant 5:984–992

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Jia Z, Qin Q, Darvill AG, York WS (2003) Structure of the xyloglucan produced by suspension-cultured tomato cells. Carbohydr Res 338:1197–1208

    Article  CAS  PubMed  Google Scholar 

  • Jia Z, Cash M, Darvill AG, York WS (2005) NMR characterization of endogenously O-acetylated oligosaccharides isolated from tomato (Lycopersicon esculentum) xyloglucan. Carbohydr Res 340:1818–1825

    Article  CAS  PubMed  Google Scholar 

  • Keegstra K, Cavalier D (2010) Glycosyltransferases of the GT34 and GT37 Families. Annu Plant Rev 41:235–249 (Wiley-Blackwell)

    Article  Google Scholar 

  • Lampugnani ER, Moller IE, Cassin A, Jones DF, Koh PL, Ratnayake S, Beahan CT, Wilson SM, Bacic A, Newbigin E (2013) In Vitro grown pollen tubes of Nicotiana alata actively synthesise a fucosylated xyloglucan. PLoS ONE 8:e77140

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Lerouxel O, Choo TS, Seveno M, Usadel B, Faye L, Lerouge P, Pauly M (2002) Rapid structural phenotyping of plant cell wall mutants by enzymatic oligosaccharide fingerprinting. Plant Physiol 130:1754–1763

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Liepman AH, Wightman R, Geshi N, Turner SR, Scheller HV (2010) Arabidopsis—a powerful model system for plant cell wall research. Plant J 61(6):1107–1121

    Article  CAS  PubMed  Google Scholar 

  • Matsunaga T, Ishii T, Matsumoto S, Higuchi M, Darvill A, Albersheim P, O’Neill MA (2004) Occurrence of the primary cell wall polysaccharide rhamnogalacturonan II in pteridophytes, lycophytes, and bryophytes. Implications for the evolution of vascular plants. Plant Physiol 134:339–351

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • McDougall GJ, Fry SC (1989) Structure-activity relationships for xyloglucan oligosaccharides with antiauxin activity. Plant Physiol 89:883–887

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plantarum 15:473–497

    Article  CAS  Google Scholar 

  • Nakagawa T, Kurose T, Hino T, Tanaka K, Kawamukai M, Niwa Y, Toyooka K, Matsuoka K, Jinbo T, Kimura T (2007) Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation. J Biosci Bioeng 104:34–41

    Article  CAS  PubMed  Google Scholar 

  • O’Neill RA, Albersheim P, Darvill AG (1989) Purification and characterization of a xyloglucan oligosaccharide-specific xylosidase from pea seedlings. J Biol Chem 264:20430–20437

    PubMed  Google Scholar 

  • Pauly M, Keegstra K (2008) Cell-wall carbohydrates and their modification as a resource for biofuels. Plant J 54(4):559–568

    Article  CAS  PubMed  Google Scholar 

  • Pauly M, Andersen LN, Kauppinen S, Kofod LV, York WS, Albersheim P, Darvill A (1999) A xyloglucan-specific endo-b-1,4-glucanase from Aspergillus aculeatus: expression cloning in yeast, purification and characterization of the recombinant enzyme. Glycobiology 9:93–100

    Article  CAS  PubMed  Google Scholar 

  • Pauly M, Qin Q, Greene H, Albersheim P, Darvill A, York WS (2001) Changes in the structure of xyloglucan during cell elongation. Planta 212:842–850

    Article  CAS  PubMed  Google Scholar 

  • Pauly M, Gille S, Liu L, Mansoori N, de Souza A, Schultink A, Xiong G (2013) Hemicellulose biosynthesis. Planta 238:627–642

    Article  CAS  PubMed  Google Scholar 

  • Pena MJ, Darvill AG, Eberhard S, York WS, O’Neill MA (2008) Moss and liverwort xyloglucans contain galacturonic acid and are structurally distinct from the xyloglucans synthesized by hornworts and vascular plants. Glycobiology 18:891–904

    Article  CAS  PubMed  Google Scholar 

  • Popper ZA, Fry SC (2003) Primary cell wall composition of bryophytes and charophytes. Ann Bot 91:1–12

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Pre M, Caillet V, Sobilo J, McCarthy J (2008) Characterization and expression analysis of genes directing galactomannan synthesis in coffee. Ann Bot 102:207–220

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Ray B, Loutelier-Bourhis C, Lange C, Condamine E, Driouich A, Lerouge P (2004) Structural investigation of hemicellulosic polysaccharides from Argania spinosa: characterisation of a novel xyloglucan motif. Carbohydr Res 339:201–208

    Article  CAS  PubMed  Google Scholar 

  • Sampedro J, Sieiro C, Revilla G, Gonzalez-Villa T, Zarra I (2001) Cloning and expression pattern of a gene encoding an alpha-xylosidase active against xyloglucan oligosaccharides from Arabidopsis. Plant Physiol 126:910–920

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Scheller HV, Ulvskov P (2010) Hemicelluloses. Annu Rev Plant Biol 61:263–289

    Article  CAS  PubMed  Google Scholar 

  • Schultink A, Liu L, Zhu L, Pauly M (2014) Structural diversity and function of xyloglucan sidechain substituents. Plants 3:526–542

    Article  Google Scholar 

  • Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W, Lopez R, McWilliam H, Remmert M, Soding J, Thompson JD, Higgins DG (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Molecular systems biology 7:539

    Article  PubMed Central  PubMed  Google Scholar 

  • Suresh BV, Roy R, Sahu K, Misra G, Chattopadhyay D (2014) Tomato genomic resources database: an integrated repository of useful tomato genomic information for basic and applied research. PLoS One 9:e86387

    Article  PubMed Central  PubMed  Google Scholar 

  • Takizawa A, Hyodo H, Wada K, Ishii T, Satoh S, Iwai H (2014) Regulatory specialization of xyloglucan (XG) and glucuronoarabinoxylan (GAX) in pericarp cell walls during fruit ripening in tomato (Solanum lycopersicum). PLoS ONE 9:e89871

    Article  PubMed Central  PubMed  Google Scholar 

  • Varki A, Cummings RD, Esko JD, Freeze HH, Stanley P, Marth JD, Bertozzi CR, Hart GW, Etzler ME (2009) Symbol nomenclature for glycan representation. Proteomics 9:5398–5399

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Vuttipongchaikij S, Brocklehurst D, Steele-King C, Ashford DA, Gomez LD, McQueen-Mason SJ (2012) Arabidopsis GT34 family contains five xyloglucan alpha-1,6-xylosyltransferases. New Phytol 195:585–595

    Article  CAS  PubMed  Google Scholar 

  • Zabotina OA (2012) Xyloglucan and its biosynthesis. Frontiers in plant science 3:134

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Zabotina OA, van de Ven WT, Freshour G, Drakakaki G, Cavalier D, Mouille G, Hahn MG, Keegstra K, Raikhel NV (2008) Arabidopsis XXT5 gene encodes a putative alpha-1,6-xylosyltransferase that is involved in xyloglucan biosynthesis. Plant J 56(1):101–115

    Article  CAS  PubMed  Google Scholar 

  • Zabotina OA, Avci U, Cavalier D, Pattathil S, Chou YH, Eberhard S, Danhof L, Keegstra K, Hahn MG (2012) Mutations in multiple XXT genes of Arabidopsis reveal the complexity of xyloglucan biosynthesis. Plant Physiol 159:1367–1384

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Ken Keegstra, Plant Research Lab at Michigan State University, for providing seeds of the Arabidopsis xxt1/2/5 triple mutant and Kirk Schnorr, Novozymes, Bagsvaerd, Denmark, for the gift of recombinant XEG. JS was funded by DAAD-RISE.

Author information

Author notes

  1. Julia Schubert

    Present address: University of Potsdam, 14469, Potsdam, BB, Germany

Authors and Affiliations

  1. Energy Biosciences Institute, University of California-Berkeley, Berkeley, CA, 94704, USA

    Nasim Mansoori, Julia Schubert & Markus Pauly

  2. Department of Plant and Microbial Biology, Energy Biosciences Institute, University of California, Energy Biosciences Building 212C, 2151 Berkeley Way, Berkeley, CA, 94720-5230, USA

    Alex Schultink & Markus Pauly

Authors
  1. Nasim Mansoori
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Alex Schultink
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Julia Schubert
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Markus Pauly
    View author publications

    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence to Markus Pauly.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 1679 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mansoori, N., Schultink, A., Schubert, J. et al. Expression of heterologous xyloglucan xylosyltransferases in Arabidopsis to investigate their role in determining xyloglucan xylosylation substitution patterns. Planta 241, 1145–1158 (2015). https://doi.org/10.1007/s00425-015-2243-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-015-2243-2

Keywords