Skip to main content
SpringerLink
Log in

Induced somatic sector analysis of cellulose synthase (CesA) promoter regions in woody stem tissues

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

Abstract

The increasing focus on plantation forestry as a renewable source of cellulosic biomass has emphasized the need for tools to study the unique biology of woody genera such as Eucalyptus, Populus and Pinus. The domestication of these woody crops is hampered by long generation times, and breeders are now looking to molecular approaches such as marker-assisted breeding and genetic modification to accelerate tree improvement. Much of what is known about genes involved in the growth and development of plants has come from studies of herbaceous models such as Arabidopsis and rice. However, transferring this information to woody plants often proves difficult, especially for genes expressed in woody stems. Here we report the use of induced somatic sector analysis (ISSA) for characterization of promoter expression patterns directly in the stems of Populus and Eucalyptus trees. As a case study, we used previously characterized primary and secondary cell wall-related cellulose synthase (CesA) promoters cloned from Eucalyptus grandis. We show that ISSA can be used to elucidate the phloem and xylem expression patterns of the CesA genes in Eucalyptus and Populus stems and also show that the staining patterns differ in Eucalyptus and Populus stems. These findings show that ISSA is an efficient approach to investigate promoter function in the developmental context of woody plant tissues and raise questions about the suitability of heterologous promoters for genetic manipulation in plant species.

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

Access this article

Log in via an institution

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

Similar content being viewed by others

Abbreviations

ATS−2 :

Average of transformed sectors per cm2

ISSA:

Induced somatic sector analysis

References

  • Alwen A, Moreno RMB, Vincente O, Heberle-Bors E (1992) Plant endogenous β-glucuronidase activity: how to avoid interference with the use of the E. coli β-glucuronidase as a reporter gene in transgenic plants. Transgenic Res 1:63–70

    Article  PubMed  CAS  Google Scholar 

  • Arioli T, Peng L, Betzner AS, Burn J, Wittke W, Herth W, Camilleri C, Hofte H, Plazinski J, Birch R, Cork A, Glover J, Redmond J, Williamson RE (1998) Molecular analysis of cellulose biosynthesis in Arabidopsis. Science 279:717–720

    Article  PubMed  CAS  Google Scholar 

  • Beeckman T, Przemeck Gerhard KH, Stamatiou G, Lau R, Terryn N, De Rycke R, Inze D, Berleth T (2002) Genetic complexity of CESA gene function in Arabidopsis embryogenesis. Plant Physiol 130:1883–1893

    Article  PubMed  CAS  Google Scholar 

  • Benfey PN, Chua N-H (1990) The cauliflower mosaic virus 35S promoter: combinatorial regulation of transcription in plants. Science 250:959–966

    Article  PubMed  CAS  Google Scholar 

  • Carroll A, Specht CD (2011) Understanding plant cellulose synthases through a comprehensive investigation of the cellulose synthase family sequences. Front Plant Sci 2:5

    Article  PubMed  CAS  Google Scholar 

  • Creux NM, Ranik M, Berger DK, Myburg AA (2008) Comparative analysis of orthologous cellulose synthase promoters from Arabidopsis, Populus and Eucalyptus: evidence of conserved regulatory elements in angiosperms. New Phytol 179:722–737

    Article  PubMed  CAS  Google Scholar 

  • Curtis MD, Grossniklaus U (2003) A gateway cloning vector set for high-throughput functional analysis of genes in planta. Plant Physiol 133:462–469

    Article  PubMed  CAS  Google Scholar 

  • Desprez T, Vernhettes S, Fagard M, Refrégier G, Desnos T, Aletti E, Py N, Pelletier S, Höfte H (2002) Resistance against herbicide isoxaben and cellulose deficiency caused by distinct mutations in same cellulose synthase isoform CESA6. Plant Physiol 128:482–490

    Article  PubMed  CAS  Google Scholar 

  • Djerbi S, Lindskog M, Arvestad L, Sterky F, Teeri TT (2005) The genome sequence of black cottonwood (Populus trichocarpa) reveals 18 conserved cellulose synthase (CesA) genes. Planta 221:739–746

    Article  PubMed  CAS  Google Scholar 

  • Dowell RD (2010) Transcription factor binding variation in the evolution of gene regulation. Trends Genet 26:468–475

    Article  PubMed  CAS  Google Scholar 

  • Fei H, Chaillou S, Hirel B, Polowick P, Mahon JD, Vessey JK (2006) Effects of the overexpression of a soybean cytosolic glutamine synthetase gene (GS15) linked to organ-specific promoters on growth and nitrogen accumulation of pea plants supplied with ammonium. Plant Physiol Biochem 44:543–550

    Article  PubMed  CAS  Google Scholar 

  • Grattapaglia D, Plomion C, Kirst M, Sederoff RR (2009) Genomics of growth traits in forest trees. Curr Opin Plant Biol 12:148–156

    Article  PubMed  CAS  Google Scholar 

  • Hamann T, Osborne E, Youngs HL, Misson J, Nussaume L, Somerville C (2004) Global expression analysis of CESA and CSL genes in Arabidopsis. Cellulose 11:279–286

    Article  CAS  Google Scholar 

  • Hinchee M, Rottmann W, Mullinax L, Zhang C, Chang S, Cunningham M, Pearson L, Nehra N, Tomes D, Lakshmanan P, Songstad D (2011) Short-rotation woody crops for bioenergy and biofuels applications. In: Dwight T, Lakshmanan P, songstad D (eds) Biofuels. Global impact on renewable energy, production agriculture, and technological advancements. Springer, New York, pp 139-156

  • Ho MCW, Johnsen H, Goetz SE, Schiller BJ, Bae E, Tran DA, Shur AS, Allen JM, Rau C, Bender W, Fisher WW, Celniker SE, Drewell RA (2009) Functional evolution of cis-regulatory modules at a homeotic gene in Drosophila. PLoS Genet 5:e1000709

    Article  PubMed  Google Scholar 

  • Hu R, Qi G, Kong Y, Kong D, Gao Q, Zhou G (2010) Comprehensive analysis of NAC domain transcription factor gene family in Populus trichocarpa. BMC Plant Biol 10:145

    Article  PubMed  Google Scholar 

  • Hussey S, Mizrachi E, Spokevicius A, Bossinger G, Berger D, Myburg A (2011) SND2, a NAC transcription factor gene, regulates genes involved in secondary cell wall development in Arabidopsis fibres and increases fibre cell area in Eucalyptus. BMC Plant Biol 11:173

    Article  PubMed  CAS  Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    PubMed  CAS  Google Scholar 

  • Kimura S, Laosinchai W, Itoh T, Cui X, Linder CR, Brown RM Jr (1999) Immunogold labeling of rosette terminal cellulose-synthesizing complexes in the vascular plant Vigna angularis. Plant Cell 11:2075–2086

    PubMed  CAS  Google Scholar 

  • Ko J-H, Kim H-T, Hwang I, Han K-H (2012) Tissue-type-specific transcriptome analysis identifies developing xylem-specific promoters in poplar. Plant Biotechnol 10:587–596

    Article  CAS  Google Scholar 

  • Kumar M, Thammannagowda S, Bulone V, Chiang V, Han K-H, Joshi CP, Mansfield SD, Mellerowicz E, Sundberg B, Teeri T, Ellis BE (2009) An update on the nomenclature for the cellulose synthase genes in Populus. Trends Plant Sci 14:248–254

    Article  PubMed  CAS  Google Scholar 

  • Lazo G, Stein P, Ludwig R (1991) A DNA transformation-competent Arabidopsis genomic library in Agrobacterium. Biotechnology 9:963–967

    Article  PubMed  CAS  Google Scholar 

  • Mansfield SD (2009) Solutions for dissolution-engineering cell walls for deconstruction. Curr Opin Biotech 20:286–294

    Article  PubMed  CAS  Google Scholar 

  • Mizrachi E, Mansfield SD, Myburg AA (2012) Cellulose factories: advancing bioenergy production from forest trees. New Phytol 194:54–62

    Article  PubMed  CAS  Google Scholar 

  • Mu XJ, Lu ZJ, Kong Y, Lam HYK, Gerstein MB (2011) Analysis of genomic variation in non-coding elements using population-scale sequencing data from the 1000 Genomes Project. Nucleic Acids Res 39:7058–7076

    Article  PubMed  CAS  Google Scholar 

  • Odell JT, Nagy F, Chua N-H (1985) Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313:810–812

    Article  PubMed  CAS  Google Scholar 

  • Plomion C, Leprovost G, Stokes A (2001) Wood formation in trees. Plant Physiol 127:1513–1523

    Article  PubMed  CAS  Google Scholar 

  • Popper ZA, Michel G, Herve C, Domozych DS, Willats WGT, Tuohy MG, Kloareg B, Stengel DB (2011) Evolution and diversity of plant cell walls: from algae to flowering plants. Annu Rev Plant Biol 62:567–590

    Article  PubMed  CAS  Google Scholar 

  • Qin JY, Zhang L, Clift KL, Hulur I, Xiang AP, Ren B-Z, Lahn BT (2010) Systematic comparison of constitutive promoters and the doxycycline-inducible promoter. PLoS ONE 5:e10611

    Article  PubMed  Google Scholar 

  • Ranik M, Myburg AA (2006) Six new cellulose synthase genes from Eucalyptus are associated with primary and secondary cell wall biosynthesis. Tree Physiol 26:545–556

    Article  PubMed  CAS  Google Scholar 

  • Rathmann R, Szklo A, Schaeffer R (2010) Land use competition for production of food and liquid biofuels: an analysis of the arguments in the current debate. Renew Energ 35:14–22

    Article  Google Scholar 

  • Richmond TA, Somerville CR (2000) The cellulose synthase superfamily. Plant Physiol 124:495–498

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbour Press, New York

    Google Scholar 

  • Samuga A, Joshi CP (2004) Differential expression patterns of two new primary cell wall-related cellulose synthase cDNAs, PtrCesA6 and PtrCesA7 from aspen trees. Gene 334:73–82

    Article  PubMed  CAS  Google Scholar 

  • Scheible WR, Eshed R, Richmond T, Delmer D, Somerville C (2001) Modifications of cellulose synthase confer resistance to isoxaben and thiazolidinone herbicides in Arabidopsis ixr1 mutants. Proc Natl Acad Sci USA 98:10079–10084

    Article  PubMed  CAS  Google Scholar 

  • Seguin A (2011) How could forest trees play an important role as feedstock for bioenergy production? Curr Opin Environ Sustain 3:90–94

    Article  Google Scholar 

  • Song D, Shen J, Li L (2010) Characterization of cellulose synthase complexes in Populus xylem differentiation. New Phytol 187:777–790

    Article  PubMed  CAS  Google Scholar 

  • Spokevicius AV, Van Beveren K, Leitch MM, Bossinger G (2005) Agrobacterium-mediated in vitro transformation of wood-producing stem segments in eucalypts. Plant Cell Rep 23:617–624

    Article  PubMed  CAS  Google Scholar 

  • Spokevicius AV, Southerton SG, MacMillan CP, Qiu D, Gan S, Tibbits JFG, Moran GF, Bossinger G (2007) β-Tubulin affects cellulose microfibril orientation in plant secondary fibre cell walls. Plant J 51:717–726

    Article  PubMed  CAS  Google Scholar 

  • Stork J, Harris D, Griffiths J, Williams B, Beisson F, Li-Beisson Y, Mendu V, Haughn G, DeBolt S (2010) CELLULOSE SYNTHASE9 serves a nonredundant role in secondary cell wall synthesis in Arabidopsis epidermal testa cells. Plant Physiol 153:580–589

    Article  PubMed  CAS  Google Scholar 

  • Suzuki S, Li LG, Sun YH, Chiang VL (2006) The cellulose synthase gene superfamily and biochemical functions of xylem-specific cellulose synthase-like genes in Populus trichocarpa. Plant Physiol 142:1233–1245

    Article  PubMed  CAS  Google Scholar 

  • Taylor NG (2008) Cellulose biosynthesis and deposition in higher plants. New Phytol 178:239–252

    Article  PubMed  CAS  Google Scholar 

  • Taylor NG, Laurie S, Turner SR (2000) Multiple cellulose synthase catalytic subunits are required for cellulose synthesis in Arabidopsis. Plant Cell 12:2529–2539

    PubMed  CAS  Google Scholar 

  • Taylor NG, Howells RM, Huttly AK, Vickers K, Turner SR (2003) Interactions among three distinct CESA proteins essential for cellulose synthesis. Proc Natl Acad Sci USA 100:1450–1455

    Article  PubMed  CAS  Google Scholar 

  • Turner SR, Somerville CR (1997) Collapsed xylem phenotype of Arabidopsis identifies mutants deficient in cellulose deposition in the secondary cell wall. Plant Cell 9:689–701

    PubMed  CAS  Google Scholar 

  • Tuskan GA, DiFazio S, Jansson S et al (2006) The genome of black cottonwood, Populus trichocarpa (Torr. & Gray). Science 313:1596–1604

    Article  PubMed  CAS  Google Scholar 

  • Van Beveren K, Spokevicius AV, Tibbits J, Wang Q, Bossinger G (2006) Transformation of cambial tissue in vivo provides efficient means for Induced Somatic Sector Analysis (ISSA) and gene testing in stems of woody plants species. Funct Plant Biol 33:629–638

    Article  Google Scholar 

  • Yin Y, Huang J, Xu Y (2009) The cellulose synthase superfamily in fully sequenced plants and algae. BMC Plant Biol 9:99

    Article  PubMed  Google Scholar 

  • Zhang P, Bohl-Zenger S, Puonti-Kaerlas J, Potrykus I, Gruissem W (2003) Two cassava promoters related to vascular expression and storage root formation. Planta 218:192–203

    Article  PubMed  CAS  Google Scholar 

  • Zhong R, Lee C, Ye Z-H (2010) Evolutionary conservation of the transcriptional network regulating secondary cell wall biosynthesis. Trends Plant Sci 15:625–632

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We are grateful to Martin Ranik for providing the CesA-promoter::GUS constructs which made this study possible and Minique De Castro for aiding in the construction of the second CAMV35S-promoter::GUS construct. We also thank Julio Najera, Valerie Frassiant and Angelique Manuel for laboratory assistance. This work was supported through funding provided by Mondi and Sappi to the Forest Molecular Genetics (FMG) Programme, the Technology and Human Resources for Industry Programme (THRIP) and the National Research Foundation (NRF) of South Africa as well as a Linkage Grant from the Australian Research Council (LP0776563) to GB, AAM and AVS.

Author information

Authors and Affiliations

  1. Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Private Bag X20, Pretoria, 0028, South Africa

    Nicky M. Creux & Alexander A. Myburg

  2. Department of Forest and Ecosystem Science, Melbourne School of Land and Environment, The University of Melbourne, Creswick, VIC, Australia

    Gerd Bossinger & Antanas V. Spokevicius

Authors
  1. Nicky M. Creux
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gerd Bossinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alexander A. Myburg
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Antanas V. Spokevicius
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gerd Bossinger.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Creux, N.M., Bossinger, G., Myburg, A.A. et al. Induced somatic sector analysis of cellulose synthase (CesA) promoter regions in woody stem tissues. Planta 237, 799–812 (2013). https://doi.org/10.1007/s00425-012-1792-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-012-1792-x

Keywords

Search

Navigation