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RESEARCH ARTICLE
Contents Vol 33(7)

Transformation of cambial tissue in vivo provides an efficient means for induced somatic sector analysis and gene testing in stems of woody plant species

Kim S. Van Beveren A , Antanas V. Spokevicius A , Josquin Tibbits A , Qing Wang A and Gerd Bossinger A B
+ Author Affiliations
- Author Affiliations

A School of Forest and Ecosystem Science, The University of Melbourne, Water Street, Creswick, Vic. 3363, Australia.

B Corresponding author. Email: gerd@unimelb.edu.au

Functional Plant Biology 33(7) 629-638 https://doi.org/10.1071/FP06057
Submitted: 17 March 2006  Accepted: 28 April 2006   Published: 3 July 2006

Abstract

Large-scale functional analysis of genes and transgenes suspected to be involved in wood development in trees is hindered by long generation times, low transformation and regeneration efficiencies and difficulties with phenotypic assessment of traits, especially those that appear late in a tree’s development. To avoid such obstacles many researchers have turned to model plants such as Arabidopsis thaliana (L.) Heynh., Zinnia elegans Jacq. and Nicotiana ssp., or have focused their attention on in vitro wood formation systems or in vivo approaches targeting primary meristems for transformation. Complementing such efforts, we report the use of Agrobacterium to introduce transgenes directly into cambial cells of glasshouse-grown trees in order to create transgenic somatic tissue sectors. These sectors are suitable for phenotypic evaluation and analysis of target gene function. In our experiments the wood formation zone containing the cambium of Eucalyptus, Populus and Pinus species of varying age was inoculated with Agrobacterium containing a CaMV 35S::GUS construct. Following an initial wound response, frequent and stable transformation was observed in the form of distinct GUS-staining patterns (sectors) in newly formed secondary tissues. Sector size and extent depended on the cell type transformed, the species and the length of time treated plants were allowed to grow (more than two years in some cases). Induced somatic sector analysis (ISSA) can now be efficiently used to study cell fate and gene function during secondary growth in stems of forest tree species.

Keywords: Agrobacterium, cambium, Eucalyptus, in vivo transformation, Pinus, Populus, secondary growth, wood formation.


Acknowledgments

We thank Simon Southerton, Zander Myburg and Rick Meilan for critically reading our manuscript and for many helpful comments.


References


Allona I, Quinn M, Shoop E, Swope K, St Cyr S , et al. (1998) Analysis of xylem formation in pine by cDNA sequencing. Proceedings of the National Academy of Sciences USA 95, 9693–9698.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Baima S, Possenti M, Matteucci A, Wisman E, Altamura MM, Ruberti I, Morelli G (2001) The Arabidopsis ATHB-8 HD-zip protein acts as a differentiation-promoting transcription factor of the vascular meristems. Plant Physiology 126, 643–655.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Barker WG (1954) A contribution to the concept of wound repair in woody stems. Canadian Journal of Botany 32, 486–490. open url image1

Bossinger G (2002) Plant chimeras and mosaics. In ‘Encyclopedia of life sciences. Vol. 1’. pp. 387–391. (Nature Publishing Group: London)

Bossinger G , Leitch M (2000) Isolation of cambial-specific genes from Eucalyptus globulus Labill. In ‘Cell and molecular biology of wood’. (Eds RA Savidge, JR Barnett, R Napier) pp. 203–207. (BIOS Scientific Publishers Ltd: Oxford)

Brown CL, Sax K (1962) Influence of pressure on differentiation of secondary tissues. American Journal of Botany 49, 683–691.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Catesson AM (1974) Cambial cells. In ‘Dynamic aspects of plant ultrastructure’. (Ed. AW Robards) pp. 358–390. (McGraw-Hill Book Co: London)

Catesson AM (1980) The vascular cambium. In ‘Control of shoot growth in trees. Proceedings of the joint workshop of IUFRO working parties on xylem and shoot growth physiology’. (Ed. CHA Little) pp. 12–40. (IUFRO: Fredericton, New Brunswick)

Catesson AM (1984) Cambial dynamics. Annales des Sciences Naturelles-Botanique et Biologie Vegetale 6, 23–43. open url image1

Chaffey N, Cholewa E, Regan S, Sundberg B (2002) Secondary xylem development in Arabidopsis: a model for wood formation. Physiologia Plantarum 114, 594–600.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Chang S, Puryear J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Molecular Biology Reporter 11, 113–116. open url image1

Confalonieri M, Balestrazzi A, Bisoffi S, Carbonera D (2003) In vitro culture and genetic engineering of Populus spp.: synergy for forest tree improvement. Plant Cell, Tissue and Organ Culture 72, 109–138.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Decleene M, Deley J (1976) Host range of crown gall. Botanical Review 42, 389–466. open url image1

Déjardin A, Leplé JC, Lesage-Descauses MC, Costa G, Pilate G (2004) Expressed sequence tags from poplar wood tissues — a comparative analysis from multiple libraries. Plant Biology 6, 55–64.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Dwivedi UN, Campbell WH, Yu J, Datla RSS, Bugos RC, Chiang VL, Podila GK (1994) Modification of lignin biosynthesis in transgenic Nicotiana through expression of an antisense O-methyltransferase gene from Populus. Plant Molecular Biology 26, 61–71.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Ellis D, Roberts D, Sutton B, Lazaroff W, Webb D, Flinn B (1989) Transformation of white spruce and other conifer species by Agrobacterium tumefaciens. Plant Cell Reports 8, 16–20.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Fillatti JJ, Sellmer J, McCown B, Haissig B, Comai L (1987) Agrobacterium-mediated transformation and regeneration of Populus. Molecular and General Genetics 206, 192–199.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Goujon T, Sibout R, Pollet B, Maba B, Nussaume L , et al. (2003) A new Arabidopsis thaliana mutant deficient in the expression of O-methyltransferase impacts lignins and sinapoyl esters. Plant Molecular Biology 51, 973–989.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Harcourt RL, Kyozuka J, Floyd RB, Bateman KS, Tanaka H, Decroocq V, Llewellyn DJ, Zhu X, Peacock WJ, Dennis ES , et al. (2000) Insect- and herbicide-resistant transgenic eucalypts. Molecular Breeding 6, 307–315.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Hertzberg M, Aspeborg H, Schrader J, Andersson A, Erlandsson R , et al. (2001) A transcriptional roadmap to wood formation. Proceedings of the National Academy of Sciences USA 98, 14 732–14 737.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Ho CK, Chang SH, Tsay JY, Tsai CJ, Chiang VL, Chen ZZ (1998) Agrobacterium tumefaciens-mediated transformation of Eucalyptus camaldulensis and production of transgenic plants. Plant Cell Reports 17, 675–680.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Klopfenstein NB, Shi NQ, Kernan A, McNabb HS, Hall RB, Hart ER, Thornburg RW (1991) Transgenic Populus hybrid express a wound-inducible potato proteinase inhibitor–Ii–cat gene fusion. Canadian Journal of Forest Research 21, 1321–1328. open url image1

Lazo GR, Stein PA, Ludwig RA (1991) A DNA transformation-competent Arabidopsis genomic library. Agrobacterium 9, 963–967. open url image1

Leitch MA , Bossinger G (2004) In vitro systems for the study of wood formation. In ‘Molecular genetics and breeding of forest trees’. (Eds S Kumar, M Fladung) pp. 193–211. (Haworth Press: New York)

Li W, Guo GQ, Zheng GC (2000) Agrobacterium-mediated transformation: state of the art and future prospect. Chinese Science Bulletin 45, 1537–1546. open url image1

Loopstra CA, Stomp AM, Sederoff RR (1990) Agrobacterium-mediated DNA transfer in sugar pine. Plant Molecular Biology 15, 1–9.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Lorenz WW, Dean JFD (2002) SAGE Profiling and demonstration of differential gene expression along the axial developmental gradient of lignifying xylem in loblolly pine (Pinus taeda). Tree Physiology 22, 301–310.
| PubMed |
open url image1

Mahmood A (1968) Cell grouping and primary wall generations in cambial zone xylem and phloem in Pinus. Australian Journal of Botany 16, 177–195.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Mahmood A (1990) The parental cell wall. In ‘The vascular cambium’. (Ed. M Iqbal) pp. 113–125. (Research Studies Press: Somerset)

Moralejo M, Rochange F, Boudet AM, Teulieres C (1998) Generation of transgenic Eucalyptus globulus plantlets through Agrobacterium tumefaciens mediated transformation. Australian Journal of Plant Physiology 25, 207–212. open url image1

Mullins KV, Llewellyn DJ, Hartney VJ, Strauss S, Dennis ES (1997) Regeneration and transformation of Eucalyptus camaldulensis. Plant Cell Reports 16, 787–791.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Murmanis L (1970) Locating the initial in vascular cambium of Pinus strobus by electron microscopy. Wood Science and Technology 4, 1–14.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Nester EW (1987) Analysis of host range in transformation of higher plants by Agrobacterium tumefaciens. In ‘Genetic manipulation of woody plants’. (Eds JW Hanover, DE Keathley, A Hollaender) pp. 219–229. (Plenum Press: New York)

Newman IV (1956) Pattern in the meristem of vascular plants. I. Cell partition in living apices and in the cambial zone in relation to concepts of initial cells and apical cells. Phytomorphology 6, 1–19. open url image1

Noel ARA (1968) Callus formation and differentiation at an exposed cambial surface. Annals of Botany 32, 347–359. open url image1

Parsons TJ, Sinkar VP, Stettler RF, Nester EW, Gordon MP (1986) Transformation of poplar by Agrobacterium tumefaciens. Bio–Technology 4, 533–536.
| Crossref |
open url image1

Ralph J, Hatfield RD, Piquemal J, Yahiaoui N, Pean M, Lapierre C, Boudet AM (1998) NMR characterization of altered lignins extracted from tobacco plants down-regulated for lignification enzymes cinnamyl-alcohol dehydrogenase and cinnamoyl-CoA reductase. Proceedings of the National Academy of Sciences USA 95, 12 803–12 808.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Savidge RA (2000) Biochemistry of seasonal cambial growth and wood formation — an overview of the challenges. In ‘Cell and molecular biology of wood formation’. (Eds RA Savidge, JR Barnett, R Napier) pp. 1–28. (BIOS Scientific Publishers Ltd: Oxford)

Sharples A, Gunnery H (1933) Callus formation in Hibiscus rosa-sinensis L. and Hevea brasiliensis mull. arg. Annals of Botany 47, 827–839. open url image1

Spena A , Salamini F (1995) Genetic tagging of cells and cell layers for studies of plant development. In ‘Methods in cell biology’. (Eds DW Galbraith, HJ Bohnert, DP Bourque) pp. 331–354. (Academic Press: New York)

Spokevicius AV, Van Beveren K, Leitch MM, Bossinger G (2005) Agrobacterium-mediated in vitro transformation of wood-producing stem segments in eucalypts. Plant Cell Reports 23, 617–624.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Spokevicius AV, Van Beveren K, Bossinger G (2006) Agrobacterium-mediated transformation of dormant lateral buds in poplar trees reveals developmental patterns in secondary stem tissues. Functional Plant Biology 33, 133–139.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Sterky F, Regan S, Karlsson J, Hertzberg M, Rohde A , et al. (1998) Gene discovery in the wood-forming tissues of poplar: analysis of 5692 expressed sequence tags. Proceedings of the National Academy of Sciences USA 95, 13 330–13 335.
| Crossref | GoogleScholarGoogle Scholar | open url image1

Stomp AM, Loopstra CA, Chilton WS, Sederoff RR, Moore LW (1990) Extended host range of Agrobacterium tumerfaciens in the genus Pinus. Plant Physiology 92, 1226–1232.
| PubMed |
open url image1

Szymkowiak EJ, Sussex IM (1996) What chimeras can tell us about plant development. Annual Review of Plant Physiology and Plant Molecular Biology 47, 351–376.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Tang W, Sederoff RR, Whetten R (2001) Regeneration of transgenic loblolly pine (Pinus taeda L.) from zygotic embryos transformed with Agrobacterium tumefaciens. Planta 213, 981–989.
| PubMed |
open url image1

Timell TE (1980) Organization and ultrastructure of the dormant cambial zone in compression wood of Picea abies. Wood Science and Technology 14, 161–179. open url image1

van Raemdonck D, Pesquet E, Cloquet S, Beeckman H, Boerjan W, Goffner D, El Jaziri M, Baucher M (2005) Molecular changes associated with the setting up of secondary growth in aspen. Journal of Experimental Botany 56, 2211–2227.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

von Schwartzenberg K, Doumas P, Jouanin L, Pilate G (1994) Enhancement of the endogenous cytokinin concentration in poplar by transformation with Agrobacterium T-DNA gene Ipt. Tree Physiology 14, 27–35.
| PubMed |
open url image1

Whetten R, Sun YH, Zhang Y, Sederoff R (2001) Functional genomics and cell wall biosynthesis in loblolly pine. Plant Molecular Biology 47, 275–291.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Yang JM, Park S, Kamdem DP, Keathley DE, Retzel E, Paule C, Kapur V, Han KH (2003) Novel gene expression profiles define the metabolic and physiological processes characteristic of wood and its extractive formation in a hardwood tree species, Robinia pseudoacacia. Plant Molecular Biology 52, 935–956.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Ye ZH, Droste DL (1996) Isolation and characterization of cDNAs encoding xylogenesis-associated and wounding-induced ribonucleases in Zinnia elegans. Plant Molecular Biology 30, 697–709.
| Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1