Abstract
Tracheid and sclereid differentiation was induced in callus cultures of Pinus radiata D. Don by culturing on a basal medium containing activated charcoal but no phytohormones; sclereids differentiated in callus derived from xylem strips, but not in callus derived from hypocotyl segments. The tracheids differentiated in hypocotyl-derived callus had helical, scalariform, reticulated or pitted secondary cell-wall patterns, but those differentiated in xylem-derived callus had a reticulate or pitted pattern. The thickened tracheid and sclereid walls contained lignin as indicated by the red colour reaction given with phloroglucinol–HCl. The presence of lignin in the cell walls of differentiated callus was confirmed using pyrolysis gas chromatography–mass spectrometry by the detection of phenylpropanoid components derived from lignin. Lignin was also detected using solid-state 13C cross-polarisation/magic-angle spinning nuclear magnetic resonance spectroscopy and quantified as thioglycolic acid lignin. Monosaccharide analyses of the cell walls isolated from differentiated and undifferentiated calli showed that the cell walls of the differentiated calli contained higher proportions of glucose and mannose, consistent with the presence of greater proportions of gluco- and/or galactogluco-mannans in the secondary cell walls of the differentiated cells. A protocol for the stable transformation of undifferentiated, xylem-derived cultures was successfully developed. Transgenic cell lines were established following Biolistic particle bombardment with a plasmid containing the coding region of the nptII gene and the coding region of the cad gene from P. radiata. Expression of the nptII gene in transgenic lines was confirmed by an NPTII–enzyme-linked immunosorbent assay. The overexpression of cad in the transgenic lines resulted in a down-regulation of cinnamyl alcohol dehydrogenase (EC 1.1.1.195) expression.
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Abbreviations
- BAP:
-
6-benzylaminopurine
- CAD:
-
cinnamyl alcohol dehydrogenase
- CP/MAS:
-
cross-polarisation/magic-angle spinning
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- NAA:
-
1-naphthalene acetic acid
- nptII:
-
neomycin phosphotransferase II
- PCIB:
-
p-chlorophenoxyisobutyric acid
- PVPP:
-
polyvinyl polypyrrolidone
- SPE:
-
single-pulse excitation
- TGAL:
-
thioglycolic acid lignin
- TFA:
-
trifluoroacetic acid
- TIBA:
-
2,3,5-triiodobenzoic acid.
References
Aitken-Christie J, Singh AP, Davies H (1988) Multiplication of meristematic tissue: a new tissue culture system for radiata pine. In: Hanover JW, Keathley DE (eds) Genetic manipulation of woody plants. Plenum, New York, pp 413–432
Andrew IG, Du Z, Bright J, Fenemor DR, McKee JW, O'Donoghue EM (2000) Pectic arabinans of Pinus radiata. American Society of Plant Physiologists plant biology meeting, San Diego, California, 2000. Abstract 309
Blee KA, Wheatley ER, Bonham VA, Mitchell GP, Robertson D, Slabas AR, Burrell MM, Wojtaszek P, Bolwell GP (2001) Proteomic analysis reveals a novel set of cell wall proteins in a transformed tobacco cell culture that synthesises secondary walls as determined by biochemical and morphological parameters. Planta 212:404–415
Booker FL, Maier CA (2001) Atmospheric carbon dioxide, irrigation, and fertilization effects on phenolic and nitrogen concentrations in loblolly pine (Pinus taeda) needles. Tree Physiol 21:609–616
Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254
Campbell MA, Kinlaw CS, Neale DB (1992) Expression of luciferase and β-glucuronidase in Pinus radiata suspension cells using electroporation and particle bombardment. Can J For Res 22:2014–2018
Capek P, Toman R, Kardosová A, Rosík J (1983) Polysaccharides from the roots of the marsh mallow (Althaea officinalis L.): structure of an arabinan. Carbohydr Res 117:133–140
Carnachan SM, Harris PJ (2000a) Polysaccharide compositions of primary cell walls of the palms Phoenix canariensis and Rhopalostylis sapida. Plant Physiol Biochem 38:699–708
Carnachan SM, Harris PJ (2000b) Ferulic acid is bound to the primary cell walls of all gymnosperm families. Biochem Syst Ecol 28:865–879
Depicker A, van Montagu M (1997) Post-transcriptional gene silencing in plants. Curr Opin Cell Biol 9:373–382
Eberhardt TL, Bernards MA, Lanfang H, Laurence BD, Wooten JB, Lewis NG (1993) Lignification in cell suspension cultures of Pinus taeda. J Biol Chem 268:21088–21096
Faix O, Meier D, Fortmann I (1990a) Thermal degradation products of wood. A collection of electron-impact (EI) mass spectra of monomeric lignin derived products. Holz Roh Werkst 48:351–354
Faix O, Meier D, Fortmann I (1990b) Thermal degradation products of wood. Gas chromatographic separation and mass spectrometric characterization of monomeric lignin-derived products. Holz Roh Werkst 48:281–285
Falconer MM, Seagull RW (1988) Xylogenesis in tissue culture III: continuing wall deposition during tracheary element development. Protoplasma 144:10–16
FAO (2000) The global outlook for future wood supply from forest plantations. Global forest products outlook study, Working paper series, Working paper No: GFPOS/WP/03. http://www.fao.org/DOCREP/003/X8423E/X8423E00.HTM
Fukuda H (1992) Tracheary element formation as a model system of cell differentiation. Int Rev Cytol 136:289–332
George EF (1993) Plant propagation by tissue culture. Exegetics, Edington
Harwood VD (1972) Studies on the cell wall polysaccharides of Pinus radiata I. Isolation and structure of a xylan. Sven Papperstidn 75:207–212
Harwood VD (1973) Studies on the cell wall polysaccharides of Pinus Radiata II. Structure of a glucomannan. Sven Papperstidn 76:377–379
Havel L, Scarano MT, Durzan DJ (1997) Xylogenesis in Cupressus callus involves apoptosis. Adv Hort Sci 11:37–40
Hawkes GE, Smith CZ, Utley JHP, Vargas RR, Viertler H (1993) A comparison of solution state 13C NMR spectra of lignins and lignin model compounds. Holzforschung 47:302–312
Hotter GS (1997) Elicitor-induced oxidative burst and phenylpropanoid metabolism in Pinus radiata cell suspension cultures. Aust J Plant Physiol 24:797–804
Jarvis MC, Fenwick KM, Apperley DC (1996) Cross-polarisation kinetics and proton NMR relaxation in polymers of Citrus cell walls. Carbohydr Res 288:1-14
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6 :3901–3907
Karácsonyi S, Kovácik V, Kákoniová D (2002) Isolation and characterisation of cell wall polysaccharides from Picea abies L. Cellul Chem Technol 30:359–370
Khan A, Chauhan YS (1993) In vitro studies on xylogenesis in pummelo juice vesicles: effect of pH and the nutrient medium on cytodifferentiation. Phytomorphology 43:35–39
Kuroda K, Shimaji K (1984) Wound effects on xylem cell differentiation in a conifer. IAWA Bull 5:295–305
MacKay JJ, Liu W, Whetten R, Sederoff RR, O'Malley DM (1995) Genetic analysis of cinnamyl alcohol dehydrogenase in loblolly pine: single gene inheritance, molecular characterization and evolution. Mol Gen Genet 247:537–545
Mankarios AT, Jones CFG, Jarvis MC, Threlfall DR, Friend J (1979) Hydrolysis of plant polysaccharides and GLC analysis of their constituent neutral sugars. Phytochemistry 18:419–422
Milioni D, Sado PE, Stacey NJ, Domingo C, Roberts K, McCann MC (2001) Differential expression of cell-wall-related genes during the formation of tracheary elements in the Zinnia mesophyll cell system. Plant Mol Biol 47:221–238
Newman RH, Davies LM, Harris PJ (1996) Solid-state 13C nuclear magnetic resonance characterization of cellulose in the cell walls of Arabidopsis thaliana leaves. Plant Physiol 111:475–485
O'Connell A, Bolwell P, Schuch W (1998) Impact of forest tree biotechnology on the pulp and paper-making processes in the 21st century. In: Lindsey K (ed) Transgenic plant research. Harwood, Amsterdam, pp 175–186
Pan MJ, Staden van J (1998) The use of charcoal in in vitro culture — a review. Plant Growth Regul 26:155–163
Ralph J, Hatfield RD (1991) Pyrolysis-GC-MS characterization of forage materials. J Agric Food Chem 39:1426–1437
Ramsden L, Northcote DH (1987) Tracheid formation in cultures of pine (Pinus sylvestris). J Cell Sci 88:467–474
Renard CM, Jarvis MC (1999) A cross-polarization, magic-angle-spinning, 13C-nuclear-magnetic-resonance study of polysaccharides in sugar beet cell walls. Plant Physiol 119:1315–1322
Sanford JC, Smith FD, Russell JA (1993) Optimizing the Biolistic process for different biological applications. Methods Enzymol 217:483–509
Savidge RA (1983) The role of plant hormones in higher plant cellular differentiation. II. Experiments with the vascular cambium, and sclereid and tracheid differentiation in the pine, Pinus contorta. Histochem J 15:447–466
Singh AP (1996) Ultrastructural features of compression wood cells in relation to bacterial decay in Pinus radiata. In: Donaldson LA, Singh AP, Butterfield BG, Whitehouse LJ (eds) Recent advances in wood anatomy. New Zealand Forest Research Institute, Rotorua, pp 400–407
Smith BG, Harris PJ (1995) Polysaccharide composition of unlignified cell walls of pineapple [Ananas comosus (L.) Merr.] fruit. Plant Physiol 107:1399–1409
Smith BG, Harris PJ, Melton LD, Newman RH (1998) The range of mobility of the non-cellulosic polysaccharides is similar in primary cell walls with different polysaccharide compositions. Physiol Plant 103:233–246
Sosanwo OA, Fawcett AH, Apperley D (1995) 13C CP-MAS NMR spectra of tropical hardwoods. Polym Int 36:247–259
Stam M, Mol JNM, Kooter JM (1997) The silence of genes in transgenic plants. Ann Bot 79:3–12
Thornber JP, Northcote DH (1962) Changes in the chemical composition of a cambial cell during its differentiation into xylem and phloem tissue in trees 3. Xylan, glucomannan and α-cellulose fractions. Biochem J 82:340–346
Vaucheret H, Béclin C, Elmayan T, Feuerbach F, Godon C, Morel J-B, Mourrain P, Palauqui J-C, Vernhettes S (1998) Transgene-induced gene silencing in plants. Plant J 16:651–659
Von Aderkas P, Label P, Lelu A-M (2002) Charcoal affects early development and hormonal concentrations of somatic embryos of hybrid larch. Tree Physiol 22:431–434
Walter C, Grace LJ (2000) Genetic engineering of conifers for plantation forestry: Pinus radiata transformation. In: Jain SM, Minocha SC (eds) Molecular biology of woody plants. Kluwer, Dordrecht, pp 79–104
Walter C, Grace L, Wagner A, White DWR, Walden AR, Donaldson SS, Hinton H, Gardner RC, Smith DR (1998) Stable transformation and regeneration of transgenic plants of Pinus radiata D. Don. Plant Cell Rep 17:460–468
Washer J, Reilly KJ, Barnett JR (1977) Differentiation in Pinus radiata callus culture: the effect of nutrients. N Z J For Sci 7:321–328
Wyrambik D, Grisebach H (1975) Purification and properties of isoenzymes of cinnamyl-alcohol dehydrogenase from soybean-cell-suspension cultures. Eur J Biochem 59:9–15
Acknowledgements
R.M. acknowledges Forest Research, Rotorua, New Zealand for financial support. We thank N. Cranshaw, L. Donaldson, L. Grace, S. Hill, A. Singh, J. Smith, D. Steward, and A. Wagner for their help and advice.
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Möller, R., McDonald, A.G., Walter, C. et al. Cell differentiation, secondary cell-wall formation and transformation of callus tissue of Pinus radiata D. Don. Planta 217, 736–747 (2003). https://doi.org/10.1007/s00425-003-1053-0
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DOI: https://doi.org/10.1007/s00425-003-1053-0