Abstract
An efficient procedure for producing transgenic Hevea brasiliensis callus and plant lines from clone PB 260 was established with Agrobacterium tumefaciens using strain EHA105 harbouring the vector pCAMBIA2301. Transformation capacity and competence of the embryogenic calli were improved after two cycles of cryopreservation. When the cocultivation temperature was reduced from 27 to 20°C, the duration of this phase could be increased up to 7 days, promoting an increase in GUS activity. These transformation conditions led to the isolation of 24 callus lines resistant to paromomycin, which is used as a selection agent. Nineteen of these lines revealed the existence of one to four copies of T-DNA by Southern-blot analysis. Nine of them were transferred for regeneration by somatic embryogenesis. Three hundred seventy-four transgenic plants have thus been generated from six independent lines bearing 1, 2 or 3 copies of T-DNA. The efficiency and reproducibility of this method means that functional characterization of genes involved in natural rubber production can be envisaged.
Similar content being viewed by others
Abbreviations
- 3,4-D :
-
3,4-Dichlorophenoxyacetic acid
- ABA :
-
Abscisic acid
- BAP :
-
Benzylaminopurine
- DEV :
-
Development medium
- EXP :
-
Expression medium
- FM :
-
Fresh matter
- GR :
-
Growth regulator
- GUS :
-
β-Glucuronidase
- MM :
-
Maintenance medium
References
Arokiaraj P, Jones H, Jaafar H, Coomber S, Charlwood BV (1996) Agrobacterium mediated transformation of Hevea anther calli and their regeneration into plantlets. J Nat Rubber Res 11:77
Arokiaraj P, Rueker F, Obermayr E, Shamsul Bahri AR, Hafsah J, Carter DC, Yeang HY (2002) Expression of human serum albumin in transgenic Hevea brasiliensis. J Rubber Res 5:157–166
Arokiaraj P, Wan Abdul Rahaman WY (1991) Agrobacterium-mediated transformation of hevea cells derived from in vitro and in vivo seedling cultures. J Nat Rubber Res 6:55–61
Arokiaraj P, Yeang HY, Cheong KF, Hamzah S, Jones H, Coomber S, Charlwood BV (1998) CaMV 35S promoter directs β-glucuronidase expression in the laticiferous system of transgenic Hevea brasiliensis (rubber tree). Plant Cell Rep 17:621–625
Baron C, Domke N, Beinhofer M, Hapfelmeier S (2001) Elevated temperature differentially affects virulence, VirB protein accumulation, and T-pilus formation in different Agrobacterium tumefaciens and Agrobacterium vitis strains. J Bacteriol 183:6852–6861
Blanc G, Lardet L, Martin A, Jacob J-L, Carron M-P (2002) Differential carbohydrate metabolism conducts morphogenesis in embryogenic callus of Hevea brasiliensis (Müll. Arg.). J Exp Bot 53:1–10
Carron MP, Enjalric F, Lardet L, Deschamps A (1989) Rubber (Hevea brasiliensis Müll. Arg.). In: Biotechnology in agriculture and forestry, vol. 5. Springer-Verlag, Berlin Heidelberg, pp 222–245
Carron MP, Etienne H, Lardet L, Campagna S, Perrin Y, Leconte A, Chaine C (1995) Somatic embryogenesis in rubber (Hevea brasiliensis Müll. Arg.). In: Jain SM, Gupta PK, Newton RJ (eds) Somatic embryogenesis in woody plants, vol 2. Kluwer Academic Publishers, Dordrecht, pp 117–136
Cornejo MJ, Wong VL, Blechl AE (1995) Cryopreserved callus: a source of protoplasts for rice transformation. Plant Cell Rep 14:210–214
De Clercq J, Zambre M, Van Montagu M, Dillen W, Angenon G (2002) An optimized Agrobacterium-mediated transformation procedure for Phaseolus acutifolius A. Gray Plant Cell Rep 21:333–340
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version 2. Plant Mol Biol Rep 1:19–22
Dillen W, De Clercq J, Kapila J, Zambre M, Van Montagu M, Angenon G (1997) The effect of temperature on Agrobacterium tumefaciens-mediated gene transfer to plants. Plant J 12:1459–1463
Elleuch H, Gazeau C, David H, David A (1998) Cryopreservation does not affect the expression of a foreign sam gene in transgenic Papaver somniferum cells. Plant Cell Rep 18:94–98
Engelmann F, Lartaud M, Chabrillange N, Carron MP, Etienne H (1997) Cryopreservation of embryogenic calluses of two commercial clones of Hevea brasiliensis. Cryo-Letters 18:107–116
Etienne H, Lartaud M, Michauxferriere N, Carron MP, Berthouly M, Teisson C (1997) Improvement of somatic embryogenesis in Hevea Brasiliensis (Mull Arg) using the temporary immersion technique. In Vitro Cellular Dev Biol-Plant 33:81–87
Fullner KJ, Lara JC, Nester EW (1996) Pilus assembly by Agrobacterium T-DNA transfer genes. Science 273:1107–1109
Fullner KJ, Nester EW (1996) Temperature affects the T-DNA transfer machinery of Agrobacterium tumefaciens. J Bacteriol 178:1498–1504
Gheysen G, Herman L, Breyne P, Van Montagu M, Depicker A (1989) Agrobacterium tumefaciens as a tool for the genetic transformation of plants. In: Butler LO, Harwood C, Moseley BEB (eds) Genetic transformation and expression. Intercept, Andover, pp 151–174
Grant JE, Thomson LMJ, Pither-Joyce MD, Dale TM, Cooper PA (2003) Influence of Agrobacterium tumefaciens strain on the production of transgenic peas (Pisum sativum L.). Plant Cell Rep 21:1207–1210
Hajdukiewicz PSZ, Maliga P (1994) The small versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25:989–994
Hood EE, Gelvin SB, Melchers LS, Hoekema A (1993) New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res 2:208–218
Jayashree R, Rekha K, Venkatachalam P, Uratsu SL, Dandekar AM, Kumari Jayasree P, Kala RG, Priya P, Sushma Kumari S, Sobha S, Ashokan MP, Sethuraj MR, Thulaseedharan A (2003) Genetic transformation and regeneration of rubber tree (Hevea brasiliensis Muell. Arg) transgenic plants with a constitutive version of an anti-oxidative stress superoxide dismutase gene. Plant Cell Rep 22:201–209
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
Jokipii S, Ryynanen L, Kallio PT, Aronen T, Haggman H (2004) A cryopreservation method maintaining the genetic fidelity of a model forest tree, Populus tremula L. × Populus tremuloides Michx. Plant Sci 166:799–806
Montoro P, Etienne H, Carron M-P, Nougarède A (1992) Effect of cytokinins on the induction of embryogenesis and the quality of somatic embryos in Hevea brasiliensis Müll. Arg C R Acad Paris 315:567–574
Montoro PHE, Michaux-Ferriere N, Carron MP (1993) Callus friability and somatic embryogenesis in Hevea brasiliensis. Plant Cell, Tissue Organ Cult 33:331–338
Montoro P, Rattana W, Pujade-Renaud V, Michaux-Ferriere N, Monkolsook Y, Kanthapura R, Adunsadthapong S (2003) Production of Hevea brasiliensis transgenic embryogenic callus lines by Agrobacterium tumefaciens:roles of calcium. Plant Cell Rep 21:1095–1102
Montoro P, Teinseree N, Rattana W, Kongsawadworakul P, Michaux-Ferriere N (2000) Effect of exogenous calcium on Agrobacterium tumefaciens-mediated gene transfer in Hevea brasiliensis (rubber tree) friable calli. Plant Cell Rep 19:851–855
Rattana W, Teinseree N, Tadakittisarn S, Pujade-Renaud V, Monkolsook Y, Montoro P (2001) Characterisation of factors involved in tissue growth recovery and stability of GUS activity in rubber tree (Hevea brasiliensis) friable calli transformed by Agrobacterium tumefaciens. Thai J Agric Sci 34:195–204
Salas MG, Park SH, Srivatanakul M, Smith RH (2001) Temperature influence on stable T-DNA integration in plant cells. Plant Cell Rep 20:701–705
Sambrook J, Fritsch E, Maniatis T (1989) Molecular cloning, a laboratory manual. CHS PressCalifornia
Sunilkumar G, Rathore KS (2001) Transgenic cotton: factors influencing Agrobacterium-mediated transformation and regeneration. Mol Breed 8:37–52
Vancanneyt G, Schimdt R, O'Connor-Sanchez A, Wilmitzer L, Rocha-Sosa M (1990) Construction of an intron-containing marker gene:splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacterium-mediated plant transformation. Mol Gen Genet 220:245–250
Yeang H-Y, Arokiaraj P, Jaafar H, Hamzah S, Arif SMA, Jones H (1998) Rubber latex as an expression system for high-value proteins. In: Shewry PR, Napier JA, Davis PJ (eds) Engineering crop plants for industrial end uses. Portland Press, Portland, pp 55–64
Acknowledgements
The authors thank Dr Marc-Philippe Carron and Dr Ludovic Lardet (CIRAD – France) for providing friable callus lines, Dr Julie Leclercq (CIRAD – France) for careful reading of this manuscript, Dr Richard Jefferson (CAMBIA – Australia) for providing the pCAMBIA2301 vector and the EHA105 A. tumefaciens strain, and Peter Biggins for the English translation
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by P. Debergh
Rights and permissions
About this article
Cite this article
Blanc, G., Baptiste, C., Oliver, G. et al. Efficient Agrobacterium tumefaciens-mediated transformation of embryogenic calli and regeneration of Hevea brasiliensis Müll Arg. plants. Plant Cell Rep 24, 724–733 (2006). https://doi.org/10.1007/s00299-005-0023-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-005-0023-3