Abstract
Chinese leymus [Leymus chinensis (Trin.) Tzvel.] is a perennial grass (tribe Gramineae) that is widely distributed throughout northern China and Mongolia where it is produced as a forage product. Severe production losses due to weed growth have serious economic consequences, and as non-selective herbicides not only kill the weeds but are also harmful to this forage grass, the introduction of a foreign gene for resistance to the herbicide Basta is necessary since this species lacks herbicide resistance. We have investigated the transformation of a gene for phosphinothricin acetyltransferase (PAT) through microprojectile bombardment in Chinese leymus. Calli from immature inflorescences cultured on N6 medium supplemented with 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 5.0 mg/l of glutamine were bombarded. The bombarded calli survived on selection medium with 1.0 mg/l of phosphinothricin (PPT). Twenty-three plantlets regenerated from resistant calli on differentiation medium supplemented with 1.0 mg/l 6-benzylaminopurine, 1.0 mg/l kinetin, and 1.0 mg/l PPT, and five of these regenerated plantlets survived on rooting medium with 1.0 mg/l of PPT. PCR and Southern blotting analyses indicated that the PAT gene had been integrated into the genomes of two Chinese leymus plantlets and that the gene was stably transferred to its clonal offsprings. There were no other phenotypic effects associated with transgene expression during vegetative growth except tolerance to the herbicide Basta.
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Abbreviations
- BAP:
-
6-Benzylaminopurine
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- CaMV 35S:
-
Cauliflower mosaic virus
- KIN:
-
Kinetin
- PAT:
-
Phosphinothricin acetyltransferase
- PPT:
-
Phosphinothricin
References
Anonymous (1976) In: Institute of Botany, the Chinese Academy of Sciences (eds) Iconographia Cormophytorum Sinicorum, Tomus. Science Publ, Beijing7
Chen M, Li ZH, Pu SJ (1988) The observation and research on reproductive characteristics of Aneurolepidium chinensis. Res Grassl Ecosyst 2:193–208
Choi YE, Jeong JH, In JK, Yang DC (2003) Production of herbicide-resistant transgenic Panax ginseng through the introduction of the phosphinothricin acetyl transferase gene and successful soil transfer. Plant Cell Rep 21:563–568
Christou P, Ford TL, Kofron M (1991) Production of transgenic rice (Oryza sativa L.) from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Biotechnology 9:957–962
Chu CC, Wang CC, Sun CS, Hsu C, Yin KC, Chu CY (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sin 18:659–668
Clayton WD, Renboize SA (1999) Genera Graminum grasses of the world. Kew Bull Additional Ser 13. Royal Botanic Gardens, Kew
Finer JJ, Philippe V, Johes MW (1992) Development of the particle inflow gene for DNA delivery to plant cell. Plant Cell Rep 11:323–328
Gamborg OL, Miller RA, Okajima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–156
Ghosh Biswas GC, Chen DF, Elliott MC (1998) A routine system for generation of transgenic rice (Oryza sativa L.) plants by microprojectile bombardment of embryogenic cell clusters. Plant Sci 133:203–210
Junghans H, Metzlaff MA (1990) A simple and rapid method for preparation of total plant DNA. Biotechniques 8:176
Leason M, Cunliffe D, Parkin D, Lea PJ, Miflin J (1982) Inhibition of pea leaf glutamine synthetase by sulphoximine, phosphinothricin and other glutamate analogues. Phytochemistry 21:855–857
Lemaux PG, Cho MJ, Zhang S, Bregitzer P (1999) Transgenic cereals: Hordeum vulgare (barley). In: Vasil IK (ed) Molecular improvement of cereal crops. Kluwer, Dordrecht, pp 255–316
Li CH (1990) Studies on the chromosome karyotype and G-banded chromosomes of Leymus chinensis and Roegneria pendulina. Acta Pratacul Sin 1:55–62
Liu GS, Liu JS, Qi DM, Chu CC, Li HJ (2004) Factors affecting plant regeneration from tissue culture of Chinese leymus (Leymus chinensis). Plant Cell Tissue Organ Cult 76:175–178
Mallik MAB (2001) Selective isolation and screening of soil microorganisms for metabolites with herbicidal potential. J Crop Prot 4:219–236
Maniatis T, Fritsch EF, Sambrook J (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Press, New York
Marchant R, Power JB, Lucas JA, Davey MR (1998) Biolistic transformation of Rose (Rosa hybrida L. Ann Bot 81:109–114
Merkel U, Rathke GW, Schuster C, Warnstorff K, Diepenbrock W (2004) Use of glufosinate-ammonium to control cruciferous weed species in glufosinate-resistant winter oilseed rape. Field Crops Res 85:237–249
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Plant Physiol 15:473–497
Rogers SO, Bendich AJ (1985) Extraction of DNA from milligram amounts of fresh, herbatium and mummified plant tissues. Plant Mol Biol 5:69–76
Sripaoraya S, Marchant R, Power JB, Davey MR (2001) Herbicide-tolerant transgenic pineapple (Ananas comosus) produced by microprojectile bombardment. Ann Bot 88:597–603
Vain P, McMullen MD, Finer JJ (1993) Osmotic treatment enhances particle bombardment mediated transient and stable transformation of maize. Plant Cell Rep 12:84–88
Vasil V, Castillo AM, Fromm ME, Vasil IK (1992) Herbicide resistant fertile transgenic wheat plants obtained by microprojectile bombardment of regenerable embryogenic callus. Biotechnology 10:667–674
Acknowledgements
We are grateful to Prof. C.C. Chu for his advice. Our special thanks are given to Jiang-shu Liu for her technical assistance and Dr. Shi-hua Shen for providing plasmids with the PAT gene. The Biotechnology of Pasture Plant Program is funded by the Key Project of the Chinese Academy of Sciences (KSCXI-08)
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Communicated by M. Beckert
The Biotechnology of Pasture Plant Program is funded by the Key Project of the Chinese Academy of Sciences (KSCX1-08)
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Shu, Q.Y., Liu, G.S., Xu, S.X. et al. Genetic transformation of Leymus chinensis with the PAT gene through microprojectile bombardment to improve resistance to the herbicide Basta. Plant Cell Rep 24, 36–44 (2005). https://doi.org/10.1007/s00299-004-0908-6
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DOI: https://doi.org/10.1007/s00299-004-0908-6