Abstract
A transformation system for triploid bermudagrass (Cynodon dactylon x C. transvaalensis cv. TifEagle) was established with a biolistic bombardment delivery system. Embryogenic callus was induced from stolons and maintained on Murashige and Skoog's medium supplemented with 30 μM dicamba, 20 μM benzylaminopurine, and 100 mg/l myo-inositol. Using the hygromycin phosphotransferase (hpt) gene as the selectable marker gene, we obtained 75 transgenic lines from 18 petri dishes bombarded. Integration of the hpt gene into genomic DNA and transcription of hpt was confirmed by Southern and Northern blot analyses, respectively. Through suspension culture screening, we obtained homogeneously transformed plants showing stable transcription of the hpt gene.
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Abbreviations
- BAP :
-
Benzylaminopurine
- hpt :
-
Hygromycin phosphotransferase gene
- Hyg :
-
Hygromycin B
- GUS :
-
β-Glucuronidase
References
Ahn BJ, Hung FH, King JW (1985) Plant regeneration through somatic embryogenesis in common bermudagrass tissue culture. Crop Sci 25:1107–1109
Ahn BJ, Hung FH, King JW (1987) Regeneration of bermudagrass cultivars and evidence of somatic embryogenesis. Crop Sci 27:594–97
Bettany AJE, Dalton SJ, Timms E, Morris P (1998) Stability of transgene expression during vegetative propagation of protoplast-derived tall fescue (Festuca arundinaceous Schreb.) plants. J Exp Bot 49:1797–1804
Bilang R, Iida S, Peterhans A (1991) The 3′-terminal region of the hygromycin-B resistance gene is important for its activity in E. coli and Nicotiana tabacum. Gene 100:247–25
Christensen AH, Sharrock RA, Quail PH (1992) Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Mol Biol 18:675–689
Croughan SS, Quisenberry SS (1989) Enhancement of fall armyworm (Lepidoptera: Noctuidae) resistance in bermudagrass through cell culture. J Econ Entomol 82:236–38
Dibble MS, Griffin JD (1998) Stable-transformed Kentucky bluegrass plants expressing GUS or BAR. ASA Annu Meet Abstr 1998:143–144
Hartman CL, Lee L, Day PR, Tumer NE (1994) Herbicide resistant turfgrass (Agrostis palustris Huds.) by biolistic transformation. Biotechnology 12:919–23
Hirochika H, Sugimoto K, Otsuki Y, Tsugawa H, Kanda M (1996) Retrotransposons of rice involved in mutations induced by tissue culture. Proc Natl Acad Sci USA 93:7783–88
Inokuma C, Sugiura K, Imaizumi N, Cho C, Kaneko S (1997) Transgenic zoysia (Zoysia japonica Steud.) plants regenerated from protoplasts. Int Turfgrass Soc Res J 8:297–303
Liao CH, Chen TA (1981) Isolation, culture and pathogenicity to Sudan grass of a corynebacterium associated with ratoon stunting of sugarcane and with Bermuda grass. Phytopathology 71:1303–06
McElroy D, Blowers AD, Jenes B (1991) Construction of expression vectors based on the rice actin (Act1) 5′ region for use in monocot transformation. Mol Gen Genet 231:150–160
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–497
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning—a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Spangenberg G, Wang ZY, Wu X, Nagel J, Potrykus I (1995) Transgenic perennial ryegrass (Lolium perenne) plants from microprojectile bombardment of embryogenic suspension cells. Plant Sci 108:209–217
Tai TH, Tanksley SD (1990) A rapid and inexpensive method for isolation of total DNA from dehydrated plant tissue. Plant Mol Biol Rep 8:297–303
Vain P, McAullen MD, Finer JJ (1993) Osmotic treatment particle bombardment-mediated transient and stable transformation of maize. Plant Cell Rep 12:84–88
Wang ZY, Takamizo T, Iglesias VA, Osusky M, Nagel J, Potrykus I, Spangenberg G (1992) Transgenic plants of tall fescue (Festuca arundinacea schreb.) obtained by direct gene transfer to protoplasts. Biotechnology 10:691–896
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Communicated by E.D. Earle
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Zhang, G., Lu, S., Chen, T.A. et al. Transformation of triploid bermudagrass (Cynodon dactylon x C. transvaalensis cv. TifEagle) by means of biolistic bombardment. Plant Cell Rep 21, 860–864 (2003). https://doi.org/10.1007/s00299-003-0585-x
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DOI: https://doi.org/10.1007/s00299-003-0585-x