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Ploidy variation among herbicide-resistant bermudagrass plants of cv. TifEagle transformed with the bar gene

  • Genetic Transformation and Hybridization
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Abstract

A protocol was developed for biolistic transformation of hybrid bermudagrass cv. TifEagle using the bar gene. TifEagle is an ultradwarf used exclusively on golf greens. Herbicide resistance should serve as a useful management tool, especially if methyl-bromide is unavailable for fumigation prior to plant establishment. Hybrid bermudagrass is completely sterile, which should limit the chance of gene escape via out-crossing. Sliced nodes were used to initiate embryogenic tissue cultures on MS medium supplemented with 1 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.01 mg/l 6-benzylaminopurine (BA). Embryogenic tissue was bombarded with the bar gene, and herbicide-resistant tissue was selected in the dark on medium supplemented with 0.75 mg/l 2,4-D, 0.01 mg/l BA and 5–15 mg/l phosphinothricin (PPT). Resistant somatic embryos were induced to germinate in the light on MS medium supplemented with 0.13 mg/l 2,4-D and 0.5 mg/l BA. Plants were transferred to the greenhouse after rooting in the presence of 10–15 mg/l PPT and testing positive in a chlorophenol red assay. A total of 89 herbicide-resistant plants were recovered from at least nine independent events from six separate bombardments, although the number of independent transformation events was not confirmed for the entire group. Flow cytometry indicated that most of the plants (82/89) were hexaploid and that the remaining seven plants were triploid. The hexaploid plants were a darker green than the triploids or TifEagle control. Other variation, present only in the hexaploids, included an increased leaf width and length. Southern blot hybridization confirmed genomic integration of the bar gene in triploid and a subset of hexaploid herbicide-resistant plants. AFLP analysis did not indicate changes in DNA profiles using [33P] and a sample of 32 hexaploid plants recovered from a single bombardment. DNA profiles were very similar to that of the TifEagle control with a semi-automated fluorescence-based AFLP.

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Abbreviations

BA::

6-Benzylaminopurine

2,4-D::

2,4-Dichlorophenoxyacetic acid

GFP::

Green fluorescent protein

GUS::

β-Glucuronidase

PAT::

Phosphinothricin acetyl transferase

PPM::

Plant preservative mixture

PPT::

Phosphinothricin

References

  • Bregitzer P, Halbert SE, Lemaux PG (1998) Somaclonal variation in the progeny of transgenic barley. Theor Appl Genet 96:421–425

    Article  Google Scholar 

  • Chaudhury A, Qu R (2000) Somatic embryogenesis and plant regeneration of turf-type bermudagrass: effects of 6-benzyladenine in callus induction medium. Plant Cell Tissue Organ Cult 60:113–120

    Article  CAS  Google Scholar 

  • Cho MJ, Ha CD, Lemaux PG (2000) Production of transgenic tall fescue and red fescue plants by particle bombardment of mature seed-derived highly regenerative tissues. Plant Cell Rep 19:1084–1089

    CAS  Google Scholar 

  • Christensen AH, Quail PH (1996) Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transgenic Res 5:1–6

    Google Scholar 

  • Datla R, Bekkaoui F, Hammerlindl JK, Pilate G, Dunstan DI, Crosby WL (1993) Improved high-level constitutive foreign gene expression in plants using an AMV RNA4 untranslated leader sequence. Plant Sci 94:139–149

    CAS  Google Scholar 

  • Fleming GH, Olivares-Fuster O, Fatta Del-Bosco S, Grosser JW (2000) An alternative method for the genetic transformation of sweet orange. In Vitro Cell Dev Biol Plant 36:450–455

    Google Scholar 

  • Goldman JJ, Hanna WW, Fleming G, Ozias-Akins P (2003) Fertile transgenic pearl millet [Pennisetum glaucum (L.) R. Br.] plants recovered through microprojectile bombardment and phosphinothricin selection of apical meristem-,inflorescence-, and immature embryo-derived embryogenic tissues. Plant Cell Rep 21:999–1099

    Article  CAS  PubMed  Google Scholar 

  • Hanna WW, Elsner JE (1999) TifEagle bermudagrass. Crop Sci 39:1258

    Google Scholar 

  • Hanna WW, Burton GW, Johnson AW (1990) Registration of Tifton 10 turf bermudagrass. Crop Sci 30:1355–1356

    Google Scholar 

  • Hartman CL, Lee L, Day PR, Tumer NE (1994) Herbicide resistant turfgrass (Agrostis palustris Huds.) by biolistic transformation. Biotechnology 12:919–23

    CAS  Google Scholar 

  • Karp A (1994) Origins, causes, and uses of variation in plant tissue cultures. In: Vasil IK, Thorpe TA (eds) Plant cell and tissue culture. Kluwer, Dordrecht, pp 139–151

  • Karp A (1995) Somaclonal variation as a tool for crop improvement. Euphytica 85:295–302

    Google Scholar 

  • Kramer K, DiMaio J, Carswell GK, Shillito RD (1993) Selection of transformed protoplast-derived Zea mays colonies with phosphinothricin and a novel assay using the pH indicator chlorophenol red. Planta 190:454–458

    CAS  Google Scholar 

  • Moffett AA, Hurcombe R (1949) Chromosome numbers of South African grasses. Heredity 3:369–373

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    CAS  Google Scholar 

  • Ozias-Akins P, Lubbers EL, Hanna WW, McNay JW (1993) Transmission of the apomictic mode of reproduction in Pennisetum:co-inheritance of the trait and molecular markers. Theor Appl Genet 85:632–638

    Google Scholar 

  • Powell JB, Burton GW, Taliaferro CM (1968) A hexaploid clone from a tetraploid x diploid cross in Cynodon. Crop Sci 8:184–185

    Google Scholar 

  • Rogers SO, Bendich AJ (1985) Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Mol Biol 5:69–7

    CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

  • SAS Institute (1999) The SAS system for Windows. Release 8.0. SAS Institute, Cary, N.C.

  • 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

    Google Scholar 

  • Taylor MG, Vasil IK (1996) The ultrastructure of somatic embryo development in pearl millet (Pennisetum glaucum; Poaceae). Am J Bot 83:28–44

    Google Scholar 

  • Vos P, Hogers R, Bleeker M, Reijans M, van der Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414

    CAS  PubMed  Google Scholar 

  • Wang Y, Browning M, Ruemmele BA, Chandlee JM, Kausch AP, Jackson N (2003) Glufosinate reduces fungal disease in transgenic glufosinate-resistant bentgrasses (Agrostis spp.). Weed Sci 51:130–137

    CAS  Google Scholar 

  • Zabeau M, Vos P (1993) Selective restriction fragment amplification: a general method for DNA fingerprinting. Eur Patent Application No. 92402629.7

    Google Scholar 

  • Zhang GY, Lu S, Chen TA, Chen SY, Funk R, Meyer W (2001) Transformation of triploid bermudagrass with the BADH gene for stress tolerance. In: Am Soc Agron (ed) Annu Meet Abstr 2001. American Society of Agronomy, Madison, Wis., p

  • Zhang GY, Lu S, Chen TA, Funk CR, Meyer M (2002) Application of somaclonal variation in triploid bermudagrass breeding. In: Am Soc Agron (ed) Annu Meet Abstr 2002. American Society of Agronomy, Madison, Wis., p

  • Zhang GY, Lu S, Chen TA, Funk CR, Meyer WA (2003) Transformation of triploid bermudagrass (Cynodon dactylon × C. transvaalensis cv. TifEagle) by means of biolistic bombardment. Plant Cell Rep 21:860–864

    Google Scholar 

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Acknowledgements

We gratefully thank Anne Bell, Jacolyn Merriman, Gunawati Gunawan, and the turf genetics group at the University of Georgia Coastal Plain Experiment Station at Tifton for excellent technical assistance. We also thank Drs. Zhenbang Chen and Shailendra Goel for molecular assistance and Dr. Benjamin Mullinix for statistical assistance. Funding was provided by the University of Georgia Research Foundation and the Georgia Seed Development Commission.

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Authors and Affiliations

  1. Department of Crop and Soil Sciences, University of Georgia Tifton Campus, Tifton, GA 31793-0748, USA

    J. J. Goldman & W. W. Hanna

  2. Department of Horticulture, University of Georgia Tifton Campus, Tifton, GA 31793-0748, USA

    G. H. Fleming & P. Ozias-Akins

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  1. J. J. Goldman
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  2. W. W. Hanna
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  3. G. H. Fleming
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  4. P. Ozias-Akins
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Correspondence to P. Ozias-Akins.

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Communicated by M.E. Horn

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Goldman, J.J., Hanna, W.W., Fleming, G.H. et al. Ploidy variation among herbicide-resistant bermudagrass plants of cv. TifEagle transformed with the bar gene. Plant Cell Rep 22, 553–560 (2004). https://doi.org/10.1007/s00299-003-0736-0

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