Skip to main content
SpringerLink
Log in

Production of transgenic creeping bentgrass Agrostis stolonifera var. palustris plants by Agrobacterium tumefaciens-mediated transformation using hygromycin selection

  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

A protocol was developed for Agrobacterium tumefaciens-mediated transformation of creeping bentgrass [Agrostis stolonifera L. var. palustris (Huds) Farw]. The transformation was performed using the vector pCAMBIA 1301 which contains the reporter (uidA) gene and the selectable marker hygromycin phosphotransferase (hph) gene. Embryogenic calli initiated from mature seeds were infected with A. tumefaciens strain EHA105 followed by hygromycin selection. Effects of some variables influencing efficiency of T-DNA delivery and selection were examined, and the protocol was successfully employed in the production of transgenic creeping bentgrass. Southern blot analysis demonstrated the integration of a single or two copies of foreign gene, and RT-PCR confirmed the transcription of the genes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BA:

6-benzylaminopurine

MS:

Murashige and Skoog media

T-DNA:

DNA region transferred to plant genome from the Agrobacterium tumefaciens Ti plasmid

2,4-D:

2,4-dichlorophenoxyacetic acid

SAAT:

sonication-assisted Agrobacterium-mediated transformation

GUS:

β-glucuronidase

References

  • F Altpeter JP Xu (2000) ArticleTitleRapid production of transgenic turfgrass (Festuca rubra L.) plants J. Plant Physiol. 157 441–448

    Google Scholar 

  • Y Asano Y Ito M Fukami K Sugiura A Fujiie (1998) ArticleTitleHerbicide-resistant transgenic creeping bentgrass plants obtained by eletroporation using an altered buffer Plant Cell Rep. 17 963–967

    Google Scholar 

  • C Basu H Luo Ap Kausch JM Chandlee (2003) ArticleTitlePromoter analysis in transient assays using a GUS reporter gene construct in creeping bentgrass (Agrostis palustris L.) J. Plant Physiol. 160 1233–1239

    Google Scholar 

  • AJE Bettany SJ Dalton E Timms B Manderyck MS Dhanoa P Morris (2003) ArticleTitleAgrobacterium tumefaciens-mediated transformation of Festuca arundinacea (Schreb.) and Lolium multiflorum (Lam.) Plant Cell Rep. 21 437–444

    Google Scholar 

  • ML Chai KK Senthil DH Kim (2004) ArticleTitleTransgenic plants of colonial bentgrass from embryogenic callus via Agrobacterium-mediated transformation Plant Cell, Tissue Organ Cult. 77 165–171

    Google Scholar 

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

    Google Scholar 

  • SJ Dalton AJE Bettany E Timms P Morris (1998) ArticleTitleTransgenic plants of Lolium multiflorum, Lolium perenne, Festuca arundinacea and Agrostis stolonifera by silicon carbide fibre mediated transformation of cell suspension cultures Plant Sci. 132 31–43

    Google Scholar 

  • M Fagard H Vaucheret (2000) ArticleTitle(Trans)gene silencing in plants: How many mechanisms Annu. Rev. Plant Physiol. Plant Mol. Biol. 51 167–194

    Google Scholar 

  • JJ Goldman WW Hanna GH Fleming P Ozias-Akins (2004) ArticleTitlePloidy variation among herbicide-resistance bermudagrass plants of cv Tif Eagle transformed with the bar gene. Plant Cell Rep. 22 553–560

    Google Scholar 

  • SB Ha FS Wu TK Thorne (1992) ArticleTitleTransgenic turf-type tall fescue (Festuca arundinacea Schreb.) plants regenerated from protoplasts Plant Cell Rep. 11 601–604

    Google Scholar 

  • Y Hiei S Ohta T Komari T Kumashiro (1994) ArticleTitleEfficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA Plant J. 6 271–282

    Google Scholar 

  • EE Hood SB Gelvin LS Melchers A Hoekema (1993) ArticleTitleNew Agrobacterium helper plasmids for gene transfer to plants Transgenic Res. 2 208–218

    Google Scholar 

  • Y Ishida H Saito S Ohta Y Hiei T Komari T Kumashiro (1996) ArticleTitleHigh efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens Nat. Biotechnol. 14 745–750

    Google Scholar 

  • RA Jefferson (1987) ArticleTitleAssaying chimeric genes in plants: the GUS gene fusion system Plant Mol. Biol. Rep. 5 387–405

    Google Scholar 

  • L Li R Qu (2004) ArticleTitleDevelopment of highly regenerable callus lines and biolistic transformation of turf-type common bermudagrass (Cynodon dactylon (L.) Pers.) Plant Cell Rep. 22 403–407

    Google Scholar 

  • H Luo Q Hu C Longo AP Kausch JM Chandlee JK Wipff CR Fricker (2003) ArticleTitleAgrobacterium tumefaciens-mediated creeping bentgrass (Agrostis stolonifera L.) transformation using phosphinothricin selection results in a high frequency of single-copy transgene integration Plant Cell Rep. 22 645–652

    Google Scholar 

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

    Google Scholar 

  • M Murray WF Thompson (1980) ArticleTitleRapid isolation of high molecular weight plant DNA Nucleic Acids Res. 8 4321–4325

    Google Scholar 

  • J Sambrook EF Fritsch T Maniatis (1989) Molecular Cloning A Laboratory Manual 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • D Shibata YG Liu (2000) ArticleTitleAgrobacterium-mediated plant transformation with large DNA fragments Trends Plant Sci. 5 354–357

    Google Scholar 

  • G Spangenberg ZY Wang XL Wu J Nagel VA Iglesias I Potrykus (1995) ArticleTitleTransgenic tall fescue (Festuca arundinacea) and red fescue (F rubra) plants from microprojectile bombardment of embryogenic suspension cells. J. Plant Physiol. 145 693–701

    Google Scholar 

  • THN Trick JJ Finer (1998) ArticleTitleSonication-assisted Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill] embryogenic suspension culture tissue Plant Cell Rep. 17 482–488

    Google Scholar 

  • ZY Wang T Takamizo VA Iglesias M Osusky J Nagel I Potrykus G Spangenberg (1992) ArticleTitleTransgenic plants of tall fescue (Festuca Arundiacea Schreb.) obtained by direct gene transfer to protoplasts Biotechnology 10 691–696

    Google Scholar 

  • L Xiao SB Ha (1997) ArticleTitleEfficient selection and regeneration of creeping bentgrass transformations following particle bombardment Plant Cell Rep. 16 874–878

    Google Scholar 

  • TT Yu DZ Skinner GH Liang HN Trick S Muthukrishnan (2000) ArticleTitleAgrobacterium-mediated transformation of creeping bentgrass using GFP as a reporter gene Hareditas 133 229–233

    Google Scholar 

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

    Google Scholar 

  • H Zhong MG Bolyard C Srinivasan MB Sticklen (1993) ArticleTitleTransgenic plants of turfgrass (Agrostis palustris Huds.) from microprojectile bombardment of embryogenic callus Plant Cell Rep. 13 1–6

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. State Key laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, 310012, Hangzhou, China

    Ning Han, Dong Chen, Hong-Wu Bian, Min-Juan Deng & Mu-Yuan Zhu

Authors
  1. Ning Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dong Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hong-Wu Bian
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Min-Juan Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mu-Yuan Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mu-Yuan Zhu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Han, N., Chen, D., Bian, HW. et al. Production of transgenic creeping bentgrass Agrostis stolonifera var. palustris plants by Agrobacterium tumefaciens-mediated transformation using hygromycin selection. Plant Cell Tiss Organ Cult 81, 131–138 (2005). https://doi.org/10.1007/s11240-004-4042-5

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-004-4042-5

Keywords

Search

Navigation