Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Rapid and efficient production of transgenic bermudagrass and creeping bentgrass bypassing the callus formation phase

Zeng-Yu Wang A B and Yaxin Ge A
+ Author Affiliations
- Author Affiliations

A Forage Improvement Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401, USA.

B Corresponding author. Email: zywang@noble.org

Functional Plant Biology 32(9) 769-776 https://doi.org/10.1071/FP05083
Submitted: 7 April 2005  Accepted: 27 May 2005   Published: 26 August 2005

Abstract

Callus culture has been an inevitable step in genetic transformation of monocotyledonous (monocot) species. The induction and maintenance of embryogenic calluses is time-consuming, laborious and also requires experience. A straightforward and callus-free transformation procedure was developed and demonstrated for two monocot species, bermudagrass (Cynodon spp.) and creeping bentgrass (Agrostis stolonifera). Stolon nodes were infected and co-cultivated with Agrobacterium tumefaciens harboring pCAMBIA or pTOK233 binary vectors. Green shoots were directly produced from infected stolon nodes 4–5 weeks after hygromycin selection. Without callus formation and with minimum tissue culture, this procedure allowed us to obtain well-rooted transgenic plantlets in only 7 weeks and greenhouse-grown plants in only 9 weeks. The established plants were screened by PCR; the transgenic nature of the plants was demonstrated by Southern hybridisation analysis. Expression of the transgenes was confirmed by northern hybridisation analysis and GUS staining. Based on the number of transgenic plants obtained and the number of stolon nodes inoculated, transformation frequencies of 4.8%–6.1% and 6.3%–11.3% were achieved for bermudagrass and creeping bentgrass, respectively. The rapid and efficient production of transgenic plants without callus induction is a significant improvement for genetic transformation of monocot species.

Keywords: Agrobacterium, Agrostis, bermudagrass, creeping bentgrass, Cynodon, forage and turf grass, genetic transformation, transgenic plant.


Acknowledgments

We thank Kathy Spohn for editing the manuscript. The research was supported by the Samuel Roberts Noble Foundation.


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