CSIRO Publishing Books Journals About Us Shopping Cart You are here: Journals > Functional Plant Biology   
Functional Plant Biology
  Plant Function & Evolutionary Biology
 
Search
 
 
  Advanced Search
   

Journal Home
About the Journal
Editorial Board
Contacts
Content
Online Early
Current Issue
Just Accepted
All Issues
Special Issues
Research Fronts
Reviews
Evolutionary Reviews
Sample Issue
For Authors
General Information
Notice to Authors
Submit Article
Open Access
For Referees
General Information
Review Article
Annual Referee Index
Referee Guidelines
For Subscribers
Subscription Prices
Customer Service
Print Publication Dates

 Early Alert
Subscribe to our email Early Alert or RSS feeds for the latest journal papers.

 Connect with us
facebook   youtube

 PrometheusWiki
PrometheusWiki
Protocols in ecological and environmental plant physiology

 

Article << Previous     |     Next >>   Contents Vol 32(9)

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

Zeng-Yu Wang A B, Yaxin Ge A

A Forage Improvement Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401, USA.
B Corresponding author. Email: zywang@noble.org
 
PDF (1 MB) $25
 Export Citation
 Print
  


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.


   
Subscriber Login
Username:
Password:  

    


 
Top  Email this page
 
Legal & Privacy | Contact Us | Help

CSIRO

© CSIRO 1996-2012