Plant Biotechnology
Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 13 No. 3, Issue of May 15, 2010
© 2010 by Pontificia Universidad Católica de Valparaíso -- Chile Received June 23, 2009 / Accepted December 3, 2009
DOI: 10.2225/vol13-issue3-fulltext-6
RESEARCH ARTICLE

Genotypic variation of Agrobacterium-mediated transformation of Italian ryegrass

Ki-Won Lee
 Grassland and Forages Division
National Institute of Animal Science
Rural Development Administration
Cheonan, Chungnam, 330-801, South Korea

Gi Jun Choi
 Grassland and Forages Division
National Institute of Animal Science
Rural Development Administration
Cheonan, Chungnam, 330-801, South Korea

Ki-Yong Kim
 Grassland and Forages Division
National Institute of Animal Science
Rural Development Administration
Cheonan, Chungnam, 330-801, South Korea

Sei Hyung Yoon
 Grassland and Forages Division
National Institute of Animal Science
Rural Development Administration
Cheonan, Chungnam, 330-801, South Korea

Hee Chung JI
 Grassland and Forages Division
National Institute of Animal Science
Rural Development Administration
Cheonan, Chungnam, 330-801, South Korea

Hyung Soo Park
 Grassland and Forages Division
National Institute of Animal Science
Rural Development Administration
Cheonan, Chungnam, 330-801, South Korea

Young Chul Lim
 Grassland and Forages Division
National Institute of Animal Science
Rural Development Administration
Cheonan, Chungnam, 330-801, South Korea

Sang-Hoon Lee*
 Grassland and Forages Division
National Institute of Animal Science
Rural Development Administration
Cheonan, Chungnam, 330-801, South Korea
E-mail: sanghoon@korea.kr

*Corresponding author

Financial support: This work was supported by grants from the National Institute of Animal Science (Project no. 200901OFT082558123) and the BioGreen 21 Program (Project no. 20070301-034-015), Rural Development Administration (RDA), Korea.

Keywords: Agrobacterium, forage crop, Italian ryegrass, transgenic plants.

Abbreviations:

BA:  6-benzyladenine
GUS: β-glucuronidase
HPT: hygromycin phosphotransferase
X-gluc: 5-bromo-4-chloro-3-indolyl-b-glucuronide
Abstract   Full Text

In the present study, genotypic variation of Agrobacterium-mediated transformation of Korean Italian ryegrass has been evaluated. Mature seed-derived calli of seven cultivars were inoculated and co-cultured with Agrobacterium tumefaciens carrying the binary vector pCAMBIA1301, which contains a reporter gene (gus) and a plant selectable marker gene conferring resistance to hygromycin (hpt) in the T-DNA region. The effects of several factors such as callus type and callus age on transformation effectiveness and the expression of the GUS gene were investigated. The highest transformation effectiveness (6.7%) was obtained with the Hwasan 101 cultivar when 9-week-old calli (type-I) were inoculated with Agrobacterium. The overall transformation rates of the examined cultivars ranged from 0.4% to 6.7%. GUS histochemical assays, PCR, and southern analysis of transgenic plants demonstrated that transgenes were successfully integrated into the genome of Italian ryegrass. Thus, evaluation of transformation effectiveness and selection of a suitable cultivar of Italian ryegrass may improve molecular breeding of this species.
Supported by UNESCO / MIRCEN network