Journal of Zhejiang University-SCIENCE A

, Volume 4, Issue 3, pp 346–351

Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass

  • Chai Ming-liang 
  • Wang Bing-liang 
  • Kim Jae-yeoul 
  • Lee Jong-min 
  • Kim Doo-hwan 
Chemical and Biotechnology

DOI: 10.1631/jzus.2003.0346

Cite this article as:
Chai, M., Wang, B., Kim, J. et al. J. Zhejiang Univ. Sci. A (2003) 4: 346. doi:10.1631/jzus.2003.0346

Abstract

Embryogenic calli were induced from the seeds of creeping bentgrass (Agrostis palustris Huds.) cv. Regent and colonial bentgrass (Agrostis Tenuis Sibth. Fl. Oxen.) cv. Tiger. The embryogenic calli were precultured on fresh medium for 4–7 days and then co-cultivated withAgrobacterium tumefaciens, LBA4404. which contains plasmid vector-pSBGM harboring bar coding region, synthetic green fluorescent protein (sGFP) coding region and matrix attachment region (MAR). After 3 days of co-cultivation, the calli were washed thoroughly and transferred to MS medium containing 2 mg/L of 2, 4-D, 12–15 mg/L phosphinothricin (PPT) and 250 mg/L of cefotaxime. After 2–3 months of selection, the actively growing calli of ‘Regent’ and ‘Tiger’ were transferred to MS medium with 12–15 mg/L PPT and 250 mg/L cefotazime for regeneration. The putative transformants were maintained on MS medium with 3 mg/L PPT for long period but control died within 1 month. After establishing in greenhouse, the transformants also showed strong resistance to 0.4% of herbicide Basta but control plants died within 2 weeks. Under confocal microscope, both young leaves and roots showed significant GFP expression. PCR analysis revealed the presence of a DNA fragment of GFP gene at the expected size (380 bp) in the transformants and its absence in a randomly selected control plant.

Key words

Agrostis palustris Agrostis tenuis Agrobacterium Herbicide resistance sGFP (synthetic green fluorescent protein) 

Document code

CLC number

S603.6 

Copyright information

© Zhejiang University Press 2003

Authors and Affiliations

  • Chai Ming-liang 
    • 1
  • Wang Bing-liang 
    • 1
  • Kim Jae-yeoul 
    • 2
  • Lee Jong-min 
    • 3
  • Kim Doo-hwan 
    • 2
  1. 1.Department of HorticultureZhejiang UniversityHangzhouChina
  2. 2.Department of Horticultural ScienceKonkuk UniversitySeoulKorea
  3. 3.Department of Biology ScienceThe University of AucklandNew Zealand

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