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Journal of Zhejiang University SCIENCE A 2003 Vol.4 No.3 P.346-351


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

Author(s):  CHAI Ming-liang, WANG Bing-liang, KIM Jae-yeoul, LEE Jong-min, KIM Doo-hwan

Affiliation(s):  Department of Horticulture, Zhejiang University, Hangzhou 310029, China; more

Corresponding email(s):   mlchai@zju.edu.cn

Key Words:  Agrostis palustris, Agrostis tenuis, Agrobacterium, Herbicide resistance, sGFP(synthetic green fluorescent protein)

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CHAI Ming-liang, WANG Bing-liang, KIM Jae-yeoul, LEE Jong-min, KIM Doo-hwan. Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass[J]. Journal of Zhejiang University Science A, 2003, 4(3): 346-351.

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journal="Journal of Zhejiang University Science A",
publisher="Zhejiang University Press & Springer",

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%A CHAI Ming-liang
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%A KIM Jae-yeoul
%A LEE Jong-min
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%J Journal of Zhejiang University SCIENCE A
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2003.0346

T1 - Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass
A1 - CHAI Ming-liang
A1 - WANG Bing-liang
A1 - KIM Jae-yeoul
A1 - LEE Jong-min
A1 - KIM Doo-hwan
J0 - Journal of Zhejiang University Science A
VL - 4
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SP - 346
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.2003.0346

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 with agrobacterium 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 cefotaxime 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.

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