| Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass |
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| 作者姓名： | 柴明良 汪炳良 KIM Jae-yeoul LEE Jong-min KIM Doo-hwan |
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| 作者单位： | Department of Horticulture，Zhejiang University，Department of Horticulture，Zhejiang University，Department of Horticultural Science，Konkuk University，Department of Biology Science，The University of Auckland，New Zealand，Department of Horticultural Science，Konkuk University Hangzhou 310029，China，Hangzhou 310029，China，Seoul 143 701，Korea，Seoul 143 701，Korea |
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| 摘 要： | INTRODUCTIONCreepingbentgrass (AgrostispalustrisHuds.)isanoutstandingcool seasonspeciesavailableforuseongolfcourseputtinggreens,tees,andcloselymowedfairways.Althoughco lonialbentgrass (AgrostistenuisSibth .Fl.Ox en .)isnotwelladaptedtotheverylowmowingheig…
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Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass |
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| CHAI Ming liang,WANG Bing liang,KIM Jae yeoul ,LEE Jong min ,KIM Doo hwan WT.,.BZ.Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass[J].Journal of Zhejiang University Science,2003(3). |
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| Authors: | CHAI Ming liang WANG Bing liang KIM Jae yeoul LEE Jong min KIM Doo hwan [WT BZ] |
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| Institution: | CHAI Ming liang,WANG Bing liang,KIM Jae yeoul 2,LEE Jong min 3,KIM Doo hwan 2 [WT9.,10.BZ] |
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| 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 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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| Keywords: | Agrostis palustris Agrostis tenuis Agrobacterium Herbicide resistance sGFP(synthetic green fluorescent protein) |
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