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《Acta Prataculturae Sinica》 2011-02
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Establishment of an Agrobacterium mediated genetic transformation system for Eremochloa ophiuroides

ZHANG Fang,WANG Zhou,ZONG Jun-qin,LIU Jian-xiu,SHE Jian-ming(Institute of Botany,Jiangsu Province and Chinese Academy of Sciences,Nanjing 210014,China)  
Agrobacterium tumefaciens strain LBA4404 harboring the plasmid pCAMBIA1301 which contains stress tolerance-related transcription factor ZjDREB1 gene and reporter gene HPT was used to transform axillary bud-derived embryogenic calluses of Eremochloa ophiuroides 'E126'.To systematically optimize the conditions of centipedegrass transformation,several factors which could influence Agrobacterium-mediated DNA transfer were examined,including concentration of selective agent and bacteriostat,bacterial concentration,duration of infection and co-cultivation.Consequently,a transformation system of transgenic centipedegrass plants was established.It was found that the efficiency of transformation highly relied on the following optimal conditions: The concentration of hygromycin for selection was 30 mg/L;the cefotaxime concentration for restraining the Agrobacterium was 400 mg/L;OD600 as 0.3,30 min of infection time and 3 days.Co-cultivation time was the optimized condition for genetic transformation system in E.ophiuroides.Ten transgenic plants were identified by PCR detection from initial 44 regenerated plants.
【Fund】: 江苏省科技支撑项目(BE2008403);; 江苏省科技公共服务平台项目(BM2009905)资助
【CateGory Index】: S688.4
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