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학술저널
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한국잡초학회·한국잔디학회 Weed&Turfgrass Science 한국잔디학회지 제18권 제3호
발행연도
2004.9
수록면
141 - 148 (8page)

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In this report, several factors such as infection time and concentration of bacterial suspension, influencing on transient gene expression in Agrobacterium-mediated transformation were evaluated. An appropriate concentration (O.D600nm = 1.0-1.2) of bateria and 30 min of infection time showed a higher level of GUS expression. To improve transformation efficiency (TE), friable embryogenic calli (FEC) were bombarded by tungsten particles without plasmid DNA, and then co-cultivated with A tumefaciens LBA4404 which contains pTOK233 super binary vector, carrying neomycin phosphotransferase (NPTII), hygromycin phosphotransferase (hpt) and ß-glucuronidase (GUS) genes. Three days after co-cultivation with A tumefaciens and particle bombardment, FEC cultures were transferred to the selection medium (SM' MS medium supplemented with BA l mg/l, hygromycin 100 mg/l, cefotaxime 250 mg/l and vancomycin 200 mg/l). They were cultured for 2 weeks and then transferred to the second SM containing hygromycin 5O mg/l, cefotaxime 200 mg/I and vancomycin 100 mg/l. Later, stable GUS expression was detected 4 to 6 weeks after transfer to the SM. Further, TE from Agrobacterium-mediated transformation after particle bombardment increased to about 3-folds compared with Agrobacterium-mediated transformation without particle bombardment. In the present study, we established an efficient transformation protocol of zoysiagrass by using A tumefaciens in the combination with particle bombardment for the first time.

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