• Title/Summary/Keyword: agrobacterium tumefaciens

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Improved in vitro Regeneration of Potato (Solanum tuberosum cv. Superior) Transformed by Agrobacterium Expressing $\beta-Glucuronidase$

  • Park, Yoon-Kyung;Park, Gene-Sue;Yang, Young-Ki;Cheong, Hyeon-Sook
    • Journal of Plant Biology
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    • v.39 no.2
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    • pp.93-98
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    • 1996

  • In order to enhance the system of potato transformation and further regeneration, potato was transformed using the Agrobacterium tumefaciens harboring $\beta$-glucuronidase (GUS) gene. We found that a series fo modified medium ttained 100% shoot regeneration within 5 weeks after the preincubated explants on stage I medium were infected with Agrobacterium. Callus appeared at the cut edges of stem segments on stage II medium, mainly at the basal parts. Some explants started to form shoots after two to three weeks on stage III medium containing kanamycin (50 mg/L). When transferred to MS medium containing 200 mg/L kanamycin, 81% of the transformed shoots formed roots at the cut edge of the plantlets. In contrast, untrasformed shoots never rooted and became yellowish after few weeks under the same conditions. Southern and northern analysis indicated in vitro shoot regeneration on the callus derived from the potato explants, which were incubated with Agrobacteria. The regeneration cycle was shortened after the transformatin and finally the transformation efficiency was highly enhanced.

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Construction of a Plant Expression Vector for the Coat Protein Gene of Cucumber Mosaic Virus-As Strain for Plant Transformation (오이 모자이크 바이러스 As계통 외피단백질 유전자의 식물체 형질질환을 위한 발현벡타의 구축)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.66-72
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    • 1995

  • The coat protein (CP) gene of cucumber mosaic virus-As (CMV-As) strain was engineered for expression in the plant by using the cauliflower mosaic virus 35S transcript regulatory sequences. The CP gene was cloned into an Agrobacterium-derived binary vector. A chimeric gene was constructed by the cDNA of CMV-As CP and plant expression vector pBI121. The clone, pCMAS66, was first introduced into the phagemid vector pSPORT1 for situating sense orientation for translation and making restriction sites in order to re-introduce plant expression vector, pHI121. The resulting subclone pCASCP02 and plant expression vector pBI121 were treated with BamHI-SacI for excising the target gene and removing GUS gene, respectively. After Agrobacterium transformation by freeze-thaw technique, the clone, pCMASCP121-123 which contains sense orientation of the target gene, was selected and confirmed by restriction endonuclease analysis. The CMV-As CP gene was introduced into A. tumefaciens. The results on tobacco plant transformation with the vector system revealed that the system could be successfully introduced and showed high frequency of selection to putative transformations.

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