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<p>Carnation (<i>Dianthus caryophyllus</i>, L.) is a major ornamental plant, and efficient transformation methods are desired. The purpose of this report is to provide an efficient transformation method using agrobacterium (<i>Rhyzobium radiobactor</i>). In vitro multiple shoots were induced, and the leaves were used as the explants. Agrobacterium strains, binary vectors, compositions of the co-cultivation medium, concentrations of sucrose in the regeneration and selection medium, and tissue culture vessels were studied and showed significant improvements. A highly virulent agrobacterium strain, AGL0 showed higher transformation efficiency than a strain, LBA4404. A binary vector with the neomycin phosphotransferase II (NPTII) gene driven by the cauliflower mosaic virus 35S (35S) promoter was more effective than a vector with the NPTII gene driven by the nopaline synthase (Nos) promoter. Removal of the total components of Murashige and Skoog (MS) medium from the co-cultivation medium was important for efficient transformation. The best sucrose concentration in the regeneration and selection medium was 10&#xa0;g/L. Plastic boxes during the regeneration and selection step gave higher transformation efficiency than petri dishes. Combination of these factors led to transformation efficiency of more than 20%, which means 20 out of 100 explants produced transgenic plants in this study. The transformed plants expressed the β-glucuronidase (GUS) reporter gene in the whole plants, and the reporter gene was transmitted to the next generations by cross-pollination.</p>

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Agrobacterium strain, promoter of selection marker gene, medium for co-cultivation and regeneration, and culture vessel were important for efficient transformation in carnation

  • 期刊论文

  • 发表日期：

    2026年2月5日

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  • Kanji Mamiya,
  • Eiko Mori

摘要

Carnation (Dianthus caryophyllus, L.) is a major ornamental plant, and efficient transformation methods are desired. The purpose of this report is to provide an efficient transformation method using agrobacterium (Rhyzobium radiobactor). In vitro multiple shoots were induced, and the leaves were used as the explants. Agrobacterium strains, binary vectors, compositions of the co-cultivation medium, concentrations of sucrose in the regeneration and selection medium, and tissue culture vessels were studied and showed significant improvements. A highly virulent agrobacterium strain, AGL0 showed higher transformation efficiency than a strain, LBA4404. A binary vector with the neomycin phosphotransferase II (NPTII) gene driven by the cauliflower mosaic virus 35S (35S) promoter was more effective than a vector with the NPTII gene driven by the nopaline synthase (Nos) promoter. Removal of the total components of Murashige and Skoog (MS) medium from the co-cultivation medium was important for efficient transformation. The best sucrose concentration in the regeneration and selection medium was 10 g/L. Plastic boxes during the regeneration and selection step gave higher transformation efficiency than petri dishes. Combination of these factors led to transformation efficiency of more than 20%, which means 20 out of 100 explants produced transgenic plants in this study. The transformed plants expressed the β-glucuronidase (GUS) reporter gene in the whole plants, and the reporter gene was transmitted to the next generations by cross-pollination.