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Highly efficient Agrobacterium-based transformation system for callus cells of the C3 halophyte Suaeda salsa

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Abstract

Genetic manipulation technologies have been limited in the halophyte Suaeda salsa L. due to the lack of an efficient transformation system. Here, we examined factors affecting transformation and developed an efficient transformation system at the cell level using S. salsa hypocotyl as starting material. S. salsa hypocotyl explants from 10-day-old seedlings were precultured for 2 days on a hygromycin (hyg)-free callus induction medium (CIM) and then inoculated with Agrobacterium tumefaciens suspension at a concentration of 0.5 at OD600 for 5–10 min. After cocultivation with A. tumefaciens for 4 days in the dark, followed by selection on carbenicillin (carb) for 3 days, explants were placed on CIM containing 10 mg l−1 hyg and 500 mg l−1 carb with three to four consecutive subcultures for up to 45 days. β-Glucuronidase assays showed an average transformation frequency of 62.89%. Gene integration was confirmed by polymerase chain reaction analysis and Northern blot analysis. To our knowledge, this is the first study to show Agrobacterium-mediated transformation in the C3 halophyte S. salsa.

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Acknowledgments

We are grateful for financial support by the NSFC (National Natural Science Research Foundation of China, Project Nos. 30670177 and 30270793) and Specialized Research Fund for the Doctoral Program of Higher Education (SRFDP, 20050445003). We are also grateful to Prof. Juren Zhang (College of Life Science, Shandong University, China) for providing plasmids pCAMBIA1301 and LBA4404. We thank Prof. Hans J. Bohnert (Department of Plant Biology and Department of Crop Sciences University of Illinois at Urbana-Champaign, USA) and Ken C. Sink (Department of Horticulture, Michigan State University, USA) for a critical reading of the manuscript.

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Authors and Affiliations

  1. Key Laboratory of Plant Stress Research, College of Life Science, Shandong Normal University, No. 88, East Wenhua Road, Jinan, 250014, People’s Republic of China

    Shu-Zhen Zhao, Yuan Ruan & Bao-Shan Wang

  2. Department of Biological Science and Technology, Shandong Institute of Education, No. 36, Lishan Road, Jinan, 250013, People’s Republic of China

    Hong-Zhao Sun

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  1. Shu-Zhen Zhao
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  2. Yuan Ruan
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  4. Bao-Shan Wang
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Correspondence to Bao-Shan Wang.

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Communicated by J. Sadowski.

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Zhao, SZ., Ruan, Y., Sun, HZ. et al. Highly efficient Agrobacterium-based transformation system for callus cells of the C3 halophyte Suaeda salsa . Acta Physiol Plant 30, 729–736 (2008). https://doi.org/10.1007/s11738-008-0174-2

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  • DOI: https://doi.org/10.1007/s11738-008-0174-2

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