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Agrobacterium-mediated genetic transformation of grapevine (Vitis vinifera L.) with a novel stilbene synthase gene from Chinese wild Vitis pseudoreticulata

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Abstract

A novel stilbene synthase gene (STS), cloned from Chinese wild Vitis pseudoreticulata (W. T. Wang) and responsible for synthesis of the phytoalexin resveratrol in grapevine, was successfully transferred into V. vinifera L. cv. Thompson Seedless via Agrobacterium tumefaciens-mediated transformation. Using transformation procedures developed in the present study, 72% GFP-positive germinated embryos were produced with about 38% of transformed embryos regenerated into normal plantlets. Integration of the STS gene into the transgenic plants was verified by PCR and Southern blot analysis. Expression of the STS gene was detected by high performance liquid chromatography (HPLC), which showed that the resveratrol concentration in the transgenic plants was 5.5 times higher than that in non-transformed control plants.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

BAP:

6-Benzylaminopurine

CTAB:

Cetyltrimethylammonium bromide

GFP:

Green fluorescent protein

GUS:

β-Glucuronidase

Hyg:

Hygromycin B

HPLC:

High-performance liquid chromatography

HPT II:

Hygromycin phosphotransferase

NOA:

2-Naphthoxyacetic acid

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Acknowledgements

The authors acknowledge Dr. Zongrang Liu, USDA-ARS, Appalachian Fruit Research Station, USA, for providing the base plant expression vector pCAMBIA and Prof. Qiaochun Wang for his critical reading of the manuscript. This work was supported by the National Natural Science Foundation of China (NO. 30370993 & NO. 30571280), Ministry of Science and Technology of China for Transgenic Plant Research and Commercialization Project (JY03-A-19-02), and Ministry of Education of China for Trans-century Talent-training Program at the Northwest A&F University.

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Authors and Affiliations

  1. Key Laboratory of Horticultural Plant Germplasm Resources & Genetic Improvement in Northwest China, Ministry of Agriculture of China, Yangling, Shaanxi, P. R. China

    Chaohong Fan, Ni Pu, Xiping Wang, Yuejin Wang, Li Fang, Weirong Xu & Jianxia Zhang

  2. Key Laboratory of Molecular Biology of Agriculture, Yangling, Shaanxi, P. R. China

    Chaohong Fan, Ni Pu, Xiping Wang, Yuejin Wang, Li Fang, Weirong Xu & Jianxia Zhang

  3. College of Horticulture, Northwest A & F University, Yangling, Shaanxi, 712100, P. R. China

    Chaohong Fan, Ni Pu, Xiping Wang, Yuejin Wang, Li Fang, Weirong Xu & Jianxia Zhang

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  1. Chaohong Fan
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  2. Ni Pu
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  3. Xiping Wang
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  4. Yuejin Wang
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  5. Li Fang
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Correspondence to Yuejin Wang.

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Chaohong Fan and Ni Pu contributed equally to this work.

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Fan, C., Pu, N., Wang, X. et al. Agrobacterium-mediated genetic transformation of grapevine (Vitis vinifera L.) with a novel stilbene synthase gene from Chinese wild Vitis pseudoreticulata . Plant Cell Tiss Organ Cult 92, 197–206 (2008). https://doi.org/10.1007/s11240-007-9324-2

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  • DOI: https://doi.org/10.1007/s11240-007-9324-2

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