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Sprout vacuum-infiltration: a simple and efficient agroinoculation method for virus-induced gene silencing in diverse solanaceous species

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Abstract

Virus-induced gene silencing (VIGS) is a robust technique for identifying the functions of plant genes. Tobacco rattle virus (TRV)-mediated VIGS has been commonly used in many plants. In order to overcome the limitations of existing agroinoculation methods, we report an easy and effective method of agroinoculation for virus-induced gene silencing—sprout vacuum-infiltration (SVI). Using sprout vacuum-infiltration, we have successfully silenced the expression of phytoene desaturase and Mg-protoporphyrin chelatase genes in four important solanaceous crops, including tomato, eggplant, pepper, and Nicotiana benthamiana. The gene-silenced phenotypes are conspicuous in 1-week-old plants. The method is simple, low cost and rapid compared to other techniques such as leaf infiltration or agrodrench. It may be more practical for studying gene function in the early stages of plant growth. An important aspect of SVI is that it will be used for high-throughput VIGS screens in the future. SVI will be an effective tool to overcome the limitations of current inoculation methods and to facilitate large-scale VIGS analysis of cDNA libraries.

Key message SVI is a simple, low cost agroinoculation method for VIGS. It is practical for studying the function of genes expressed in early stages of plant growth and high-throughput VIGS screens.

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Acknowledgments

We would like to thank Dr S.P. Dinesh-Kumar (Yale University) for offering pTRV1 and pTRV2 vector. This work was supported by grants (No. 31000925, No. 30871741 and No. 30972037) of the National Nature Science Foundation of China, National one hundred outstanding doctoral dissertation special fund (No.201062) and Innovation Fund for Graduate Student of China Agricultural University (No. KYCX2011062).

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Authors and Affiliations

  1. Laboratory of Food Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Qinghua Donglu Road, HaiDian District, Beijing, 100083, China

    Hua-xue Yan, Da-qi Fu, Ben-zhong Zhu, Hai-ping Liu, Xiao-ying Shen & Yun-bo Luo

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  1. Hua-xue Yan
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  2. Da-qi Fu
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  3. Ben-zhong Zhu
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  4. Hai-ping Liu
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  5. Xiao-ying Shen
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  6. Yun-bo Luo
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Corresponding author

Correspondence to Yun-bo Luo.

Additional information

Communicated by Q. Zhao.

H. Yan and D. Fu contributed equally.

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Yan, Hx., Fu, Dq., Zhu, Bz. et al. Sprout vacuum-infiltration: a simple and efficient agroinoculation method for virus-induced gene silencing in diverse solanaceous species. Plant Cell Rep 31, 1713–1722 (2012). https://doi.org/10.1007/s00299-012-1285-1

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  • DOI: https://doi.org/10.1007/s00299-012-1285-1

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