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A comparison study of Agrobacterium-mediated transformation methods for root-specific promoter analysis in soybean

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Abstract

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Both in vitro and in vivo hairy root transformation systems could not replace whole plant transformation for promoter analysis of root-specific and low-P induced genes in soybean.

Abstract

An efficient genetic transformation system is crucial for promoter analysis in plants. Agrobacterium-mediated transformation is the most popular method to produce transgenic hairy roots or plants. In the present study, first, we compared the two different Agrobacterium rhizogenes-mediated hairy root transformation methods using either constitutive CaMV35S or the promoters of root-preferential genes, GmEXPB2 and GmPAP21, in soybean, and found the efficiency of in vitro hairy root transformation was significantly higher than that of in vivo transformation. We compared Agrobacterium rhizogenes-mediated hairy root and Agrobacterium tumefaciens-mediated whole plant transformation systems. The results showed that low-phosphorous (P) inducible GmEXPB2 and GmPAP21 promoters could not induce the increased expression of the GUS reporter gene under low P stress in both in vivo and in vitro transgenic hairy roots. Conversely, GUS activity of GmPAP21 promoter was significantly higher at low P than high P in whole plant transformation. Therefore, both in vitro and in vivo hairy root transformation systems could not replace whole plant transformation for promoter analysis of root-specific and low-P induced genes in soybean.

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Acknowledgments

We are grateful to Dr. Kan Wang for the generous gift of the pTF102 vector, and Dr. Peter M. Gresshoff for A. rhizogenes strain K599. We thank CC Li and XX Li for GmPAP21 and GmEXPB2 vector constructs. We also thank Mrs./Mr. Jun Fang, Jiao Zhou, Shuang Zhang and Tao Yang for technical help in soybean whole plant transformation. We are also grateful to the anonymous referees for their valuable comments on two previous versions. This research is supported by funds from the National Natural Science Foundation of China (31372126), and National Key Basic Research Special Funds of China (2011CB100301).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Root Biology Center, South China Agricultural University, Guangzhou, 510642, China

    Caifeng Li, Haiyan Zhang, Xiurong Wang & Hong Liao

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  1. Caifeng Li
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  2. Haiyan Zhang
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  3. Xiurong Wang
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  4. Hong Liao
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Corresponding author

Correspondence to Xiurong Wang.

Additional information

Communicated by Howard S. Judelson.

C. Li and H. Zhang contributed equally to this work.

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Li, C., Zhang, H., Wang, X. et al. A comparison study of Agrobacterium-mediated transformation methods for root-specific promoter analysis in soybean. Plant Cell Rep 33, 1921–1932 (2014). https://doi.org/10.1007/s00299-014-1669-5

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  • DOI: https://doi.org/10.1007/s00299-014-1669-5

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