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A synthetic antimicrobial peptide BTD-S expressed in Arabidopsis thaliana confers enhanced resistance to Verticillium dahliae

Abstract

BTD-S is a synthetic non-cyclic θ-defensin derivative which was previously designed in our laboratory based on baboon θ-defensins (BTDs). It shows robust antimicrobial activity against economically important phytopathogen, Verticillium dahliae. Here, we deduced the coding nucleotide sequence of BTD-S and introduced the gene into wild-type (ecotype Columbia-0) Arabidopsis thaliana plants. Results demonstrated that BTD-S-transgenic lines displayed in bioassays inhibitory effects on the growth of V. dahliae in vivo and in vitro. Based on symptom severity, enhanced resistance was found in a survey of BTD-S-transgenic lines. Besides, crude protein extracts from root tissues of BTD-S-transformed plants significantly restricted the growth of fungal hyphae and the germination of conidia. Also, fungal biomass over time determined by real-time PCR demonstrated the overgrowth of V. dahliae in wild-type plants 2–3 weeks after inoculation, while almost no fungal DNA was detected in aerial tissues of their transgenic progenitors. The result suggested that fungus failed to invade and progress acropetally up to establish a systemic infection in BTD-S-transgenic plants. Moreover, the assessment of basal defense responses was performed in the leaves of WT and BTD-S-transgenic plants. The mitigated oxidative stress and low antioxidase level in BTD-S-transgenic plants revealed that BTD-S acts via permeabilizing target microbial membranes, which is in a category different from hypersensitive response-dependent defense. Taken together, our results demonstrate that BTD-S is a promising gene to be explored for transgenic engineering for plant protection against Verticillium wilt.

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Acknowledgments

The authors would like to thank Prof. Zhigang Zhou of College of Fisheries and Life Science, Shanghai Ocean University, China for kindly providing the Arabidopsis (Col-0) seeds transformed with empty vector pCAMBIA 1301.

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Correspondence to Xuede Wang.

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This work was financially supported by the National Natural Science Foundation of China (31171616 and 31471567) and the Public Technology Application Research Project of Zhejiang Province (2014C32007).

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Li, F., Shen, H., Wang, M. et al. A synthetic antimicrobial peptide BTD-S expressed in Arabidopsis thaliana confers enhanced resistance to Verticillium dahliae . Mol Genet Genomics 291, 1647–1661 (2016). https://doi.org/10.1007/s00438-016-1209-9

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Keywords

  • Synthetic antimicrobial peptide
  • BTD-S
  • Verticillium dahliae
  • Arabidopsis thaliana