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Seed-derived defensins from Scots pine: structural and functional features

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Abstract

Main conclusion

The recombinant Ps Def5.1 defensin inhibits the growth of phytopathogenic fungi, Gram-positive and Gram-negative bacteria, and human pathogen Candida albicans . Expression of seed-derived Scots pine defensins is tissue-specific and developmentally regulated.

Abstract

Plant defensins are ubiquitous antimicrobial peptides that possess a broad spectrum of activities and multi-functionality. The genes for these antimicrobial proteins form a multigenic family in the plant genome and are expressed in every organ. Most of the known defensins have been isolated from seeds of various monocot and dicot species, but seed-derived defensins have not yet been characterized in gymnosperms. This study presents the isolation of two new 249 bp cDNA sequences from Scots pine seeds with 97.9% nucleotide homology named PsDef5.1 and PsDef5.2. Their deduced amino acid sequences have typical plant defensin features, including an endoplasmic reticulum signal sequence of 31 amino acids (aa), followed by a characteristic defensin domain of 51 aa. To elucidate the functional activity of new defensins, we expressed the mature form of PsDef5.1 in a prokaryotic system. The purified recombinant peptide exhibited activity against the phytopathogenic fungi and Gram-negative and Gram-positive bacteria with the IC50 of 5–18 µM. Moreover, it inhibited the growth of the human pathogen Candida albicans with the IC50 of 6.0 µM. Expression analysis showed that transcripts of PsDef5.1–2 genes were present in immature and mature pine seeds and different parts of seedlings at the early stage of germination. In addition, unlike the PsDef5.2, the PsDef5.1 gene was expressed in the reproductive organs. Our findings indicate that novel defensins are promising candidates for transgenic application and the development of new antimicrobial drugs.

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All data generated or analyzed during this study are included in this published article and its supplementary information files.

Abbreviations

EST:

Expressed sequence tag

IC50 :

Concentration at which 50% growth inhibition is reached

Trx:

Thioredoxin

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Acknowledgements

The authors thank Zabolotny Institute of Microbiology and Virology (Kyiv, Ukraine) and the Department of Genetics and Biotechnology, Ivan Franko National University of Lviv (Ukraine). YIS was supported by a doctoral fellowship from the Ministry of Education and Science of Ukraine.

Funding

This work was supported by the U.S. Civilian Research and Development Foundation (CRDF Global) grants FSA-19-65490-0 and FSA-19-65490-1 to IVN and VAK, respectively, and Ministry of Education and Science of Ukraine (Grant # M52-2020).

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Correspondence to Valentina A. Kovaleva.

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Communicated by Anastasios Melis.

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Shalovylo, Y.I., Yusypovych, Y.M., Hrunyk, N.I. et al. Seed-derived defensins from Scots pine: structural and functional features. Planta 254, 129 (2021). https://doi.org/10.1007/s00425-021-03788-w

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  • DOI: https://doi.org/10.1007/s00425-021-03788-w

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