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Obtainment and Analysis of Marker-Free Oil Plants Camelina sativa (L.) Expressing of Antimicrobial Peptide Cecropin P1 Gene

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Abstract

Marker-free transgenic Camelina sativa (L.) plants carrying a synthetic gene for cecropin P1, an antimicrobial peptide, under the control of the cauliflower mosaic virus 35S RNA promoter have been obtained and analyzed. The plants were transformed with an agrobacterial binary vector free of selective genes of antibiotic and herbicide resistance. The marker-free transformants were screened via measurement of the antibacterial activity of cecropin P1 and enzyme immunoassay. The obtained plants exhibited an increased resistance to infection with the bacteria Erwinia carotovora, the fungi Fusarium graminearum, and oxidative stress during infection. Analysis of the fatty acid composition of seed oil showed an increased amount of α-linolenic acid in the transgenic Camelina lines as compared to unmodified plants. The results indicate that the cecropin P1 gene can be included in an integral antistress plant-protective system.

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ACKNOWLEDGMENTS

The authors are grateful to I.L. Rodionov for the synthesis of the cecropin P1 peptide and to A.G. Laman for the production of antibodies to cecropin P1 and KLH-CEC conjugates.

Funding

The work was financially supported by the State Assignments no. 0101-2014-0046 and RK 01201352439 with partial financing from the Russian Foundation for Basic Research (projects 18-08-00752 and 16-04-00623).

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Authors and Affiliations

  1. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino Branch, 142290, Pushchino, Moscow oblast, Russia

    N. S. Zakharchenko, O. V. Furs, S. V. Pigoleva, O. V. Dyachenko, Ya. I. Buryanov & T. V. Shevchuk

  2. State Research Center for Applied Microbiology and Biotechnology, 142279, Obolensk, Moscow oblast, Russia

    A. V. Aripovskii

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  1. N. S. Zakharchenko
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  2. O. V. Furs
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  3. S. V. Pigoleva
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Correspondence to N. S. Zakharchenko.

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This article does not contain any studies involving animals or human participants performed by any of the authors.

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Translated by I. Gordon

Abbreviations: AMPs—antimicrobial peptides; сесР1—cecropin P1; cecP1—cecP1 gene; CaMV 35S—35S RNA promoter of cauliflower mosaic virus; dNTP—deoxynucleoside triphosphate(s); LB medium—nutrient medium of Luriya Bertani; MS medium—hormone-free Murashige—Skoog medium; OD600, optical density at wavelength of 600 nm; PAGE—polyacrylamide gel electrophoresis; PGA—potato-glucose agar; SOD—superoxide dismutase.

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Zakharchenko, N.S., Furs, O.V., Pigoleva, S.V. et al. Obtainment and Analysis of Marker-Free Oil Plants Camelina sativa (L.) Expressing of Antimicrobial Peptide Cecropin P1 Gene. Appl Biochem Microbiol 55, 888–898 (2019). https://doi.org/10.1134/S0003683819090096

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