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Abstract

Effective control of Leptinotarsa decemlineata remains an urgent problem for agriculture worldwide. Minimization of the use of non-selective neonicotinoid insecticides, such as thiomethoxam, is an actual vector of development of potato cultivation. In this rapid communication, we show the prospect of the topical use of short unmodified antisense fragment of L. decemlineata CYP6B gene as a DNA insecticide. Investigated parameters, namely, number of larvae per plant, aboveground biomass, yield and number of potatoes produced per plant indicate the possibility of this post-genomic approach as a safe and effective method of L. decemlineata control.
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Authors and Affiliations

Volodymyr Oberemok
Kateryna Laikova
Maksym Shumskykh
Igor Kenyo
Igor Kasich
Deri Karim
Eleonora Seidosmanova
Alisa Krasnodubets
Viktoriya Bekirova
Gal’chinsky Nikita
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Abstract

In this short communication describing experiments carried out on the larvae of two insects, Unaspis euonymi Comstock (feeding on Euonymus japonicus Thunb.) and Dynaspidiotus britannicus Newstead (feeding on Laurus nobilis L.), we evaluate for the first time the efficiency of using DNA insecticides in the control of sap-sucking insects, including armored scale insects. Over a period of 10 days, high insect mortality was detected in both U. euonymi and D. britannicus, accompanied by a significant decrease in the concentration of target RNAs. At the same time, no visible changes were observed when the leaves of the host plants were subjected to treatment with DNA insecticides for one month. The results show the high efficiency of DNA insecticides used against hemipteran insect pests. It is noteworthy that the high efficiency of DNA insecticides and their low cost in comparison with RNA preparations provides a safe and extremely promising potential vehicle for the control of sap-sucking insects.

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Authors and Affiliations

Nikita Gal’chinsky
Refat Useinov
Ekaterina Yatskova
Kateryna Laikova
Ilya Novikov
Mikhail Gorlov
Natalya Trikoz
Alexander Sharmagiy
Yuri Plugatar
Volodymyr Oberemok
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Abstract

5.8S ribosomal RNA plays an important role in protein synthesis and eukaryotic ribosome translocation. Contact DNA insecticides based on antisense fragments of 5.8S ribosomal RNA gene of gypsy moth Lymantria dispar L. showed prospective insecticidal activity on its larvae. The most pronounced insecticidal effect was found for antisense fragments 10 and 11 nucleotides long (oligoRIBO-10 and oligoRIBO-11), whereas 12 nucleotides long fragment (oligoRIBO-12) caused the lowest level of insect mortality. This data corresponds to results obtained earlier using rabbit reticulocyte and wheat germ extracts, where maximum inhibition of protein synthesis was observed when a relevant oligomer 10-11 nucleotides long was used, whilst longer chain lengths resulted in reduced inhibition. Using oligoRIBO-11 fragment we have shown penetration of antisense oligonucleotides to insect cells through insects’ exoskeletons. MALDI technique registered the penetration of the oligoRIBO-11 fragment into insect cells after 30 min and a significant response of insect cells to the applied oligonucleotide after 60 min, which indicates not only that the oligonucleotide enters the insect cells, but also the synthesis of new substances in response to the applied DNA fragment. Contact DNA insecticides developed from the L. dispar 5.8S ribosomal RNA gene provide a novel biotechnology for plant protection using unmodified antisense oligonucleotides.

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Authors and Affiliations

Volodymyr V. Oberemok
Kateryna V. Laikova
Refat Z. Useinov
Nikita V. Gal’chinsky
Ilya A. Novikov
Kseniya A. Yurchenko
Mikhail E. Volkov
Mikhail V. Gorlov
Valentina A. Brailko
Yuri V. Plugatar

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