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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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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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