Insecticidal activity of three 10–12 nucleotides long antisense sequences from 5.8S ribosomal RNA gene of gypsy moth Lymantria dispar L. against its larvae
Volodymyr V. Oberemok 1, 2, A-F,   Kateryna V. Laikova 1, 3, A-F,   Refat Z. Useinov 1, A-F  
,   Nikita V. Gal'chinsky 1, A-F,   Ilya A. Novikov 1, 3, A-F
,   Kseniya A. Yurchenko 1, A-F,   Mikhail E. Volkov 4, A-F,   Mikhail V. Gorlov 5, A-F
,   Valentina A. Brailko 2, A-F,   Yuri V. Plugatar 6, A-F
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Biochemistry Department, Vernadsky Crimean Federal University, Simferopol, Russia
Laboratory of Plant Genomics and Bioinformatics, Nikita Botanical Gardens – National Scientific Centre Russian Academy of Sciences, Yalta, Russia
Department of Essential Oil and Medicinal Crops, Research Institute of Agriculture of Crimea, Russia
Department of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry techniques, Ltd “NPF Syntol”, Moscow, Russia
Department of Chemical Technology of Plastics, Mendeleev University of Chemical Technology of Russia, Moscow, Russia
Department of Natural Ecosystems, Nikita Botanical Gardens – National Scientific Centre Russian Academy of Sciences, Yalta, Russia
A - Research concept and design; B - Collection and/or assembly of data; C - Data analysis and interpretation; D - Writing the article; E - Critical revision of the article; F - Final approval of article
Refat Z. Useinov   

Biochemistry Department, Vernadsky Crimean Federal University, Simferopol, Russia
Submission date: 2019-08-28
Acceptance date: 2019-11-13
Online publication date: 2019-12-06
Journal of Plant Protection Research 2019;59(4):561–564
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.
The authors have declared that no conflict of interests exist.
The reported study was funded by RFBR (The Russian Foundation for Basic Research) as research project No. 19-03-01048.
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