DNA insecticide developed from the Lymantria dispar 5.8S ribosomal RNA gene provides a novel biotechnology for plant protection

Yuri I. Gninenko,Iryna I. Fomochkina,Maksym N. Shumskykh,Alisa M. Krasnodubets,Evgenia Y. Bessalova,Tatiana P. Makalish,Zenure Z. Temirova,Anatoly V. Kubyshkin,Nikita V. Gal’chinsky,Refat Z. Useinov,Ilya A. Novikov,Volodymyr V. Oberemok,Kateryna V. Laikova,Valeriy V. Dyadichev,Anna I. Repetskaya

doi:10.1038/s41598-019-42688-8

Abstract

Having observed how botanicals and other natural compounds are used by nature to control pests in the environment, we began investigating natural polymers, DNA and RNA, as promising tools for insect pest management. Over the last decade, unmodified short antisense DNA oligonucleotides have shown a clear potential for use as insecticides. Our research has concentrated mainly on Lymantria dispar larvae using an antisense oligoRING sequence from its inhibitor-of-apoptosis gene. In this article, we propose a novel biotechnology to protect plants from insect pests using DNA insecticide with improved insecticidal activity based on a new antisense oligoRIBO-11 sequence from the 5.8S ribosomal RNA gene. This investigational oligoRIBO-11 insecticide causes higher mortality among both L. dispar larvae grown in the lab and those collected from the forest; in addition, it is more affordable and faster acting, which makes it a prospective candidate for use in the development of a ready-to-use preparation.

Highlights

As the approaches used to develop preparations to protect plants from insect degradation have evolved, a pivotal moment has arrived: the beginning of the end of the hegemony held by chemical insecticides as they are replaced by better, more efficient preparations based on nucleic acids
Our investigation of unmodified antisense DNA oligonucleotides developed from the RING domain of the LdMNPV (Lymantria dispar multiple nucleopolyhedrovirus) IAP-3 gene has recently shown that these oligonucleotides can cause high mortality among LdMNPV-free larvae[9] and LdMNPV-infected gypsy moth larvae[5,10]
The observation that healthy LdMNPV-free gypsy moth larvae grown under lab conditions are not affected by the oligoRING, unlike larvae exposed to stress factors[6,9] in natural habitats, are similar to those of researchers examining the effects of RNA interference (RNAi) in lepidopterans[11], and provides an explanation why the oligoRING does not always exert its insecticidal effect

Summary

Introduction

As the approaches used to develop preparations to protect plants from insect degradation have evolved, a pivotal moment has arrived: the beginning of the end of the hegemony held by chemical insecticides as they are replaced by better, more efficient preparations based on nucleic acids. Www.nature.com/scientificreports outbreak levels in many areas of the world, causing economic losses to forests in Europe, Asia, Africa, North America[3], and even New Zealand[4] This led us to investigate how to create DNA-based insecticides to help address this damage[5,6,7], because phytophagous insects, such as the gypsy moth, present an ideal insect for testing topical application of insecticides produced from nucleic acids[8]. In a complex natural habitat, such as a forest, the presence of many stress factors (baculoviruses and bacterial infections, UV radiation, air pollution, etc.) may activate an apoptosis– anti-apoptosis system[10,12] This provides compelling evidence explaining why earlier-stage larvae collected from forests experienced significant mortality as the result of the down-regulation of the target host IAP gene, unlike LdMNPV-free larvae grown under laboratory conditions. We designed an 11 nucleotide long (5′-TGCGTTCGAAA-3′) antisense oligonucleotide (oligoRIBO-11) from the L. dispar 5.8S ribosomal RNA gene that includes the universally conserved antisense 5′-GTTC-3′ sequence and applied it as a contact DNA insecticide in our experiments

Methods

Results

Conclusion