DNA insecticides- Use in Insect pest management, Advantages and Disadvantages.pptx

DNA insecticides are a novel pest control method that use specific genetic sequences to disrupt vital processes in pests, such as reproduction or development. They offer precise targeting of pests without harming beneficial organisms or the environment. Additionally, they are biodegradable and help manage resistance by focusing on unique genetic pathways.

1. University Of Agricultural Sciences, Bangalore Department Of Agricultural Entomology Arpitha, K. C. - PAMB 3030 ENT 606: Insect Toxicology & Residues (2+1) DNA insecticides -An Emerging Tool In Pest Management

2. Oligonucleotide Insecticides Olinscides or DNA insecticides Short, artificially synthesized fragments of DNA or RNA Analog of DNA insecticides found in nature is extracellular DNA (ex DNA) (Oberemok, 2022) 2

3. 1978 2008 1979 2020 2019 Paul Zamechnik and Mary Stephenson Rous sarcoma virus Donis -Keller RNase H cleavage Oberemok Oligonucleotide insecticides to be used for plant protection Target: IAP genes of NPV Oberemok Target: rRNAs Length of oligonucleotide insecticides - 11 Oberemok Sternorrhyncha -sensitive to oligonucleotide insecticides Developmental timeline 3

4. How do DNA insecticides target insect pests? DNA contamination ( DNAc ) mechanism 1. Interaction of DNA Oligonucleotide with target site: 60S 40S “Arrested” ribosomes 5′-CCA-TCT-TTC-GG-3′ Target rRNA Oberemok et al ., 2024 4

5. 2. RNase H Cleavage RNase H Removal of RNA primers during DNA replication Degradation of RNA in RNA-DNA hybrids Mechanism in the Context of Oligonucleotide Insecticides Oberemok et al ., 2024 60S 40S 5′-CCA-TCT-TTC-GG-3′ Antisense DNA oligonucleotide RNA- DNA hybrids 5

6. Galchinsky et al ., 2023 Control of Icerya purchasi  Maskell (Hemiptera: Monophlebidae ) Using Oligonucleotide Insecticides Location: Nikita Botanical Garden (Yalta, Crimea) Concentration : 100 ng/µL in nuclease-free water Hand sprayer to  P ittosporum tobira  leaves (mg of olinscide per m 2  of leaves) 6

7. Day Control (Water) oligoICER-11 3rd 14.02 ± 2.79 42.92 ± 2.25 * 7th 20.08 ± 6.25 45.05 ± 4.74 * 10th 23.7 ± 8.87 70.55 ± 0.77 * Mortality (%) of  I. purchasi  larvae pH of leaves  Pittosporum tobira Day Control (Water) oligoICER-11 1st 6.01 ± 0.06 6.02 ± 0.04 7th 6.04 ± 0.04 6.05 ± 0.04 Galchinsky et al ., 2023 7

8. Oberemok et al ., 2016 Transgenerational effect of Oligonucleotides 8

9. Effect of the ssDNA insecticide Coccus-11 on  C. hesperidium  larvae (a) Survival rate Oberemok et al ., 2022 Coccus-11 (5′-CCA-TCT-TTC-GG-3′) 9

10. (b) Dose-effect curve for Coccus-11 (c) Expression of 28S rRNA was analyzed by PCR Oberemok et al ., 2022 10

11. 1—control larva with intact integuments 2—larva from ACTG-11 group with intact integuments 3—larva from Coccus-11 group with tissue necrosis at the edges of larva body 4—larva from Actara group with noticeable darkening of the body 11

12. Insect Oligonucleotide insecticide Target Mortality Reference Unaspis euonymi oligoUE-11 (5′-AGA-CCGACG- AC-3′) 28S rRNA 99.24 ± 1.32% Gal'chinsky et al ., 2020 Dynaspidiotus britannicus oligoDB-11 (5′-ATA-CCGACG- AT-3′) 28S rRNA 82.44± 15.62% Gal'chinsky et al ., 2020). Ceroplastes japonicus oligoCJ-11 (5′-CGA-CCGACG- AA-3′) 28S rRNA 78.82 ± 18.60% Useinov et al ., 2020 Diaspis echinocacti Cactus-NBG (5′-ATC-GCTGCG- GA-3′) 28S rRNA 84.2± 2.2% Plugatar et al ., 2021 Coccus hesperidum Coccus-11 (5′-CCA-TCT-TTCGG- 3′) 28S rRNA 95.59 ± 1.63% Oberemok et al ., 2022 Macrosiphoniella sanborni Macsan-11 (5′-TGT-GTTCGT- TA-3′) ITS2 of polycistronic rRNA 97.38 ± 2.49% Puzanova et al ., 2023 Tetranychus urticae Turka (5′-AGC-GAC-GTC-GC- 3′) 28S rRNA 77 ± 0.4% Novikov et al ., 2023 List of species that can be controlled using DNA insecticides 12

13. Advanced features of Oligonucleotide insecticides Low carbon footprint Insecticides against I. purchasi Corresponding compound Reagents and Solvents t CO2/t Ratio Oligonucleotide insecticide (oligoICER-11) Antisense oligonucleotide Amidites, tetrazole, 1-methylimidazole, triethylamine, acetic or propionic anhydride, pyridine, iodine, acetic acid, trichloroacetic acid, dichloromethane, acetonitrile ~0 Chlorothiazole (Thiamethoxam) Thiamethoxam S-methyl-N- nitroisothiourea , methylamine, N-methyl-N′-nitroguanidine, formaldehyde, formic acid, tetrahydro-1,3,5-oxadiazine, 2-chloro-5-chloromethylthiazole, dimethylformamide, potassium carbonate 0.351 Galchinsky et al ., 2023 13

14. Rapid biodegradability High selectivity in action and safety for non-target organisms Avoidance of target site resistance Advanced features of Oligonucleotide insecticides Galchinsky et al ., 2023 14

15. How can oligonucleotide insecticides be developed? DNA Insector program - Developed by Volodymyr Oberemok & Nikita Marochkin in 2023 15

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17. Disadvantages of using DNA Insecticides Hard to penetrate cryptic feeding insects and adult beetles They may not be successful in controlling insect pests in an emergency situation like pest outbreak. The carriers used are very expensive. 17

18. Thank You 18