International Research & Citation Index
IRCI Article ID: IRCI-AR-0000000953

Synergistic action of the gut microbiota in environmental RNA interference in a leaf beetle

Authors:
Letian XuShijing XuLiuwei SunYiqiu ZhangJing LuoRalph BockJiang Zhang

Journal: Microbiome

Publication: 2021-12-01 · Vol. 9 No. 1

DOI: 10.1186/s40168-021-01066-1

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Abstract
Abstract

                Background

                RNA interference (RNAi) has emerged as an efficient tool to control insect pests. When insects ingest double-stranded RNAs (dsRNAs) targeted against essential genes, strong gene silencing and mortality can be induced. To exert their function, dsRNA molecules must pass through the insect’s gut and enter epithelial cells and/or the hemolymph. Gut bacteria are known to play multifarious roles in food digestion and nutrition, and confer protection against pathogens and parasites. Whether there is a cross talk between gut bacteria and ingested dsRNAs and whether the microbiome affects RNAi efficiency are unknown.

              

                Results

                Here, using a leaf beetle gut microbiota system, we investigated whether gut bacteria interact with dsRNA molecules and how the gut microbiota affects RNAi responses in insects. We first showed that the leaf beetle Plagiodera versicolora (Coleoptera) is highly susceptible to RNAi. We then demonstrated that ingestion of dsRNAs by non-axenic P. versicolora larvae results in (i) significantly accelerated mortality compared with axenic larvae, and (ii) overgrowth and dysbiosis of the gut microbiota. The latter may be caused by bacterial utilization of dsRNA degradation products. Furthermore, we found that Pseudomonas putida, a gut bacterium of P. versicolora, acts as major accelerator of the death of P. versicolora larvae by transitioning from commensal to pathogenic lifestyle.

              

                Conclusions

                The present study illuminates the complex interplay between lethal dsRNA, the insect host, and its gut microbiota. The ingestion of dsRNA by the leaf beetle caused a dysbiosis of gut bacterial community, and the dsRNA degradation products by host insect preferentially promoted the growth of an entomopathogenic bacterium, which accelerated dsRNA lethality to the insect. Our findings reveal a synergistic role of the gut microbiota in dsRNA-induced mortality of pest insects, and provide new insights in the mechanisms of RNAi-based pest control.

                

              


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Authors
Letian Xu
Affiliation: Hubei University
ORCID: 0000-0003-3336-9907
Shijing Xu
Affiliation: Hubei University
Liuwei Sun
Affiliation: Hubei University
Yiqiu Zhang
Affiliation: Hubei University
ORCID: 0000-0002-0206-0061
Jing Luo
Affiliation: Hubei University
ORCID: 0000-0002-1566-6644
Ralph Bock
Affiliation: Max Planck Institute of Molecular Plant Physiology; Hubei University
ORCID: 0000-0001-7502-6940
Jiang Zhang
Affiliation: Hubei University
ORCID: 0000-0003-3949-3765


References
Zhang J, Khan S, Heckel D, Bock R. Next-generation insect-resistant plants: RNAi-mediated crop protection. Trends Biotechnol. 2017;35(9):871–82. https://doi.org/10.1016/j.tibtech.2017.04.009.
Garbutt JS, Bellés X, Richards EH, Reynolds SE. Persistence of double-stranded RNA in insect hemolymph as a potential determiner of RNA interference success: evidence from Manduca sexta and Blattella germanica. J Insect Physiol. 2013;59(2):171–8. https://doi.org/10.1016/j.jinsphys.2012.05.013.
Zhu F, Xu J, Palli R, Ferguson J, Palli SR. Ingested RNA interference for managing the populations of the Colorado potato beetle, Leptinotarsa decemlineata. Pest Manag Sci. 2011;67(2):175–82. https://doi.org/10.1002/ps.2048.
Baum JA, Bogaert T, Clinton W, Heck GR, Feldmann P, Ilagan O, et al. Control of coleopteran insect pests through RNA interference. Nat Biotechnol. 2007;25(11):1322–6. https://doi.org/10.1038/nbt1359.
Terenius O, Papanicolaou A, Garbutt JS, Eleftherianos I, Huvenne H, Kanginakudru S, et al. RNA interference in Lepidoptera: an overview of successful and unsuccessful studies and implications for experimental design. J Insect Physiol. 2011;57(2):231–45. https://doi.org/10.1016/j.jinsphys.2010.11.006.
Cooper AM, Silver K, Zhang J, Park Y, Zhu KY. Molecular mechanisms influencing efficiency of RNA interference in insects. Pest Manag Sci. 2018;75:18–28.
Shukla JN, Kalsi M, Sethi A, Narva KE, Fishilevich E, Singh S, et al. Reduced stability and intracellular transport of dsRNA contribute to poor RNAi response in lepidopteran insects. RNA Biol. 2016;13(7):656–69. https://doi.org/10.1080/15476286.2016.1191728.
Yoon JS, Gurusamy D, Palli SR. Accumulation of dsRNA in endosomes contributes to inefficient RNA interference in the fall armyworm, Spodoptera frugiperda. Insect Biochem Mol Biol. 2017;90:53–60. https://doi.org/10.1016/j.ibmb.2017.09.011.
Sugahara R, Tanaka S, Jouraku A, Shiotsuki T. Geographic variation in RNAi sensitivity in the migratory locust. Gene. 2017;605:5–11. https://doi.org/10.1016/j.gene.2016.12.028.
Engel P, Moran NA. The gut microbiota of insects - diversity in structure and function. FEMS Microbiol Rev. 2013;37(5):699–735. https://doi.org/10.1111/1574-6976.12025.
Broderick NA, Raffa KF, Handelsman J. Midgut bacteria required for Bacillus thuringiensis insecticidal activity. Proc Natl Acad Sci USA. 2006;103(41):15196–9. https://doi.org/10.1073/pnas.0604865103.
Caccia S, Lelio ID, Storia AL, Marinelli A, Varricchio P, Franzetti E, et al. Midgut microbiota and host immunocompetence underlie Bacillus thuringiensis killing mechanism. Proc Natl Acad Sci USA. 2016;113(34):9486–91. https://doi.org/10.1073/pnas.1521741113.
Johnston PR, Crickmore N. Gut bacteria are not required for the insecticidal activity of Bacillus thuringiensis toward the tobacco hornworm, Manduca sexta. Appl Environ Microbiol. 2009;75(15):5094–9. https://doi.org/10.1128/AEM.00966-09.
Broderick NA, Robinson CJ, McMahon MD, Holt J, Handelsman J, Raffa KF. Contributions of gut bacteria to Bacillus thuringiensis-induced mortality vary across a range of Lepidoptera. BMC Biol. 2009;7(1):11. https://doi.org/10.1186/1741-7007-7-11.
Mason KL, Stepien TA, Blum JE, Holt JF, Labbe NH, Rush JS, et al. From commensal to pathogen: translocation of Enterococcus faecalis from the midgut to the hemocoel of Manduca sexta. MBio. 2011;2(3):e00065–11. https://doi.org/10.1128/mBio.00065-11.
Xu L, Deng J, Zhou F, Cheng C, Zhang L, Zhang J, et al. Gut microbiota in an invasive bark beetle infected by a pathogenic fungus accelerates beetle mortality. J Pest Sci. 2019;92(1):343–51. https://doi.org/10.1007/s10340-018-0999-4.
Wei G, Lai Y, Wang G, Chen H, Li F, Wang S. Insect pathogenic fungus interacts with the gut microbiota to accelerate mosquito mortality. Proc Natl Acad Sci USA. 2017;114(23):5994–9. https://doi.org/10.1073/pnas.1703546114.
Urban J. Contribution to the knowledge of development and harmfulness of imported willow leaf beetle (Plagiodera versicolora) (Coleoptera, Chrysomelidae). J Forest Sci. 2005;51:481–507.
Demirci M, Sevim E, Demir İ, Sevim A. Culturable bacterial microbiota of Plagiodera versicolora (L.) (Coleoptera: Chrysomelidae) and virulence of the isolated strains. Folia Microbiol. 2013;58:201–10.
Wu Y, Xu L, Chang L, Ma M, You L, Jiang C, et al. Bacillus thuringiensis (Bt) cry1C expression from the plastid genome of poplar leads to high mortality of leaf eating caterpillars. Tree Physiol. 2019;39:1523–32.
Xu L, Lou Q, Cheng C, Lu M, Sun J. Gut-associated bacteria of Dendroctonus valens and their involvement in verbenone production. Microbial Ecology. 2015;70(4):1012–23. https://doi.org/10.1007/s00248-015-0625-4.
Gallup J. qPCR inhibition and amplification of difficult templates. In Kennedy S, Oswald N (ed), PCR troubleshooting and optimization: the essential guide. Norfolk: Caister Academic Press; 2011. p 23– 65.
Kesnerova L, Emery O, Troilo M, Liberti J, Erkosar B, Engel P. Gut microbiota structure differs between honeybees in winter and summer. ISME J. 2019;14(3):801–14. https://doi.org/10.1038/s41396-019-0568-8.
Oksanen JF, Blanchet G, Friendly M, Kindt R, Legendre P, McGlinn P, et al. Vegan: Community Ecology Package. R package version 2.5-3.2018.
Wynant N, Santos D, Verdonck R, Spit J, Van Wielendaele P, Vanden BJ. Identification, functional characterization and phylogenetic analysis of double stranded RNA degrading enzymes present in the gut of the desert locust, Schistocerca gregaria. Insect Biochem Mol Biol. 2014;46:1–8. https://doi.org/10.1016/j.ibmb.2013.12.008.
Stanier RY. Simultaneous adaption: a new technique for the study of metabolic pathways. J Bacteriol. 1947;64:339–48.
Newell PD, Douglas AE. Interspecies interactions determine the impact of the gut microbiota on nutrient allocation in Drosophila melanogaster. Appl Environ Microbiol. 2014;80(2):788–96. https://doi.org/10.1128/AEM.02742-13.
Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method. Nat Protoc. 2008;3(6):1101–8. https://doi.org/10.1038/nprot.2008.73.
Bergmark L, Poulsen PH, Al-Soud WA, Norman A, Hansen LH, Sørensen SJ. Assessment of the specificity of Burkholderia and Pseudomonas qPCR assays for detection of these genera in soil using 454 pyrosequencing. FEMS Microbiol Lett. 2012;333(1):77–84. https://doi.org/10.1111/j.1574-6968.2012.02601.x.
Haugland RA, Siefring SC, Wymer LJ, Brenner KP, Dufour AP. Comparison of Enterococcus measurements in freshwater at two recreational beaches by quantitative polymerase chain reaction and membrane filter culture analysis. Water Res. 2005;39(4):559–68. https://doi.org/10.1016/j.watres.2004.11.011.
Silva-Junior WP, Martins AS, Xavier PC, Appel KL, Oliveira Junior SA, Palhares DB. Etiological profile of early neonatal bacterial sepsis by multiplex qPCR. J Infect Dev Ctries. 2016;10(12):1318–24. https://doi.org/10.3855/jidc.7474.
Ulrich J, Dao VA, Majumdar U, Schmitt-Engel C, Schwirz J, Schultheis D, et al. Large scale RNAi screen in Tribolium reveals novel target genes for pest control and the proteasome as prime target. BMC Genomics. 2015;16(1):674. https://doi.org/10.1186/s12864-015-1880-y.
Zhang Y, Xu L, Li S, Zhang J. Bacteria-mediated RNA interference for management of Plagiodera versicolora (Coleoptera: Chrysomelidae). Insects. 2019;10(12):415.
Ayscough KR. In vivo functions of actin-binding proteins. Curr Opin Cell Biol. 1998;10(1):102–11. https://doi.org/10.1016/S0955-0674(98)80092-6.
Bernstein HD, Poritz MA, Strub K, Hoben PJ, Brenner S, Walter P. Model for signal sequence recognition from amino-acid sequence of 54K subunit of signal recognition particle. Nature. 1989;340(6233):482–6. https://doi.org/10.1038/340482a0.
Söllner T, Bennett MK, Whiteheart SW, Scheller RH, Rothman JE. A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion. Cell. 1993;75(3):409–18. https://doi.org/10.1016/0092-8674(93)90376-2.
van der Bliek AM, Meyerowitz EM. Dynamin-like protein encoded by the Drosophila shibire gene associated with vesicular traffic. Nature. 1991;351(6325):411–4. https://doi.org/10.1038/351411a0.
Yaman M, Erturk O, Unal S, Selek F. Isolation and identification of bacteria from four important poplar pests. Rev Col Entomol. 2017;43(1):34–7. https://doi.org/10.25100/socolen.v43i1.6644.
Chung SH, Rosa C, Scully ED, Peiffer M, Tooker JF, Hoover K, et al. Herbivore exploits orally secreted bacteria to suppress plant defenses. Proc Natl Acad Sci USA. 2013;110(39):15728–33. https://doi.org/10.1073/pnas.1308867110.
Xie J, Li S, Zhang W, Xia Y. RNAi-knockdown of the Locusta migratoria nuclear export factor protein results in insect mortality and alterations in gut microbiome. Pest Manag Sci. 2019;75(5):1383–90. https://doi.org/10.1002/ps.5258.
Rusk N. Prokaryotic RNAi. Nat Methods. 2012;9(3):220–1. https://doi.org/10.1038/nmeth.1916.
Guan RB, Li HC, Fan YJ, Hu SR, Christiaens O, Smagghe G, et al. A nuclease specific to lepidopteran insects suppresses RNAi. J Biol Chem. 2018;293(16):6011–21. https://doi.org/10.1074/jbc.RA117.001553.
Spit J, Philips A, Wynant N, Santos D, Plaetinck G, Vanden BJ. Knockdown of nuclease activity in the gut enhances RNAi efficiency in the Colorado potato beetle, Leptinotarsa decemlineata, but not in the desert locust, Schistocerca gregaria. Insect Biochem Mol Biol. 2017;81:103–16. https://doi.org/10.1016/j.ibmb.2017.01.004.
Song H, Zhang J, Li D, Cooper AMW, Silver K, Li T, et al. A double-stranded RNA degrading enzyme reduces the efficiency of oral RNA interference in migratory locust. Insect Biochem Mol Biol. 2017;86:68–80. https://doi.org/10.1016/j.ibmb.2017.05.008.
Arimatsu Y, Fruno T, Sugimura Y, Togoh M, Ishihara R, Tokizane M, et al. Purification and properties of double-stranded RNA-degrading nuclease, dsRNase, from the digestive juice of the silkworm, Bombyx mori. J Insect Biotechnol Sericol. 2007;76:57–62.
Wynant N, Duressa TF, Santos D, Van Duppen J, Proost P, Huybrechts R, et al. Lipophorins can adhere to dsRNA, bacteria and fungi present in the hemolymph of the desert locust: a role as general scavenger for pathogens in the open body cavity. J Insect Physiol. 2014;64:7–13. https://doi.org/10.1016/j.jinsphys.2014.02.010.
Huvenne H, Smagghe G. Mechanisms of dsRNA uptake in insects and potential of RNAi for pest control: A review. J Insect Physiol. 2010;56(3):227–35. https://doi.org/10.1016/j.jinsphys.2009.10.004.
Preston GM. Plant perceptions of plant growth-promoting Pseudomonas. Philos Trans R Soc Lond B Biol Sci. 2004;359(1446):907–18. https://doi.org/10.1098/rstb.2003.1384.
Lifshitz R, Kloepper JW, Kozlowski M, Simonson C, Carlson J, Tipping EM, et al. Growth promotion of canola (rapeseed) seedlings by a strain of Pseudomonas putida under gnotobiotic conditions. Can J Microbiol. 1987;33(5):390–5. https://doi.org/10.1139/m87-068.
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