Abstract
Bacillus thuringiensis (Bt), an effective entomopathogen, has been widely used for pest control. However, insect resistance risk threatens the sustainable utility of Bt products. Previous findings suggest the interactions between gut microbiota and the host probably influence the evolution of insect resistance. To understand how the microbiota affects the development of insect resistance and manage the resistance, we characterized the gut microbiota of Chilo suppressalis from five Bt-resistant or Bt-susceptible strains by 16S rRNA sequencing. The diversity, richness, and composition of gut microbial community were analyzed among these five strains by alpha and beta analyses. Gut microbiota diversity was significantly higher in Bt-resistant (BJ1Ab-R and FZ1Ca-R) than that in Bt-susceptible strains (BJ-S and FZ-S). A significantly higher abundance of the genus Enterococcus were found in BJ-S- and FZ-S-susceptible strains than that in BJ1Ab-R- and FZ1Ca-R-resistant strains. The genus Bifidobacterium significantly dominated in the FZ1Ca-R-resistant strain, compared with the other four strains. Moreover, the gut microbial community displayed significantly more complex cooccurrence patterns in Bt-resistant than in Bt-susceptible strains by network analysis. Furthermore, the BJ-S, FZ-S and FZ1Ca-R strains had significantly reduced larval mortalities in bioassays with Bt toxin after larval pretreatment with antibiotics to remove gut bacteria. This study suggests that the gut microbiota participates in regulating the Bt-induced killing mechanism in C. suppressalis, and provides insights into the impact of Bt selective pressure on microbiome composition and potential insect resistance induced by microbiome alterations.
Graphical abstract
This is a preview of subscription content, log in via an institution to check access.
Code availability
Not applicable.
Availability of data and material
All the raw reads of the 16srRNA-seq data in this study have been deposited in the NCBI SRA database under accession numbers SRR15826673 (Gut microbiome of FZ-S C. suppressalis strain), SRR15826672 (Gut microbiome of FZ1Ca-R C. suppressalis strain), SRR15826671(Gut microbiome of FZ1Ab-R C. suppressalis strain), SRR15826670 (Gut microbiome of BJ-S C. suppressalis strain), SRR15826669 (Gut microbiome of BJ1Ab- R C. suppressalis strain).
References
Adair KL, Douglas AE (2017) Making a microbiome: the many determinants of host-associated microbial community composition. Curr Opin Microbiol 35:23–29. https://doi.org/10.1016/j.mib.2016.11.002
Akami M, Njintang NY, Gbaye OA, Andongma AA, Rashid MA, Niu CY et al (2019) Gut bacteria of the cowpea beetle mediate its resistance to dichlorvos and susceptibility to Lippia adoensis essential oil. Sci Rep 9:6435. https://doi.org/10.1038/s41598-019-42843-1
Bader GD, Hogue CW (2003) An automated method for finding molecular complexes in large protein interaction networks. BMC Bioinform 4:2. https://doi.org/10.1186/1471-2105-4-2
Bastian M, Heymann S, Jacomy M (2009) Gephi: an open source software for exploring and manipulating networks, In: Proceedings of the third international conference on weblogs and social media, ICWSM, San Jose, California, USA, pp 17–20
Beegle CC, Lewis LC, Lynch RE, Martinez AJ (1981) Interaction of larval age and antibiotic on the susceptibility of three insect species to Bacillus thuringiensis. J Invert Pathol 37:143–153. https://doi.org/10.1016/0022-2011(81)90068-9
Berry D, Widder S (2014) Deciphering microbial interactions and detecting keystone species with co-occurrence networks. Front Microbiol 5:219. https://doi.org/10.3389/fmicb.2014.00219
Bokulich NA, Subramanian S, Faith JJ, Gevers D, Gordon JI, Knight R et al (2013) Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing. Nat Method 10(1):57–59. https://doi.org/10.1038/nmeth.2276
Bottacini F, Ventura M, Sinderen DV, Motherway MO (2014) Diversity, ecology and intestinal function of Bifidobacteria. Microb Cell Fact 13(Suppl 1):S4. https://doi.org/10.1186/1475-2859-13-S1-S4
Broderick NA, Raffa KF, Handelsman J (2006) Midgut bacteria required for Bacillus thuringiensis insecticidal activity. Proc Natl Acad Sci 103:15196–15199. https://doi.org/10.1073/pnas.0604865103
Broderick NA, Robinson CJ, McMahon MD, Holt J, Handelsman J, Raffa KF (2009) Contributions of gut bacteria to Bacillus thuringiensis-induced mortality vary across a range of Lepidoptera. BMC Biol 7(11):1–9. https://doi.org/10.1186/1741-7007-7-11
Caccia S, Lelio ID, Storiaa AL, Marinelli A, Varricchio P, Franzetti E et al (2016) Midgut microbiota and host immunocompetence underlie Bacillus thuringiensis killing mechanism. Proc Natl Acad Sci 113(34):9486–9491. https://doi.org/10.1073/pnas.1521741113
Caporaso J, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK et al (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Method 7:335–336. https://doi.org/10.1038/nmeth.f.303
Chen BS, Du KQ, Sun C, Vimalanathan A, Liang XL, Li Y, Wang BH, Lu XM, Li LJ, Shao YQ (2018) Gut bacterial and fungal communities of the domesticated silkworm (Bombyx mori) and Wild mulberry-feeding relatives. ISME J 12:2252–2262. https://doi.org/10.1038/s41396-018-0174-1
Chen G, Wang YH, Liu YM, Chen FJ, Han LZ (2020) Differences in midgut transcriptomes between resistant and susceptible strains of Chilo suppressalis to Cry1C toxin. BMC Genom 21:634–653. https://doi.org/10.1186/s12864-020-07051-6
Dubois NR, Dean DH (1995) Synergism between Cry1A insecticidal crystal proteins and spores of Bacillus thuringiensis, other bacterial spores, and vegetative cells against Lymantria dispar (Lepidoptera: Lymantriidae) larvae. Environ Entomol 24:1741–1747. https://doi.org/10.1093/ee/24.6.1741
Edgar RC (2013) UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat Method 10(10):996–998. https://doi.org/10.1038/nmeth.2604
Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27(16):2194–2200. https://doi.org/10.1093/bioinformatics/btr381
Fedhila S, Nel P, Lereclus D (2002) The InhA2 metalloprotease of Bacillus thuringiensis strain 407 is required for pathogenicity in insects infected via the oral route. J Bacteriol 184(12):3296–3304. https://doi.org/10.1128/JB.184.12.3296-3304.2002
Gao X, Li W, Luo J, Zhang L, Ji J, Zhu X et al (2019) Biodiversity of the microbiota in Spodoptera exigua (Lepidoptera: noctuidae). J Appl Microbiol 126(4):1199–1208. https://doi.org/10.1111/jam.14190
Gomes AFF, Omoto C, Cônsoli FL (2020) Gut bacteria of field-collected larvae of Spodoptera frugiperda undergo selection and are more diverse and active in metabolizing multiple insecticides than laboratory-selected resistant strains. J Pest Sci 93:833–851. https://doi.org/10.1007/s10340-020-01202-0
Haas BJ, Gevers D, Earl AM, Feldgarden M, Ward DV, Giannoukos G et al (2011) Chimeric 16S rRNA sequence formation and detection in sanger and 454-pyrosequenced PCR amplicons. Genome Res 21(3):494–504. https://doi.org/10.1101/gr.112730.110
Han LZ, Li SB, Liu PL, Peng YF, Hou ML (2012) New artificial diet for continuous rearing of Chilo suppressalis (Lepidoptera: Crambidae). Ann Entomol Soc Am 105(2):253–258. https://doi.org/10.1603/AN10170
Han LZ, Jiang XF, Peng YF (2016) Potential resistance management for the sustainable use of insect-resistant genetically modified corn and rice in China. Curr Opin Insect Sci 15:139–143. https://doi.org/10.1016/j.cois.2016.04.004
He YP, Zhang JF, Gao CF, Su JY, Chen JM, Shen JL (2013) Regression analysis of dynamics of insecticide resistance in field populations of Chilo suppressalis (Lepidoptera: Crambidae) during 2002–2011 in China. J Econ Entomol 106:1832–1837. https://doi.org/10.1603/ec12469
Hilbeck A, Defarge N, Bøhn T, Krautter M, Conradin C, Amiel C et al (2018) Impact of antibiotics on efficacy of Cry toxins produced in two different genetically modified Bt maize varieties in two lepidopteran herbivore species. Ostrinia Nubilalis Spod Littoralis Toxins 10:489. https://doi.org/10.3390/toxins10120489
Hofte H, Whiteley HR (1989) Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Mol Biol Rev 53:242–255. https://doi.org/10.1128/mmbr.53.2.242-255.1989
Johnston PR, Crickmore N (2009) Gut bacteria are not required for the insecticidal activity of Bacillus thuringiensis toward the tobacco hornworm, Manduca sexta. Appl Environ Microbiol 75:5094–5099. https://doi.org/10.1128/AEM.00966-09
Jurat-Fuentes JL, Crickmore N (2017) Specificity determinants for Cry insecticidal proteins: insights from their mode of action. J Invertebr Pathol 142:5–10. https://doi.org/10.1016/j.jip.2016.07.018
Khaing MM, Yang XM, Zhao M, Zhang WN, Wang BJ, Wei JZ et al (2018) Effects of antibiotics on biological activity of Cry1Ac in Bt-susceptible and Bt-resistant Helicoverpa armigera strains. J Invertebr Pathol 151:197–200. https://doi.org/10.1016/j.jip.2017.10.007
Kikuchi Y, Hayatsu M, Hosokawa T, Nagayama A, Tago K, Fukatsu T (2012) Symbiont-mediated insecticide resistance. Proc Natl Acad Sci 109(22):8618–8622. https://doi.org/10.1073/pnas.1200231109
Kwong WK, Engel P, Koch H, Moran NA (2014) Genomics and host specialization of honey bee and bumble bee gut symbionts. Proc Natl Acad Sci 111:11509–11514. https://doi.org/10.1073/pnas.1405838111
Li YH, Hallerman EM, Liu QS, Wu KM, Peng YF (2016) The development and status of Bt rice in China. Plant Biotechnol J 14:839–848. https://doi.org/10.1111/pbi.12464
Li S, Xiaoxia X, De Mandal S, Shakeel M, Hua Y, Shoukat RF, Dongran Fu, Jin F (2021) Gut microbiota mediate Plutella xylostella susceptibility to Bt Cry1Ac protoxin is associated with host immune response. Environ Pollut 271:116271. https://doi.org/10.1016/j.envpol.2020.116271
Looft T, Johnson TA, Allen HK, Bayles DO, Alt DP, Stedtfeld RD et al (2012) In-feed antibiotic effects on the swine intestinal microbiome. Proc Natl Acad Sci 109:1691–1696. https://doi.org/10.1073/pnas.1120238109
Ma ZS (2018) The P/N (positive-to-negative Links) ratio in complex networks-A promising in silico biomarker for detecting changes cccurring in the human microbiome. Microb Ecol 75:1063–1073. https://doi.org/10.1007/s00248-017-1079-7
Magoč T, Salzberg SL (2011) FLASH: fast length adjustment of short reads to improve genome assemblies. Bioinformatics 27(21):2957–2963. https://doi.org/10.1093/bioinformatics/btr507
Malathi VM, More RP, Anandham R, Gracy GR, Mohan M, Venkatesan T, Samaddar S, Jalali SK, Sa TM (2018) Gut bacterial diversity of insecticide-susceptible and -resistant nymphs of the brown planthopper Nilaparvata lugens Stål (Hemiptera: delphacidae) and Elucidation of Their Putative Functional Roles. J Microbiol Biotechnol 28(6):976–986. https://doi.org/10.4014/jmb.1711.11039
Mason KL, Stepien TA, Blum JE, Holt JF, Labbe NH, Rush JS, Raffa KF, Handelsman J (2011) From commensal to pathogen: translocation of enterococcus faecalis from the Midgut to the Hemocoel of Manduca sexta. mBio. https://doi.org/10.1128/mBio.00065-11
Paddock Kyle J, Pereira Adriano E, Finke Deborah L, Ericsson Aaron C, Hibbard Bruce E, Shelby Kent S (2021) Host resistance to Bacillus thuringiensis is linked to altered bacterial community within a specialist insect herbivore. Mol Ecol 30(21):5438–5453. https://doi.org/10.1111/mec.15875
Paramasiva I, Sharma HC, Krishnayya PV (2014) Antibiotics influence the toxicity of the delta endotoxins of Bacillus thuringiensis towards the cotton bollworm Helicoverpa armigera. BMC Microbiol 14(1):1–12
Pardo-López L, Soberón M, Bravo A (2013) Bacillus thuringiensis insecticidal three-domain Cry toxins: mode of action, insect resistance and consequences for crop protection. FEMS Microbiol Rev 37:3–22. https://doi.org/10.1111/j.1574-6976.2012.00341.x
Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P et al (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acid Res 41:590–596. https://doi.org/10.1093/nar/gks1219
Raymond B, Johnston PR, Wright DJ, Ellis RJ, Crickmore N, Bonsall MB (2009) A mid-gut microbiota is not required for the pathogenicity of Bacillus thuringiensis to diamondback moth larvae. Environ Microbiol 11:2556–2563. https://doi.org/10.1111/j.1462-2920.2009.01980.x
Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS et al (2011) Metagenomic biomarker discovery and explanation. Genome Biol 12:R60. https://doi.org/10.1186/gb-2011-12-6-r60
Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D et al (2003) Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res 13:2498–2504. https://doi.org/10.1101/gr.1239303
Somerville HT, Tanada Y, Omi EM (1970) Lethal effect of purified spore and crystalline endotoxin preparations of Bacillus thuringiensis on several lepidopterous insects. J Invert Pathol 16:241–248. https://doi.org/10.1016/0022-2011(70)90065-0
Suzuki MT, Lereclus D, Nagy Arantes OM (2004) Fate of Bacillus thuringiensis strains in different insect larvae. Can J Microbiol 50:973–975. https://doi.org/10.1139/w04-087
Tabashnik BE, Carrière Y (2017) Surge in insect resistance to transgenic crops and prospects for sustainability. Nat Biotechnol 35(10):926–935. https://doi.org/10.1038/nbt.3974
Tang JY, Wang JQ (2004) Damage and control strategy of rice stem borers. China Agricultural Press, China, Beijing
Tang H, Chen G, Chen FJ, Han LZ, Peng YF (2018) Development and relative fitness of Cry1C resistance in Chilo suppressalis. Pest Manag Sci 74(3):590–597. https://doi.org/10.1002/ps.4740
Turroni F, Ventura M, ButtÓ LF, Duranti S, O’Toole PW, Motherway MO et al (2014) Molecular dialogue between the human gut microbiota and the host: a Lactobacillus and Bifidobacterium perspective. Cell Mol Life Sci 71:183–203. https://doi.org/10.1007/s00018-013-1318-0
Visweshwar R, Sharma HC, Akbar SMD, Sreeramulu K (2015) Elimination of gut microbes with antibiotics confers resistance to Bacillus thuringiensis toxin proteins in Helicoverpa armigera (Hubner). Appl Biochem Biotechnol 177:1621–1637. https://doi.org/10.1007/s12010-015-1841-6
Xiao Y, Wu K (2019) Recent progress on the interaction between insects and Bacillus thuringiensis crops. Phil Trans R Soc Lond B Biol Sci 374(1767):20180316. https://doi.org/10.1098/rstb.2018.0316
Zhang MY, Lövgren A, Low MG, Landén R (1993) Characterization of an avirulent pleiotropic mutant of the insect pathogen Bacillus thuringiensis: reduced expression of flagellin and phospholipases. Infect Immun 61(12):4947–4954
Acknowledgements
We are grateful to Dr. Yongbo Liu, from Chinese Research Academy of Environmental Sciences, and Dr Xiaoli Bing, from Nanjing Agricultural University, for helpful comments on this manuscript
Funding
This research was funded by the National Natural Science Foundation of China (31572336 and 31871963), the National Genetically Modified Organisms Key Breeding Projects of China (2018ZX08011-01B, 2016ZX08011-001 and 2016ZX08012-005), the National Key Research and Development Program of China (2017YFD0200400), and the Scientific and Technological Innovation Project of the Chinese Academy of Agricultural Science.
Author information
Authors and Affiliations
Contributions
GC, FJC and LZH conceived and designed research. GC and QWL conducted experiments and analyzed data. XWY, YHL and WWL reviewed and revised the manuscript. GC and LZH wrote the manuscript. All authors read and approved the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no conflicts of interest.
Ethical approval
This research does not involve the use of animal or humans in experimentation.
Additional information
Communicated by Cesar Rodriguez-Saona .
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Chen, G., Li, Q., Yang, X. et al. Comparison of the co-occurrence patterns of the gut microbial community between Bt-susceptible and Bt-resistant strains of the rice stem borer, Chilo suppressalis. J Pest Sci 96, 299–315 (2023). https://doi.org/10.1007/s10340-022-01512-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10340-022-01512-5