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Bacillus subtilis expressing double-stranded RNA induced RNA interference in Tribolium castaneum.

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Title: Bacillus subtilis expressing double-stranded RNA induced RNA interference in Tribolium castaneum.
Authors: He L; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China., Zhou Y; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China., Xie J; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China., Gu Y; Syngenta Jealott's Hill International Research Centre, Berkshire, UK., Huang Y; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China., Tang X; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China.
Source: Pest management science [Pest Manag Sci] 2026 Oct; Vol. 82 (10), pp. 10073-10080. Date of Electronic Publication: 2026 Jun 24.
Publication Type: Journal Article
Language: English
Journal Info: Publisher: Published for SCI by Wiley Country of Publication: England NLM ID: 100898744 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1526-4998 (Electronic) Linking ISSN: 1526498X NLM ISO Abbreviation: Pest Manag Sci Subsets: MEDLINE
Imprint Name(s): Original Publication: West Sussex, UK : Published for SCI by Wiley, c2000-
MeSH Terms: Bacillus subtilis*/genetics , Bacillus subtilis*/metabolism , RNA, Double-Stranded*/genetics , RNA, Double-Stranded*/metabolism , Tribolium*/genetics , Tribolium*/growth & development , Pest Control, Biological*/methods , RNA Interference*, Larva/growth & development ; Larva/genetics ; Animals
Abstract: Background: RNA interference represents a promising and eco-friendly approach for pest management. As the core component of RNA biopesticides, the synthesis of double-stranded RNA (dsRNA) is the cornerstone for the further development and practical application of these biopesticides. Bacillus subtilis, with favorable characteristics (e.g., excellent safety, strong heterologous expression capability, and easy industrial fermentation), is an attractive dsRNA bio-factory for large-scale production of RNA biopesticides.
Results: An RNase-III-deficient B. subtilis strain was successfully constructed. Among the promoters screened from genes with high expression levels at each growth stage, pcotY, pcgeA, and phag exhibited high potential in driving dsRNA expression. Further optimization of the promoter phag significantly improved the yield of dsRNA. Using the mutant B. subtilis strain as the host, the optimized phag derivative phag-1 was utilized to express dsRNA targeting TcSpt5 and TcPolr2A of Tribolium castaneum. Larvae fed on an artificial diet containing these recombinant strains displayed significant reductions in survival and pupation rate, accompanied by reduced target gene expression.
Conclusion: Collectively, B. subtilis serves as an efficient dsRNA production platform that can be employed to control pests. © 2026 Society of Chemical Industry.
(© 2026 Society of Chemical Industry.)
References: Int J Mol Sci. 2024 Jun 13;25(12):. (PMID: 38928236)
Nat Biotechnol. 2007 Nov;25(11):1322-6. (PMID: 17982443)
J Bacteriol. 1975 Jul;123(1):366-71. (PMID: 806579)
Trends Biotechnol. 2026 Jan;44(1):193-219. (PMID: 40935726)
Environ Entomol. 2015 Apr;44(2):418-25. (PMID: 26313197)
FEMS Microbiol Rev. 2021 Nov 23;45(6):. (PMID: 34227665)
Microorganisms. 2020 Jul 12;8(7):. (PMID: 32664655)
Annu Rev Entomol. 2022 Jan 7;67:347-365. (PMID: 34614365)
PLoS Genet. 2012;8(12):e1003181. (PMID: 23300471)
Mol Biol Evol. 2003 Dec;20(12):2076-90. (PMID: 12949151)
Front Bioeng Biotechnol. 2022 Oct 10;10:980592. (PMID: 36299286)
Int J Insect Sci. 2019 Apr 24;11:1179543319840323. (PMID: 31040730)
Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2582-6. (PMID: 7708689)
J Bacteriol. 2003 May;185(10):3118-26. (PMID: 12730172)
Microorganisms. 2023 Jul 28;11(8):. (PMID: 37630488)
Microb Cell Fact. 2021 Aug 9;20(1):156. (PMID: 34372831)
PLoS One. 2015 Apr 30;10(4):e0124508. (PMID: 25928543)
Appl Environ Microbiol. 2016 Aug 15;82(17):5421-7. (PMID: 27342565)
DNA. 1984;3(1):17-21. (PMID: 6199171)
Front Bioeng Biotechnol. 2022 Nov 29;10:1080576. (PMID: 36524052)
Synth Syst Biotechnol. 2024 Feb 11;9(2):234-241. (PMID: 38385152)
Front Physiol. 2022 Apr 13;13:836106. (PMID: 35492592)
Microorganisms. 2023 Dec 06;11(12):. (PMID: 38138072)
Mol Gen Genet. 1998 May;258(4):385-8. (PMID: 9648743)
Mol Microbiol. 2007 Jun;64(6):1605-20. (PMID: 17555441)
J Agric Food Chem. 2025 Apr 9;73(14):8440-8447. (PMID: 40108859)
Methods. 2001 Dec;25(4):402-8. (PMID: 11846609)
J Agric Food Chem. 2022 Nov 2;70(43):13935-13944. (PMID: 36278912)
J Microbiol. 2023 Feb;61(2):199-210. (PMID: 36745334)
Biotechnol Lett. 2017 Jun;39(6):873-881. (PMID: 28238059)
Mol Microbiol. 1996 May;20(3):633-43. (PMID: 8736542)
Protein Expr Purif. 2018 Nov;151:72-77. (PMID: 29894806)
Pest Manag Sci. 2020 May;76(5):1699-1704. (PMID: 31758591)
Physiol Mol Biol Plants. 2025 Aug;31(8):1237-1257. (PMID: 41080511)
Int J Mol Sci. 2023 May 25;24(11):. (PMID: 37298215)
Nucleic Acids Res. 2008 Jan;36(Database issue):D93-6. (PMID: 17962296)
Grant Information: the National Key Research and Development Program of China (2023YFD1700700); the 2025Yangtze River Delta Science and Technology Innovation Community Joint Research (Basic Research) Project (2025CSJZN01300); the Syngenta PhD Scholarship; the Shanghai Agricultural Science and Technology Innovation Project (K2025007)
Contributed Indexing: Keywords: Bacillus subtilis; RNAi; Tribolium castaneum; dsRNA; promoter
Substance Nomenclature: 0 (RNA, Double-Stranded)
Entry Date(s): Date Created: 20260624 Date Completed: 20260911 Latest Revision: 20260913
Update Code: 20260913
PubMed Central ID: PMC13569281
DOI: 10.1002/ps.71045
PMID: 42340137
Database: MEDLINE
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  Data: Bacillus subtilis expressing double-stranded RNA induced RNA interference in Tribolium castaneum.
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  Data: <searchLink fieldCode="AU" term="%22He+L%22">He L</searchLink>; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China.<br /><searchLink fieldCode="AU" term="%22Zhou+Y%22">Zhou Y</searchLink>; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China.<br /><searchLink fieldCode="AU" term="%22Xie+J%22">Xie J</searchLink>; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China.<br /><searchLink fieldCode="AU" term="%22Gu+Y%22">Gu Y</searchLink>; Syngenta Jealott's Hill International Research Centre, Berkshire, UK.<br /><searchLink fieldCode="AU" term="%22Huang+Y%22">Huang Y</searchLink>; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China.<br /><searchLink fieldCode="AU" term="%22Tang+X%22">Tang X</searchLink>; School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.; Shanghai Yangtze River Delta Eco-Environmental Change and Management Observation and Research Station (Shanghai Urban Ecosystem Research Station), Ministry of Science and Technology, National Forestry and Grassland Administration, Shanghai, China.
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  Data: <searchLink fieldCode="JN" term="%22100898744%22">Pest management science</searchLink> [Pest Manag Sci] 2026 Oct; Vol. 82 (10), pp. 10073-10080. <i>Date of Electronic Publication: </i>2026 Jun 24.
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  Data: <i>Publisher: </i><searchLink fieldCode="PB" term="%22Published+for+SCI+by+Wiley%22">Published for SCI by Wiley </searchLink><i>Country of Publication: </i>England <i>NLM ID: </i>100898744 <i>Publication Model: </i>Print-Electronic <i>Cited Medium: </i>Internet <i>ISSN: </i>1526-4998 (Electronic) <i>Linking ISSN: </i><searchLink fieldCode="IS" term="%221526498X%22">1526498X </searchLink><i>NLM ISO Abbreviation: </i>Pest Manag Sci <i>Subsets: </i>MEDLINE
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  Data: <i>Original Publication</i>: West Sussex, UK : Published for SCI by Wiley, c2000-
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  Data: <searchLink fieldCode="MM" term="%22Bacillus+subtilis%22">Bacillus subtilis*</searchLink>/<searchLink fieldCode="MM" term="%22Bacillus+subtilis+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Bacillus+subtilis%22">Bacillus subtilis*</searchLink>/<searchLink fieldCode="MM" term="%22Bacillus+subtilis+metabolism%22">metabolism</searchLink> <br /><searchLink fieldCode="MM" term="%22RNA%2C+Double-Stranded%22">RNA, Double-Stranded*</searchLink>/<searchLink fieldCode="MM" term="%22RNA%2C+Double-Stranded+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22RNA%2C+Double-Stranded%22">RNA, Double-Stranded*</searchLink>/<searchLink fieldCode="MM" term="%22RNA%2C+Double-Stranded+metabolism%22">metabolism</searchLink> <br /><searchLink fieldCode="MM" term="%22Tribolium%22">Tribolium*</searchLink>/<searchLink fieldCode="MM" term="%22Tribolium+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Tribolium%22">Tribolium*</searchLink>/<searchLink fieldCode="MM" term="%22Tribolium+growth+%26+development%22">growth & development</searchLink> <br /><searchLink fieldCode="MM" term="%22Pest+Control%2C+Biological%22">Pest Control, Biological*</searchLink>/<searchLink fieldCode="MM" term="%22Pest+Control%2C+Biological+methods%22">methods</searchLink> <br /><searchLink fieldCode="MM" term="%22RNA+Interference%22">RNA Interference*</searchLink><br /><searchLink fieldCode="MH" term="%22Larva%22">Larva</searchLink>/<searchLink fieldCode="MH" term="%22Larva+growth+%26+development%22">growth & development</searchLink> ; <searchLink fieldCode="MH" term="%22Larva%22">Larva</searchLink>/<searchLink fieldCode="MH" term="%22Larva+genetics%22">genetics</searchLink> ; <searchLink fieldCode="MH" term="%22Animals%22">Animals</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Background: RNA interference represents a promising and eco-friendly approach for pest management. As the core component of RNA biopesticides, the synthesis of double-stranded RNA (dsRNA) is the cornerstone for the further development and practical application of these biopesticides. Bacillus subtilis, with favorable characteristics (e.g., excellent safety, strong heterologous expression capability, and easy industrial fermentation), is an attractive dsRNA bio-factory for large-scale production of RNA biopesticides.<br />Results: An RNase-III-deficient B. subtilis strain was successfully constructed. Among the promoters screened from genes with high expression levels at each growth stage, pcotY, pcgeA, and phag exhibited high potential in driving dsRNA expression. Further optimization of the promoter phag significantly improved the yield of dsRNA. Using the mutant B. subtilis strain as the host, the optimized phag derivative phag-1 was utilized to express dsRNA targeting TcSpt5 and TcPolr2A of Tribolium castaneum. Larvae fed on an artificial diet containing these recombinant strains displayed significant reductions in survival and pupation rate, accompanied by reduced target gene expression.<br />Conclusion: Collectively, B. subtilis serves as an efficient dsRNA production platform that can be employed to control pests. © 2026 Society of Chemical Industry.<br /> (© 2026 Society of Chemical Industry.)
– Name: Ref
  Label: References
  Group: RefInfo
  Data: Int J Mol Sci. 2024 Jun 13;25(12):. (PMID: <searchLink fieldCode="PM" term="%2238928236%22">38928236)</searchLink><br />Nat Biotechnol. 2007 Nov;25(11):1322-6. (PMID: <searchLink fieldCode="PM" term="%2217982443%22">17982443)</searchLink><br />J Bacteriol. 1975 Jul;123(1):366-71. (PMID: <searchLink fieldCode="PM" term="%22806579%22">806579)</searchLink><br />Trends Biotechnol. 2026 Jan;44(1):193-219. (PMID: <searchLink fieldCode="PM" term="%2240935726%22">40935726)</searchLink><br />Environ Entomol. 2015 Apr;44(2):418-25. (PMID: <searchLink fieldCode="PM" term="%2226313197%22">26313197)</searchLink><br />FEMS Microbiol Rev. 2021 Nov 23;45(6):. (PMID: <searchLink fieldCode="PM" term="%2234227665%22">34227665)</searchLink><br />Microorganisms. 2020 Jul 12;8(7):. (PMID: <searchLink fieldCode="PM" term="%2232664655%22">32664655)</searchLink><br />Annu Rev Entomol. 2022 Jan 7;67:347-365. (PMID: <searchLink fieldCode="PM" term="%2234614365%22">34614365)</searchLink><br />PLoS Genet. 2012;8(12):e1003181. (PMID: <searchLink fieldCode="PM" term="%2223300471%22">23300471)</searchLink><br />Mol Biol Evol. 2003 Dec;20(12):2076-90. (PMID: <searchLink fieldCode="PM" term="%2212949151%22">12949151)</searchLink><br />Front Bioeng Biotechnol. 2022 Oct 10;10:980592. (PMID: <searchLink fieldCode="PM" term="%2236299286%22">36299286)</searchLink><br />Int J Insect Sci. 2019 Apr 24;11:1179543319840323. (PMID: <searchLink fieldCode="PM" term="%2231040730%22">31040730)</searchLink><br />Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2582-6. (PMID: <searchLink fieldCode="PM" term="%227708689%22">7708689)</searchLink><br />J Bacteriol. 2003 May;185(10):3118-26. (PMID: <searchLink fieldCode="PM" term="%2212730172%22">12730172)</searchLink><br />Microorganisms. 2023 Jul 28;11(8):. (PMID: <searchLink fieldCode="PM" term="%2237630488%22">37630488)</searchLink><br />Microb Cell Fact. 2021 Aug 9;20(1):156. (PMID: <searchLink fieldCode="PM" term="%2234372831%22">34372831)</searchLink><br />PLoS One. 2015 Apr 30;10(4):e0124508. (PMID: <searchLink fieldCode="PM" term="%2225928543%22">25928543)</searchLink><br />Appl Environ Microbiol. 2016 Aug 15;82(17):5421-7. (PMID: <searchLink fieldCode="PM" term="%2227342565%22">27342565)</searchLink><br />DNA. 1984;3(1):17-21. (PMID: <searchLink fieldCode="PM" term="%226199171%22">6199171)</searchLink><br />Front Bioeng Biotechnol. 2022 Nov 29;10:1080576. (PMID: <searchLink fieldCode="PM" term="%2236524052%22">36524052)</searchLink><br />Synth Syst Biotechnol. 2024 Feb 11;9(2):234-241. (PMID: <searchLink fieldCode="PM" term="%2238385152%22">38385152)</searchLink><br />Front Physiol. 2022 Apr 13;13:836106. (PMID: <searchLink fieldCode="PM" term="%2235492592%22">35492592)</searchLink><br />Microorganisms. 2023 Dec 06;11(12):. (PMID: <searchLink fieldCode="PM" term="%2238138072%22">38138072)</searchLink><br />Mol Gen Genet. 1998 May;258(4):385-8. (PMID: <searchLink fieldCode="PM" term="%229648743%22">9648743)</searchLink><br />Mol Microbiol. 2007 Jun;64(6):1605-20. (PMID: <searchLink fieldCode="PM" term="%2217555441%22">17555441)</searchLink><br />J Agric Food Chem. 2025 Apr 9;73(14):8440-8447. (PMID: <searchLink fieldCode="PM" term="%2240108859%22">40108859)</searchLink><br />Methods. 2001 Dec;25(4):402-8. (PMID: <searchLink fieldCode="PM" term="%2211846609%22">11846609)</searchLink><br />J Agric Food Chem. 2022 Nov 2;70(43):13935-13944. (PMID: <searchLink fieldCode="PM" term="%2236278912%22">36278912)</searchLink><br />J Microbiol. 2023 Feb;61(2):199-210. (PMID: <searchLink fieldCode="PM" term="%2236745334%22">36745334)</searchLink><br />Biotechnol Lett. 2017 Jun;39(6):873-881. (PMID: <searchLink fieldCode="PM" term="%2228238059%22">28238059)</searchLink><br />Mol Microbiol. 1996 May;20(3):633-43. (PMID: <searchLink fieldCode="PM" term="%228736542%22">8736542)</searchLink><br />Protein Expr Purif. 2018 Nov;151:72-77. (PMID: <searchLink fieldCode="PM" term="%2229894806%22">29894806)</searchLink><br />Pest Manag Sci. 2020 May;76(5):1699-1704. (PMID: <searchLink fieldCode="PM" term="%2231758591%22">31758591)</searchLink><br />Physiol Mol Biol Plants. 2025 Aug;31(8):1237-1257. (PMID: <searchLink fieldCode="PM" term="%2241080511%22">41080511)</searchLink><br />Int J Mol Sci. 2023 May 25;24(11):. (PMID: <searchLink fieldCode="PM" term="%2237298215%22">37298215)</searchLink><br />Nucleic Acids Res. 2008 Jan;36(Database issue):D93-6. (PMID: <searchLink fieldCode="PM" term="%2217962296%22">17962296)</searchLink>
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  Data: the National Key Research and Development Program of China (2023YFD1700700); the 2025Yangtze River Delta Science and Technology Innovation Community Joint Research (Basic Research) Project (2025CSJZN01300); the Syngenta PhD Scholarship; the Shanghai Agricultural Science and Technology Innovation Project (K2025007)
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  Data: <i>Keywords: </i>Bacillus subtilis; RNAi; Tribolium castaneum; dsRNA; promoter
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        Text: English
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        StartPage: 10073
    Subjects:
      – SubjectFull: Larva growth & development
        Type: general
      – SubjectFull: Larva genetics
        Type: general
      – SubjectFull: Animals
        Type: general
      – SubjectFull: Bacillus subtilis genetics
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      – SubjectFull: RNA, Double-Stranded metabolism
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      – SubjectFull: Tribolium genetics
        Type: general
      – SubjectFull: Tribolium growth & development
        Type: general
      – SubjectFull: Pest Control, Biological methods
        Type: general
      – SubjectFull: RNA Interference
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    Titles:
      – TitleFull: Bacillus subtilis expressing double-stranded RNA induced RNA interference in Tribolium castaneum.
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              Text: 2026 Oct
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