The microbial environment influences animal physiology. However, the underlying molecular mechanisms of such functional interactions are largely undefined. Previously, we showed that during chronic undernutrition, strains of Lactobacillus plantarum, a major commensal partner of Drosophila, promote host juvenile growth and maturation partly through enhanced expression of intestinal peptidases. By screening a transposon insertion library of Lactobacillus plantarum in gnotobiotic Drosophila larvae, we identify a bacterial cell-wall-modifying machinery encoded by the pbpX2-dlt operon that is critical to enhance host digestive capabilities and promote animal growth and maturation. Deletion of this operon leads to bacterial cell wall alteration with a complete loss of d-alanylation of teichoic acids. We show that L. plantarum cell walls bearing d-alanylated teichoic acids are directly sensed by Drosophila enterocytes to ensure optimal intestinal peptidase expression and activity, juvenile growth and maturation during chronic undernutrition. We thus conclude that besides peptidoglycan, teichoic acid modifications participate in the host–commensal bacteria molecular dialogue occurring in the intestine.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $5.17 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Erkosar, B. et al. Drosophila microbiota modulates host metabolic gene expression via IMD/NF-κB signaling. PLoS ONE 9, e94729 (2014).
Hooper, L. V. & Gordon, J. I. Commensal host–bacterial relationships in the gut. Science 292, 1115–1118 (2001).
Erkosar, B. et al. Pathogen virulence impedes mutualist-mediated enhancement of host juvenile growth via inhibition of protein digestion. Cell Host Microbe 18, 445–455 (2015).
Storelli, G. et al. Lactobacillus plantarum promotes drosophila systemic growth by modulating hormonal signals through TOR-dependent nutrient sensing. Cell Metab. 14, 403–414 (2011).
McFall-Ngai, M. et al. Animals in a bacterial world, a new imperative for the life sciences. Proc. Natl Acad. Sci. USA 110, 3229–3236 (2013).
Kim, E.-K., Park, Y. M., Lee, O. Y. & Lee, W.-J. Draft genome sequence of Lactobacillus plantarum strain WJL, a Drosophila gut symbiont. Genome Announc. 1, e00937-13 (2013).
Fraune, S. & Bosch, T. C. G. Why bacteria matter in animal development and evolution. Bioessays 32, 571–580 (2010).
Martino, M. E. et al. Resequencing of the Lactobacillus plantarum strain WJL genome. Genome Announc. 3, e01382-15 (2015).
Gury, J. R. M., Barthelmebs, L. & Cavin, J.-F. O. Random transposon mutagenesis of Lactobacillus plantarum by using the pGh9:ISS1 vector to clone genes involved in the regulation of phenolic acid metabolism. Arch. Microbiol. 182, 337–345 (2004).
Schroeder, B. O. & Bäckhed, F. Signals from the gut microbiota to distant organs in physiology and disease. Nat. Med. 22, 1079–1089 (2016).
Schwarzer, M. et al. Lactobacillus plantarum strain maintains growth of infant mice during chronic undernutrition. Science 351, 854–857 (2016).
Ma, D., Storelli, G., Mitchell, M. & Leulier, F. Studying host–microbiota mutualism in Drosophila: harnessing the power of gnotobiotic flies. Biomed. J. 38, 285–293 (2015).
Licandro-Seraut, H. et al. Development of an efficient in vivo system (Pjunc-TpaseIS1223) for random transposon mutagenesis of Lactobacillus casei. Appl. Environ. Microbiol. 78, 5417–5423 (2012).
Wong, C. N. A., Ng, P. & Douglas, A. E. Low-diversity bacterial community in the gut of the fruitfly Drosophila melanogaster. Environ. Microbiol. 13, 1889–1900 (2011).
Licandro-Seraut, H., Scornec, H., Pedron, T., Cavin, J. F. & Sansonetti, P. J. Functional genomics of Lactobacillus casei establishment in the gut. Proc. Natl Acad. Sci. USA. 111, E3101–E3109 (2014).
Goh, Y. J. & Klaenhammer, T. R. Genomic features of Lactobacillus species. Front. Biosci. 14, 1362–1386 (2009).
Perpetuini, G. et al. Identification of critical genes for growth in olive brine by transposon mutagenesis of Lactobacillus pentosus C11. Appl. Environ. Microbiol. 79, 4568–4575 (2013).
Kleerebezem, M. et al. The extracellular biology of the lactobacilli. FEMS Microbiol. Rev. 34, 199–230 (2010).
Sharon, G. et al. Commensal bacteria play a role in mating preference of Drosophila melanogaster. Proc. Natl Acad. Sci. USA 107, 20051–20056 (2010).
Blum, J. E., Fischer, C. N., Miles, J. & Handelsman, J. Frequent replenishment sustains the beneficial microbiome of Drosophila melanogaster. mBio 4, e00860-13 (2013).
Neuhaus, F. C. & Baddiley, J. A continuum of anionic charge: structures and functions of d-alanyl-teichoic acids in gram-positive bacteria. Microbiol. Mol. Biol. Rev. 67, 686–723 (2003).
Jones, R. M. et al. Symbiotic lactobacilli stimulate gut epithelial proliferation via Nox-mediated generation of reactive oxygen species. EMBO J. 32, 3017–3028 (2013).
Palumbo, E. et al. d-Alanyl ester depletion of teichoic acids in Lactobacillus plantarum results in a major modification of lipoteichoic acid composition and cell wall perforations at the septum mediated by the Acm2 autolysin. J. Bacteriol. 188, 3709–3715 (2006).
Newell, P. D. & Douglas, A. E. Among-species interactions determine the impact of gut microbiota on nutrient allocation in Drosophila melanogaster. Appl. Environ. Microbiol. 80, 788–796 (2013).
Wong, A. C. N., Dobson, A. J. & Douglas, A. E. Gut microbiota dictates the metabolic response of Drosophila to diet. J. Exp. Biol. 217, 1894–1901 (2014).
Perea Vélez, M. et al. Functional analysis of d-alanylation of lipoteichoic acid in the probiotic strain Lactobacillus rhamnosus GG. Appl. Environ. Microbiol. 73, 3595–3604 (2007).
Kovács, M. et al. A functional dlt operon, encoding proteins required for incorporation of d-alanine in teichoic acids in gram-positive bacteria, confers resistance to cationic antimicrobial peptides in Streptococcus pneumoniae. J. Bacteriol. 188, 5797–5805 (2006).
Leulier, F., Rodriguez, A., Khush, R. S., Abrams, J. M. & Lemaitre, B. The Drosophila caspase Dredd is required to resist gram-negative bacterial infection. EMBO Rep. 1, 353–358 (2000).
Leulier, F. et al. The Drosophila immune system detects bacteria through specific peptidoglycan recognition. Nat. Immunol. 4, 478–484 (2003).
Lhocine, N. et al. PIMS modulates immune tolerance by negatively regulating Drosophila innate immune signaling. Cell Host Microbe 4, 147–158 (2008).
Axelsson, L. et al. Genome sequence of the naturally plasmid-free Lactobacillus plantarum strain NC8 (CCUG 61730). J. Bacteriol. 194, 2391–2392 (2012).
Reichmann, N. T., Cassona, C. P. & Grundling, A. Revised mechanism of d-alanine incorporation into cell wall polymers in Gram-positive bacteria. Microbiology 159, 1868–1877 (2013).
Martino, M. E. et al. Nomadic lifestyle of Lactobacillus plantarum revealed by comparative genomics of 54 strains isolated from different habitats. Environ. Microbiol. 18, 4974–4989 (2016).
Grangette, C. et al. Enhanced antiinflammatory capacity of a Lactobacillus plantarum mutant synthesizing modified teichoic acids. Proc. Natl Acad. Sci. USA 102, 10321–10326 (2005).
Rigaux, P. et al. Immunomodulatory properties of Lactobacillus plantarum and its use as a recombinant vaccine against mite allergy. Allergy 64, 406–414 (2009).
Tabuchi, Y. et al. Inhibitory role for d-alanylation of wall teichoic acid in activation of insect toll pathway by peptidoglycan of Staphylococcus aureus. J. Immunol. 185, 2424–2431 (2010).
Kristian, S. A. et al. Alanylation of teichoic acids protects Staphylococcus aureus against Toll-like receptor 2-dependent host defense in a mouse tissue cage infection model. J. Infect. Dis. 188, 414–423 (2003).
Kristian, S. A. et al. d-Alanylation of teichoic acids promotes group A Streptococcus antimicrobial peptide resistance, neutrophil survival, and epithelial cell invasion. J. Bacteriol. 187, 6719–6725 (2005).
Peschel, A., Vuong, C., Otto, M. & Götz, F. The d-alanine residues of Staphylococcus aureus teichoic acids alter the susceptibility to vancomycin and the activity of autolytic enzymes. Antimicrob. Agents Chemother. 44, 2845–2847 (2000).
Perego, M. et al. Incorporation of d-alanine into lipoteichoic acid and wall teichoic acid in Bacillus subtilis. Identification of genes and regulation. J. Biol. Chem. 270, 15598–15606 (1995).
Smelt, M. J. et al. The impact of Lactobacillus plantarum WCFS1 teichoic acid d-alanylation on the generation of effector and regulatory T-cells in healthy mice. PLoS ONE 8, e63099 (2013).
Scornec, H. et al. Rapid 96-well plates DNA extraction and sequencing procedures to identify genome-wide transposon insertion sites in a difficult to lyse bacterium: Lactobacillus casei. J. Microbiol. Methods 106, 1–5 (2014).
Alikhan, N.-F. et al. BLAST Ring Image Generator (BRIG): simple prokaryote genome comparisons. BMC Genomics 12, 402 (2011).
Stothard, P. & Wishart, D. S. Circular genome visualization and exploration using CGView. Bioinformatics 21, 537–539 (2005).
Maguin, E. et al. Efficient insertional mutagenesis in lactococci and other gram-positive bacteria. J. Bacteriol. 178, 931–935 (1996).
Clarke, A. J. Compositional analysis of peptidoglycan by high-performance anion-exchange chromatography. Anal. Biochem. 212, 344–350 (1993).
Schneider, C. A. et al. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671–675 (2012).
Ducret, A. et al. MicrobeJ, a tool for high throughput bacterial cell detection and quantitative analysis. Nat. Microbiol. 1, 16077 (2016).
The authors would like to thank M. Strigini and G. Storelli for critical reading and editing of the manuscript; C. Grangeasse for helpful discussions and help with bacterial cell imaging; C. Login and T. Meylheuc from MIMA2 platform at INRA Jouy-en-Josas Research Center for TEM and SEM sample preparation and observation, respectively; the Arthro-Tools and PLATIM platforms of the SFR Biosciences (UMS3444/US8) for providing Drosophila and imaging facilities; the IGFL sequencing platform for deep sequencing; P. Serror for pG+host9 and H. Licandro-Seraut for the Pjunc-TpaseIS 1223 system. R.C.M. thanks the ‘Fondation pour la Recherche Médicale’ for financial support through a postdoctoral scholarship, SPF20140129318. M.E.M. was funded by the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement N8659510. This work was funded by an ERC starting grant (FP7/2007-2013-No. 309704). The laboratory of F.L. is supported by the FINOVI foundation and the EMBO Young Investigator Program.
The authors declare no competing financial interests.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Enrichment of intestinal Lactobacillus by enhanced secretory IgA coating alters glucose homeostasis in P2rx7−/− mice
Scientific Reports (2019)
Microbial Ecology (2019)
Genome Editing with CRISPR-Cas9 in Lactobacillus plantarum Revealed That Editing Outcomes Can Vary Across Strains and Between Methods
Biotechnology Journal (2019)
The Promoting Effect of Gut Microbiota on Growth and Development of Red Palm Weevil, Rhynchophorus ferrugineus (Olivier) (Coleoptera: Dryophthoridae) by Modulating Its Nutritional Metabolism
Frontiers in Microbiology (2019)
Evolutionary “Experiments” in Symbiosis: The Study of Model Animals Provides Insights into the Mechanisms Underlying the Diversity of Host–Microbe Interactions