Asaia symbionts interfere with infection by Flavescence dorée phytoplasma in leafhoppers
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The transmission of microbial pathogens by insect vectors can be affected by the insect’s microbial symbionts, which may compete in colonizing organs, express antagonistic factors or activate host immune response. Acetic acid bacteria of the genus Asaia are symbionts of the leafhopper Scaphoideus titanus, which transmits Flavescence dorée phytoplasma. These bacteria could be used as control agents against the disease. Here, we experimentally investigated the interaction between different strains of Asaia and phytoplasma transmission in the laboratory by using the model leafhopper Euscelidius variegatus and the plant host Vicia faba. We found that uncultivable and low concentrations of Asaia phylotypes were associated with E. variegatus. When we supplied different Asaia strains isolated from other insects and exhibiting different phenotypes to E. variegatus orally, the bacteria stably colonized the leafhopper, reached relatively higher densities and could then be isolated from the host. We conducted transmission trials of Flavescence dorée phytoplasma with individuals colonized with three exogenous Asaia strains. When the phytoplasma became established in the bodies of E. variegatus, leafhoppers were able to transmit it to broad beans, with transmission rates ranging from 33 to 76% in different experiments. However, leafhoppers that were colonized by one of the Asaia strains producing an air–liquid interface biofilm exhibited significantly reduced phytoplasma acquisition, with infection rates at 5–28%, whereas they were 25–77% in control insects. Although the mechanisms regulating this interference remain to be elucidated, our results provide evidence of the potential use of Asaia as a biocontrol agent.
KeywordsFlavescence dorée Acetic acid bacteria Vector competence Symbiotic control
The authors are grateful to Federico Lessio and Luca Picciau for their essential help in insect rearing and laboratory activities. This work was supported by the ‘INTEFLAVI’ (Un approccio integrato alla lotta contro la flavescenza dorata della vite) project. D.D. thanks King Abdullah University of Science and Technology for support through baseline research funds. The authors thank Prof. Guido Favia for providing A. stephensi samples. Funding was provided by Fondazione Cassa di Risparmio di Cuneo.
Compliance with ethical standards
Conflict of interest
All authors declare that they have no conflict of interest.
All applicable international, national and/or institutional guidelines for the care and use of animals were followed.
- Ammar E-D, Ramos JE, Hall DG, Dawson WO, Shatters RG Jr (2016) Acquisition, replication and inoculation of Candidatus Liberibacter asiaticus following various acquisition periods on Huanglongbing-infected citrus by nymphs and adults of the asian citrus psyllid. PLoS ONE 11(7):e0159594CrossRefPubMedCentralGoogle Scholar
- Capone A, Ricci I, Damiani C, Mosca M, Rossi P, Scuppa P, Crotti E, Epis S, Angeletti M, Valzano M, Sacchi L, Bandi C, Daffonchio D, Mandrioli M, Favia G (2013) Interactions between Asaia, Plasmodium and Anopheles: new insights into mosquito symbiosis and implications in Malaria Symbiotic Control. Parasite Vector 6:182CrossRefGoogle Scholar
- Chouaia B, Rossi P, Montagna M, Ricci I, Crotti E, Damiani C, Epis S, Faye I, Sagnon N, Alma A, Favia G, Daffonchio D, Bandi C (2010) Molecular evidence for multiple infections as revealed by typing of Asaia bacterial symbionts of four mosquito species. Appl Environ Microbiol 76:7444–7450CrossRefPubMedPubMedCentralGoogle Scholar
- Crotti E, Damiani C, Pajoro M, Gonella E, Rizzi A, Ricci I, Negri I, Scuppa P, Rossi P, Ballarini P, Raddadi N, Marzorati M, Sacchi L, Clementi E, Genchi M, Mandrioli M, Bandi C, Favia G, Alma A, Daffonchio D (2009) Asaia, a versatile acetic acid bacterial symbiont, capable of cross-colonizing insects of phylogenetically distant genera and orders. Environ Microbiol 11:3252–3264CrossRefPubMedGoogle Scholar
- Fagen JR, Giongo A, Brown CT, Davis-Richardson AG, Gano KA, Triplett EW (2012) Characterization of the relative abundance of the citrus pathogen Ca. Liberibacter asiaticus in the microbiome of its insect vector, Diaphorina citri, using high throughput 16S rRNA sequencing. Open Microbiol J 6:29–33CrossRefPubMedPubMedCentralGoogle Scholar
- Favia G, Ricci I, Damiani C, Raddadi N, Crotti E, Marzorati M, Rizzi A, Urso R, Brusetti L, Borin S, Mora D, Scuppa P, Pasqualini L, Clementi E, Genchi M, Corona S, Negri I, Grandi G, Alma A, Kramer L, Esposito F, Bandi C, Sacchi L, Daffonchio D (2007) Bacteria of the genus Asaia stably associate with Anopheles stephensi, an Asian malarial mosquito vector. Proc Natl Acad Sci USA 104:9047–9051CrossRefPubMedPubMedCentralGoogle Scholar
- Fuchs BM, Wallner G, Beisker W, Schwippl I, Ludwig W, Amann R (1998) Flow cytometric analysis of the in situ accessibility of Escherichia coli 16S rRNA for fluorescently labeled oligonucleotide probes. Appl Environ Microbiol 42:4973–4982Google Scholar
- Galetto L, Nardi M, Saracco P, Bressan A, Marzachì C, Bosco D (2009) Host plant determines the phytoplasma transmission competence of Empoasca decipiens (Hemiptera: Cicadellidae) variation in vector competency depends on chrysanthemum yellows phytoplasma distribution within Euscelidius variegatus. Entomol Exp Appl 131:200–207CrossRefGoogle Scholar
- Gonella E, Negri I, Marzorati M, Mandrioli M, Sacchi L, Pajoro M, Crotti E, Rizzi A, Clementi E, Tedeschi R, Bandi C, Alma A, Daffonchio D (2011) Bacterial endosymbiont localization in Hyalesthes obsoletus, the insect vector of Bois Noir in Vitis vinifera. Appl Environ Microbiol 77(4):1423–1435CrossRefPubMedGoogle Scholar
- Hughes GL, Dodson BL, Johnson RM, Murdock CC, Tsujimoto H, Suzuki Y, Patt AA, Cui L, Nossa CW, Barry RM, Sakamoto JM, Hornett EA, Rasgon JL (2014) Native microbiome impedes vertical transmission of Wolbachia in Anopheles mosquitoes. Proc Natl Acad Sci USA 111(34):12498–12503CrossRefPubMedPubMedCentralGoogle Scholar
- Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, New York, pp 115–175Google Scholar
- Lessio F, Picciau L, Gonella E, Mandrioli M, Tota F, Alma A (2016) The mosaic leafhopper Orientus ishidae: host plants, spatial distribution, infectivity, and transmission of 16SrV phytoplasmas to vines. Bull Insectol 69(2):277–289Google Scholar
- Li W, Yajima T, Saito K, Nishimura H, Fushimi T, Ohshima Y, Tsukamoto Y, Yoshikai Y (2004) Immunostimulating properties of intragastrically administered Acetobacter-derived soluble branched (1,4)-beta-d-glucans decrease murine susceptibility to Listeria monocytogenes. Infect Immun 72(12):7005–7011CrossRefPubMedPubMedCentralGoogle Scholar
- Mandrioli M, Monti M, Tedeschi R (2015) Presence and conservation of the immunoglobulin superfamily in insects: current perspective and future challenges. Invertebr Surviv J 12:188–194Google Scholar
- Mapelli F, Marasco R, Rolli E, Barbato M, Cherif H, Guesmi A, Ouzari I, Daffonchio D, Borin S (2013) Potential for plant growth promotion of rhizobacteria associated with Salicornia growing in Tunisian hypersaline soils. Biomed Res Int 248078Google Scholar
- Marzachì C, Palermo S, Boarino A, Veratti F, D’Aquilio M, Loria A, Boccardo G (2001) Optimisation of one step PCR assay for the diagnosis of flavescence dorée-related phytoplasmas in field-grown grapevines and vector populations. Vitis 40:213–217Google Scholar
- Marzorati M, Alma A, Sacchi L, Pajoro M, Palermo S, Brusetti L, Raddadi N, Balloi A, Tedeschi R, Clementi E, Corona S, Quaglino F, Bianco PA, Beninati T, Bandi C, Daffonchio D (2006) A novel bacteroidetes symbiont is localized in Scaphoideus titanus, the insect vector of Flavescence Dorée in Vitis vinifera. Appl Environ Microbiol 72(2):1467–1475CrossRefPubMedPubMedCentralGoogle Scholar
- Rossi P, Ricci I, Cappelli A, Damiani C, Ulissi U, Mancini MV, Valzano M, Capone A, Epis S, Crotti E, Chouaia B, Scuppa P, Joshi D, Xi Z, Mandrioli M, Sacchi L, O’Neill SL, Favia G (2015) Mutual exclusion of Asaia and Wolbachia in the reproductive organs of mosquito vectors. Parasit Vectors 8:278CrossRefPubMedPubMedCentralGoogle Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
- Tedeschi R, Monti M, Gonella E, Melchiori G, Alma A, Mandrioli M (2017) Molecular and cellular analysis of immunity in the phytoplasma vector Euscelidius variegatus: exploiting immunity to improve biological control strategies. Invertebr Surviv J 14:63–72Google Scholar
- Vacchini V, Gonella E, Crotti E, Prosdocimi EM, Mazzetto F, Chouaia B, Callegari M, Mapelli F, Mandrioli M, Alma A, Daffonchio D (2017) Bacterial diversity shift determined by different diets in the gut of the spotted wing fly Drosophila suzukii is primarily reflected on acetic acid bacteria. Environ Microbiol Rep 9(2):91–103CrossRefPubMedGoogle Scholar
- Wangkeeree J, Miller TA, Hanboonsong Y (2011) Predominant bacteria symbionts in the leafhopper Matsumuratettix hiroglyphicus—the vector of sugarcane white leaf phytoplasma. Bull Insectol 64:S215–S216Google Scholar