Diverse bacterial symbionts of insect-pathogentic fungi and possible impact on the maintenance of virulence during infection

Symbiosis - Tập 69 - Trang 47-58 - 2015
Chun Chen1, Xiangbo Chen1, Tingna Xie1, Justin Louis Hatting2, Xiaoping Yu1, Sudan Ye3, Zhengliang Wang1, Xuping Shentu1
1Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou, China
2ARC-Small Grain Institute (ARC-SGI), Bethlehem, South Africa
3Zhejiang Economic & Trade Polytechnic, Hangzhou, China

Tóm tắt

Bacterial-fungal interactions (BFIs) which have important ramifications for the biology of the interacting partners have been demonstrated extensively. Here we show for the first time that diverse bacterial symbionts occur in insect-pathogenic fungi. We firstly detected the bacterial symbionts by fluorescence in situ hybridization in combination with confocal laser scanning microscopy in Pandora neoaphidis. We also compared P. neoaphidis with the closely related species, P. nouryi, and found that 26 OTUs of bacterial symbionts covered taxonomically six classes in bacterial lineage, most were member of Gammaproteobacteria. To make inferences about the vertical transmission of bacterial symbionts, we compared their diversity in the hyphae and primary conidia of one isolate. Nine OTUs were identified and classified into four classes. Although the frequency of class Gammaproteobacteria was higher than other classes, the Shannon-Weiner diversity index was similar. Additionally, to understand the relationship between fungal virulence and bacterial symbiont diversity within fungi, we compared the predictive 16s rRNA clone library and the virulence of two genetically comparable isolates with different diversities of symbionts (F98028+ and F98028−). Conclusively, our study revealed the diversity of potential bacterial symbionts found inside insect-fungal structures through 16s rRNA clone library construction, and the potential impact between P. neoaphidis virulence and diversity in the bacterial symbionts harbored within their hyphae and conidia.

Tài liệu tham khảo

Arumugam M et al (2011) Enterotypes of the human gut microbiome. Nature 473:174–180. doi:10.1038/nature09944 Batut J, Andersson SGE, O’Callaghan D (2004) The evolution of chronic infection strategies in the alpha-proteobacteria. Nat Rev Microbiol 2:933–945. doi:10.1038/nrmicro1044 Bertaux J, Schmid M, Prevost-Boure NC, Churin JL, Hartmann A, Garbaye J, Frey-Mett P (2003) In situ identification of intracellular bacteria related to Paenibacillus spp. in the mycelium of the ectomycorrhizal fungus Laccatia bicolor S238N. Appl Environ Microbiol 69:4243–4248. doi:10.1128/aem.69.7.4243-4248.2003 Bertaux J, Schmid M, Hutzler P, Hartmann A, Garbaye J, Frey-Klett P (2005) Occurrence and distribution of endobacteria in the plant-associated mycelium of the ectomycorrhizal fungus Laccaria bicolor S238N. Environ Microbiol 7:1786–1795. doi:10.1111/j.1462-2920.2005.00867.x Bianciotto V, Bandi C, Minerdi D, Sironi M, Tichy HV, Bonfante P (1996) An obligately endosymbiotic mycorrhizal fungus itself harbors obligately intracellular bacteria. Appl Environ Microbiol 62:3005–3010 Bianciotto V, Lumini E, Lanfranco L, Minerdi D, Bonfante P, Perotto S (2000) Detection and identification of bacterial endosymbionts in arbuscular mycorrhizal fungi belonging to the family gigasporaceae. Appl Environ Microbiol 66:4503–4509. doi:10.1128/aem.66.10.4503-4509.2000 Bianciotto V, Genre A, Jargeat P, Lumini E, Becard G, Bonfante P (2004) Vertical transmission of endobacteria in the arbuscular mycorrhizal Fungus Gigaspora margarita through generation of vegetative spores. Appl Environ Microbiol 70:3600–3608. doi:10.1128/aem.70.6.3600-3608.2004 Bonfante P, Anca IA (2009) Plants, mycorrhizal fungi, and bacteria: a network of interactions. Annu Rev Microbiol 63:363–383. doi:10.1146/annurev.micro.091208.073504 Cannone JJ et al (2002) The Comparative RNA Web (CRW) Site: an online database of comparative sequence and structure information for ribosomal, intron, and other RNAs: Correction (vol 3, pg 2, 2002). BMC Bioinf 3:1. doi:10.1186/1471-2105-3-15 Chen C, Feng MG (2006) Experimental simulation of transmission of an obligate aphid pathogen with aphid flight dispersal. Environ Microbiol 8:69–76. doi:10.1111/j.1462-2920.2005.00869.x Chen AC, Imachi H, Sekiguchi Y, Ohashi A, Harada H (2003) Archaeal community compositions at different depths (up to 30 m) of a municipal solid waste landfill in Taiwan as revealed by 16S rDNA cloning analyses. Biotechnol Lett 25:719–724. doi:10.1023/a:1023458631699 Crawford IP (1989) Evolution of a biosynthetic-pathway-the tryptophan paradigm. Annu Rev Microbiol 43:567–600 Dale C, Moran NA (2006) Molecular interactions between bacterial symbionts and their hosts. Cell 126:453–465. doi:10.1016/j.cell.2006.07.014 de Boer W, Folman LB, Summerbell RC, Boddy L (2005) Living in a fungal world: impact of fungi on soil bacterial niche development. FEMS Microbiol Rev 29:795–811. doi:10.1016/j.femsre.2004.11.005 Engel P, Moran NA (2013) The gut microbiota of insects - diversity in structure and function. FEMS Microbiol Rev 37:699–735. doi:10.1111/1574-6976.12025 Frey-Klett P, Burlinson P, Deveau A, Barret M, Tarkka M, Sarniguet A (2011) Bacterial-fungal interactions: hyphens between agricultural, clinical, environmental, and food microbiologists. Microbiol Mol Biol Rev 75:583. doi:10.1128/mmbr.00020-11 Grell MN, Jensen AB, Olsen PB, Eilenberg J, Lange L (2011) Secretome of fungus-infected aphids documents high pathogen activity and weak host response. Fungal Genet Biol 48:343–352. doi:10.1016/j.fgb.2010.12.003 Gussow D, Clackson T (1989) Direct clone characterization from plaques and colonies by the polymerase chain reaction. Nucleic Acids Res 17:4000–4000 Hatting JL, Poprawski TJ, Miller RM (2000) Prevalences of fungal pathogens and other natural enemies of cereal aphids (Homoptera: Aphididae) in wheat under dryland and irrigated conditions in South Africa. BioControl 45:179–199 Hoffman MT, Arnold AE (2010) Diverse bacteria inhabit living hyphae of phylogenetically diverse fungal endophytes. Appl Environ Microbiol 76:4063–4075. doi:10.1128/aem.02928-09 Jargeat P, Cosseau C, Ola’h B, Jauneau A, Bonfante P, Batut J, Becard G (2004) Isolation, free-living capacities, and genome structure of “Candidatus glomeribacter gigasporarum,” the endocellular bacterium of the mycorrhizal fungus Gigaspora margarita. J Bacteriol 186:6876–6884. doi:10.1128/jb.186.20.6876-6884.2004 Lackner G, Mobius N, Scherlach K, Partida-Martinez LP, Winkler R, Schmitt I, Hertweck C (2009a) Global distribution and evolution of a toxinogenic burkholderia-rhizopus symbiosis. Appl Environ Microbiol 75:2982–2986. doi:10.1128/aem.01765-08 Lackner G, Partida-Martinez LP, Hertweck C (2009b) Endofungal bacteria as producers of mycotoxins. Trends Microbiol 17:570–576. doi:10.1016/j.tim.2009.09.003 Lambert B, Joos H, Dierickx S, Vantomme R, Swings J, Kersters K, Vanmontagu M (1990) Identification and plant interaction of a phyllobacterium sp, a predominant Rhizobacterium of young sugar-beet plants. Appl Environ Microbiol 56:1093–1102 Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, Chichester, pp 115–175 Levy A, Chang BJ, Abbott LK, Kuo J, Harnett G, Inglis TJJ (2003) Invasion of spores of the arbuscular mycorrhizal fungus Gigaspora decipiens by Burkholderia spp. Appl Environ Microbiol 69:6250–6256. doi:10.1128/aem.69.10.6250-6256.2003 Lidstrom ME, Chistoserdova L (2002) Plants in the pink: cytokinin production by Methylobacterium. J Bacteriol 184:1818–1818. doi:10.1128/jb.184.7.1818.2002 Lumini E et al (2007) Presymbiotic growth and sporal morphology are affected in the arbuscular mycorrhizal fungus Gigaspora margarita cured of its endobacteria. Cell Microbiol 9:1716–1729. doi:10.1111/j.1462-5822.2007.00907.x Manz W, Amann R, Ludwig W, Vancanneyt M, Schleifer KH (1996) Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment. Microbiology-Uk 142:1097–1106 Marquez LM, Redman RS, Rodriguez RJ, Roossinck MJ (2007) A virus in a fungus in a plant: Three-way symbiosis required for thermal tolerance. Science 315:513–515. doi:10.1126/science.1136237 Moran NA, Dunbar HE (2006) Sexual acquisition of beneficial symbionts in aphids. Proc Natl Acad Sci U S A 103:12803–12806. doi:10.1073/pnas.0605772103 Moran NA, Russell JA, Koga R, Fukatsu T (2005) Evolutionary relationships of three new species of Enterobacteriaceae living as symbionts of aphids and other insects. Appl Environ Microbiol 71:3302–3310. doi:10.1128/aem.71.6.3302-3310.2005 Oliver KM, Russell JA, Moran NA, Hunter MS (2003) Facultative bacterial symbionts in aphids confer resistance to parasitic wasps. Proc Natl Acad Sci U S A 100:1803–1807. doi:10.1073/pnas.0335320100 Papierok B (1989) On the occurrence of Entomophthorales (Zygomycetes) in Finland.1. species attacking aphids (Homoptera, Aphididae). Ann Entomol Fennici 55:63–69 Partida-Martinez LP, Hertweck C (2005) Pathogenic fungus harbours endosymbiotic bacteria for toxin production. Nature 437:884–888. doi:10.1038/nature03997 Partida-Martinez LP, Monajembashi S, Greulich KO, Hertweck C (2007) Endosymbiont-dependent host reproduction maintains bacterial-fungal mutualism. Curr Biol 17:773–777. doi:10.1016/j.cub.2007.03.039 Pell JK, Eilenberg J, Hajek AE, Steinkraus DC (2001) Biology, ecology and pest management potential of Entomophthorales. In: Butt TM, Jackson C, Magan N (eds) Fungal biological control agents: progress, problems and potential. CABI Publishing, Oxon, pp 71–153 Rikkinen J, Oksanen I, Lohtander K (2002) Lichen guilds share related cyanobacterial symbionts. Science 297:357–357. doi:10.1126/science.1072961 Sato Y et al (2009) Detection of betaproteobacteria inside the mycelium of the fungus Mortierella elongata. Microbes Environ 25:321–324. doi:10.1264/jsme2.ME10134 Scarborough CL, Ferrari J, Godfray HCJ (2005) Aphid protected from pathogen by endosymbiont. Science 310:1781–1781. doi:10.1126/science.1120180 Spellerberg IF, Fedor PJ (2003) A tribute to Claude Shannon (1916–2001) and a plea for more rigorous use of species richness, species diversity and the ‘Shannon-Wiener’ Index. Glob Ecol Biogeogr 12:177–179. doi:10.1046/j.1466-822X.2003.00015.x Stackebrandt E, Goebel BM (1994) A place for DNA-DNA reassociation and 16S ribosomal-RNA sequence-analysis in the present species definition in bacteriology. Int J Syst Bacteriol 44:846–849 Tang QY, Feng MG (2002) DPS data processing system for practical statistics. Science Press, Beijing Tarkka MT, Sarniguet A, Frey-Klett P (2009) Inter-kingdom encounters: recent advances in molecular bacterium-fungus interactions. Curr Genet 55:233–243. doi:10.1007/s00294-009-0241-2 Viswanathan VK, Green JM, Nichols BP (1995) Kinetic characterization of 4-amino 4-deoxychorismate synthase for Escherichia coli. J Bacteriol 177:5918–5923 Xu JH, Feng MG (2000) The time-dose-mortality modeling and virulence indices for two entomophthoralean species, Pandora delphacis and P. neoaphidis, against the green peach aphid, Myzus persicae. Biol Control 17:29–34. doi:10.1006/bcon.1999.0767