Abstract
Pinewood nematode, Bursaphelenchus xylophilus, is one of the greatest threats to pine trees and is spreading all over the world. During the nematode’s pathogenesis, plant microorganisms play important roles. However, many microbial communities, such as that in Pinus massoniana, a major host of B. xylophilus that is widely distributed in China, are not well studied, especially the fungal communities. Here, the endophytic and rhizospheric bacterial and fungal communities associated with healthy and B. xylophilus–infected P. massoniana were analyzed. The results showed that 7639 bacterial and 3108 fungal OTUs were annotated from samples of P. massoniana, the rhizosphere, and B. xylophilus. There were significant diversity differences of endophytic microbes between healthy and infected P. massoniana. The abundances of endophytic bacteria Paenibacillus, unidentified_Burkholderiaceae, Serratia, Erwinia, and Pseudoxanthomonas and fungi Penicillifer, Zygoascus, Kirschsteiniothelia, Cyberlindnera, and Sporothrix in infected pines were greater than those in healthy pines, suggesting an association of particular microbial abundances with the pathogenesis of B. xylophilus in pines. Meanwhile, the abundances of microbes of unidentified_Burkholderiaceae, Saitozyma, and Pestalotiopsis were greater and Acidothermus and Trichoderma were lower in the rhizosphere under infected pines than those under healthy pines and the differences might be caused by B. xylophilus–induced weakening of the health of pines. Our study explored the endophytic and rhizospheric microbial community changes potentially caused by B. xylophilus infection of pines.
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References
Nickle W, Golden A, Mamiya Y, Wergin W (1981) On the taxonomy and morphology of the pine wood nematode, Bursaphelenchus xylophilus (Steiner & Buhrer 1934) Nickle 1970. J Nematol 13:385
Cheng H, Lin M, Li W, Fang Z (1983) The occurrence of a pine wilting disease caused by a nematode found in Nanjing. For Pest Dis 4:1–5
Mota MM, Braasch H, Bravo MA, Penas AC, Burgermeister W, Metge K, Sousa E (1999) First report of Bursaphelenchus xylophilus in Portugal and in Europe. Nematology 1:727–734
Yi CK, Byun BH, Park JD, Yang S, Chang KH (1989) First finding of the pine wood nematode, Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle and its insect vector in Korea. Research Reports of the Forestry Research Institute (Seoul): 141-149
Zamora P, Rodriguez V, Renedo F, Sanz AV, Dominguez JC, Perez-Escolar G, Miranda J, Alvarez B, Gonzalez-Casas A, Mayor E, Duenas M, Miravalles A, Navas A, Robertson L, Martin AB (2015) First report of Bursaphelenchus xylophilus causing pine wilt disease on Pinus radiata in Spain. Plant Dis 99:1449–1449
Zhao LL, Wei W, Kang L, Sun JH (2007) Chemotaxis of the pinewood nematode, Bursaphelenchus xylophilus, to volatiles associated with host pine, Pinus massoniana, and its vector Monochamus alternatus. J Chem Ecol 33:1207–1216
Ichihara Y, Fukuda K, Suzuki K (2000) Early symptom development and histological changes associated with migration of Bursaphelenchus xylophilus in seedling tissues of Pinus thunbergii. Plant Dis 84:675–680
Shi J, Chen F, Luo YQ, Wang Z, Xie BY, Sturrock RN (2012) First isolation of pine wood nematode from Pinus tabuliformis forests in China. For Pathol 43:59–66
Zhao B, Futai Kazuyoshi, Sutherland J, Takeuchi Yuko (2008) Pine wilt disease. Springer Japan
Guo Q, Guo D, Zhao B, Xu J, Li R (2007) Two cyclic dipeptides from Pseudomonas fluorescens GcM5-1A carried by the pine wood nematode and their toxicities to Japanese black pine suspension cells and seedlings in vitro. J Nematol 39:243–247
Han Z, Hong Y, Zhao B (2003) A study on pathogenicity of bacteria carried by pine wood nematodes. J Phytopathol 151:683–689
Vicente CSL, Ikuyo Y, Mota M, Hasegawa K (2013) Pinewood nematode-associated bacteria contribute to oxidative stress resistance of Bursaphelenchus xylophilus. BMC Microbiol 13:299
Oku H, Shiraishi T, Ouchi S, Kurozumi S, Ohta H (1980) Pine wilt toxin, the metabolite of a bacterium associated with a nematode. Naturwissenschaften 67:198–199
Dang QL, Choi GJ, Kim J-C, Cheon H-M, Jang KS, Son SW, Choi YH, Lim CH (2011) Pyochelin isolated from Burkholderia arboris KRICT1 carried by pine wood nematodes exhibits phytotoxicity in pine callus 13:521
Han ZM, Han SF, Wang HL, Zhao BG (2003) Distribution and pathogenicity of bacteria species carried by Bursaphelenchus xylophilus in China. Nematology 5:899–906
Kawazu K, Zhang H, Kanzaki H (1996) Accumulation of benzoic acid in suspension cultured cells of Pinus thunbergii Parl. in response to phenylacetic acid administration. Biosci Biotechnol Biochem 60:1410–1412
Vicente CSL, Nascimento FX, Espada M, Barbosa P, Hasegawa K, Mota M, Oliveira S (2013) Characterization of bacterial communities associated with the pine sawyer beetle Monochamus galloprovincialis, the insect vector of the pinewood nematode Bursaphelenchus xylophilus. FEMS Microbiol Lett 347:130–139
Kwon SJ, Jin HC, Lee S, Nam MH, Chung JH, Kwon SI, Ryu C-M, Park OK (2009) GDSL lipase-like 1 regulates systemic resistance associated with ethylene signaling in Arabidopsis. Plant J 58:235–245
Raffaele S, Leger A, Roby D (2009) Very long chain fatty acid and lipid signaling in the response of plants to pathogens. Plant Signal Behav 4:94–99
Wang Z, Zhang Y, Wang C, Wang Y, Sung C (2017) Esteya vermicola controls the pinewood nematode, Bursaphelenchus xylophilus, in pine seedlings. J Nematol 49:86–91
Maehara N (2008) Reduction of Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae) population by inoculating Trichoderma spp. into pine wilt-killed trees. Biol Control 44:61–66
Alves M, Pereira A, Vicente C, Matos P, Henriques J, Lopes H, Nascimento F, Mota M, Correia A, Henriques I (2018) The role of bacteria in pine wilt disease: insights from microbiome analysis. FEMS Microbiol Ecol 94:fiy077
Bal A, Anand R, Berge O, Chanway CP (2012) Isolation and identification of diazotrophic bacteria from internal tissues of Pinus contorta and Thuja plicata. Can J For Res 42:807–813
Izumi H, Anderson IC, Killham K, Moore ERB (2008) Diversity of predominant endophytic bacteria in European deciduous and coniferous trees. Can J Microbiol 54:173–179
Carper DL, Carrell AA, Kueppers LM, Frank AC (2018) Bacterial endophyte communities in Pinus flexilis are structured by host age, tissue type, and environmental factors. Plant Soil 428:335–352
Ma Y, Qu Z-L, Liu B, Tan J-J, Asiegbu FO, Sun H (2020) Bacterial community structure of Pinus thunbergii naturally infected by the nematode Bursaphelenchus xylophilus. Microorganisms 8:307
Pirttila AM, Laukkanen H, Pospiech H, Myllyla R, Hohtola A (2000) Detection of intracellular bacteria in the buds of Scotch pine (Pinus sylvestris L.) by in situ hybridization. Appl Environ Microbiol 66:3073–3077
Feng YH, Yang ZQ, Wang J, Luo QF, Li HG (2014) Development and characterization of SSR markers from Pinus massoniana and their transferability to P. elliottii, P. caribaea and P. yunnanensis. Genet Mol Res 13:1508–1513
Guo Y, Lin Q, Chen L, Carballar-Lejarazú R, Zhang A, Shao E, Liang G, Hu X, Wang R, Xu L, Zhang F, Wu S (2020) Characterization of bacterial communities associated with the pinewood nematode insect vector Monochamus alternatus Hope and the host tree Pinus massoniana. BMC Genomics:21
Fu Y-M, Liu H-B, Wu X-Q (2020) Diversity and function of endo-bacteria in Bursaphelenchus xylophilus from Pinus massoniana Lamb. in different regions. Forests:11
Plomion C, Bahrman N, Durel CE, O’Malley DM (1995) Genomic mapping in Pinus pinaster (maritime pine) using RAPD and protein markers. Heredity 74:661–668
Magoč T, Salzberg SL (2011) FLASH: fast length adjustment of short reads to improve genome assemblies. Bioinformatics 27:2957–2963
Bokulich NA, Subramanian S, Faith JJ, Gevers D, Gordon JI, Knight R, Mills DA, Caporaso JG (2013) Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing. Nat Methods 10:57–59
Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27:2194–2200
Haas BJ, Gevers D, Earl AM, Feldgarden M, Ward DV, Giannoukos G, Ciulla D, Tabbaa D, Highlander SK, Sodergren E, Methé B, DeSantis TZ, Consortium THM, Petrosino JF, Knight R, Birren BW (2011) Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons. Genome Res 21:494–504
Edgar RC (2013) UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat Methods 10:996–998
Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glöckner FO (2012) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41:D590–D596
Kõljalg U, Nilsson RH, Abarenkov K, Tedersoo L, Taylor AFS, Bahram M, Bates ST, Bruns TD, Bengtsson-Palme J, Callaghan TM, Douglas B, Drenkhan T, Eberhardt U, Dueñas M, Grebenc T, Griffith GW, Hartmann M, Kirk PM, Kohout P, Larsson E, Lindahl BD, Lücking R, Martín MP, Matheny PB, Nguyen NH, Niskanen T, Oja J, Peay KG, Peintner U, Peterson M, Põldmaa K, Saag L, Saar I, Schüßler A, Scott JA, Senés C, Smith ME, Suija A, Taylor DL, Telleria MT, Weiss M, Larsson K-H (2013) Towards a unified paradigm for sequence-based identification of fungi. Mol Ecol 22:5271–5277
Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797
Kobayashi DY, Crouch JA (2009) Bacterial/fungal interactions: from pathogens to mutualistic endosymbionts. Annu Rev Phytopathol 47:63–82
Lodewyckx C, Vangronsveld J, Porteous F, Moore ERB, Taghavi S, Mezgeay M, der Lelie D (2002) Endophytic bacteria and their potential applications. Crit Rev Plant Sci 21:583–606
Proença DN, Grass G, Morais PV (2017) Understanding pine wilt disease: roles of the pine endophytic bacteria and of the bacteria carried by the disease-causing pinewood nematode. MicrobiologyOpen 6:e00415
Li L, Tan J, Chen F (2018) Bacillus pumilus strain LYMC-3 shows nematicidal activity against Bursaphelenchus xylophilus via the production of a guanidine compound. Biocontrol Sci Tech 28:1128–1139
Proenca DN, Santo CE, Grass G, Morais PV (2012) Draft genome sequence of Serratia sp. strain M24T3, isolated from pinewood disease nematode Bursaphelenchus xylophilus. J Bacteriol 194:3764–3764
Prescott CE, Grayston SJ (2013) Tree species influence on microbial communities in litter and soil: current knowledge and research needs. For Ecol Manag 309:19–27
Finkel OM, Burch AY, Lindow SE, Post AF, Belkin S (2011) Geographical location determines the population structure in phyllosphere microbial communities of a salt-excreting desert tree. Appl Environ Microbiol 77:7647–7655
Coulon F, McKew BA, Osborn AM, McGenity TJ, Timmis KN (2007) Effects of temperature and biostimulation on oil-degrading microbial communities in temperate estuarine waters. Environ Microbiol 9:177–186
Bardgett RD, Lovell RD, Hobbs PJ, Jarvis SC (1999) Seasonal changes in soil microbial communities along a fertility gradient of temperate grasslands. Soil Biol Biochem 31:1021–1030
Belnap CP, Pan C, Denef VJ, Samatova NF, Hettich RL, Banfield JF (2011) Quantitative proteomic analyses of the response of acidophilic microbial communities to different pH conditions. ISME J 5:1152–1161
Zhang W, Zhao L, Zhou J, Yu H, Zhang C, Lv Y, Lin Z, Hu S, Zou Z, Sun J (2019) Enhancement of oxidative stress contributes to increased pathogenicity of the invasive pine wood nematode. Philos Trans Royal Soc B Biol Sci 374:20180323
Wingfield MJ (1987) Fungi associated with the pine wood nematode, Bursaphelenchus Xylophilus, and cerambycid beetles in Wisconsin. Mycologia 79:325–328
Zhang B, Zhang W, Lu M, Ahmad F, Tian H, Ning J, Liu X, Zhao L, Sun J (2017) Chemical signals of vector beetle facilitate the prevalence of a native fungus and the invasive pinewood nematode. J Nematol 49:341–347
Yang Z, Yu Z, Lei L, Xia Z, Shao L, Zhang K, Li G (2012) Nematicidal effect of volatiles produced by Trichoderma sp. J Asia-Pacif Entomol 15:647–650
Proenca DN, Francisco R, Kublik S, Schoeler A, Vestergaard G, Schloter M, Morais PV (2017) The microbiome of endophytic, wood colonizing bacteria from pine trees as affected by pine wilt disease. Sci Rep 7:4205
Vorholt JA (2012) Microbial life in the phyllosphere. Nat Rev Microbiol 10:828–840
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This work was funded by the National Key R&D Program of China (2018YFC1200400) and the National Natural Science Foundation of China (NSFC 31901315).
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Zhang, W., Wang, X., Li, Y. et al. Pinewood Nematode Alters the Endophytic and Rhizospheric Microbial Communities of Pinus massoniana. Microb Ecol 81, 807–817 (2021). https://doi.org/10.1007/s00248-020-01619-1
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DOI: https://doi.org/10.1007/s00248-020-01619-1