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Culturable Leaf-Associated Bacteria on Tomato Plants and Their Potential as Biological Control Agents

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Abstract

Culturable leaf-associated bacteria inhabiting a plant have been considered as promising biological control agent (BCA) candidates because they can survive on the plant. We investigated the relationship between bacterial groups of culturable leaf-associated bacteria on greenhouse- and field-grown tomato leaves and their antifungal activities against tomato diseases in vitro and in vivo. In addition, the isolated bacteria were analyzed for N-acyl-homoserine lactone (AHL) and indole-3-acetic acid (IAA) production, which have been reported to associate with bacterial colonization, and resistance to a tomato alkaloid (α-tomatine). Leaf washings and subsequent leaf macerates were used to estimate the population size of epiphytic and more internal bacteria. Bacterial population sizes on leaves at the same position increased as the leaves aged under both greenhouse and field conditions. Field-grown tomatoes had significantly larger population sizes than greenhouse-grown tomatoes. Analysis of 16S rRNA gene (rDNA) sequencing using 887 culturable leaf-associated bacteria revealed a predominance of the Bacillus and Pseudomonas culturable leaf-associated bacterial groups on greenhouse- and field-grown tomatoes, respectively. Curtobacterium and Sphingomonas were frequently recovered from both locations. From the 2138 bacterial strains tested, we selected several strains having in vitro antifungal activity against three fungal pathogens of tomato: Botrytis cinerea, Fulvia fulva, and Alternaria solani. Among bacterial strains with strong in vitro antifungal activities, Bacillus and Pantoea tended to show strong antifungal activities, whereas Curtobacterium and Sphingomonas were not effective. The results indicated the differences in antifungal activity among predominant bacterial groups. Analysis of α-tomatine resistance revealed that most bacterial strains in the dominant groups exhibited moderate or high resistance to α-tomatine in growth medium. Furthermore, some Sphingomonas and Pantoea strains showed AHL and IAA production activities. Strain 125NP12 (Pantoea ananatis) showed particular α-tomatine resistance, and AHL and IAA production had the highest protective value (91.7) against gray mold. Thus, the differences of these physiological properties among dominant bacteria may be associated with the disease suppression ability of BCAs on tomato plants.

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References

  1. Andrews, JH (1992) Biological control in the phyllosphere. Annu Rev Phytopathol 30: 603–635

    Article  PubMed  CAS  Google Scholar 

  2. Arwiyanto, T, Sakata, K, Goto, M, Thuyumu, S, Takikawa, Y (1994) Induction of tomatine in tomato plant by an avirulent strain of Pseudomonas solanacearum. Ann Phytopathol Soc Jpn 60: 288–294

    CAS  Google Scholar 

  3. Beattie, GA, Lindow, SE (1999) Bacterial colonization of leaves: a spectrum of strategies. Phytopathology 89: 353–359

    Article  PubMed  CAS  Google Scholar 

  4. Beattie, GA, Marcell, LM (2002) Comparative dynamics of adherent and nonadherent bacterial populations on maize leaves. Phytopathology 92: 1015–1023

    Article  PubMed  Google Scholar 

  5. Beimen, A, Bermpohl, A, Meletzus, D, Eichenlaub, R, Barz, W (1992) Accumulation of phenolic compounds in leaves of tomato plants after infection with Clavibacter michigamense subsp. michiganense strains differing in virulence. Z Naturforsch [C] 47: 898–909

    Google Scholar 

  6. Bouarab, K, Melton, R, Peart, J, Baulocombe, D, Osbourn, A (2002) A saponin-detoxifying enzyme mediates suppression of plant defences. Nature 418: 889–892

    Article  PubMed  CAS  Google Scholar 

  7. Brandl, MT, Lindow, SE (1998) Contribution of indole-3-acetic acid production to the epiphytic fitness of Erwinia herbicola. Appl Environ Microbiol 64: 3256–3263

    PubMed  CAS  Google Scholar 

  8. Brandl, MT, Quinones, B, Lindow, SE (2001) Heterogenous transcription of an indoleacetic acid biosynthetic gene in Erwinia herbicola on plant surfaces. Proc Natl Acad Sci U S A 98: 3454–3459

    Article  PubMed  CAS  Google Scholar 

  9. Cattelan, AJ, Hartel, PG, Fuhrmann, JJ (1998) Bacterial composition in the rhizosphere of nodulating and non-nodulating soybean. Soil Sci Soc Am J 62: 1549–1555

    Article  CAS  Google Scholar 

  10. Chilton, MD, Currier, TC, Farrand, SK, Bendich, AJ, Gordon, MP, Nester, EW (1974) Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detected in grown gall tumors. Proc Natl Acad Sci U S A 71: 3672–3676

    Article  PubMed  CAS  Google Scholar 

  11. Cottyn, B, Regalado, E, Lanoot, B, De Cleene, M, Mew, TW, Swings, J (2000) Bacterial populations associated with rice seed in the tropical environment. Phytopathology 91: 282–292

    Article  Google Scholar 

  12. Denton, CS, Bardgett, RD, Cook, R, Hobbs, PJ (1999) Low amounts of root herbivory positively influence the rhizosphere microbial community in a temperate grassland soil. Soil Biol Biochem 31: 155–165

    Article  CAS  Google Scholar 

  13. Ercolani, GL (1991) Distribution of epiphytic bacteria on olive leaves and the influence of leaf age and sampling time. Microb Ecol 21: 35–48

    Article  Google Scholar 

  14. Friedman, M (2002) Tomato glycoalkaloides: role in the plant and in the diet. J Agric Food Chem 50: 5751–5780

    Article  PubMed  CAS  Google Scholar 

  15. Fukui, R, Fukui, H, Alvarez, AM (1999) Suppression of bacterial blight by a bacterial community isolated from the guttation fluids of anthuriums. Appl Environ Microbiol 65: 1020–1028

    PubMed  CAS  Google Scholar 

  16. Fukui, R, Fukui, H, Alvarez, AM (1999) Comparisons of single versus multiple bacterial species on biological control of anthurium blight. Phytopathology 89: 366–373

    Article  PubMed  CAS  Google Scholar 

  17. Gardan, L, David, C, Morel, M, Glickmann, E, Abu-Ghorrah, M, Petit, A, Dessaux, Y (1992) Evidence for correlation between auxin production and host plant species among strains of Pseudomonas syringae subsp. savastanoi. Appl Environ Microbiol 58: 1780–1783

    PubMed  CAS  Google Scholar 

  18. Gardan, SA, Weber, RP (1951) Colorimetric estimation of indole acetic acid. Plant Physiol 26: 192–195

    Article  Google Scholar 

  19. Germida, JJ, Siciliano, SD, Renato de Freitas, J, Seib, AM (1998) Diversity of root-associated bacteria associated with field-grown canola (Brassica napus L.) and wheat (Triticum aestivum L.). FEMS Microbiol Ecol 26: 43–50

    Article  CAS  Google Scholar 

  20. Goodfriend, WL (1997) Microbial community patterns of potential substrate utilization: a comparison of salt marsh, sand dune, and seawater-irrigated agronomic system. Soil Biol Biochem 30: 1169–1176

    Article  Google Scholar 

  21. Hirano, SS, Baker, LS, Upper, CD (1996) Raindrop momentum triggers growth of leaf-associated populations of Pseudomonas syringae on field-grown snap bean plants. Appl Environ Microbiol 62: 2560–2566

    PubMed  CAS  Google Scholar 

  22. Hirano, SS, Nordheim, EV, Arny, DC, Upper, CD (1982) Lognormal distribution of epiphytic bacterial populations on leaf surfaces. Appl Environ Microbiol 44: 695–700

    PubMed  CAS  Google Scholar 

  23. Hirano, SS, Upper, CD (1983) Ecology and epidemiology of foliar bacterial plant pathogens. Annu Rev Phytopathol 21: 43–269

    Article  Google Scholar 

  24. Hirano, SS, Upper, CD (2000) Bacterial in the ecosystem with emphasis on Pseudomonas syringae—a pathogen, ice nucleus, and epiphyte. Microbiol Mol Biol Rev 64: 624–653

    Article  PubMed  CAS  Google Scholar 

  25. Jurkevitch, EJ, Shapira, G (2000) Structure and colonization dynamics of epiphytic bacterial communities and of selected component strains on tomato (Lycopersicon esculentum) leaves. Microb Ecol 40: 300–308

    PubMed  Google Scholar 

  26. Kirimura, K, Nakagawa, H, Tsuji, K. Matsuda, K, Kurane, R, Usami, S (1999) Selective and continuous degradation of carbazole contained in petroleum oil by resting cells of Sphingomonas sp. CDH-7. Biosci Biotechnol Biochem 63: 1563–1568

    Article  PubMed  CAS  Google Scholar 

  27. Krause, DK, Easter, RA, White, BA, Mackie, RI (1995) Effect of weaning diet on the ecology of adherent Lactobacillus in the gastrointestinal tract of the pig. J Anim Sci 73: 2347–2354

    PubMed  CAS  Google Scholar 

  28. Lairini, K, Perez-Espinosa, A, Pineda, M, Ruiz-Rubio, M (1996) Purification and characterization of tomatinase from Fusarium oxysporum f. sp. lycopersici. Appl Environ Microbiol 62: 1604–1609

    PubMed  CAS  Google Scholar 

  29. Legard, DE, McQuiken, MP, Whipps, JM, Fenlon, JS, Fermor, TR, Thompson, IP, Bailey, MJ, Lynch, JM (1994) Studies of seasonal changes in the microbial populations on the phyllosphere of spring wheat as a prelude to the release of a genetically modified microorganism. Agric Ecosyst Environ 50: 87–101

    Article  Google Scholar 

  30. Lindow, SE, Brandl, MT (2003) Microbiology of the phyllosphere. Appl Environ Microbiol 69: 1875–1883

    Article  PubMed  CAS  Google Scholar 

  31. Mahaffee, WF, Kloepper, JW (1997) Bacterial communities of the rhizosphere and endorhiza associated with field-grown cucumber plants inoculated with a plant growth-promoting rhizobacterium or its genetically modified derivative. Can J Microbiol 43: 344–353

    Article  PubMed  CAS  Google Scholar 

  32. Marchesi, JR, Sato, T, Weightman, AJ, Martin, TA, Fry, JC, Hiom, SJ, Wade, WG (1998) Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA. Appl Environ Microbiol 64: 795–799

    PubMed  CAS  Google Scholar 

  33. Monier, JM, Lindow, SE (2003) Differential survival of solitary and aggregated bacterial cells promotes aggregate formation on leaf surfaces. Proc Natl Acad Sci U S A 100: 15977–15982

    Article  PubMed  CAS  Google Scholar 

  34. Nishiyama, K, Ezuka, A (1977) Rough-colony isolates of Pseudomonas coronafaciens var. atropurpurea obtained from diseased leaves of ryegrasses. Ann Phytopathol Soc Jpn 43: 426–431 (in Japanese with English summary)

    Google Scholar 

  35. Piper, KR, Beck von Bodman, S, Farrand, SK (1993) Conjugation factor of Agrobacterium tumefaciens regulates Ti plasmid transfer by autoinduction. Nature 362: 448–450

    Article  PubMed  CAS  Google Scholar 

  36. Sandrock, RW, Dellapenna, D, VanEtten, HD (1995) Purification and characterization of β2-tomatinase, an enzyme involved in the degradation of α-tomatine and isolation of the gene encoding β2-tomatinase from Septoria lycopersici. Mol Plant Microbe Interact 8: 960–970

    PubMed  CAS  Google Scholar 

  37. Shiomi, Y, Nishiyama, M, Onizuka, T, Marumoto, T (1999) Comparison of bacterial community structures in the rhizoplane of tomato plants grown soils suppressive and conducive towards bacterial wilt. Appl Environ Microbiol 65: 3996–4001

    PubMed  CAS  Google Scholar 

  38. Siciliano, SD, Theoret, CM, de Freitas, JR, Hucl, PJ, Germida, JJ (1998) Differences in the microbial communities associated with the root of different cultivars of canola and wheat. Can J Microbiol 44: 844–851

    Article  CAS  Google Scholar 

  39. Smalla, K, Wachtendorf, U, Heuer, H, Liu, WT, Forney, L (1998) Analysis of BIOLOG GN substrate utilization patterns by microbial communities. Appl Environ Microbiol 64: 1220–1225

    PubMed  CAS  Google Scholar 

  40. Thompson, IP, Bailey, MJ, Fenlon, JS, Fermor, TR, Lilley, AK, Lynch, JM, McCormack, PJ, McQuilken, MP, Purdy, KJ, Rainey, PB, Whipps, JM (1993) Quantitative and qualitative seasonal changes in the microbial community from the phyllosphere of sugar beet (Beta vulgaris). Plant Soil 150: 177–191

    Article  Google Scholar 

  41. Weller, DM (1988) Biological control of soilborne plant pathogens in the rhizosphere with bacteria. Annu Rev Phytopathol 26: 379–407

    Article  Google Scholar 

  42. Yang, CH, Crowley, DE, Borneman, J, Keen, NT (2001) Microbial phyllosphere populations are more complex than previously realized. Proc Natl Acad Sci U S A 98: 3889–3894

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We are grateful to Dr. Naoyuki Matsumoto (National Institute for Agro-Environmental Sciences) and Dr. Takako Sakai (Kyowa Seed Co., Ltd.) for suggestions and critical reading of the manuscript. The authors thank Dr. Linda L. Kinkel (University of Minnesota) for help with the AHLs detection. Thanks are due to Rie Adachi, Yuriko Watanabe, Nobutaka Numaziri, and Machiko Imai for technical assistance.

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Authors and Affiliations

  1. Department of Agricultural Science, Tokyo University of Agriculture, 1737, Funako, Atsugi, Kanagawa, 243-0034, Japan

    Junichiro Enya,  Hiromitsu Negishi & Kazuo Suyama

  2. Research and Development Division, Kyowa Seed Co., Ltd., 377, Ozawa, Chonan, Chosei, Chiba, 297-0142, Japan

    Junichiro Enya

  3. Plant Protection Laboratory, National Agricultural Research Center for Tohoku Region, 50, Harajuku-Minami, Arai, Fukushima, Fukushima, 960-2156, Japan

    Hirosuke Shinohara

  4. Environmental Biofunction Division, National Institute for Agro-Environmental Sciences, 3-1-3, Kannondai, Tsukuba, Ibaraki, 305-8604, Japan

    Shigenobu Yoshida & Seiya Tsushima

  5. Laboratory of Plant Pathology, National Institute of Livestock and Grassland Science, 768, Senbonmatsu, Shiobara, Tochigi, 329-2793, Japan

    Takao Tsukiboshi

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  1. Junichiro Enya
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  2. Hirosuke Shinohara
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  3. Shigenobu Yoshida
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  4. Takao Tsukiboshi
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  5. Hiromitsu Negishi
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  6. Kazuo Suyama
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  7. Seiya Tsushima
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Correspondence to Seiya Tsushima.

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Enya, J., Shinohara, H., Yoshida, S. et al. Culturable Leaf-Associated Bacteria on Tomato Plants and Their Potential as Biological Control Agents. Microb Ecol 53, 524–536 (2007). https://doi.org/10.1007/s00248-006-9085-1

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  • DOI: https://doi.org/10.1007/s00248-006-9085-1

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