Microbial Ecology

, Volume 53, Issue 4, pp 524–536 | Cite as

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

  • Junichiro Enya
  • Hirosuke Shinohara
  • Shigenobu Yoshida
  • Takao Tsukiboshi
  • Hiromitsu Negishi
  • Kazuo Suyama
  • Seiya TsushimaEmail author


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.


Sphingomonas Gray Mold Tomato Leave Pantoea Strong Antifungal Activity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



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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Copyright information

© Springer Science+Business Media, Inc. 2007

Authors and Affiliations

  • Junichiro Enya
    • 1
    • 2
  • Hirosuke Shinohara
    • 3
  • Shigenobu Yoshida
    • 4
  • Takao Tsukiboshi
    • 5
  • Hiromitsu Negishi
    • 1
  • Kazuo Suyama
    • 1
  • Seiya Tsushima
    • 4
    Email author
  1. 1.Department of Agricultural ScienceTokyo University of AgricultureKanagawaJapan
  2. 2.Research and Development DivisionKyowa Seed Co., Ltd.ChibaJapan
  3. 3.Plant Protection LaboratoryNational Agricultural Research Center for Tohoku Region FukushimaJapan
  4. 4.Environmental Biofunction DivisionNational Institute for Agro-Environmental SciencesIbarakiJapan
  5. 5.Laboratory of Plant PathologyNational Institute of Livestock and Grassland ScienceTochigiJapan

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