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Mutation in cyaA in Enterobacter cloacae decreases cucumber root colonization

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Abstract

Strains of Enterobacter cloacae show promise as biological control agents for Pythium ultimum-induced damping-off on cucumber and other crops. Enterobacter cloacae M59 is a mini-Tn5 Km transposon mutant of strain 501R3. Populations of M59 were significantly lower on cucumber roots and decreased much more rapidly than those of strain 501R3 with increasing distance from the soil line. Strain M59 was decreased or deficient in growth and chemotaxis on most individual compounds detected in cucumber root exudate and on a synthetic cucumber root exudate medium. Strain M59 was also slightly less acid resistant than strain 501R3. Molecular characterization of strain M59 demonstrated that mini-Tn5 Km was inserted in cyaA, which encodes adenylate cyclase. Adenylate cyclase catalyzes the formation of cAMP and cAMP levels in cell lysates from strain M59 were approximately 2% those of strain 501R3. Addition of exogenous, nonphysiological concentrations of cAMP to strain M59 restored growth (1 mM) and chemotaxis (5 mM) on synthetic cucumber root exudate and increased cucumber seedling colonization (5 mM) by this strain without serving as a source of reduced carbon, nitrogen, or phosphorous. These results demonstrate a role for cyaA in colonization of cucumber roots by Enterobacter cloacae.

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Acknowledgments

We thank Dr. Li Wei, Ernest Williams, and Ricky Brathwaite for assistance with certain experiments. We also thank Dr. Steve Rehner, USDA-ARS, Beltsville, MD, for assistance in sequencing cyaA.

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  1. Scott M. Lohrke

    Present address: Naval Health Research Center, Environmental Health Effects Laboratory, Geo-Centers, Inc., Wright-Patterson Air Force Base, OH, 45433, USA

  2. Hye Suk Kong

    Present address: Laboratory of Methods Development, CBER, FDA, Rockville, MD, 20852, USA

  3. Jorge T. de Souza

    Present address: CEPLAC/CEPEC/SEFIT, Caixa Postal 7, Km 22, Rodovia Ilheus-Itabuna, BA, Brazil

Authors and Affiliations

  1. Sustainable Agricultural Systems Laboratory, USDA-Agricultural Research Service, Bldg. 001, Rm. 140, 10300 Baltimore Avenue, Beltsville, MD, 20705-2350, USA

    Daniel P. Roberts, Laurie F. McKenna, Scott M. Lohrke, Hye Suk Kong & John Lydon

  2. Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Baojian, Wuchang, Wuhan, 430062, People’s Republic of China

    Xiaojia Hu

  3. Wye Research and Education Center, University of Maryland, Queenstown, MD, 21658, USA

    Jorge T. de Souza

  4. Molecular Plant Pathology Laboratory, USDA – Agricultural Research Service, Bldg. 004, 10300 Baltimore Avenue, Beltsville, MD, 20705, USA

    C. Jacyn Baker

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Roberts, D.P., McKenna, L.F., Hu, X. et al. Mutation in cyaA in Enterobacter cloacae decreases cucumber root colonization. Arch Microbiol 187, 101–115 (2007). https://doi.org/10.1007/s00203-006-0177-6

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