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Applied and Environmental Microbiology, December 2002, p. 6182-6192, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.6182-6192.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Relatedness of Chromosomal and Plasmid DNAs of Erwinia pyrifoliae and Erwinia amylovora

Gayle C. McGhee,1* Elise L. Schnabel,1 Kimberly Maxson-Stein,1 Beatrix Jones,2 Verlyn K. Stromberg,3 George H. Lacy,3 and Alan L. Jones1

Department of Plant Pathology, Michigan State University, East Lansing, Michigan 48824-1312,1 Department of Statistics, Pennsylvania State University, State College, Pennsylvania 16802,2 Department of Plant Pathology, Physiology, and Weed Science, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-03303

Received 18 June 2002/ Accepted 20 September 2002

The plant pathogen Erwinia pyrifoliae has been classified as a separate species from Erwinia amylovora based in part on differences in molecular properties. In this study, these and other molecular properties were examined for E. pyrifoliae and for additional strains of E. amylovora, including strains from brambles (Rubus spp.). The nucleotide composition of the internal transcribed spacer (ITS) region was determined for six of the seven 16S-23S rRNA operons detected in these species with a 16S rRNA gene probe. Each species contained four operons with a tRNAGlu gene and two with tRNAIle and tRNAAla genes, and analysis of the operons from five strains of E. amylovora indicated a high degree of ITS variability among them. One tRNAGlu-containing operon from E. pyrifoliae Ep1/96 was identical to one in E. amylovora Ea110, but three tRNAGlu operons and two tRNAIle and tRNAAla operons from E. pyrifoliae contained unique nucleotide changes. When groEL sequences were used for species-specific identification, E. pyrifoliae and E. amylovora were the closest phylogenetic relatives among a set of 12 bacterial species. The placement of E. pyrifoliae distinct from E. amylovora corroborated molecular hybridization data indicating low DNA-DNA similarity between them. Determination of the nucleotide sequence of plasmid pEP36 from E. pyrifoliae Ep1/96 revealed a number of presumptive genes that matched genes previously found in pEA29 from E. amylovora and similar organization for the genes and origins of replication. Also, pEP36 and pEA29 were incompatible with clones containing the reciprocal origin regions. Finally, the ColE1-like plasmid pEP2.6 from strain Ep1/96 contained sequences found in small plasmids in E. amylovora strains IL-5 and IH3-1.

* Corresponding author. Mailing address: 103 Center for Integrated Plant Systems (CIPS), Department of Plant Pathology, Michigan State University, East Lansing, MI 48824. Phone: (517) 353-3272. Fax: (517) 353-5598. E-mail: mcgheeg{at}msu.edu.

Applied and Environmental Microbiology, December 2002, p. 6182-6192, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.6182-6192.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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