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Applied and Environmental Microbiology, July 2009, p. 4539-4549, Vol. 75, No. 13
0099-2240/09/$08.00+0     doi:10.1128/AEM.01336-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Phylogeny and Virulence of Naturally Occurring Type III Secretion System-Deficient Pectobacterium Strains{triangledown}

Hye-Sook Kim,1 Bing Ma,2 Nicole T. Perna,3 and Amy O. Charkowski1*

Department of Plant Pathology,1 Genome Center,2 Department of Genetics, University of Wisconsin—Madison, Madison, Wisconsin3

Received 3 July 2008/ Accepted 26 April 2009

Pectobacterium species are enterobacterial plant-pathogenic bacteria that cause soft rot disease in diverse plant species. Previous epidemiological studies of Pectobacterium species have suffered from an inability to identify most isolates to the species or subspecies level. We used three previously described DNA-based methods, 16S-23S intergenic transcribed spacer PCR-restriction fragment length polymorphism analysis, multilocus sequence analysis (MLSA), and pulsed-field gel electrophoresis, to examine isolates from diseased stems and tubers and found that MLSA provided the most reliable classification of isolates. We found that strains belonging to at least two Pectobacterium clades were present in each field examined, although representatives of only three of five Pectobacterium clades were isolated. Hypersensitive response and DNA hybridization assays revealed that strains of both Pectobacterium carotovorum and Pectobacterium wasabiae lack a type III secretion system (T3SS). Two of the T3SS-deficient strains assayed lack genes adjacent to the T3SS gene cluster, suggesting that multiple deletions occurred in Pectobacterium strains in this locus, and all strains appear to have only six rRNA operons instead of the seven operons typically found in Pectobacterium strains. The virulence of most of the T3SS-deficient strains was similar to that of T3SS-encoding strains in stems and tubers.

* Corresponding author. Mailing address: Department of Plant Pathology, University of Wisconsin—Madison, Madison, WI 53706. Phone: (608) 262-7911. Fax: (606) 263-2626. E-mail: amyc{at}plantpath.wisc.edu

{triangledown} Published ahead of print on 1 May 2009.

Applied and Environmental Microbiology, July 2009, p. 4539-4549, Vol. 75, No. 13
0099-2240/09/$08.00+0     doi:10.1128/AEM.01336-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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