Articles
RELATEDNESS OF ERWINIA AMYLOVORA STRAINS BASED ON AMPLIFIED 16S-23S RIBOSOMAL DNA RESTRICTION ENZYME ANALYSIS – ARDREA
Article number
489_5
Pages
55 – 60
Language
Abstract
The genetic diversity of 16 strains of Erwinia amylovora, isolated from different host plants and geographic regions, was investigated by amplified 16S-23S ribosomal DNA restriction enzyme analysis (ARDREA). Erwinia herbicola and Agrobacterium tumefaciens were used as outgroups.
Intergenic spacer (IGS) region of rDNA between the 16S and 23S rRNA genes were amplified by PCR and amplicons were digested with six different restriction endonucleases.
The resulting patterns were analyzed to determine similarity coefficients between strains that were used in the UPGMA cluster analysis procedure.
Cluster analysis, based on the number of ARDREA fragments shared between strains, showed that strains of E. amylovora isolated from subfamily Maloideae (Pomoideae) formed a single group, whereas three strains from Rubus (subfamily Rosoideae) formed a second group.
Two strains isolated from cotoneaster in the Royal Botanic Gardens, Melbourne, Australia and obtained from the International Collection of Micro-organisms from Plant, Auckland, New Zealand, were very similar to Maloideae strains of E. amylovora. One strain isolated from Asian pear in Hokkaido, Japan was related to Maloideae strains, whereas a different strain from Hokkaido was closely related to Rubus strains.
This study shows that E. amylovora exhibit genetic diversity detectable by ARDREA. Molecular and statistical analysis of ARDREA fragments can be used to differentiate groups of strains and to determine relatedness between them.
Grouping of E. amylovora strains based on RAPD (Momol et al. Journal of Applied Microbiology 1997, 82:389–398) and ARDREA fingerprintings were similar except for the Japanese strains which did not cluster into a specific subgroup by ARDREA. Resolution of Maloideae strains was greater with RAPD analysis than with to ARDREA. Maloideae strains are pathogenic to apple, to European and Asian pear, and to many other members of the Rosaceae family; Rubus strains are pathogenic only to Rubus spp.; and Japanese strains are primarily pathogenic to Asian and European Pears, with limited pathogenecity to apple (Aldwinckle and Gustafson, unpublished). Even though the IGS region is a small part of the bacterial genome, two of the three pathogenicity-specific subgroups (Maloideae, Rubus and Hokkaido) could be identified by ARDREA. Both ARDREA and RAPD fingerprinting analysis allowed us to differentiate Hokkaido strains from other E. amylovora strains.
Intergenic spacer (IGS) region of rDNA between the 16S and 23S rRNA genes were amplified by PCR and amplicons were digested with six different restriction endonucleases.
The resulting patterns were analyzed to determine similarity coefficients between strains that were used in the UPGMA cluster analysis procedure.
Cluster analysis, based on the number of ARDREA fragments shared between strains, showed that strains of E. amylovora isolated from subfamily Maloideae (Pomoideae) formed a single group, whereas three strains from Rubus (subfamily Rosoideae) formed a second group.
Two strains isolated from cotoneaster in the Royal Botanic Gardens, Melbourne, Australia and obtained from the International Collection of Micro-organisms from Plant, Auckland, New Zealand, were very similar to Maloideae strains of E. amylovora. One strain isolated from Asian pear in Hokkaido, Japan was related to Maloideae strains, whereas a different strain from Hokkaido was closely related to Rubus strains.
This study shows that E. amylovora exhibit genetic diversity detectable by ARDREA. Molecular and statistical analysis of ARDREA fragments can be used to differentiate groups of strains and to determine relatedness between them.
Grouping of E. amylovora strains based on RAPD (Momol et al. Journal of Applied Microbiology 1997, 82:389–398) and ARDREA fingerprintings were similar except for the Japanese strains which did not cluster into a specific subgroup by ARDREA. Resolution of Maloideae strains was greater with RAPD analysis than with to ARDREA. Maloideae strains are pathogenic to apple, to European and Asian pear, and to many other members of the Rosaceae family; Rubus strains are pathogenic only to Rubus spp.; and Japanese strains are primarily pathogenic to Asian and European Pears, with limited pathogenecity to apple (Aldwinckle and Gustafson, unpublished). Even though the IGS region is a small part of the bacterial genome, two of the three pathogenicity-specific subgroups (Maloideae, Rubus and Hokkaido) could be identified by ARDREA. Both ARDREA and RAPD fingerprinting analysis allowed us to differentiate Hokkaido strains from other E. amylovora strains.
Publication
Authors
E.A. Momol, M.T. Momol, J.L. Norelli, S.V. Beer, T.J. Burr, H.S. Aldwinckle
Keywords
Fire blight, fingerprinting, genetic diversity
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