Significance and impact of the study the current

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Significance and Impact of the Study: The current detection techniques are mainly based on the genetic similarities observed within the strains from the cultivated tree-fruit crops. For a more reliable detection of the fire blight pathogen also in wild and ornamentals Rosaceous plants the genetic features of deviating E. amylovora strains have to be studied in detail. Journal of Applied Microbiology ISSN 1364-5072 1084 Journal compilation ª 2006 The Society for Applied Microbiology, Journal of Applied Microbiology 100 (2006) 1084–1094 ª 2006 The Authors
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protocols. Recently, several molecular techniques have revealed unexpected genetic variability among E. amylo- vora strains. Both repetitive-sequence PCR (rep-PCR) and random amplified polymorphic DNA (RAPD) analysis allowed the differentiation of strains isolated from Malo- ideae plant species such as pear ( Pyrus communis L.), apple ( Malus domestica Borkh.), quince ( Cydonia oblonga L.), hawthorn ( Crataegus spp.) from the strains obtained from Rosoideae such as raspberry ( Rubus spp.) (McManus and Jones 1995a; Momol et al. 1997). Differentiation of Rubus strains was also possible using 23S rDNA sequence analysis (Maes et al. 1996), DNA restriction enzyme analysis of the 16S–23S intergenic spacer regions (Momol et al. 1999) and amplified fragment length polymorphism (AFLP) ana- lysis (Rico et al. 2004). In addition, relationship between some E. amylovora strains and their geographical origins has been inferred using pulsed-field gel electrophoresis (Zhang and Geider 1997; Jock et al. 2002). An important feature of the bacterium utilized both for characterization and detection is plasmid pEA29. With few exceptions, all E. amylovora strains carry a similar plasmid of c. 29 kb that can be detected by PCR amplifi- cation of 900 bp Pst I fragment (Bereswill et al. 1992). However, the Pst I fragment of some strains was larger than the expected size (McManus and Jones 1995b). Moreover, variability in the length of the DNA fragment was observed after its digestion with Msp I and Sau 3A restriction enzymes, and strains were classified into three groups according to the length of the PCR products (Lecomte et al. 1997). The fragment length variability is due to the presence of different numbers of short- sequence DNA repeats (SSR) (Schnabel and Jones 1998). The number of SSR units has been recently used to try to type E. amylovora strains (Kim and Geider 1999; Jock et al. 2003; Ruppitsch et al. 2004), and, in case of a very low or very high number of SSR repeats the traceability of certain strains was possible (Ruppitsch et al. 2004). Recently, an E. amylovora strain isolated from Amelan- chier sp. (Maloideae) grown in Canada, showed a restric- ted pathogenicity upon artificial inoculation to apple, pear, raspberry and Amelanchier sp. plants. In fact, only the latter plant showed an extensive progression of the lesion along the twig (Giorgi and Scortichini 2005). In addition, also the assessment of the hrpN and dspA/E genes involved in the pathogenicity of E. amylovora poin- ted out some missense point mutations that could have implications for the particular pathogenicity of the strain (Giorgi and
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  • Spring '08
  • Devartanian
  • DNA, DNA sequencing, Restriction enzyme, Greece Greece Greece Greece Greece Greece Greece Greece Hungary Hungary Hungary Hungary Hungary Hungary Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy Italy USA USA Italy Italy Italy Italy, E. amylovora

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