An Investigation on Strains of Clavibacter michiganensis subsp. michiganensis in North and North West of Iran

Abstract

Tomato bacterial canker disease was first reported from Urmiyeh in West Azerbaijan province in Iran. The disease causes lesion (canker), wilting and dryness of infected plants, leaf and fruit spots and the decline of the whole plant. Out of 102 isolates obtained from the fields in the major tomato producing areas of understudy regions, 98 were found Gram positive, yellow-pigmented isolates, identified as Clavibacter michiganensis subsp. michiganensis based on the morphological and biochemical characteristics described in previous studies. Among these strains, 64 were virulent and 34 showed poor virulence. A strain of Cmm (NCPPB382) was used as a check (standard) in all steps of this study. DNA fingerprinting with repetitive-sequence-based PCR (rep-PCR) (BOX primer) carried out among 11 representative strains (eight strains from West Azerbaijan, two from Golestan and one as standard). The most virulent strain was chosen as representative in each location. Dendrograms were prepared using NTSYSpc version 2⁄o2e software, unweighted pair group with arithmetic average method and simple matching similarity coefficient. According to the site of cut-off line, three groups (clusters) with 82⁄5% similarity and six groups with 55% similarity were separated based on biochemical and SDS-PAGE data, and rep-PCR reactions respectively. Low similarity among groups (55%) can be explained as high genetic diversity among the strains. One strain of west Azerbaijan and the strains of Golestan, clustered in the same group suggesting that they may have been originated from a common source. Other strains of west Azerbaijan were clustered into different groups including II, III, IV, V and VI, suggesting the possibility of occurrence of different populations in a geographical region.www.blackwell-synergy.com

Polymorphism analysis of housekeeping genes for identification and differentiation of Clavibacter michiganensis subspecies

“…BOX PCR revealed four distinct groups of Cmm strains from four regions in the USA (Louws et al 1998) and from four regions in Japan (Kawaguchi et al 2010). A similar analysis of Cmm strains from Iran produced six fingerprint patterns (Nazari et al 2007). When Cmm strains from the Canary Islands were characterized and compared with strains from seven other countries using BOX-PCR, 12 genotypes were detected, while the use of RAPD techniques increased the number of different profiles to 18, with five AFLP types (De León et al 2009).…”

“…The exception is subspecies Cmm: strains from different countries displayed significant diversity and could be divided into several clusters. Strains were successfully distinguished from other subspecies by analysis of repetitive sequences like rep, BOX, or ERIC (Louws et al 1998;Nazari et al 2007;Kleitman et al 2008;De León et al 2009;Kawaguchi et al 2010); by random DNA amplification techniques such as RAPD (Pastrik and Rainey 1999;De León et al 2009); by pulsed field gel electrophoresis (PFGE) (Kleitman et al 2008), by PCR-RFLP (De León et al 2009), and by amplified fragment length polymorphisms (AFLP), (De León et al 2009). …”

Characterization of phenotypic variants of Clavibacter michiganensis subsp. michiganensis isolated from Capsicum annuum

“…Genetic variations between populations of Cmm have consistently been reported from various countries, and these results have been used to estimate their origins (De Leon et al 2009;Ignatov et al 2004;Kaneshiro et al 2006;Nazari et al 2007). Pepper isolates could have long existed in pepper fields undiscovered because they were not usually a major disease problem.…”

“…It was also reported that Cmm can be subdivided into four or six distinct groups on basis of the diversity in repetitive sequence-based PCR (rep-PCR) fingerprint patterns (Louws et al 1998;Nazari et al 2007;Kleitman et al 2008). More recently, genetic diversity related to geographical area was observed within the Cmm population (De Leon et al 2009;Ignatov et al 2004;Kaneshiro et al 2006;Nazari et al 2007). Among the five Cm subspecies, the relatively high genetic diversity of Cmm strains was confirmed by PCR-RFLP and PFGE analysis (Kleitman et al 2008;Waleron et al 2011).…”

Diversity of Plant Associated Actinobacteria

“…rep-PCR, provides genomic fingerprint of chromosome structure (Rademaker and De Brujin 1997). Thus, it has been used for differentiation at species and subspecies level (Sadowsky et al 1996;Clark et al 1998;Louws et al 1998;Jeong and Myrold 1999;Smith et al 2001;Lanoot et al 2004;Nazari et al 2007;de Leon et al 2009;Trujillo et al 2010). This technique is simple, rapid and inexpensive and allows grouping closely related organisms according to their fingerprints and separating them from distant groups.…”