Molecular Characterization of Copper Resistance Genes from Xanthomonas citri subsp.
            <i>citri</i>
            and Xanthomonas alfalfae subsp. citrumelonis

Behlau, Franklin; Canteros, Blanca Isabel; Minsavage, Gerald V.; Jones, Jeffrey B.; Graham, James H.

doi:10.1128/aem.03043-10

Cited by 139 publications

(186 citation statements)

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“…actinidiae in New Zealand (2). Continual use of copper can lead to a toxic buildup of copper residues within the environment (27,28), and bacterial resistance to copper has been documented (29)(30)(31)). …”

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Identification of Bacteriophages for Biocontrol of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae

Frampton

Taylor

Moreno

et al. 2014

Appl Environ Microbiol

107

105

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c Pseudomonas syringae pv. actinidiae is a reemerging pathogen which causes bacterial canker of kiwifruit (Actinidia sp.). Since 2008, a global outbreak of P. syringae pv. actinidiae has occurred, and in 2010 this pathogen was detected in New Zealand. The economic impact and the development of resistance in P. syringae pv. actinidiae and other pathovars against antibiotics and copper sprays have led to a search for alternative management strategies. We isolated 275 phages, 258 of which were active against P. syringae pv. actinidiae. Extensive host range testing on P. syringae pv. actinidiae, other pseudomonads, and bacteria isolated from kiwifruit orchards showed that most phages have a narrow host range. Twenty-four were analyzed by electron microscopy, pulse-field gel electrophoresis, and restriction digestion. Their suitability for biocontrol was tested by assessing stability and the absence of lysogeny and transduction. A detailed host range was performed, phage-resistant bacteria were isolated, and resistance to other phages was examined. The phages belonged to the Caudovirales and were analyzed based on morphology and genome size, which showed them to be representatives of Myoviridae, Podoviridae, and Siphoviridae. Twenty-one Myoviridae members have similar morphologies and genome sizes yet differ in restriction patterns, host range, and resistance, indicating a closely related group. Nine of these Myoviridae members were sequenced, and each was unique. The most closely related sequenced phages were a group infecting Pseudomonas aeruginosa and characterized by phages JG004 and PAK_P1. In summary, this study reports the isolation and characterization of P. syringae pv. actinidiae phages and provides a framework for the intelligent formulation of phage biocontrol agents against kiwifruit bacterial canker.

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Identification of Bacteriophages for Biocontrol of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae

Frampton

Taylor

Moreno

et al. 2014

Appl Environ Microbiol

107

105

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“…The copABCD genes, which were first described in a plasmid of P. syringae pv. tomato PT23 (8), is an important determinant of copper resistance in P. syringae and other phytopathogenic bacteria (6,8,9,(12)(13)(14). However, additional studies have shown that P. syringae may harbor other variants of Cu r determinants (3, 7, 15).…”

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Recruitment and Rearrangement of Three Different Genetic Determinants into a Conjugative Plasmid Increase Copper Resistance in Pseudomonas syringae

Gutiérrez‐Barranquero

Vicente

Carrión

et al. 2013

Appl Environ Microbiol

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We describe the genetic organization of a copper-resistant plasmid containing copG and cusCBA genes in the plant pathogen Pseudomonas syringae. Chromosomal variants of czcCBA and a plasmid variant of cusCBA were present in different P. syringae pathovar strains. Transformation of the copper-sensitive Pseudomonas syringae pv. syringae FF5 strain with copG or cusCBA conferred copper resistance, and quantitative real-time PCR (qRT-PCR) experiments confirmed their induction by copper. Pseudomonas syringae is a common foliar bacterium and causal agent of plant diseases in many different hosts worldwide, affecting both woody trees and herbaceous plants (1).Copper bactericides have been widely used for decades to control bacterial infections in crop plants (2). However, the extensive use of copper bactericides can lead to many problems, among them the reduction of efficacy of this antimicrobial agent due to the selection of copper-resistant (Cu r ) strains (3, 4). Copper resistance determinants have been detected and described in several pathovars of P. syringae (3, 5-7) and are frequently encoded within native plasmids (3,8,9). These native Cu r plasmids contribute to the dissemination of resistance genes among P. syringae strains from different pathovars (5, 10, 11). The copABCD genes, which were first described in a plasmid of P. syringae pv. tomato PT23 (8), is an important determinant of copper resistance in P. syringae and other phytopathogenic bacteria (6,8,9,(12)(13)(14). However, additional studies have shown that P. syringae may harbor other variants of Cu r determinants (3, 7, 15). The efflux system czcCBA and the analogous system cusCBA are probably the best-characterized members of the heavy metal efflux (HME)-RND (resistance-nodulation-cell division) family. The czcCBA system functions in the detoxification of cadmiun, zinc, and cobalt (16,17), and the cusCBA system works in detoxifying monovalent cations, such as silver and copper (16,18). These efflux systems have been widely studied in Cupriavidus metallidurans (formerly Ralstonia metallidurans or Alcaligenes eutrophus) (16,(19)(20)(21)(22), Escherichia coli (18, 23), and Pseudomonas putida KT2440 (24-26), but no further studies have been carried out for the plant pathogen P. syringae pv. syringae.In this work, we detected that the majority of the cusCBA genes from different Pseudomonas species could be wrongly annotated as czcCBA genes, based on the analysis of the conserved motifs of the RND transporter domains (27). This inaccurate annotation creates problems in the database regarding czcCBA or cusCBA genes in the Pseudomonas genus.Our goal was to identify and characterize plasmid-encoded Cu r genes in the P. syringae pv. syringae UMAF0081 strain. The bacterial strains used, and sizes of native plasmids harbored by the strains, are listed in Table 1. Each native plasmid examined is a member of the pPT23A plasmid family, a group of related plasmids broadly distributed within P. syringae and the related phytopathogen Pseudomonas savastanoi (32). These...

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“…Under these conditions, frequent sprays of copper‐based bactericides are required, increasing production costs as well as environmental contamination, without necessarily providing adequate protection. Moreover, frequent sprays with relatively high concentrations of Cu might cause phytotoxicity and often lead to emergence of Cu‐resistant strains (Hancock, 2001; Behlau et al ., 2011; Worthington et al ., 2012). Therefore, there is an urgent need to develop novel technologies to manage and prevent bacterial plant diseases and prevent food loss.…”

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confidence: 99%

Random peptide mixtures as new crop protection agents

Topman

Tamir‐Ariel

Bochnic-Tamir

et al. 2018

Microbial Biotechnology

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SummaryMany types of crops are severely affected by at least one important bacterial disease. Chemical control of bacterial plant diseases in the field vastly relies on copper‐based bactericides, yet with limited efficacy. In this study, we explored the potential of two random peptide mixture (RPM) models as novel crop protection agents. These unique peptide mixtures consist of random combination of l‐phenylalanine and l‐ or d‐lysine (FK‐20 and FdK‐20, respectively) along the 20 mer chain length of the peptides. Both RPMs displayed powerful bacteriostatic and bactericidal activities towards strains belonging to several plant pathogenic bacterial genera, for example, Xanthomonas, Clavibacter and Pseudomonas. In planta studies in the glasshouse revealed that RPMs significantly reduced disease severity of tomato and kohlrabi plants infected with Xanthomonas perforans and Xanthomonas campestris pv. campestris respectively. Moreover, RPM effects on reduction in disease severity were similar to those exerted by the commercial copper‐based bactericide Kocide 2000 that was applied at a 12‐fold higher concentration of the active compound relative to the RPM treatments. Importantly, the two tested RPM compounds had no toxic effect on survival of bees and Caco‐2 mammalian cells. This study demonstrates the potential of these innovative RPMs to serve as crop protection agents against crop diseases caused by phytopathogenic bacteria.

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Molecular Characterization of Copper Resistance Genes from Xanthomonas citri subsp. citri and Xanthomonas alfalfae subsp. citrumelonis

Cited by 139 publications

References 39 publications

Identification of Bacteriophages for Biocontrol of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae

Identification of Bacteriophages for Biocontrol of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae

Recruitment and Rearrangement of Three Different Genetic Determinants into a Conjugative Plasmid Increase Copper Resistance in Pseudomonas syringae

Random peptide mixtures as new crop protection agents

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