1. Isolation and characterization of five Erwinia amylovora bacteriophages and assessment of phage resistance in strains of Erwinia amylovora
E L Schnabel, A L Jones Appl Environ Microbiol. 2001 Jan;67(1):59-64. doi: 10.1128/AEM.67.1.59-64.2001.
Phages able to infect the fire blight pathogen Erwinia amylovora were isolated from apple, pear, and raspberry tissues and from soil samples collected at sites displaying fire blight symptoms. Among a collection of 50 phage isolates, 5 distinct phages, including relatives of the previously described phages phiEa1 and phiEa7 and 3 novel phages named phiEa100, phiEa125, and phiEa116C, were identified based on differences in genome size and restriction fragment pattern. phiEa1, the phage distributed most widely, had an approximately 46-kb genome which exhibited some restriction site variability between isolates. Phages phiEa100, phiEa7, and phiEa125 each had genomes of approximately 35 kb and could be distinguished by their EcoRI restriction fragment patterns. phiEa116C contained an approximately 75-kb genome. phiEa1, phiEa7, phiEa100, phiEa125, and phiEa116C were able to infect 39, 36, 16, 20, and 40, respectively, of 40 E. amylovora strains isolated from apple orchards in Michigan and 8, 12, 10, 10, and 12, respectively, of 12 E. amylovora strains isolated from raspberry fields (Rubus spp.) in Michigan. Only 22 of 52 strains were sensitive to all five phages, and 23 strains exhibited resistance to more than one phage. phiEa116C was more effective than the other phages at lysing E. amylovora strain Ea110 in liquid culture, reducing the final titer of Ea110 by >95% when added at a ratio of 1 PFU per 10 CFU and by 58 to 90% at 1 PFU per 10(5) CFU.
2. Bacteriocin Serratine-P as a biological tool in the control of fire blight Erwinia amylovora
H Schoofs, K Vandebroek, A Pierrard, P Thonart, P Lepoivre, T Beaudry, T Deckers Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2002;67(2):361-8.
Fire blight, caused by the bacterium Erwinia amylovora (Burill Winslow et al.), is the most important bacterial disease in European pear growing. It can cause a lot of damage in some countries on apple and on pear trees in orchards and also in the fruit tree nurseries. In Belgium, the disease is present since 1972. Control of fire blight in Belgian fruit orchards is made on a broad basis of measurements in and around the fruit trees. The use of an antibiotic is allowed for application only during the primary blossom period under strict controlled regulations. The use of antobiotics in agriculture is strongly discussed on the European level today and will probably disappear in the near future. Therefore, the research on fire blight control concentrates on the possibilities of biological control with antagonistic bacteria such as Pantoea agglomerans (Erwinia herbicola), Bacillus subtilis or Pseudomonas syringae strain A 506. The use of Serratine-P, a phage tail-like bacteriocin, produced by Serratia plymiticum, shows an interesting antibacterial activity against Erwinia amylovora. Its mode of action consists in the perforation of the cytoplasmic membrane of the target cell, inducing perturbations in cellular exchanges and a final lysis of the bacterial cell. In this paper some trials are discussed on the use of Serratine-P at different doses and on different infection types on pear trees. The results indicate interesting protection possibilities on blossom- and fruit infections.
3. Molecular comparison of pathogenic bacteria from pear trees in Japan and the fire blight pathogen Erwinia amylovora
W S Kim, M Hildebrand, S Jock, K Geider Microbiology (Reading). 2001 Nov;147(Pt 11):2951-9. doi: 10.1099/00221287-147-11-2951.
Several strains of the genus Erwinia, which were isolated in Japan from pear trees with necrotic symptoms that resembled fire blight, and tentatively identified as Erwinia amylovora, were reinvestigated for their relationship to the fire blight pathogen. These isolates produced ooze on slices of immature pears and were mucoid on MM2Cu agar plates, but did not synthesize levan and did not give the expected PCR signals with several primer pairs specific for Erwinia amylovora. The isolates tested positive with PCR primers designed to detect the novel pear pathogen Erwinia pyrifoliae, which was isolated from Nashi pear trees in South Korea. The nucleotide sequence analysis of a DNA fragment preceding the gene cluster for exopolysaccharide synthesis revealed a closer relationship to Erwinia pyrifoliae than to Erwinia amylovora. Plasmid profiles, protein patterns and genomic DNA analysed by PFGE after XbaI and SpeI digestion were different than Erwinia amylovora. Experiments with strains of Erwinia amylovora isolated from raspberry (Rubus sp.), Erwinia mallotivora and Enterobacter pyrinus also did not reveal a relationship between these bacteria and the Japanese Erwinia strains. The latter are not identical to Erwinia pyrifoliae, but possess many similar features to this pathogen that causes Asian pear blight. It is concluded that pathogenic bacteria isolated in Japan from pear trees with symptoms resembling fire blight are possibly different from Erwinia amylovora.
