A novel pathogen from Asian pears (Pyruspyrifolia Nakai) was analysed by sequencing the 16s rDNA and the adjacent intergenic region, and the data were compared t o related Enterobacteriaceae. The 165 rDNA of the Asian pear pathogen was almost identical with the sequence of Erwinia amylovora, in contrast t o the 165-235 rRNA intergenic transcribed spacer region of both species. A dendrogram was deduced from determined sequences of the spacer regions including those of several related species such as Erwinia amylovora, Enterobacter pyrinus, Pantoea stewartii subsp. stewartii and Escherichia coli. Dendrograms derived from 121 biochemical characteristics including Biotype 100 data placed the Asian pear pathogen close to Erwinia amylovora and more distantly t o other members of the species Erwinia and t o the species Pantoea and Enterobacter. Another DNA relatedness study was performed by DNA hybridizations and estimation of AT, , , values. The Asian pear strains constituted a tight DNA hybridization group (89-100%) and were barely related t o strains of Erwinia amylovora (40-50%) with a AT, , , in the range of 5-2-6.8. The G+C content of DNA from the novel pathogen is 52 mol%. Therefore, it is proposed that strains isolated from Asian pears constitute a new species and the name Erwinia pyrifoliae is suggested; the type strain is strain E~1 6 / 9 6~ (= CFBP 4172T = DSM 121633.
Bacteria from necrotic branches of Asian pear trees (Pyrus pyrifolia) in Korea were consistently isolated as white colonies on nutrient agar and formed mucoid, slightly yellow colonies on a minimal medium with copper sulphate. Isolates with this colony morphology were studied in a series of microbiological, molecular and pathological tests. Most isolates allowed the verification of Koch's postulate on P. pyrifolia seedlings and on slices from immature pear (Pyrus communis) fruits and were also positive in hypersensitivity tests on tobacco leaves. They showed characteristics common to species in the genus Erwinia, but were different from Erwinia amylovora, the agent of fire blight. A relationship between the novel pathogen and E. amylovora was found in microbiological and serological tests. Both organisms had similar but not identical protein patterns in 2‐D gel electrophoresis, and in growth morphology the new pathogen produced colonies on MM2 Cu medium that were mucoid and slightly yellow, compared with the clearly yellow colonies of E. amylovora. No similarity was found in the plasmid profiles, and consequently no PCR signal was obtained with primers from the E. amylovora plasmid pEA29. REP‐PCR also produced bands differing for the two organisms.
The recently described pathogen Erwinia pyrifoliae, isolated from Nashi pear fruit trees in Korea, resembles the fire blight pathogen Erwinia amylovora in some of its properties. The two pathogens were classified into different species by DNA hybridization kinetics and microbiological criteria. From the nucleotide sequences of the 16S rRNA and the internal transcribed spacer (ITS) region as well as extracellular polysaccharide (EPS)-encoding genes, polymerase chain reaction (PCR) primers were designed that specifically detect E. pyrifoliae but not the fire blight pathogen Erwinia amylovora, and these primers were also applied to identify E. pyrifoliae in necrotic plant material. The genomes of several strains were digested with the restriction enzyme SpeI, and the DNA fragments were analyzed by pulsed-field gel electrophoresis (PFGE). Three groups of patterns could be distinguished for the isolated E. pyrifoliae strains, all different from various E. amylovora strains, which produce a relatively homogeneous PFGE pattern after SpeI digests. Typical fire blight host plants were assayed in a growth chamber or an experimental field for their susceptibility to E. pyrifoliae. A strong preference was found for pear varieties, whereas apple, cotoneaster, hawthorn, or raspberry rarely produced necrotic symptoms. E. pyrifoliae was readily detected in samples from pear orchards in South Korea during 1995 to 1998; however, the Asian pear pathogen was not recovered in necrotic plant tissue from 1999 and 2000.
Errata Validation of publication of new names and new combinationspreviously effectively published outside the IJSB Owing to a printing error the final pagt of the paper was blank. The omitted text is reproduced below.
A crystal ~-endotoxin gene of Bacillus thuringiensis subsp, tenebrionis (B.t.t.) encoding a coleopteran insect-specific toxin was used to construct a chimeric gene which expressed the toxin in plant cells. Via an Agrobacterium tumefaciens binary vector system, the toxin gene was transferred into tomato cells. From leaf disks recombinant plants were regenerated. Hybridization experiments demonstrated that these plants synthesized toxin-specific mRNA of the expected size. Transgenic tomato plants with the chimeric B.t.t. toxin gene contained a 74 kDa protein which cross-reacted with toxin antibodies. The expression caused a significant insecticidal activity of the transgenic tomato plants against Colorado potato beetle larvae.
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