Classification, nomenclature, and plant pathogenic bacteria - a clarification

ABSTRACT In a recent Letter to the Editor of Phytopathology, proposals were made for endorsement and for rejection of selected names of plant pathogenic Pseudomonas spp. and Xanthomonas spp. We believe that support for, and rejection of, several names was based on misconceptions concerning the Approved Lists of Bacterial Names and entails misinterpretations of several Rules of the International Code of Nomenclature of Bacteria. This letter aims to clarify those misconceptions and misinterpretations.     

The Relationship of Host Range, Physiology, and Genotype to Virulence on Cantaloupe in Pseudomonas syringae from Cantaloupe Blight Epidemics in France

ABSTRACT In 1993, a bacterial blight caused important losses of cantaloupe (Cucumis melo var. cantalupensis) in southwestern France and has now been reported in all cantaloupe-growing regions of France. The causal agent of this blight is Pseudomonas syringae, although on a worldwide basis this bacterium has not been a major pathogen of melon for over 50 years. To identify the pathovar of the cantaloupe pathogen, we employed biochemical tests, plasmid and chromosomal profiling, and host range studies for 23 strains from cantaloupe and 47 reference strains of 14 pathovars of P. syringae. Numerical analysis of 119 traits, serological typing, syringomycin production, and BOX-polymerase chain reaction profiles did not allow us to differentiate among pathovars related to P. syringae pv. syringae. Host range studies of cantaloupe and references strains on 18 plant species showed that virulence to sugar beet was a common feature of strains virulent on cantaloupe, but was not common to strains avirulent on cantaloupe. Virulence to other species of plants varied among strains, but the overall extent of the host range was proportional to aggressiveness to cantaloupe. We propose that the strains attacking cantaloupe in France be considered P. syringae pv. aptata and that adequate host range testing may reveal that this pathovar is the cause of cantaloupe blight reported in other parts of the world.     

Description of a new disease on Erythrina sp. in Martinique (French West Indies) and preliminary characterization of the causal agent as a novel Erwinia species

In spring 1995, symptoms of partial defoliation were observed on Erythrina indica var. fastigiata trees, commonly used as windbreaks in banana plantations in Martinique (French West Indies). Browning of the bark surface was consistently observed at the base of defoliated branches. Bacteria were isolated as nearly pure cultures from typical necrotic lesions on the bark. Results of Gram stain, staining of flagella and biochemical tests indicated that all isolates belonged to the Enterobacteriaceae, and to the genus Erwinia . Numerical analysis of the results of 48 biochemical properties showed that Erwinia isolates from diseased erythrina trees constituted a homogenous phenotypic cluster, and that, although weakly pectolytic, these isolates were closely related to pectolytic Erwinia species: E. carotovora ssp. betavasculorum and ssp. wasabiae , and E. cacticida . However, they could be clearly distinguished by different biochemical characteristics from type strains of known plant pathogenic species of Erwinia , Pantoea and Enterobacter . Under greenhouse conditions, the isolates were pathogenic to Erythrina indica var. fastigiata cuttings, but not to banana and pineapple, which are major crops in Martinique.     

Erwinia pyrifoliae, an Erwinia species different from Erwinia amylovora, causes a necrotic disease of Asian pear trees

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.     

Description of the Bacterium Causing Blight of Leek as Pseudomonas syringae pv. porri (pv. nov.)

ABSTRACT Forty bacterial strains isolated from leek blight (Allium porrum) in France and other countries were studied by conventional biochemical methods, serological reactions, numerical taxonomy, DNA-DNA hybridization, and ice nucleation activity, as well as by pathogenicity on leek and other host plants. They were compared with reference strains of Pseudomonas, mainly pathotype strains of P. syringae pathovars and strains of P. syringae pv. syringae isolated from various host plants including onions. Leek strains sorted with P. syringae species (sensu lato) by LOPAT tests (production of levan-sucrase, oxidase, pectinase, arginine dihydrolase, and hypersensitive reaction on tobacco). Leek strains were pathogenic to leek and produced symptoms identical to those observed in the field. They were the only strains in our study that could cause blight of leek. Thus, our results justify the creation of a new pathovar. Leek strains constituted a highly homogeneous DNA group and a discrete phenon by numerical taxonomy, and they belonged to O-serogroup POR. The name of P. syringae pv. porri is proposed for the bacterium causing leek blight. Criteria for routine identification are presented and taxonomic status is discussed.     

Lipid metabolism and cellular features of skeletal muscle and subcutaneous adipose tissue in pigs differing in IGF-II genotype

In pigs, a paternally (pat) imprinted mutation in the IGF-II gene is associated with increased muscle mass and decreased backfat thickness. The aim of this study was to determine whether this mutation influenced cellular, biochemical and metabolic features of skeletal muscle and adipose tissue. Muscle (trapezius) and subcutaneous adipose tissue (SCAT) were collected from pigs (106kg) carrying (Qpat, n=6) or not carrying (qpat, n=7) the mutation. Adipocytes were isolated from those tissues by collagenase treatment. Lipid content and activity of lipogenic enzymes were determined using standard assays. Gene expression levels were determined by real-time PCR. Levels of IGF-II mRNA were higher (P<0.01) in muscle of Qpat than in that of qpat pigs, but they did not differ significantly between the two groups in SCAT. Whereas levels of IGF-I mRNA in muscle were similar in both groups, they were higher (P<0.05) in SCAT of Qpat pigs than in that of qpat pigs. Muscle lipid content and intramuscular adipocyte diameters were not influenced significantly by the IGF-II genotype. In SCAT, the reduction of backfat thickness in Qpat pigs compared with qpat pigs was associated with lower (P<0.05) lipid content and smaller (P<0.05) adipocytes, with no significant genotype-effects on expressions and/or activities of lipogenic enzymes. In summary, our results suggest that the IGF-II mutation altered body composition in pigs by favoring myofiber hypertrophy and repressing adipose cell development in SCAT.     

Pseudomonas syringae pv. avii (pv. nov.), the Causal Agent of Bacterial Canker of Wild Cherries (Prunus avium) in France

Bacterial strains isolated from cankers of wild cherry trees (Prunus avium) in France were characterized using numerical taxonomy of biochemical tests, DNA–DNA hybridization, repeat sequence primed-PCR (rep-PCR) based on REP, ERIC and BOX sequences, heteroduplex mobility assay (HMA) of internal transcribed spacer (ITS) as well as pathogenicity on wild cherry trees and other species of Prunus. They were compared to reference strains of Pseudomonas syringae pathovars isolated from wild and sweet cherry and various host plants. Wild cherry strains were closely related to P. syringae (sensu lato) in LOPAT group Ia (+ - - - +). Wild cherry strains were pathogenic to wild cherry trees and produced symptoms similar to those observed in orchards. They were pathogenic also, but at a lesser extent, to sweet cherry trees (cv. Napoléon). The wild cherry strains were collected from five different areas in France and appeared to constitute a very homogeneous group. They showed an homogenous profile of a biochemical and physiological characteristics. They were closely related by DNA–DNA hybridization and belonged to genomospecies 3 `tomato'. Rep-PCR showed that wild cherry strains constitute a tight group distinct from P. s. pv. morsprunorum races 1 and 2 and from other P. syringae pathovars. HMA profiles indicated that the ITS of all wild cherry strains were identical but different from P. s. pv. persicae strains since the two heteroduplex bands with reduced mobility were generated by hybridization with the P. s. pv. persicae pathotype strain CFBP 1573. The 8 genomospecies of Gardan et al. (1999) have not been converted into formal species as they cannot be differentiated by biochemical tests. Therefore, the pathovar system within P. syringae was currently used. P. syringae pv. avii is proposed for this bacterium causing a wild cherry bacterial canker and strain CFBP 3846 (NCPPB 4290, ICMP 14479) is designated as the pathotype.     

Multiphasic Approach for the Identification of the Different Classification Levels Of Pseudomonas savastanoi Pv. Phaseolicola

The relationships between strains of Pseudomonas savastanoi pv. phaseolicola (P. sav. phaseolicola), P. syringae pv. tabaci (P. syr. tabaci) and P. syr. syringae which all cause disease on bean; the related species P. sav. glycinea and P. syr. actinidiae, and reference bacteria, were evaluated by studying the phenotypic and genetic diversity of a collection of 62 strains. All the P. sav. phaseolicola strains tested produced characteristic watersoaked lesions on bean pods. Other pathovars produced varying combinations of symptoms including necrotic lesions, with or without watersoaked centres and sunken tissue collapse of the lesion (P. syr. tabaci) and necrotic lesions with or without sunken collapse (P. syr. syringae). At the genomospecies level, all the strains of P. sav. phaseolicola, P. sav. glycinea and P. syr. tabaci, belonging to genomospecies 2, could be separated from P. syr. syringae strains (genomospecies 1) and P. syr. actinidiae strains (unknown genomospecies) by BOX-PCR and DNA/DNA hybridisation. To distinguish P. sav. phaseolicola, within genomospecies 2, from P. sav. glycinea and P. syr. tabaci, it was necessary to perform nutritional characterisations myo-inositol negative and p-hydroxy benzoate positive for P. sav. phaseolicola strains), PCR with specific primers designed from the tox region (positive for all of the P. sav. phaseolicola strains) and serotyping, as 71% of the P. sav. phaseolicola strains reacted as O-serogroup PHA1. Important intrapathovar variation was seen by genomic fingerprinting with REP and ERIC primers, as well as with RAPD primers (AE7 and AE10) and esterase profilings. While RAPD fingerprinting detected variability correlated with two race-associated evolutionary lines, REP, ERIC and esterase profiles revealed intrapathovar variation linked to some host origins, that separated the kudzu isolates, and the mungbean isolates, from the other P. sav. phaseolicola strains.     

Epiphytic Occurrence of Pseudomonas syringae pv. papulans (Rose) in France, Where Blister Spot has Never Been Seen

Pseudomonas syringae pv. papulans (PSP) the causal agent of blister spot, on the apple cultivar Mutsu in the USA, Canada and Italy, has not been described in France. A study on epiphytic populations of P. syringae isolated from French apple orchards revealed two isolates called KA54 and E121, whose biochemical characterisation showed high similarities with PSP strains. Identical symptoms were obtained with KA54, E121 and PSP strains, after vacuum inoculation of detached immature fruits of the cultivar Fuji, and young leaves of the cultivars Fuji, Mutsu, Gala and Golden Delicious. Koch's postulate was verified. These results indicate the presence of PSP in France. Differential characterisation criteria including serological, molecular and pathogenicity tests are proposed.