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.     

植物病原细菌的VBNC状态研究进展

 有活力但不可培养状态（Viable But Non-Culturable,VBNC）被认为是细菌对不良环境条件的一种抗逆反应,包括5种植物病原细菌在内的数十种细菌已报道具有VBNC状态。这种特定的生理状态已经导致细菌学研究中一些方法的探索和改进,对基于分离培养的植物病原细菌学研究产生了重大影响。本文对已经报道VBNC状态的植物病原细菌进行了综述,介绍了VBNC状态细菌的诱导因素、恢复条件、致病性、检测方法等方面的研究进展,对未来植物病原细菌的VBNC研究提出了展望。     

Analysis of the toxicity of purothionins and hordothionins for plant pathogenic bacteria

Purothionins (PTHs) and hordothionins (HTHs) were purified by cation-exchange chromatography from petroleum-ether extracts of wheat and barley flour respectively. The HTHs could be separated into two fractions, HTH-1 and HTH-2. Radial diffusion assays and micro-plate broth dilution assays with a number of plant pathogenic bacteria showed that these proteins were toxic forClavibacter michiganensis subsp.michiganensis, the causal agent of bacterial canker on tomato,C. m. subsp.sepedonicus, the causal agent of ring rot on potato, andXanthomonas campestris pv.vesicatoria, the causal agent of a spot disease on tomato and pepper. Only minor differences in toxicity between PTHs and HTHs, and between HTH-1 and HTH-2, were detected. Minor differences in toxicity of these thionins were also detected for different strains of these bacteria. The use of these plant proteins for engineering bacterial disease resistance into solanaceous crops will be discussed.     

Hypersensitive response suppression by type III effectors of plant pathogenic bacteria

The suppressor activity of four representative avirulence (avr) genes from the Pseudomonas syringae group against elicitors of a general hypersensitive response (HR) was examined in tobacco leaves inoculated with double transformants of Pseudomonas fluorescens containing both a cosmid plasmid (pHIR11) carrying the hrp gene cluster and a plasmid carrying each avr gene. The double transformants Pf (pHIR11) containing avrB, avrRps4, or avrPto failed to induce an HR, but that carrying avrRpt2 did induce an HR as Pf (pHIR11 + empty vector) did. Thus, some Avr proteins seem to have suppressor activity against a general HR and should promote aggressiveness of the pathogens.     

PM 7/101 (1): ELISA tests for plant pathogenic bacteria

Specific scope

Enzyme Linked Immuno Sorbent Assay (ELISA) tests for bacteria may be used for screening of large numbers of samples as an alternative to immunofluorescence (IF) in certain cases (see PM 7/97 Indirect Immunofluorescence test for plant pathogenic bacteria). They can also be used as part of the identification of pure cultures. This standard describes how to perform an ELISA test for detection and/or identification in bacterial diagnostics using: (i) Indirect; (ii) Double antibody sandwich‐DAS; (iii) Double antibody sandwich indirect‐DASI also named triple antibody sandwich‐TAS; and (iv) Direct tissue‐print, squash or colony‐dot.

Specific approval and amendment

Approved as an EPPO Standard in 2010–09     

Long-term maintenance of nonculturable apple-proliferation phytoplasmas in their micropropagated natural host plant

Phytoplasmas associated with apple proliferation (AP) disease of apple trees have been maintained in their micropropagated natural host plant Malus pumila since 1985. Different isolates of these nonculturable plant pathogens could thus be studied in vitro . Amplification of a pathogen-specific DNA fragment by polymerase chain reaction (PCR) confirmed the presence of AP phytoplasmas in the diseased plants even after 10 years of in-vitro propagation. Restriction fragment length polymorphism analysis of the amplified chromosomal DNA fragments revealed no genetic difference between the AP phytoplasma isolates. Growth parameters, symptom expression and phytoplasma concentration were examined to compare the in-vitro behaviour of four different AP phytoplasma isolates and to compare different subculture conditions. A comparison of these data obtained after 2 or 8 years of micropropagation revealed no essential differences. Eight years after culture initiation, diseased shoots still exhibited typical symptoms like witches' broom, small leaves with large stipules and stunted growth. The use of phytoplasma-diseased micropropagated plants to establish a 'type culture collection' of these otherwise nonculturable plant pathogens is discussed.     

PM 7/97 (1): Indirect immunofluorescence test for plant pathogenic bacteria

Specific scope

This standard describes how to perform an indirect immunofluorescence test (IF) for plant pathogenic bacteria. ¹ 1 Use of brand names of chemicals or equipments in these EPPO standards implies no approval of them to the exclusion of others that may also be suitable.

Specific approval and amendment

Approved as an EPPO Standard in 2009–09.

     

八株番茄内生细菌对12种病原真菌拮抗活性的测定

正内生细菌普遍存在于高等植物中,已发现的种类超过80个属(Lodewyckx et al.,2002),可通过产生抗生素、营养竞争和生态位排斥、触发植物的诱导性系统抗性等机制有效减轻或控制植物病害,从而减轻化学杀菌剂使用带来的残留、环境污染、病原菌抗药性及生态平衡破坏等问题。番茄在全世界种植历史悠久,在与病原菌长期的协同进化过程中,必然     

A note on the use of Merckoquant test strips for detection of nitrate reduction in plant pathogenic bacteria

Merckoquant nitrate test strips gave reactions identical to standard sulphanilamide N-I-naphthylethyl-enediamine reagents for determining nitrate reduction in several bacterial genera. The strips obviate the need for fresh reagents and are useful as an off-the-shelf test.     

Evidence for the physical integrity of flavescence dorée phytoplasmas purified by immunoaffinity from infected plants or leafhoppers and the plant pathogenicity of phytoplasmas from leafhoppers

Phytoplasmas were extracted from flavescence dorée-infected broadbean ( Vicia faba ) using a vacuole isolation medium, and were immunoaffinity purified from infected leafhoppers. Purified phytoplasmas from both host sources were immunolabelled and observed under the electron microscope. The infectivity of the purified phytoplasmas from leafhoppers was checked by injecting healthy leafhoppers which were then allowed to feed on healthy V. faba seedlings. The appearance of typical symptoms in these plants, and the positive results obtained in ELISA with extracts of some of the injected leafhoppers and with symptomatic V. faba , indicated that the purified phytoplasmas had retained their infectivity and had multiplied in the injected leafhoppers which had become infective. These results support a previous report that phytoplasmas purified by immunoaffinity chromatography are well preserved.     

Activity of the ilicicolins against plant pathogenic fungi

Ilicicolins D, E, F, dechloroilicicolin D, ascofuranone and arthrichitin were isolated from the fermentation broth of Nectria sp (HIL Y 90 3333). The ilicicolins showed good fungicidal activity in planta.     

Taxonomic study of plant pathogenic oomycetes

Journal of General Plant Pathology -     

The potential for using cyanobacteria (blue-green algae) and algae in the biological control of plant pathogenic bacteria and fungi

Cyanobacteria (blue-green algae) and eukaryote algae occur in freshwater, marine, and terrestrial (soil) habitats. In fact, these microorganisms comprise most of the world's biomass. Although the cyanobacteria are mostly photoautotrophic, some are facultative heterotrophs, capable of growing on certain substrates in darkness. Also, some are non-phototrophic and hence, are obligate heterotrophs. A number of cyanobacteria and eukaryote algae, particularly macroalgae, produce various, biologically active compounds. These include antibiotics which in laboratory tests inhibited bacteria and fungi that incite diseases of humans. In addition, the following fungi which are of interest to plant pathologists, were inhibitedin vitro by substances produced by various cyanobacteria: The saprophytesChaetomium globosum, Cunninghamella blakesleeana, andAspergillus oryzae and the plant pathogensRhizoctonia solani andSclerotinia sclerotiorum. Extracts from seaweeds (macroalgae) sprayed on plants have been reported to reduce the incidence ofBotrytis cinerea (gray mold) on strawberries,Erysiphe polygoni (powdery mildew) on turnips, and damping-off of tomato seedlings. Because many cyanobacteria and algae produce a large number of antibacterial and antifungal materials, are almost never a threat to the environment, and many can be grown in quantity in mass culture, they are suitable candidates for exploitation as biocontrol agents of plant pathogenic bacteria and fungi. Much additional work remains to be done however, to thoroughly evaluate cyanobacteria and algae and their products for this role.     

Reaction of saprophytic bacteria from potato peel extracts and plant pathogenic bacteria in ELISA with antisera to Erwinia chrysanthemi (serogroup O1Ha)

The specificity of two antisera raised to whole cells ofErwinia chrysanthemi (Ech), serogroup O₁H_a, was studied in double antibody sandwich (DAS-) ELISA with 100 strains of different plant pathogenic bacteria (PPB), including 39 Ech strains, and of one of these antisera with 900 saprophytic bacteria isolated from extracts of potato peelings of Dutch seed potatoes grown in several production areas.All tested European Ech strains from potato reacted positively while no reactions were observed with any of the other plant pathogenic bacterial species. Two saprophytes (A254 and A256), both identified as pectinolyticPseudomonas fluorescens species, cross-reacted strongly with polyclonal antibodies against Ech. Non-specific reactions were found in DAS-ELISA with 16 saprophytes. The detection limits for the individual saprophytes varied between c. 10⁵ and 10⁹ cells.ml^–1. The non-specific reactions were also found with monoclonal antibodies (mca 2A4) against a proteinase K resistent epitope of Ech and with antisera against other plant pathogens including an antiserum against potato virus Y^N. The non-specific reactions were observed in DAS-ELISA, but not in Ouchterlony double diffusion or immunofluorescence colonystaining, whereas A254 and A256 reacted in all tests, but only with antibodies against Ech. When in making dilution series potato peel extracts were used instead of phosphate buffered saline with 0.1% Tween 20, the 14 non-specifically reacting saprophytes only reacted at concentrations of 10⁹ cells.ml^–1 or higher. Only one of these 14 saprophytes was able to multiply on injured potato tuber tissue.In contrast to most saprophytic strains, the saprophytes A254 and A256 reacted strongly in ELISA in dilutions series made with potato peel extracts. A256 was able to grow on potato tuber tissue but only under low oxygen conditions; A254 did not grow at all on potato tissue.Defatted milk powder or bovine serum albumin added to the dilution buffer for the enzymeconjugated antibodies, drastically reduced the non-specific reactions, but not the reactions with A254 and A256.To reduce the cross-reaction with A254, an Ech antiserum was absorbed with A254. This resulted in a substantial drop in antibody reaction with the homologous antigen in Ouchterlony double diffusion.     

蝴蝶兰细菌性褐斑病病原菌初步鉴定

对蝴蝶兰褐斑病病原细菌进行了分离、致病性测定、形态观察、染色反应、培养性状观察和生理生化特性测定。结果表明,该菌与黄单胞菌属野油菜黄单胞菌种性状、特征完全相同。因此,可以初步断定,引起蝴蝶兰褐斑病的病原菌是黄单胞菌属野油菜黄单胞菌[Xamthomonas campestris(Pammel)Dowson.]。