Some phytotoxic glycopeptides from Ceratocystis ulmi, the Dutch Elm Disease pathogen

Strobel, Gary A.; Alfen, Neal Van; Hapner, Kenneth D.; McNeil, Michael; Albersheim, Peter

doi:10.1016/0304-4165(78)90252-0

Cited by 55 publications

(29 citation statements)

References 22 publications

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“…A typical preparation contained 77 to 80%7o neutral hexose on a weight basis. This value is somewhat lower than the 85% value reported earlier (19). Gas chromatographic analyses of the component sugars (as their alditol acetates) usually revealed only mannose and rhamnose (data not shown).…”

Section: Methodscontrasting

confidence: 50%

“…Dialyzed culture filtrates were routinely fractionated as previously described (19). The filtrate was subjected to Dowex 50 (H+) ion exchange chromatography and acetone precipitation prior to fractionation with Con A-Sepharose affinity chromatography.…”

Section: Methodsmentioning

confidence: 99%

“…Fluorescamine labeling of protein was accomplished by the procedure of Bohlen et al (5). C. ulmi peptidorhamnomannan was purified as previously described (19). Acid hydrolysis of toxin and saccharide fragments was carried out in a 2 N trifluoroacetic acid at 120 C for 90 min (1).…”

Section: Methodsmentioning

confidence: 99%

“…For instance, Corynebacterium insidiosum, the bacterial wilt pathogen of alfalfa, produces a large fucose-rich peptidopolysaccharide which has also been isolated from diseased plants (18). A previous report from this laboratory described the isolation and partial characterization of a peptidorhamnomannan toxin prepared from culture filtrates of Ceratocystis ulmi, the Dutch Elm disease pathogen (19). This polymer also induces wilt symptoms in stem cuttings.…”

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confidence: 99%

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Structural and Immunochemical Studies on the Phytotoxic Peptidorhamnomannan of Ceratocystis ulmi

Nordin¹,

Strobel²

1981

Plant Physiol.

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A wilt-inducing peptidorhamnomannan produced by Ceratocystis ubni, the causative agent In Dutch Elm disease, has been subjected to additional chemical and physical characterization. Gel filtration, reductive 13 elimination, hydrofluoric acid deglycosylation, and ultracentrifugation experiments provide evidence that the wilt-inducing polymer is Vascular wilt diseases in plants result from impaired fluid transport and are caused by both bacteria and fungi (8, 11,21). In several cases, the causative organism produces in culture, large carbohydrate-containing macromolecules which are suggested as potentiators of the impaired transport, because they induce wilt symptoms in stem cuttings. For instance, Corynebacterium insidiosum, the bacterial wilt pathogen of alfalfa, produces a large fucose-rich peptidopolysaccharide which has also been isolated from diseased plants (18). A previous report from this laboratory described the isolation and partial characterization of a peptidorhamnomannan toxin prepared from culture filtrates of Ceratocystis ulmi, the Dutch Elm disease pathogen (19). This polymer also induces wilt symptoms in stem cuttings. Purification of the toxin was clearly correlated on a specific activity basis with wiltinducing capability (21). The present report extends previous information on the structure and describes the production of antibodies directed against both C. ulmi and the peptidorhamno- Supelcoport (80-100 mesh) was purchased from Supelco, Inc. D-[U-14CJGlucose was obtained from New England Nuclear. BioGel polyacrylamide gels were from Bio-Rad Laboratories. Fluorescamine was obtained from Hoffman-LaRoche and scintillation fluid (Biofluor) from Packard Instruments. Potato dextrose agar was purchased from Difco, Inc. METHODSCarbohydrate was determined by the phenolsulfuric acid assay (9) with a standard composed of mannose and rhamnose in a 2:1 molar ratio and protein by the method of Lowry et al. (14) using a BSA standard or by measurement of A of peptide bonds at 208 nm. The amino acid analysis was conducted by AAA Laboratories, Seattle, WA. Sedimentation equilibrium centrifugation for molecular weight determinations was conducted according to Yphantis (24) using a double sector cell in a Beckman model E ultracentrifuge equipped with interference optics.Reductive f8 elimination of the peptidorhamnomannan was accomplished with alkaline borohydride (6). Twenty mg toxin was incubated in 4 ml 50 mm NaOH, 1 M NaBH4 for 16 h at 55 C. The reaction mixture was neutralized with Dowex 50 (H+) filtered and evaporated to dryness under reduced pressure. Boric acid was then removed by repeated co-distillation of methylborate with methanol also under reduced pressure. Deglycosylation was performed using anhydrous hydrofluoric acid according to Mort and Lamport (17). SDS gel electrophoresis ofproteins was carried out on 10o gel slabs essentially as described by Weber et al. (23) using Coomassie Blue to detect the standards. Fluorescamine labeling of protein was accomplished by the procedure of Bohlen ...

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Section: Methodscontrasting

confidence: 50%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Structural and Immunochemical Studies on the Phytotoxic Peptidorhamnomannan of Ceratocystis ulmi

Nordin¹,

Strobel²

1981

Plant Physiol.

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“…The toxin is proposed to be host spécifie (S. Takai, unpublished data) as is the fungus (Takai 1974), although the examination of the effects of the fungus on nonhost plants has provided some insight as to the prévention of the perpétuation of the disease Richards and Takai 1984;Takail974 Strobel et al 1978 Scheffer er a/. Strobel et al 1978Ysaigetal. 1989Duchesne <??#/.…”

Section: Pathogenmentioning

confidence: 99%

Molecular and cellular aspects of Dutch elm disease

Sticklen¹,

Bolyard²,

Hajela³

et al. 2005

phyto

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Cet article de synthèse présente une revue des recherches concernant les interactions moléculaires et cellulaires de la maladie hollandaise de l'orme. Cette maladie, causée par le champignon Ophiostoma ulmi, provoque un flétrissement vasculaire et elle est transmise d'ormes infectés à des ormes sains par des scolytes. Des toxines fongiques sont décrites en relation avec la pathogénèse, l'une d'elles, la cérato-ulmine, étant sous investigation au niveau moléculaire, plus particulièrement en ce qui concerne son mode d 'action et sa localisation. Le champignon a aussi été examiné au niveau moléculaire pour différencier des isolats agressifs et non agressifs sur la base de profils protéiques et d'acides nucléiques. Des cartes de lien génétique sont en développement afin de corréler la perturbation de certains gènes avec la perte du pouvoir pathogène. Des antagonistes viraux et bactériens du champignon, qui peuvent être utilisés comme mécanisme de lutte biologique contre la maladie hollandaise de l'orme, ont été caractérisés ainsi que plusieurs des molécules actives intervenant dans cette lutte. Les réponses de l'hôte sont discutées aux niveaux moléculaire et biochimique, incluant les phy toalexines et les éliciteurs des mécanismes de défense. Plusieurs avenues de recherche sont discutées afin de fournir une revue des approches moléculaires visant à comprendre et à manipuler les organismes impliqués en vue du but ultime de lutter contre la maladie hollandaise de l'orme.The folio wing review gives an overview of current research in the area of molecular and cellular interactions in Dutch elm disease. This vascular wilt disease is caused by the fungus Ophiostoma ulmi and is transmitted from diseased to healthy trees by the elm bark beetles. Fungal toxins are described which are associated with pathogenesis, one of which, ceratoulmin, is under investigation at the molecular level, particularly regarding its mode of action and localization. The fungus has also been examined at the molecular level to differentiate between aggressive and non-aggressive isolates on the basis of protein and nucleic acid profiles. Genetic linkage maps are being developed to correlate disruption of certain genes with the loss of pathogenicity. Viral and bacterial antagonists of the fungus, which may serve as biological control mechanisms for Dutch elm disease, have been characterized, as have several of the active molecules responsible for control. Host responses are also discussed at the molecular and biochemical level, including phytoalexins and defense mechanism elicitors. Several Unes of investigation are discussed to provide an overview of molecular approaches to understanding and manipulating the organisms involved with the ultimate goal of controlling Dutch elm disease

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Neue Entwicklungen in der Erforschung des Ulmensterbens

Strobel

Matern

1982

Biologie in unserer Zeit

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Gary A. Strobe1 und Ulrich Matern Die Ulmenkrankheit ist eine der schlimmsten Pflanzenkrankheiten unseres Jahrhunderts. Durch sie wurden Millionen von Ulmen in Europa, Nordamerika und Asien vernichtet. Der finanzielle Schaden erreicht Milliardenhohe. Die englische Bezeichnung dutch elm disease ist ein Tribut an die Wissenschaftler in Holland, die als erste die Krankheit untersuchten (dutch = hollandisch). Ihr wichtigster Wirt in Europa ist die Feldulme, Ulmus carpinifolia bzw. U . minor und andere U1menarten, in Nordamerika die amerikanische Ulme, U. americana. Viele der asiatischen Ulmen, wie z.B. U . pamiflora und U. pumila, sind dagegen vergleichsweise resistent. Dina Spierenburg beschrieb 1919 diese Krankheit zunachst aus Holland. Bald darauf (1921) wurde sie auch in Deutschland beobachtet, und auch aus anderen europaischen Landern wurde berichtet, dai3 die Ulmen abzusterben begannen. 1930 erreichte die Krankheit schlieglich die USA [ 3 ] . Einige Wissenschaftler vermuteten zunachst einen Zusammenhang zwischen dem Ulmensterben in Europa und dem Giftgas, das wahrend des ersten Weltkrieges angewendet worden war. Andere sahen die Ursache in der Durre, die zu jener Zeit geherrscht hatte, oder sie Ilachten an die Beteiligung eines virulenten bakteriellen Erregers.

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Some phytotoxic glycopeptides from Ceratocystis ulmi, the Dutch Elm Disease pathogen

Cited by 55 publications

References 22 publications

Structural and Immunochemical Studies on the Phytotoxic Peptidorhamnomannan of Ceratocystis ulmi

Structural and Immunochemical Studies on the Phytotoxic Peptidorhamnomannan of Ceratocystis ulmi

Molecular and cellular aspects of Dutch elm disease

Neue Entwicklungen in der Erforschung des Ulmensterbens

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