Pectin methylesterase from Botrytis cinerea: physiological, biochemical and immunochemical studies

Pectin methylesterase (PME) was purified from the supernatant of Botrytis cinerea strain Bd90. SDS-PAGE showed a single band at 42 kDa, but this band corresponded to two distinct isoforms observed by IEF-PAGE at pl values of 7.0 and 7.4. PME was produced during the exponential phase of fungal growth and independently of the carbon source. Unlike other pectinases of B. cinerea, which are polymorphic, no differences were observed between the PME profiles of 25 strains of different origins. Polyclonal antibodies were raised against purified PME from B. cinerea, and immunochemical comparisons with PMEs from Erwinia chrysanthemi, Vigna radiata and Glycine max showed the presence of common epitopes between these different enzymes.

[1]  V. Ramanathan,et al.  A Protein from Immature Raspberry Fruits which Inhibits Endopolygalacturonases from Botrytis cinerea and other Micro-organisms , 1993 .

[2]  D. J. Johnston,et al.  An immunological study of the induction of polygalacturonases in Botrytis cinerea , 1992 .

[3]  D. Tieman,et al.  An Antisense Pectin Methylesterase Gene Alters Pectin Chemistry and Soluble Solids in Tomato Fruit. , 1992, The Plant cell.

[4]  A. Bennett,et al.  ROLE OF CELL WALL HYDROLASES IN FRUIT RIPENING , 1991 .

[5]  J. Walton,et al.  Endopolygalacturonase is not required for pathogenicity of Cochliobolus carbonum on maize. , 1990, The Plant cell.

[6]  S. Movahedi,et al.  The roles of aspartic proteinase and endo-pectin lyase enzymes in the primary stages of infection and pathogenesis of various host tissues by different isolates of Botrytis cinerea Pers ex. Pers , 1990 .

[7]  W. Y. Wang,et al.  Purification and Characterization of Pectinmethylesterase from Ficus awkeotsang Makino Achenes. , 1989, Plant physiology.

[8]  J. M. Dow,et al.  Molecular cloning of a polygalacturonate lyase gene from Xanthomonas campestris pv. campestris and role of the gene product in pathogenicity , 1989 .

[9]  A. Hagerman,et al.  Continuous spectrophotometric assay for plant pectin methyl esterase , 1986 .

[10]  A. Hagerman,et al.  Plate assay for determining the time of production of protease, cellulase, and pectinases by germinating fungal spores. , 1985, Analytical biochemistry.

[11]  A. Kotoujansky,et al.  Detection of depolymerase isoenzymes after electrophoresis or electrofocusing, or in titration curves. , 1984, Analytical biochemistry.

[12]  R. Goldberg Changes in the properties of cell wall pectin methylesterase along the Vigna radiata hypocotyl , 1984 .

[13]  A. Schejter,et al.  Single step chromatographic purification and characterization of the endopolygalacturonases and pectinesterases of the fungus, Botrytis cinerea Pers , 1983 .

[14]  R. Lahoz,et al.  Cell wall plant polysaccharide-degrading enzymes in autolysis of Botrytis cinerea , 1982 .

[15]  W. N. Burnette,et al.  "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. , 1981, Analytical biochemistry.

[16]  C. Merril,et al.  Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. , 1981, Science.

[17]  C. Pallavicini,et al.  Variability of Polygalacturonase and Protein Isoelectric Focusing Patterns in Botrytis cinerea Isolates , 1980 .

[18]  F. Rombouts,et al.  Chromatographic separation of orange pectinesterase isoenzymes on pectates with different degrees of cross-linking , 1979 .

[19]  M. Tarabieh,et al.  Studies on Enzymes of Different Strains of Botrytis cinerea , 1978 .

[20]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[21]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[22]  M. Boccara,et al.  Pathogenic behavior of pectinase-defective Erwinia chrysanthemi mutants on different plants , 1993 .

[23]  A. Schejter,et al.  Isozymes of pectinesterase and polygalacturonase from Botrytis cinerea Pers , 1988 .

[24]  A. Collmer,et al.  Construction and characterization of an Erwinia chrysanthemi mutant with directed deletions in all of the pectate lyase structural genes , 1988 .

[25]  N. Keen,et al.  The Role of Pectic Enzymes in Plant Pathogenesis , 1986 .

[26]  E. Gotschlich,et al.  A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. , 1984, Analytical biochemistry.

[27]  F. Drawert,et al.  Charakterisierung extrazellulärer proteine und enzyme aus pektinkulturfiltraten von Botrytis cinerea , 1978 .

[28]  H. Urbanek,et al.  Purification of polygalacturonases produced by Botrytis cinerea E-200. , 1975, Bulletin de l'Academie polonaise des sciences. Serie des sciences biologiques.

[29]  F. Smith,et al.  COLORIMETRIC METHOD FOR DETER-MINATION OF SUGAR AND RELATED SUBSTANCE , 1956 .