Detection of pathogenesis-related proteins (PR or b) and of other proteins in the intercellular fluid of hypersensitive plants infected with tobacco mosaic virus

1984 ◽  
Vol 62 (3) ◽  
pp. 564-569 ◽  
Author(s):  
Jean-Guy Parent ◽  
Alain Asselin
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Chenopodium Quinoa ◽  
Leaf Tissue ◽  
Electrophoretic Analysis ◽  
Intercellular Fluid ◽  
Molecular Weights ◽  
Pathogenesis Related ◽  
The One ◽  
Related Proteins

In tobacco mosaic virus infected hypersensitive Nicotiana species, the pathogenesis-related (PR) or b proteins were found with other proteins in the intercellular fluid of leaf tissue. Analysis of leaves from mock-inoculated plants did not reveal the presence of detectable amounts of proteins in the intercellular fluid. The presence of proteins in the intercellular fluid seems to be widespread since 10 proteins were detected in tobacco mosaic virus infected Chenopodium quinoa Willd. These proteins were found not only in inoculated tissue but also in the intercellular fluid of uninoculated upper leaves. A two-dimensional gel electrophoretic analysis of intercellular fluid proteins from N. tabacum L. cv. Xanthi-nc infected leaves showed that the relative molecular weight of protein b2 is larger (14 700) than the one of b1 and b3 (14 200). Other proteins, ranging in molecular weights from 12 500 to 36 300, could also be detected. We postulate that the presence of rather large amounts of proteins, including the well-characterized b proteins, in the intercellular fluid of infected hypersensitive leaves reflects overall changes in cell-to-cell interactions and in the cell wall metabolism.

Comparative study of two groups of b proteins (pathogenesis related) from the intercellular fluid of Nicotiana leaf tissue infected by tobacco mosaic virus

1985 ◽  
Vol 63 (5) ◽  
pp. 932-937 ◽  
Author(s):  
Marc G. Fortin ◽  
Jean-Guy Parent ◽  
Alain Asselin
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Time Course ◽  
Leaf Tissue ◽  
Antigenic Determinants ◽  
Peptide Fragments ◽  
Intercellular Fluid ◽  
Pathogenesis Related ◽  
Time Course Study ◽  
Related Proteins

A time-course study showed that the major extracellular b (pathogenesis-related) proteins were detected approximately 37 h after tobacco mosaic virus infection of Nicotiana tabacum L. cv. Xanthi-nc tobacco leaf tissue. Silver staining of b1b2 and b3 proteins in native polyacrylamide gels required previous staining with Coomassie blue. Elution profiles of b4, b5, and b6b proteins from DEAE-Sephacel columns were similar to those of b1b2, and b3 proteins. Relationships among b proteins accumulating in the intercellular fluid of hypersensitive cv. Xanthi-nc tobacco leaves infected by tobacco mosaic virus were examined on the basis of serological relationships and by comparing peptide fragments. Results showed that b4, b5, and b6b proteins share common antigenic determinants and polypeptide fragments different from those shared by b1b2, and b3 proteins. Together with the close similarity of the molecular weights of the proteins within each group, these data suggest that at least two groups of related proteins are found in the intercellular fluid extracts of tobacco plants infected by tobacco mosaic virus. It is also shown that the antigenic determinants shared within the b4, b5, and b6b group are found with the same b proteins from systemically infected N. tabacum L. cv. Samsun and with some b proteins from N. sylvestris Speg. and Comes.

Analysis of extracellular proteins by native polyacrylamide gel electrophoresis with infiltrated leaf tissue: application to pathogenesis-related proteins

1988 ◽  
Vol 66 (6) ◽  
pp. 1227-1229 ◽  
Author(s):  
Jean Grenier ◽  
François Côté ◽  
Alain Asselin
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Leaf Tissue ◽  
Polyacrylamide Gel ◽  
Simple Method ◽  
Pathogenesis Related Proteins ◽  
Intercellular Fluid ◽  
Pathogenesis Related ◽  
Native Polyacrylamide Gel Electrophoresis ◽  
Related Proteins

In addition to polyacrylamide gel electrophoretic analysis of intercellular fluid extracts, a simple method of detection of extracellular pathogenesis-related proteins was based on direct native polyacrylamide gel electrophoresis for acidic and basic proteins with leaf tissue infiltrated with 150 mM sucrose. This technique allowed for the detection of the complete set of tobacco pathogenesis-related proteins without having to extract the intercellular fluid by low-speed centrifugation. A major advantage of the technique is the capacity to observe the distribution of extracellular endogenous or exogenous proteins in the tissue directly subjected to electrophoresis.

Detection of 10 additional pathogenesis-related (b) proteins in intercellular fluid extracts from stressed 'Xanthi-nc' tobacco leaf tissue

1987 ◽  
Vol 65 (3) ◽  
pp. 476-481 ◽  
Author(s):  
Richard Hogue ◽  
Alain Asselin
Keyword(s):  
Gel Electrophoresis ◽  
Stress Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Leaf Tissue ◽  
Polyacrylamide Gel ◽  
Basic Proteins ◽  
Pathogenesis Related Proteins ◽  
Intercellular Fluid ◽  
Pathogenesis Related ◽  
Related Proteins

By using two-dimensional polyacrylamide gel electrophoresis, 10 additional pathogenesis-related proteins (b6c, b9c, b10a, b10b, b11a, b11b, b12, b13, b14, b15) were found in intercellular fluid extracts of stressed 'Xanthi-nc' tobacco leaf tissue. Proteins were identified as extracellular pathogenesis-related stress proteins by polyacrylamide gel electrophoresis analysis of three successive intercellular fluid extracts compared with homogenates before and after making intercellular fluid extracts. Four proteins (b12, b13, b14, b15) were only resolved by using native polyacrylamide gel electrophoresis for basic proteins in the first dimension gel and they were best extracted in 0.05 M Tris–HCl (pH 7.5) and 0.05 M CaCl2 as the infiltration buffer. The four basic proteins were found in intercellular fluid extracts from leaf tissue subjected to the same types of chemical inducers as previously described for tobacco pathogenesis-related proteins. Their accumulation was inhibited by basic amino acids or spermidine (1 mM) and they were resistant to endogenous and exogenous (trypsin, subtilisin) proteolysis. They did not bind to concanavalin A – Sepharose. These findings indicate that at least 23 proteins accumulate extracellularly after various types of stress in 'Xanthi-nc' tobacco green tissue. These proteins probably represent several groups or families of plant stress proteins.

Glycoproteins, enzymatic activities, and b proteins in intercellular fluid extracts from hypersensitive Nicotiana species infected with tobacco mosaic virus

1985 ◽  
Vol 63 (5) ◽  
pp. 928-931 ◽  
Author(s):  
Jean-Guy Parent ◽  
Richard Hogue ◽  
Alain Asselin
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Electrophoretic Pattern ◽  
Protein Band ◽  
Polyacrylamide Gel ◽  
Intercellular Fluid ◽  
Nicotiana Tabacum L ◽  
The One

Intercellular fluid b proteins from hypersensitive Nicotiana tabacum L. cv. Xanthi-nc and N. sylvestris Speg. and Comes infected with tobacco mosaic virus were compared by two-dimensional (2-D) polyacrylamide gel electrophoresis. Except for missing bands b2, b6a, b6b, and b7b, the overall 2-D electrophoretic pattern of N. sylvestris intercellular fluid proteins was similar to the one observed with 'Xanthi-nc' tobacco. Intercellular proteins were also studied by chromatography on con-canavalin A. Glycoproteins corresponding to b6a and b7a proteins of N. tabacum and the [Formula: see text] analog of N. sylvestris were identified. These proteins are probably peroxidase isozymes, as peroxidase activities with the same electrophoretic mobility were detected after polyacrylamide gel electrophoresis. No esterase activity was associated with any b protein band in gels. Esterase activities decreased upon virus infection, but accumulation of b proteins and peroxidase activities increased.

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