Interactions of the nonhost French bean plant (Phaseolus vulgaris) with parasitic and saprophytic fungi. III. Cytologically detectable responses

1989 ◽  
Vol 67 (3) ◽  
pp. 676-686 ◽  
Author(s):  
Myriam R. Fernandez ◽  
Michèle C. Heath

Cytologically detectable responses of the nonhost French bean (Phaseolus vulgaris L. cv. Pinto) to saprophytic and parasitic fungi were examined when fungal spores were introduced into heated or unheated leaves via wounds or by injection. Although similar types of responses were observed in interactions with all the fungi, some of these responses were characteristic of each group (saprophytes vs. parasites) in the frequency and (or) extent with which they were elicited. Differences in responses between and within each of these groups of fungi were more related to their degree of adaptation for parasitism than to their taxonomic relationships. Certain responses that were typically elicited by the saprophytes occurred to a lesser extent in tissue responding to the parasites, suggesting that the ability to not trigger, or suppress, these responses may be a general feature of parasitic fungi. None of the fungi elicited significant levels of plant cell necrosis, and for two of the saprophytes, dead spores elicited a lower frequency of responses than live ones. The data indicate that many of the responses of a nonhost plant to living fungi may be the result of reactions to fungal activity rather than to constitutive recognition molecules such as components of the fungal cell wall.

1995 ◽  
Vol 306 (3) ◽  
pp. 745-750 ◽  
Author(s):  
D Robertson ◽  
B A McCormack ◽  
G P Bolwell

Enzyme activities involved in quantitative and qualitative flux of sugars into cell wall polysaccharides were determined following elicitor treatment of suspension cultured cells of French bean (Phaseolus vulgaris L.). Two subsets of activities were examined: the first were involved in synthesis and metabolism of UDP-glucose and the provision of the pool of UDP-sugars, and the second a selection of membrane-bound glycosyltransferases involved in the synthesis of pectins, hemicelluloses and glucans of the primary cell wall. Of the first group, only UDP-glucose dehydrogenase (EC 1.1.1.22) showed any significant induction in response to elicitor treatment, sucrose synthase (EC 2.4.1.13), UDP-glucuronate decarboxylase (EC 4.1.1.35), UDP-glucose and UDP-xylose 4-epimerases (EC 5.1.3.2 and EC 5.1.3.5 respectively) did not change in activity significantly over the time course. In contrast, enzymes of the second group showed a more complex response. Callose synthase (glucan synthase II, EC 2.4.1.12) increased in activity, as has been shown in other systems, while arabinan synthase (EC 2.4.1.-), xylan synthase (EC 2.4.1.72), xyloglucan synthase (EC 2.4.1.72) and glucan synthase I (EC 2.4.1.12) activities were rapidly depleted from membranes within 3 h following elicitor action. This rapid turnover of activity was striking, indicating that the half-life of such enzymes can be short and that elicitor action causes substantial perturbation of some membrane activities. Glucan synthase I activity appears to increase in the later stages over the time period measured, indicating some recovery of this metabolism.


1994 ◽  
Vol 299 (3) ◽  
pp. 747-753 ◽  
Author(s):  
A Zimmerlin ◽  
P Wojtaszek ◽  
G P Bolwell

A cationic (pI 8.3) wall-bound peroxidase has been purified to homogeneity from suspension-cultured cells of French bean (Phaseolus vulgaris L.). The enzyme was a glycoprotein and its M(r) was 46,000 as determined by SDS/Page and h.p.l.c. gel filtration. It was localized biochemically to microsomes and the cell wall, and the latter subcellular distribution was confirmed by immunogold techniques. The native enzyme showed absorption maxima at 403, 500 and 640 nm, with an RZ (A405/A280) of 3.3. The peroxidase oxidized guaïacol and natural phenolic acids. By desorption-chemical-ionization mass spectrometry the enzyme was found to oxidize the model compound, ferulic acid, into dehydrodiferulic acid. Kinetics studies indicated an apparent Km of 113.3 +/- 22.9 microM and a Vmax of 144 mumol.min-1.nmol-1 of protein at an H2O2 concentration of 100 microM. In comparison with a second French-bean peroxidase (FBP) and horseradish peroxidase, as a model, it acted with a 6-10-fold higher specificity in this capacity. It is a member of the peroxidase superfamily of bacterial, fungal and plant haem proteins by virtue of its highly conserved amino acid sequence within the proximal and distal haem-binding sites. This is good evidence that this particular FBP may function in constructing covalent cross-linkages in the wall during development and response to pathogens.


1989 ◽  
Vol 67 (3) ◽  
pp. 670-675
Author(s):  
Myriam R. Fernandez ◽  
Michèle C. Heath

Germination and growth of four saprophytic and five parasitic fungi nonpathogenic on French bean (Phaseolus vulgaris L. cv. Pinto) were examined in leaves after the epidermis was bypassed by injection or wounding. In unheated leaves, only the parasites had the ability to spread from the wound site and to germinate and grow inside the tissue after injection. The saprophytes behaved similarly only after the leaves were given a preinoculation heat treatment, suggesting a sensitivity to heat-sensitive inhibitors. For two of the saprophytes, evidence for the presence of heat-insensitive inhibitors also was found. Unlike the intercellular growth following stomatal penetration, growth of all of the parasites was affected by heat-sensitive defenses once the epidermis was bypassed. The greater intercellular growth after injection as compared with stomatal penetration suggested that the guard cells play a significant role in the resistance of intact, unheated leaves to Cladosporium fulvum Cooke. Of the saprophytes examined, Neurospora crassa Shear & Dodge appeared to posses a number of attributes that might be considered as potential pathogenicity factors.


1989 ◽  
Vol 67 (3) ◽  
pp. 661-669 ◽  
Author(s):  
Myriam R. Fernandez ◽  
Michèle C. Heath

Germination, penetration, and subsequent growth of four saprophytic and five parasitic fungi nonpathogenic on French bean (Phaseolus vulgaris L. cv. Pinto) were examined on, or in, killed, untreated, and heat-treated or blasticidin S treated leaves in the presence (saprophytes only) or absence of additional nutrients. The saprophytes required either an external supply of nutrients or a diffusate from autoclaved leaves to germinate, and they subsequently did not attempt to penetrate untreated or heat-treated living leaves. In contrast, the parasites germinated well, and penetrated untreated tissue almost exclusively through stomata. For some parasites, preinoculation heat treatment increased the incidence of direct penetration and the degree of fungal growth in the tissue. The saprophytes and nonbiotrophic parasites all penetrated and grew well in leaves that had been killed by autoclaving or freezing. The data suggest that living, intact bean leaves are resistant to the fungal saprophytes tested because these fungi lack pathogenicity factors necessary for germination on, and penetration of, the leaf. Resistance to the parasites, in contrast, appears to reside in heat-sensitive and heat- or blasticidin S insensitive defenses in the leaf that may differ with the fungus.


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