Peroxidase activity and induced lignification in rusted flax interactions varying in their degree of incompatibility

1984 ◽  
Vol 62 (1) ◽  
pp. 134-141 ◽  
Author(s):  
M. D. Coffey ◽  
D. S. M. Cassidy

A histological comparison of the L9, K, P, P1, P2, and P3 genotypes in flax revealed differences in their expression of incompatibility. K and P1 were only moderately incompatible, whereas P, P2, and P3 were highly incompatible, restricting fungal development completely by the 4th day after inoculation. Host cell necrosis was delayed until day 3 in P3 and day 2 in P2; it was present at day 1 in P. Peroxidase, using guaiacol as a proton donor, was detected in P after 2 days, in K at 5 days, and in L9 after 7 days. Soluble peroxidase activity increased in the rusted P genotype from day 3 and from day 7 in K. Gel electrophoresis of infected P revealed a new peroxidase isozyme at 6 days after inoculation. Three new protein bands were also detected at this stage. In 3- and 4-day-old rusted P, peroxidase was detected in the host cell walls of tissue surrounding necrotic infection sites. It was not located in the host cell walls of the compatible L9 genotype at this stage. Lignin was detected histochemically in P after 2 days, in K after 5 days, and in L9 after 9 days. The Wiesner, Maiile, and KMnO4 tests gave equivalent results. Suberin was detected in P, K, and L9 at the same time that lignin became observable.

1997 ◽  
Vol 87 (1) ◽  
pp. 108-122 ◽  
Author(s):  
Nicole Benhamou ◽  
Patrice Rey ◽  
Mohamed Chérif ◽  
John Hockenhull ◽  
Yves Tirilly

The influence exerted by the mycoparasite Pythium oligandrum in triggering plant defense reactions was investigated using an experimental system in which tomato plants were infected with the crown and root rot pathogen Fusarium oxysporum f. sp. radicis-lycopersici. To assess the antagonistic potential of P. oligandrum against F. oxysporum f. sp. radicis-lycopersici, the interaction between the two fungi was studied by scanning and transmission electron microscopy (SEM and TEM, respectively). SEM investigations of the interaction region between the fungi demonstrated that collapse and loss of turgor of F. oxysporum f. sp. radicis-lycopersici hyphae began soon after close contact was established with P. oligandrum. Ultrastructural observations confirmed that intimate contact between hyphae of P. oligandrum and cells of the pathogen resulted in a series of disturbances, including generalized disorganization of the host cytoplasm, retraction of the plasmalemma, and, finally, complete loss of the protoplasm. Cytochemical labeling of chitin with wheat germ agglutinin (WGA)/ovomucoid-gold complex showed that, except in the area of hyphal penetration, the chitin component of the host cell walls was structurally preserved at a time when the host cytoplasm had undergone complete disorganization. Interestingly, the same antagonistic process was observed in planta. The specific labeling patterns obtained with the exoglucanase-gold and WGA-ovomucoid-gold complexes confirmed that P. oligandrum successfully penetrated invading cells of the pathogen without causing substantial cell wall alterations, shown by the intense labeling of chitin. Cytological investigations of samples from P. oligandrum-inoculated tomato roots revealed that the fungus was able to colonize root tissues without inducing extensive cell damage. However, there was a novel finding concerning the structural alteration of the invading hyphae, evidenced by the frequent occurrence of empty fungal shells in root tissues. Pythium ingress in root tissues was associated with host metabolic changes, culminating in the elaboration of structural barriers at sites of potential fungal penetration. Striking differences in the extent of F. oxysporum f. sp. radicis-lycopersici colonization were observed between P. oligandrum-inoculated and control tomato plants. In control roots, the pathogen multiplied abundantly through much of the tissues, whereas in P. oligandrum-colonized roots pathogen growth was restricted to the outermost root tissues. This restricted pattern of pathogen colonization was accompanied by deposition of newly formed barriers beyond the infection sites. These host reactions appeared to be amplified compared to those seen in nonchallenged P. oligandrum-infected plants. Most hyphae of the pathogen that penetrated the epidermis exhibited considerable changes. Wall appositions contained large amounts of callose, in addition to be infiltrated with phenolic compounds. The labeling pattern obtained with gold-complexed laccase showed that phenolics were widely distributed in Fusarium-challenged P. oligandrum-inoculated tomato roots. Such compounds accumulated in the host cell walls and intercellular spaces. The wall-bound chitin component in Fusarium hyphae colonizing P. oligandrum-inoculated roots was preserved at a time when hyphae had undergone substantial degradation. These observations provide the first convincing evidence that P. oligandrum has the potential to induce plant defense reactions in addition to acting as a mycoparasite.


1975 ◽  
Vol 53 (9) ◽  
pp. 852-860 ◽  
Author(s):  
B. Truelove ◽  
R. Rodriguez-Kabana ◽  
Larry R. Jones

Changes in nitrogen contents and peroxidase activities of fractions isolated from hypocotyl tissue of Black Valentine bean (Phaseolus vulgaris L.) of increasing age were studied. As beans aged in darkness, a decreasing percentage of their nitrogen content was recovered in the isolated particulate fractions. Peroxidase activity of particulate fractions from dark-grown beans accounted for 49% of the total activity of both 3-day-old seedlings and 16-day-old senescing plants. Peroxidase specific activity of dark-grown tissue homogenates did not increase with plant age; however, after a certain period of growth, further aging resulted in increased peroxidase specific activity associated with the particulate fractions. Between day 3 and day 8 the patterns of peroxidase activity of the different fractions varied, but over the period day 9 to day 16, the patterns of all fractions were correlated. The nitrogen contents and peroxidase activities of fractions isolated from beans transferred from dark to light were different from those of fractions from beans of similar chronological age kept in darkness. Transfer of plants to light resulted in increased soluble peroxidase activity and prevention of the steep increase in particulate fraction activity recorded for dark-grown plants.


mBio ◽  
2017 ◽  
Vol 8 (2) ◽  
Author(s):  
Jeff Quigley ◽  
V. Keith Hughitt ◽  
Carlos A. Velikovsky ◽  
Roy A. Mariuzza ◽  
Najib M. El-Sayed ◽  
...  

ABSTRACT The cell wall of Mycobacterium tuberculosis is composed of unique lipids that are important for pathogenesis. Indeed, the first-ever genetic screen in M. tuberculosis identified genes involved in the biosynthesis and transport of the cell wall lipid PDIM (phthiocerol dimycocerosates) as crucial for the survival of M. tuberculosis in mice. Here we show evidence for a novel molecular mechanism of the PDIM-mediated virulence in M. tuberculosis. We characterized the DNA interaction and the regulon of Rv3167c, a transcriptional repressor that is involved in virulence regulation of M. tuberculosis, and discovered that it controls the PDIM operon. A loss-of-function genetic approach showed that PDIM levels directly correlate with the capacity of M. tuberculosis to escape the phagosome and induce host cell necrosis and macroautophagy. In conclusion, our study attributes a novel role of the cell wall lipid PDIM in intracellular host cell modulation, which is important for host cell exit and dissemination of M. tuberculosis. IMPORTANCE Mycobacterium tuberculosis is a major human pathogen that has coevolved with its host for thousands of years. The complex and unique cell wall of M. tuberculosis contains the lipid PDIM (phthiocerol dimycocerosates), which is crucial for virulence of the bacterium, but its function is not well understood. Here we show that PDIM expression by M. tuberculosis is negatively regulated by a novel transcriptional repressor, Rv3167c. In addition, we discovered that the escape of M. tuberculosis from its intracellular vacuole was greatly augmented by the presence of PDIM. The increased release of M. tuberculosis into the cytosol led to increased host cell necrosis. The discovery of a link between the cell wall lipid PDIM and a major pathogenesis pathway of M. tuberculosis provides important insights into the molecular mechanisms of host cell manipulation by M. tuberculosis. IMPORTANCE Mycobacterium tuberculosis is a major human pathogen that has coevolved with its host for thousands of years. The complex and unique cell wall of M. tuberculosis contains the lipid PDIM (phthiocerol dimycocerosates), which is crucial for virulence of the bacterium, but its function is not well understood. Here we show that PDIM expression by M. tuberculosis is negatively regulated by a novel transcriptional repressor, Rv3167c. In addition, we discovered that the escape of M. tuberculosis from its intracellular vacuole was greatly augmented by the presence of PDIM. The increased release of M. tuberculosis into the cytosol led to increased host cell necrosis. The discovery of a link between the cell wall lipid PDIM and a major pathogenesis pathway of M. tuberculosis provides important insights into the molecular mechanisms of host cell manipulation by M. tuberculosis.


1984 ◽  
Vol 62 (12) ◽  
pp. 2644-2649 ◽  
Author(s):  
M. Harche

Using diaminobenzidine as substrate, peroxidase activity was localized in the walls of parenchyma cells and differentiating fibres. In mature fibres and parenchyma a slight activity could be recognized in primary walls only. In parenchyma cells, peroxidase activity was fairly inhibited with heat, potassium cyanide, and aminotriazole, which could indicate the presence of catalase within the cell walls. However, in plasmodesmatal regions peroxidases were- resistant to the above inhibitors. Syringaldazine oxidase activity was present only in the primary wall and the outer part of the secondary wall of differentiating fibres. The parallelism between lignification and peroxidase activity in the secondary walls supports the hypothesis of the involvement of these enzymes in the lignification process.


Biologia ◽  
2011 ◽  
Vol 66 (1) ◽  
Author(s):  
Yasushi Sato ◽  
Youko Yajima ◽  
Naohito Tokunaga ◽  
Ross Whetten

AbstractLignin is synthesized not only during morphogenesis of vascular plants but also in response to various stresses. Isolated Zinnia elegans mesophyll cells can differentiate into tracheary elements (TEs), and deposit lignin into cell walls in TE-inductive medium (D medium). Meanwhile isolated mesophyll cells cultured in hormone-free medium (Co medium) accumulate stress lignin-like substance during culture. Therefore this culture system is suitable for study of lignin and lignin-like substance formation.In D medium lignin was deposited in TEs, but in Co medium, extracellular lignin-like substance accumulated. Analysis of the culture media indicated the presence of dilignols in D culture, but not in Co culture. To investigate the fate of lignin precursors, we added coniferyl alcohol (CA) in each culture. In Co medium, CA was polymerized into dilignols rapidly but they were present only temporarily, and in D medium CA was polymerized into dilignols relatively slowly but their content increased continually.Meanwhile, in Co culture, peroxidase activity in the medium was much higher than the peroxidase activity bound ionically to the cell walls. In D culture, ionically bound peroxidase activity was higher than that in the medium. These results may suggest that lignin deposition in TEs is related to ionically bound peroxidases in D culture, and lignin-like substance deposition in the medium is related to peroxidases in the medium in Co culture.


1997 ◽  
Vol 10 (7) ◽  
pp. 803-811 ◽  
Author(s):  
B. Boher ◽  
M. Nicole ◽  
M. Potin ◽  
J. P. Geiger

The location of lipopolysaccharides produced by Xanthomonas axonopodis pv. manihotis during pathogenesis on cassava (Manihot esculenta) was determined by fluorescence and electron microscopy immunolabeling with monoclonal antibodies. During the early stages of infection, pathogen lipopolysaccharides were detected on the outer surface of the bacterial envelope and in areas of the plant middle lamellae in the vicinity of the pathogen. Later in the infection process, lipopolysaccharide-specific antibodies bound to areas where the plant cell wall was heavily degraded. Lipopolysaccharides were not detected in the fibrillar matrix filling intercellular spaces of infected cassava leaves. Monoclonal antibodies specific for the exopolysaccharide xanthan side chain labeled the bacteria, the fibrillar matrix, and portions of the host cell wall. The association of Xanthomonas lipopolysaccharides with host cell walls during plant infection is consistent with a role of these bacterial extracellular polysaccharides in the infection process.


1985 ◽  
Vol 63 (4) ◽  
pp. 757-761 ◽  
Author(s):  
E. Untiedt ◽  
K. Müller

Lyophyllum palustre (Peck) Singer, a basidiomycete (Tricholomataceae) parasitizing Sphagnum, was examined for points of contact between hyphae and Sphagnum cells with the help of light microscopy, scanning electron microscopy, and transmission electron microscopy. Results indicate that the fungus attacks Sphagnum cells by penetrating cell walls and altering host cell protosplasm. In addition, the formation of additional partitioning cell walls in attacked living Sphagnum cells was observed.


1999 ◽  
Vol 26 (4) ◽  
pp. 387 ◽  
Author(s):  
Francisco J. Pérez ◽  
Verónica Morales

Soluble peroxidase activity from pedicels of seedless table grape cv. Sultana was highly stimulated by post-bloom applications of gibberellic acid (GA3) to vines. The increase in peroxidase activity was mainly due to the induction of a basic peroxidase isoenzyme (pI > 9; BPrx-HpI). The activity of two other peroxidase isoenzymes of pI 6.5 and 3.2 was not altered by the hormone treatment. BPrx-HpI was induced by GA3 in pedicels and rachis but not in berries, although in berries peroxidase activity was also stimulated by post-bloom GA3 applications. BPrx-HpI oxidised guaiacol and ortho-phenylenediamine (o-PDA), while the others peroxidases found in the pedicel and in the berry oxidised only o-PDA. Hence, BPrx-HpI was characterised as a guaiacol-peroxidase showing no activity towards ascorbic acid (ASC). The possible role of BPrx-HpI in pedicel lignification and berry-drop caused by GA3 applications to cv. Sultana vines is discussed.


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