Functional analysis of Botrytis cinerea pectin methylesterase genes by PCR‐based targeted mutagenesis: Bcpme1 and Bcpme2 are dispensable for virulence of strain B05.10

Kars, I.; McCalman, M.T.; Wagemakers, L.; Kan, Jan A.L. van

doi:10.1111/j.1364-3703.2005.00312.x

Cited by 77 publications

(67 citation statements)

References 37 publications

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“…There are some reports that gene deletions can have diverse impacts on growth and virulence in B. cinerea in distinct strain backgrounds. For example, deletion of the pectin methylesterase bcpme1 caused reduced virulence in strain Bd90 (64) but not in the B05.10 background (65). Another example involves the loss of the phytotoxin botrydial, which leads to reduced virulence in T4 but did not affect virulence in SAS56 (66).…”

Section: Discussionmentioning

confidence: 99%

Unraveling the Function of the Response Regulator BcSkn7 in the Stress Signaling Network of Botrytis cinerea

et al. 2015

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Important for the lifestyle and survival of every organism is the ability to respond to changing environmental conditions. The necrotrophic plant pathogen Botrytis cinerea triggers an oxidative burst in the course of plant infection and therefore needs efficient signal transduction to cope with this stress. The factors involved in this process and their precise roles are still not well known. Here, we show that the transcription factor Bap1 and the response regulator (RR) B. cinerea Skn7 (BcSkn7) are two key players in the oxidative stress response (OSR) of B. cinerea; both have a major influence on the regulation of classical OSR genes. A yeast-one-hybrid (Y1H) approach proved direct binding to the promoters of gsh1 and grx1 by Bap1 and of glr1 by BcSkn7. While the function of Bap1 is restricted to the regulation of oxidative stress, analyses of ⌬bcskn7 mutants revealed functions beyond the OSR. Involvement of BcSkn7 in development and virulence could be demonstrated, indicated by reduced vegetative growth, impaired formation of reproductive structures, and reduced infection cushion-mediated penetration of the host by the mutants. Furthermore, ⌬bcskn7 mutants were highly sensitive to oxidative, osmotic, and cell wall stress. Analyses of ⌬bap1 bcskn7 double mutants indicated that loss of BcSkn7 uncovers an underlying phenotype of Bap1. In contrast to Saccharomyces cerevisiae, the ortholog of the glutathione peroxidase Gpx3p is not required for nuclear translocation of Bap1. The presented results contribute to the understanding of the OSR in B. cinerea and prove that it differs substantially from that of yeast, demonstrating the complexity and versatility of components involved in signaling pathways.

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Section: Discussionmentioning

confidence: 99%

Unraveling the Function of the Response Regulator BcSkn7 in the Stress Signaling Network of Botrytis cinerea

et al. 2015

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“…B. cinerea isolates are also known to be genetically quite diverse in their polygalacturonase loci, and genetic variability in PG2 is associated with varying growth on pectin containing media (Rowe and Kliebenstein, 2007). Furthermore, the contribution of PG1 and PG2 to B. cinerea virulence seems to depend on the specific B. cinerea isolate and the host species (Kars et al, 2005a;Zhang and van Kan, 2013). B. cinerea isolates are also known to be differently sensitive to the antimicrobial secondary metabolite camalexin (Kliebenstein et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Pectin Biosynthesis Is Critical for Cell Wall Integrity and Immunity in Arabidopsis thaliana

et al. 2016

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(D.J.K.); 0000-0001-5167-736X (J.G.)Plant cell walls are important barriers against microbial pathogens. Cell walls of Arabidopsis thaliana leaves contain three major types of polysaccharides: cellulose, various hemicelluloses, and pectins. UDP-D-galacturonic acid, the key building block of pectins, is produced from the precursor UDP-D-glucuronic acid by the action of glucuronate 4-epimerases (GAEs). Pseudomonas syringae pv maculicola ES4326 (Pma ES4326) repressed expression of GAE1 and GAE6 in Arabidopsis, and immunity to Pma ES4326 was compromised in gae6 and gae1 gae6 mutant plants. These plants had brittle leaves and cell walls of leaves had less galacturonic acid. Resistance to specific Botrytis cinerea isolates was also compromised in gae1 gae6 double mutant plants. Although oligogalacturonide (OG)-induced immune signaling was unaltered in gae1 gae6 mutant plants, immune signaling induced by a commercial pectinase, macerozyme, was reduced. Macerozyme treatment or infection with B. cinerea released less soluble uronic acid, likely reflecting fewer OGs, from gae1 gae6 cell walls than from wild-type Col-0. Although both OGs and macerozyme-induced immunity to B. cinerea in Col-0, only OGs also induced immunity in gae1 gae6. Pectin is thus an important contributor to plant immunity, and this is due at least in part to the induction of immune responses by soluble pectin, likely OGs, that are released during plant-pathogen interactions.

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“…The PCR mixture contained the three template fragments and primers GACTGCTACTGAGTATT-CGGT and CTACTCAAACACCATCCGCGA. The resulting PCR fragment was excised from the gel and directly transformed to B. cinerea protoplasts using the published procedure (23).…”

Section: Targeted Mutagenesis Of B Cinereamentioning

confidence: 99%

Oxaloacetate Hydrolase, the C–C Bond Lyase of Oxalate Secreting Fungi

Han

Joosten

Niu

et al. 2007

Journal of Biological Chemistry

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Oxalate secretion by fungi is known to be associated with fungal pathogenesis. In addition, oxalate toxicity is a concern for the commercial application of fungi in the food and drug industries. Although oxalate is generated through several different biochemical pathways, oxaloacetate acetylhydrolase (OAH)-catalyzed hydrolytic cleavage of oxaloacetate appears to be an especially important route. Below, we report the cloning of the Botrytis cinerea oahA gene and the demonstration that the disruption of this gene results in the loss of oxalate formation. In addition, through complementation we have shown that the intact B. cinerea oahA gene restores oxalate production in an Aspergillus niger mutant strain, lacking a functional oahA gene. These observations clearly indicate that oxalate production in A. niger and B. cinerea is solely dependent on the hydrolytic cleavage of oxaloacetate catalyzed by OAH. In addition, the B. cinera oahA gene was overexpressed in Escherichia coli and the purified OAH was used to define catalytic efficiency, substrate specificity, and metal ion activation. These results are reported along with the discovery of the mechanism-based, tight binding OAH inhibitor 3,3-difluorooxaloacetate (K i ‫؍‬ 68 nM). Finally, we propose that cellular uptake of this inhibitor could reduce oxalate production.Numerous filamentous fungi, including the food biotechnology fungus Aspergillus niger, the opportunistic human pathogen Aspergillus fumigatus, the phytopathogenic fungi Botrytis cinerea and Sclerotinia sclerotiorum, as well as many brown-rot and white-rot basidiomycetes, are able to efficiently produce large quantities of oxalate (1, 2). It is known that oxalate secretion is associated with fungal pathogenesis (1, 3-6). In the wood-rotting fungus Fomitopsis palustris oxalate is formed as the product of glucose metabolism (7). We recently initiated investigations of the oxalate biosynthetic pathway to develop a genomic-based method for distinguishing between oxalate producing and non-producing fungi. An additional goal of this effort was to identify enzyme inhibitors that could be used to arrest oxalate formation in targeted fungi.To attenuate oxalate production in fungi, it is necessary to first identify the major pathway responsible for oxalate formation. There are three potential routes for production of oxalate in fungi: oxidation of glyoxylate (8, 9), oxidation of glycolaldehyde (10), and hydrolysis of oxaloacetate (11). The results of studies of [ 14 C]CO 2 incorporation into the metabolite pools of A. niger indicate that oxalate is derived from oxaloacetate (12). This finding parallels the results of earlier work on the purification of an enzyme "oxalacetalase" (now known as oxaloacetate acetylhydrolase or OAH) 4 that catalyzes the hydrolytic cleavage of oxaloacetate to form acetate and oxalate (11). In a subsequent study, a mutant A. niger strain, NW228 (13), was found to be deficient in both oxalate production and in the synthesis of active OAH (14). These observations suggest that oxalate is ...

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Functional analysis of Botrytis cinerea pectin methylesterase genes by PCR‐based targeted mutagenesis: Bcpme1 and Bcpme2 are dispensable for virulence of strain B05.10

Cited by 77 publications

References 37 publications

Unraveling the Function of the Response Regulator BcSkn7 in the Stress Signaling Network of Botrytis cinerea

Unraveling the Function of the Response Regulator BcSkn7 in the Stress Signaling Network of Botrytis cinerea

Pectin Biosynthesis Is Critical for Cell Wall Integrity and Immunity in Arabidopsis thaliana

Oxaloacetate Hydrolase, the C–C Bond Lyase of Oxalate Secreting Fungi

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