Determination of pathogen-related enzyme action by mass spectrometry analysis of pectin breakdown products of plant cell walls

An, Hyun Joo; Lurie, Susan; Greve, L. Carl; Rosenquist, Danielle; Kirmiz, Crystal; Labavitch, John M.; Lebrilla, Carlito B.

doi:10.1016/j.ab.2004.11.004

Cited by 42 publications

(42 citation statements)

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“…A candidate exo-PG ( BC1G_13137 ) gene that is only detected when in Botrytis -infected fruit had been detected when this pathogen was grown in minimal medium supplemented with pectins as the sole carbon source, which suggests that this enzyme is important in the degradation of host cell walls that are rich in pectins (Shah et al, 2009a). The molecules or signals lead to the expression of host-specific enzymes are not known, but some could result from the degradation of host cell walls by core CAZymes (e.g., pectin derived oligosaccharides; Körner et al, 1998; An et al, 2005). …”

Section: Discussionmentioning

confidence: 99%

Genome-wide transcriptional profiling of Botrytis cinerea genes targeting plant cell walls during infections of different hosts

et al. 2014

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Cell walls are barriers that impair colonization of host tissues, but also are important reservoirs of energy-rich sugars. Growing hyphae of necrotrophic fungal pathogens, such as Botrytis cinerea (Botrytis, henceforth), secrete enzymes that disassemble cell wall polysaccharides. In this work we describe the annotation of 275 putative secreted Carbohydrate-Active enZymes (CAZymes) identified in the Botrytis B05.10 genome. Using RNAseq we determined which Botrytis CAZymes were expressed during infections of lettuce leaves, ripe tomato fruit, and grape berries. On the three hosts, Botrytis expressed a common group of 229 potentially secreted CAZymes, including 28 pectin backbone-modifying enzymes, 21 hemicellulose-modifying proteins, 18 enzymes that might target pectin and hemicellulose side-branches, and 16 enzymes predicted to degrade cellulose. The diversity of the Botrytis CAZymes may be partly responsible for its wide host range. Thirty-six candidate CAZymes with secretion signals were found exclusively when Botrytis interacted with ripe tomato fruit and grape berries. Pectin polysaccharides are notably abundant in grape and tomato cell walls, but lettuce leaf walls have less pectin and are richer in hemicelluloses and cellulose. The results of this study not only suggest that Botrytis targets similar wall polysaccharide networks on fruit and leaves, but also that it may selectively attack host wall polysaccharide substrates depending on the host tissue.

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Section: Discussionmentioning

confidence: 99%

Genome-wide transcriptional profiling of Botrytis cinerea genes targeting plant cell walls during infections of different hosts

et al. 2014

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“…The observation that TomQ'a expression is induced in unripe wild-type fruit and in all ripening mutants at both 31 and 42 dpa but is down-regulated by B. cinerea in wild-type ripe fruit supports the conclusion that ripening affects the way fruit respond to the pathogen and what defenses are deployed. How fruit cells perceive and respond to fungal toxins or elicitors, such as cell wall fragments generated by fungal digestion of host cell walls, may also change during ripening (Cô té and Hahn, 1994;Collado et al, 2000;An et al, 2005;van Kan, 2006;Staats et al, 2007). Although B. cinerea is a necrotroph and hypersensitive responses and cell death have been thought to increase its aggressiveness (Govrin and Levine, 2000;Dickman et al, 2001;van Baarlen et al, 2004van Baarlen et al, , 2007, the timely and localized accumulation of antimicrobial substances, hydrogen peroxide, and lignin, all features of a hypersensitive response, effectively prevent expanding infections; the responses are evident in resistant unripe fruit but not in susceptible ripe fruit.…”

Section: Discussionmentioning

confidence: 99%

Ripening-Regulated Susceptibility of Tomato Fruit toBotrytis cinereaRequiresNORBut NotRINor Ethylene

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et al. 2009

Plant Physiology

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Fruit ripening is a developmental process that is associated with increased susceptibility to the necrotrophic pathogen Botrytis cinerea. Histochemical observations demonstrate that unripe tomato (Solanum lycopersicum) fruit activate pathogen defense responses, but these responses are attenuated in ripe fruit infected by B. cinerea. Tomato fruit ripening is regulated independently and cooperatively by ethylene and transcription factors, including NON-RIPENING (NOR) and RIPENING-INHIBITOR (RIN). Mutations in NOR or RIN or interference with ethylene perception prevent fruit from ripening and, thereby, would be expected to influence susceptibility. We show, however, that the susceptibility of ripe fruit is dependent on NOR but not on RIN and only partially on ethylene perception, leading to the conclusion that not all of the pathways and events that constitute ripening render fruit susceptible. Additionally, on unripe fruit, B. cinerea induces the expression of genes also expressed as uninfected fruit ripen. Among the ripening-associated genes induced by B. cinerea are LePG (for polygalacturonase) and LeExp1 (for expansin), which encode cell wall-modifying proteins and have been shown to facilitate susceptibility. LePG and LeExp1 are induced only in susceptible rin fruit and not in resistant nor fruit. Thus, to infect fruit, B. cinerea relies on some of the processes and events that occur during ripening, and the fungus induces these pathways in unripe fruit, suggesting that the pathogen itself can initiate the induction of susceptibility by exploiting endogenous developmental programs. These results demonstrate the developmental plasticity of plant responses to the fungus and indicate how known regulators of fruit ripening participate in regulating ripening-associated pathogen susceptibility.

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“…MALDI-TOFMS has also been used to characterise the pectin-derived oligosaccharides that accumulate in ripening and pathogeninfected tomato fruits. 20 …”

Section: Rapid Characterisation Of Changes In Wall Composition and Armentioning

confidence: 99%

“…21 Both structural and quantitative information can be obtained with this technique. It has been applied to characterise Arabidopsis pectic polysaccharides 20 and could also be useful to study the specificity and kinetics of plant polysaccharide hydrolases. 22 Most traditional chemical analyses of plant cell walls and the two strategies described above are destructive, in that they involve irreversible breakage of chemical bonds: wall structure is disrupted during extraction and so positional information is lost during sample preparation.…”

Section: Polysaccharide Analysis By Carbohydrate Gel Electrophoresis mentioning

confidence: 99%

“…3, 58 The expression of PEL genes in ripening tomato fruit has been reported 59 and the unsaturated oligosaccharides characteristic of PEL action have been identified in healthy tissues of Botrytis cinerea-infected tomato fruit 20 and in ripening, uninfected tomatoes (An H, Lurie S, Lebrilla C and Labavitch J, unpublished), contradicting a previous report suggesting the absence of PEL in tomato, 60 and demonstrating the value of techniques for isolating and characterizing wall breakdown products in ripening fruit. Suppression of PEL gene expression in transgenic strawberry was reported to result in substantially firmer fruits and reduced cell wall swelling, 61 and whether or not this effect can be reproduced in other fruits is an exciting question.…”

Section: 3mentioning

confidence: 99%

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The linkage between cell wall metabolism and fruit softening: looking to the future

et al. 2007

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The softening that accompanies ripening of commercially important fruits exacerbates damage incurred during shipping and handling and increases pathogen susceptibility. Thus, postharvest biologists have studied fruit softening to identify ways to manage ripening and optimise fruit quality. Studies, generally based on the premise that cell wall polysaccharide breakdown causes ripening-associated softening, have not provided the insights needed to genetically engineer, or selectively breed for, fruits whose softening can be adequately controlled. Herein it is argued that a more holistic view of fruit softening is required. Polysaccharide metabolism is undoubtedly important, but understanding this requires a full appreciation of wall structure and how wall components interact to provide strength. Consideration must be given to wall assembly as well as to wall disassembly. Furthermore, the apoplast must be considered as a developmentally and biochemically distinct, dynamic 'compartment', not just the location of the cell wall structural matrix. New analytical approaches for enhancing the ability to understand wall structure and metabolism are discussed. Fruit cells regulate their turgor pressure as well as cell wall integrity as they ripen, and it is proposed that future studies of fruit softening should include attempts to understand the bases of cell-and tissue-level turgor regulation if the goal of optimising softening control is to be reached. Finally, recent studies show that cell wall breakdown provides sugar substrates that fuel other important cellular pathways and processes. These connections must be explored so that optimisation of softening does not lead to decreases in other aspects of fruit quality.

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Determination of pathogen-related enzyme action by mass spectrometry analysis of pectin breakdown products of plant cell walls

Cited by 42 publications

References 46 publications

Genome-wide transcriptional profiling of Botrytis cinerea genes targeting plant cell walls during infections of different hosts

Genome-wide transcriptional profiling of Botrytis cinerea genes targeting plant cell walls during infections of different hosts

Ripening-Regulated Susceptibility of Tomato Fruit toBotrytis cinereaRequiresNORBut NotRINor Ethylene

The linkage between cell wall metabolism and fruit softening: looking to the future

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