Abstract
Five Botrytis cinerea endopolygalacturonase enzymes (BcPGs) were individually expressed in Pichia pastoris, purified to homogeneity and biochemically characterized. While the pH optima of the five enzymes were similar (approximately pH 4.5) the maximum activity of individual enzymes differed significantly. For hydrolysis of polygalacturonic acid (PGA), the V(max,app) ranged from 10 to 900 U mg(-1), while the K(m,app) ranged from 0.16 to 0.6 mg ml(-1). Although all BcPGs are true endopolygalacturonases, they apparently have different modes of action. PGA hydrolysis by BcPG1, BcPG2 and BcPG4 leads to the transient accumulation of oligomers with DP < 7, whereas PGA hydrolysis by BcPG3 and BcPG6 leads to the immediate accumulation of monomers and dimers. The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana. They showed different NAs on these plants. BcPG1 and BcPG2 possessed the strongest NA as tissue collapse was observed within 10 min after infiltration of broad bean leaves. The amino acid (aa) D192A substitution in the active site of BcPG2 not only abolished enzyme activity but also the NA, indicating that the NA is dependent on enzyme activity. Furthermore, deletion of the Bcpg2 gene in B. cinerea resulted in a strong reduction in virulence on tomato and broad bean. Primary lesion formation was delayed by approximately 24 h and the lesion expansion rate was reduced by 50-85%. These data indicate that BcPG2 is an important virulence factor for B. cinerea.
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Claims 13
Claim · model-written summary
Deletion of the Bcpg2 gene in B. cinerea strongly reduced its virulence on tomato and broad bean.
- Taxon
- tomato and broad bean
Vicia faba
Solanum lycopersicume5 - Treatment
- deletion of the Bcpg2 genee5
- Interactor
- B. cinerea
Botrytis cinereae5pathogen - Gene
- Bcpg2e5
- Outcome
- virulencee5
- Direction
- reductione5
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e5“Furthermore, deletion of the Bcpg2 gene in B. cinerea resulted in a strong reduction in virulence on tomato and broad bean”
Claim · model-written summary
Deletion of the Bcpg2 gene in B. cinerea delayed primary lesion formation by approximately 24 h.
- Taxon
- tomato and broad beanVicia fabaSolanum lycopersicume5
- Treatment
- deletion of the Bcpg2 genee5
- Interactor
- B. cinereaBotrytis cinereae5pathogen
- Outcome
- Primary lesion formatione6
- Direction
- delayede6
- Value
- approximately 24 he6
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e5“Furthermore, deletion of the Bcpg2 gene in B. cinerea resulted in a strong reduction in virulence on tomato and broad bean”
e6“Primary lesion formation was delayed by approximately 24 h”
Claim · model-written summary
Deletion of the Bcpg2 gene in B. cinerea reduced the lesion expansion rate by 50-85%.
- Taxon
- tomato and broad beanVicia fabaSolanum lycopersicume5
- Treatment
- deletion of the Bcpg2 genee5
- Interactor
- B. cinereaBotrytis cinereae5pathogen
- Outcome
- lesion expansion ratee7
- Direction
- reducede7
- Value
- 50-85%e7
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e5“Furthermore, deletion of the Bcpg2 gene in B. cinerea resulted in a strong reduction in virulence on tomato and broad bean”
e7“the lesion expansion rate was reduced by 50-85%”
Claim · model-written summary
BcPG1 and BcPG2 had the strongest necrotizing activity, causing tissue collapse within 10 min in broad bean leaves.
- Taxon
- broad beanVicia fabae1
- Treatment
- infiltratione1
- Gene
- BcPG1 and BcPG2e1
- Outcome
- strongest NAe1
- Direction
- strongeste1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“BcPG1 and BcPG2 possessed the strongest NA as tissue collapse was observed within 10 min after infiltration of broad bean leaves”
Claim · model-written summary
Substitution D192A in the active site of BcPG2 abolished both its enzyme activity and its necrotizing activity, so necrosis depends on enzyme activity.
- Taxon
- tomato, broad bean and Arabidopsis thaliana
Arabidopsis thalianaVicia fabaSolanum lycopersicume0 - Treatment
- D192A substitution in the active site of BcPG2e1
- Gene
- BcPG2e1
- Outcome
- enzyme activity but also the NAe1
- Direction
- abolishede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“The amino acid (aa) D192A substitution in the active site of BcPG2 not only abolished enzyme activity but also the NA”
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
Claim · model-written summary
PGA hydrolysis by BcPG1, BcPG2 and BcPG4 transiently accumulates oligomers with DP < 7.
- Taxon
- tomato, broad bean and Arabidopsis thalianaArabidopsis thalianaVicia fabaSolanum lycopersicume0
- Gene
- BcPG1, BcPG2 and BcPG4e1
- Outcome
- transient accumulation of oligomers with DP < 7e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
e1“PGA hydrolysis by BcPG1, BcPG2 and BcPG4 leads to the transient accumulation of oligomers with DP < 7”
Claim · model-written summary
PGA hydrolysis by BcPG3 and BcPG6 immediately accumulates monomers and dimers.
- Taxon
- tomato, broad bean and Arabidopsis thalianaArabidopsis thalianaVicia fabaSolanum lycopersicume0
- Gene
- BcPG3 and BcPG6e1
- Outcome
- immediate accumulation of monomers and dimerse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
e1“PGA hydrolysis by BcPG3 and BcPG6 leads to the immediate accumulation of monomers and dimers”
Claim · model-written summary
The maximum apparent Vmax of hydrolysis of polygalacturonic acid ranged from 10 to 900 U mg(-1) among the five BcPGs.
- Taxon
- tomato, broad bean and Arabidopsis thalianaArabidopsis thalianaVicia fabaSolanum lycopersicume0
- Outcome
- V(max,app)e1
- Value
- 10 to 900 U mg(-1)e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
e1“the V(max,app) ranged from 10 to 900 U mg(-1)”
Claim · model-written summary
The apparent Km of hydrolysis of polygalacturonic acid ranged from 0.16 to 0.6 mg ml(-1) among the five BcPGs.
- Taxon
- tomato, broad bean and Arabidopsis thalianaArabidopsis thalianaVicia fabaSolanum lycopersicume0
- Outcome
- K(m,app)e1
- Value
- 0.16 to 0.6 mg ml(-1)e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
e1“while the K(m,app) ranged from 0.16 to 0.6 mg ml(-1)”
Claim · model-written summary
The five BcPGs had similar pH optima of approximately pH 4.5.
- Taxon
- tomato, broad bean and Arabidopsis thalianaArabidopsis thalianaVicia fabaSolanum lycopersicume0
- Outcome
- pH optimae1
- Value
- approximately pH 4.5e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
e1“the pH optima of the five enzymes were similar (approximately pH 4.5)”
Claim · model-written summary
The maximum activity of individual BcPG enzymes differed significantly.
- Taxon
- tomato, broad bean and Arabidopsis thalianaArabidopsis thalianaVicia fabaSolanum lycopersicume0
- Outcome
- maximum activity of individual enzymese1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
e1“the maximum activity of individual enzymes differed significantly”
Claim · model-written summary
All five BcPGs are true endopolygalacturonases that nonetheless differ in mode of action.
- Taxon
- tomato, broad bean and Arabidopsis thalianaArabidopsis thalianaVicia fabaSolanum lycopersicume0
- Gene
- BcPGse1
- Outcome
- true endopolygalacturonasese1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
e1“Although all BcPGs are true endopolygalacturonases, they apparently have different modes of action”
Claim · model-written summary
The five BcPGs showed different necrotizing activities on the test plants.
- Taxon
- tomato, broad bean and Arabidopsis thalianaArabidopsis thalianaVicia fabaSolanum lycopersicume0
- Outcome
- different NAse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana”
e1“They showed different NAs on these plants”
The same questions in other papers
loss of function · Botrytis cinerea · Solanum lycopersicum (tomato)Positive and negative results
7 papers in the same direction as this paper · 18 in the opposite direction
- negative
- negative
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See all 26 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.