A polygalacturonase-inhibiting protein from grapevine reduces the symptoms of the endopolygalacturonase BcPG2 from Botrytis cinerea in Nicotiana benthamiana leaves without any evidence for in vitro interaction

D.A. Joubert, I. Kars, L. Wagemakers, C. Bergmann, G. Kemp, M.A. Vivier, J.A.L. van Kan

Research output: Contribution to journalArticleAcademicpeer-review

45 Citations (Scopus)

Abstract

Six endopolygalacturonases from Botrytis cinerea (BcPG1 to BcPG6) as well as mutated forms of BcPG1 and BcPG2 were expressed transiently in leaves of Nicotiana benthamiana using agroinfiltration. Expression of BcPG1, BcPG2, BcPG4, BcPG5, and mutant BcPG1-D203A caused symptoms, whereas BcPG3, BcPG6, and mutant BcPG2-D192A caused no symptoms. Expression of BcPG2 caused the most severe symptoms, including wilting and necrosis. BcPG2 previously has been shown to be essential for B. cinerea virulence. The in vivo effect of this enzyme and the inhibition by a polygalacturonase-inhibiting protein (PGIP) was examined by coexpressing Bcpg2 and the Vvpgip1 gene from Vitis vinifera in N. benthamiana. Coinfiltration resulted in a substantial reduction of the symptoms inflicted by the activity of BcPG2 in planta, as evidenced by quantifying the variable chlorophyll fluorescence yield. In vitro, however, no interaction between pure VvPGIP1 and pure BcPG2 was detected. Specifically, VvPGIP1 neither inhibited BcPG2 activity nor altered the degradation profile of polygalacturonic acid by BcPG2. Furthermore, using surface plasmon resonance technology, no physical interaction between VvPGIP1 and BcPG2 was detected in vitro. The data suggest that the in planta environment provided a context to support the interaction between BcPG2 and VvPGIP1, leading to a reduction in symptom development, whereas neither of the in vitro assays detected any interaction between these proteins
Original languageEnglish
Pages (from-to)392-402
JournalMolecular Plant-Microbe Interactions
Volume20
Issue number4
DOIs
Publication statusPublished - 2007

Fingerprint

Botrytis
Polygalacturonase
Nicotiana benthamiana
polygalacturonase
Botrytis cinerea
signs and symptoms (plants)
Tobacco
leaves
Plantae
Proteins
Surface Plasmon Resonance
proteins
Vitis
Chlorophyll
Virulence
agroinfiltration
Necrosis
surface plasmon resonance
Fluorescence
mutants

Keywords

  • phaseolus-vulgaris l
  • fungal polygalacturonases
  • cladosporium-fulvum
  • transgenic expression
  • cell-walls
  • pear pgip
  • gene
  • defense
  • infection
  • tomato

Cite this

@article{af318158974b4ec78cdfea3f2b747615,
title = "A polygalacturonase-inhibiting protein from grapevine reduces the symptoms of the endopolygalacturonase BcPG2 from Botrytis cinerea in Nicotiana benthamiana leaves without any evidence for in vitro interaction",
abstract = "Six endopolygalacturonases from Botrytis cinerea (BcPG1 to BcPG6) as well as mutated forms of BcPG1 and BcPG2 were expressed transiently in leaves of Nicotiana benthamiana using agroinfiltration. Expression of BcPG1, BcPG2, BcPG4, BcPG5, and mutant BcPG1-D203A caused symptoms, whereas BcPG3, BcPG6, and mutant BcPG2-D192A caused no symptoms. Expression of BcPG2 caused the most severe symptoms, including wilting and necrosis. BcPG2 previously has been shown to be essential for B. cinerea virulence. The in vivo effect of this enzyme and the inhibition by a polygalacturonase-inhibiting protein (PGIP) was examined by coexpressing Bcpg2 and the Vvpgip1 gene from Vitis vinifera in N. benthamiana. Coinfiltration resulted in a substantial reduction of the symptoms inflicted by the activity of BcPG2 in planta, as evidenced by quantifying the variable chlorophyll fluorescence yield. In vitro, however, no interaction between pure VvPGIP1 and pure BcPG2 was detected. Specifically, VvPGIP1 neither inhibited BcPG2 activity nor altered the degradation profile of polygalacturonic acid by BcPG2. Furthermore, using surface plasmon resonance technology, no physical interaction between VvPGIP1 and BcPG2 was detected in vitro. The data suggest that the in planta environment provided a context to support the interaction between BcPG2 and VvPGIP1, leading to a reduction in symptom development, whereas neither of the in vitro assays detected any interaction between these proteins",
keywords = "phaseolus-vulgaris l, fungal polygalacturonases, cladosporium-fulvum, transgenic expression, cell-walls, pear pgip, gene, defense, infection, tomato",
author = "D.A. Joubert and I. Kars and L. Wagemakers and C. Bergmann and G. Kemp and M.A. Vivier and {van Kan}, J.A.L.",
year = "2007",
doi = "10.1094/MPMI-20-4-0392",
language = "English",
volume = "20",
pages = "392--402",
journal = "Molecular Plant-Microbe Interactions",
issn = "0894-0282",
publisher = "American Phytopathological Society",
number = "4",

}

A polygalacturonase-inhibiting protein from grapevine reduces the symptoms of the endopolygalacturonase BcPG2 from Botrytis cinerea in Nicotiana benthamiana leaves without any evidence for in vitro interaction. / Joubert, D.A.; Kars, I.; Wagemakers, L.; Bergmann, C.; Kemp, G.; Vivier, M.A.; van Kan, J.A.L.

In: Molecular Plant-Microbe Interactions, Vol. 20, No. 4, 2007, p. 392-402.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - A polygalacturonase-inhibiting protein from grapevine reduces the symptoms of the endopolygalacturonase BcPG2 from Botrytis cinerea in Nicotiana benthamiana leaves without any evidence for in vitro interaction

AU - Joubert, D.A.

AU - Kars, I.

AU - Wagemakers, L.

AU - Bergmann, C.

AU - Kemp, G.

AU - Vivier, M.A.

AU - van Kan, J.A.L.

PY - 2007

Y1 - 2007

N2 - Six endopolygalacturonases from Botrytis cinerea (BcPG1 to BcPG6) as well as mutated forms of BcPG1 and BcPG2 were expressed transiently in leaves of Nicotiana benthamiana using agroinfiltration. Expression of BcPG1, BcPG2, BcPG4, BcPG5, and mutant BcPG1-D203A caused symptoms, whereas BcPG3, BcPG6, and mutant BcPG2-D192A caused no symptoms. Expression of BcPG2 caused the most severe symptoms, including wilting and necrosis. BcPG2 previously has been shown to be essential for B. cinerea virulence. The in vivo effect of this enzyme and the inhibition by a polygalacturonase-inhibiting protein (PGIP) was examined by coexpressing Bcpg2 and the Vvpgip1 gene from Vitis vinifera in N. benthamiana. Coinfiltration resulted in a substantial reduction of the symptoms inflicted by the activity of BcPG2 in planta, as evidenced by quantifying the variable chlorophyll fluorescence yield. In vitro, however, no interaction between pure VvPGIP1 and pure BcPG2 was detected. Specifically, VvPGIP1 neither inhibited BcPG2 activity nor altered the degradation profile of polygalacturonic acid by BcPG2. Furthermore, using surface plasmon resonance technology, no physical interaction between VvPGIP1 and BcPG2 was detected in vitro. The data suggest that the in planta environment provided a context to support the interaction between BcPG2 and VvPGIP1, leading to a reduction in symptom development, whereas neither of the in vitro assays detected any interaction between these proteins

AB - Six endopolygalacturonases from Botrytis cinerea (BcPG1 to BcPG6) as well as mutated forms of BcPG1 and BcPG2 were expressed transiently in leaves of Nicotiana benthamiana using agroinfiltration. Expression of BcPG1, BcPG2, BcPG4, BcPG5, and mutant BcPG1-D203A caused symptoms, whereas BcPG3, BcPG6, and mutant BcPG2-D192A caused no symptoms. Expression of BcPG2 caused the most severe symptoms, including wilting and necrosis. BcPG2 previously has been shown to be essential for B. cinerea virulence. The in vivo effect of this enzyme and the inhibition by a polygalacturonase-inhibiting protein (PGIP) was examined by coexpressing Bcpg2 and the Vvpgip1 gene from Vitis vinifera in N. benthamiana. Coinfiltration resulted in a substantial reduction of the symptoms inflicted by the activity of BcPG2 in planta, as evidenced by quantifying the variable chlorophyll fluorescence yield. In vitro, however, no interaction between pure VvPGIP1 and pure BcPG2 was detected. Specifically, VvPGIP1 neither inhibited BcPG2 activity nor altered the degradation profile of polygalacturonic acid by BcPG2. Furthermore, using surface plasmon resonance technology, no physical interaction between VvPGIP1 and BcPG2 was detected in vitro. The data suggest that the in planta environment provided a context to support the interaction between BcPG2 and VvPGIP1, leading to a reduction in symptom development, whereas neither of the in vitro assays detected any interaction between these proteins

KW - phaseolus-vulgaris l

KW - fungal polygalacturonases

KW - cladosporium-fulvum

KW - transgenic expression

KW - cell-walls

KW - pear pgip

KW - gene

KW - defense

KW - infection

KW - tomato

U2 - 10.1094/MPMI-20-4-0392

DO - 10.1094/MPMI-20-4-0392

M3 - Article

VL - 20

SP - 392

EP - 402

JO - Molecular Plant-Microbe Interactions

JF - Molecular Plant-Microbe Interactions

SN - 0894-0282

IS - 4

ER -