Disruption of Botrytis cinerea pectin methylesterase gene Bcpme1 reduces virulence on several host plants.
The pectinolytic enzyme pectin methylesterase (PME) hydrolyses pectin in methanol and polygalacturonic acid. In the expressed sequence tag library of Botrytis cinerea T4, we identified a 1,041 bp Bcpme1 cDNA potentially encoding a 346-amino acid protein of 37 kDa showing 46.8% identity with Aspergillus sp. PMEs. Bcpme1 is a single copy gene and is similarly expressed in glucose and pectin containing media. To evaluate the role of Bcpme1 in Botrytis cinerea virulence, a mutant in Bcpme1 was generated by gene disruption. The Bcpme1 mutant showed similar growth on rich medium but reduced growth on pectin medium. Two isozymes of pI 7.4 and 7.1 were detected in pectin liquid-culture supernatants of wild-type strain Bd90 analyzed by isoelectric focusing-polyacrylamide gel electrophoresis, while those of Bcpme1 mutant possessed only the pI 7.1 isozyme. BCPME1, the pI 7.4 isozyme, is the major PME activity, as PME activity is 75% reduced in Bcpme1 mutant. Moreover, the Bcpme1 mutant was less virulent on apple fruits, grapevine, and Arabidopsis thaliana leaves. Those phenotypes were complemented by reintroducing a Bcpme1 copy in the Bcpme1 mutant. These results showed that B. cinerea possessed more than one PME-encoding gene and that BCPME1 is an important determinant of B. cinerea virulence.
Pathologie Végétale UMR217, Institut National Agronomique 75231 Paris, France.
Odile
O
Valette-Collet
Agnès
A
Cimerman
Philippe
P
Reignault
Caroline
C
Levis
Martine
M
Boccara
D000595:Amino Acid Sequence; D001483:Base Sequence; D020171:Botrytis / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity*; D002265:Carboxylic Ester Hydrolases / Q000235:genetics*; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology; D010944:Plants / Q000382:microbiology*; D015183:Restriction Mapping
D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase