Ectopic expression of an esterase, which is a candidate for the unidentified plant cutinase, causes cuticular defects in Arabidopsis thaliana.
Cutinase is an esterase that degrades the polyester cutin, a major component of the plant cuticle. Although cutinase activity has been detected in pollen, the genes encoding this enzyme have not been identified. Here, we report the identification and characterization of Arabidopsis CDEF1 (cuticle destructing factor 1), a novel candidate gene encoding cutinase. CDEF1 encodes a member of the GDSL lipase/esterase family of proteins, although fungal and bacterial cutinases belong to the alpha/beta hydrolase superfamily which is different from the GDSL lipase/esterase family. According to the AtGenExpress microarray data, CDEF1 is predominantly expressed in pollen. The ectopic expression of CDEF1 driven by the 35S promoter caused fusion of organs, including leaves, stems and flowers, and increased surface permeability. Ultrastructural analysis revealed that the cuticle of the transgenic plants was often disrupted and became discontinuous. Subcellular analysis with green fluorescent protein (GFP)-tagged CDEF1 showed that the protein is secreted to the extracellular space in leaves. The recombinant CDEF1 protein has esterase activity. These results are consistent with cutinase being secreted from cells and directly degrading the polyester in the cuticle. CDEF1 promoter activity was detected in mature pollen and pollen tubes, suggesting that CDEF1 is involved in the penetration of the stigma by pollen tubes. Additionally, we found CDEF1 expression at the zone of lateral root emergence, which suggests that CDEF1 degrades cell wall components to facilitate the emergence of the lateral roots. Our findings suggest that CDEF1 is a candidate gene for the unidentified plant cutinase.
Takahashi
Kentaro
K
Department of Botany, Graduate School of Science, Kyoto University, Kyoto, Japan.
Shimada
Tomoo
T
Kondo
Maki
M
Tamai
Atsushi
A
Mori
Masashi
M
Nishimura
Mikio
M
Hara-Nishimura
Ikuko
I
D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D002265:Carboxylic Ester Hydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism; D004950:Esterases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D005110:Extracellular Space / Q000201:enzymology; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D049452:Green Fluorescent Proteins; D046529:Microscopy, Electron, Transmission; D009154:Mutation / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D019441:Plant Epidermis / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D018517:Plant Roots / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D011058:Pollen / Q000201:enzymology / Q000235:genetics; D011091:Polyesters / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics
D011091:Polyesters; D049452:Green Fluorescent Proteins; D004950:Esterases; D002265:Carboxylic Ester Hydrolases; C019952:cutinase