Journal of Experimental Botany, Vol. 54, No. 383, pp. 633-645,
February 1, 2003
© 2003 Oxford University Press
Cloning and characterization of two ripening-related strawberry (Fragaria x ananassa cv. Chandler) pectate lyase genes
Received 10 June 2002; Accepted 26 September 2002
Departamento de Bioquímica y Biología Molecular, Edificio Severo Ochoa (Edificio C-6), Campus Universitario de Rabanales, Universidad de Córdoba, 14071 Córdoba, Spain
1 These authors contributed equally to this paper.
2 To whom correspondence should be addressed. Fax: +34 957 211079. E-mail: bb1mublj{at}uco.es
Two genomic clones corresponding to putative pectate lyase genes (plA and plB) were isolated and characterized in strawberry (Fragariaxananassa cv. Chandler). The corresponding ORFs for the plA and plB genes revealed deduced proteins of 451 and 439 amino acids, respectively, that differ from that of the previously isolated strawberry plC gene. Southern blot analysis has shown that while the plB gene is a single copy gene, the plA gene is probably encoded by a small multigene family. By using specific probes corresponding to the untranslated 3' terminal region of the pl genes, and QRT-PCR methodology, the spatio-temporal expression pattern of both strawberry pl genes have been compared with that of the plC gene. The three transcripts were specifically expressed only in fruit and mainly during the ripening stages. Moreover, the expression of the plA and plB genes was induced in green de-achened fruit, but this increase was reduced by the external application of auxins as was the expression of plC. The expression of both pl genes was also strongly reduced in harvested fruit kept in controlled atmosphere (CA) containing high CO2 levels. Immunolocalization studies using antibodies raised against the strawberry PL proteins placed the proteins in the cell wall of parenchymatic cells of the fruit receptacle. The role of pl genes in cell-wall disassembly and fruit ripening softening is discussed.
Key words: Fruit-ripening, pectate lyase, softening, strawberry.
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