Premium
Characterization of a defensin in bark and fruit tissues of peach and antimicrobial activity of a recombinant defensin in the yeast, Pichia pastoris
Author(s) -
Wisniewski Michael E.,
Bassett Carole L.,
Artlip Timothy S.,
Webb Robert P.,
Janisiewicz Wojciech J.,
Norelli John L.,
Goldway Martin,
Droby Samir
Publication year - 2003
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1046/j.1399-3054.2003.00204.x
Subject(s) - pichia pastoris , biology , defensin , cdna library , complementary dna , prunus , bark (sound) , recombinant dna , microbiology and biotechnology , yeast , pichia , antimicrobial , gene , botany , biochemistry , ecology
Herein the cloning and characterization of a defensin gene ( PpDfn1 ) from a cDNA library made from peach ( Prunus persica [L.] Batsch) winter bark tissues is described. A partial clone obtained from the library was extended to full length by 5′ Rapid Amplification of cDNA Ends (RACE). The open reading frame of 237 bp codes for a 79 amino acid peptide related to the defensin family of proteins. Sequence comparison of the encoded protein using blast analysis revealed significant homology to defensins from other plant species. RNA gel blot analysis indicated that the gene is seasonally expressed in bark tissues of 1‐year‐old shoots, and is also expressed in early fruit development. Results of quantitative RT‐PCR and protein blot analysis were similar to those of RNA gel blot analyses for the bark tissues. A recombinant version, rDFN1 was expressed in the yeast, Pichia pastoris . It was found that rDFN1 inhibited germination of the fungal pathogens Penicillium expansum and Botrytis cinerea , but not the Gram‐negative bacterium Erwinia amylovora . The potential physiological role of PpDFN1 and its antimicrobial properties are discussed.