In situ development and application of cDNA-AFLP to isolate genes of Candida oleophila (strain O) potentially involved in antagonistic properties against Botrytis cinerea.
The yeast Candida oleophila (strain O) presents a high level of protective activity against Botrytis cinerea (gray mold) on postharvest apples. The cDNA-AFLP technique allows the comparison of mRNA populations extracted from cells grown in different conditions. In order to isolate yeast genes potentially involved in biological control properties, that technique was applied on strain O cells growing on apple wounds. The biological control properties of 8 C. oleophila strains and strain O were assessed in order to compare the gene expression of a non antagonistic strain against gene expression of strain O. In the absence of a non-antagonistic strain, an other comparison model was designed. It was based on the growth of strain O in different in situ conditions: strain O applied on apple wounds (O), strain O applied on apple wounds in presence of B. cinerea (B) and B. cinerea alone on apple wounds (F). A recovering technique, based on the washing of cells in the wound and a RNA extraction method followed by a DNase treatment were optimised before cDNA-AFLP application. Thirteen primer pairs were used. Their application resulted in an average of 54 and 55 bands for O and B respectively whereas no bands were observed for F. Among these bands, 8 were expressed more intensely in presence of the pathogen (1.1% of the fragments).
Massart
S
S
Agricultural University of Gembloux Passage des Déportés 2, B-5030 Gembloux, Belgium. massart.s@fsagx.ac.be
Luna-Guarda
M
M
Jijakli
M H
MH
D020171:Botrytis / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D002175:Candida / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics*; D027845:Malus / Q000382:microbiology*; D010935:Plant Diseases / Q000382:microbiology*; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D012331:RNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping
D017931:DNA Primers; D018076:DNA, Complementary; D004271:DNA, Fungal; D012331:RNA, Fungal