Identification of genes expressed during the interaction between cv. Florina and V. inaequalis by differential display-Rt-Pcr

Abstract

In order to identify genes that are specifically regulated during the expression of the resistance mechanism of the apple cv. Florina towards the apple scab fungus, the genetic expression pattern in plants inoculated with a conidial suspension of Venturia inaequalis was compared with the expression pattern in plants inoculated with water, by differential display RT-PCR. Amplified products derived from the combination of one oligodT (dT_VG) with 20 10-mer primers were separated in polyacrylamide denaturing gel and detected with silver nitrate. 14 cDNA fragments that were only visible in the sample inoculated with fungal suspension, or more abundant in this one, were purified from the gel for reamplification and cloning. Three fragments were already sequenced and one sequence showed very strong homology with a plant phytochelatin synthetase, that might be involved in stress responses. Differential accumulation of this cDNA upon fungal inoculation shall be verified by reverse-transcription PCR with specific primers.