10th INTERNATIONAL VERTICILLIUM SYMPOSIUM 16-20 ...

IN VITRO ELICITATION OF PATHOGENICITY-RELATEDGENES IN VERTICILLIUM DAHLIAE USING POTATO ROOTEXTRACTSAHMED F. EL-BEBANY, MARIA A. HENRIQUEZ, MOHAMED A.BADAWI, LORNE R. ADAM, ABDELBASSET EL HADRAMI * AND FOUADDAAYF *University of Manitoba, Department of Plant Science, 222 Agriculture Building, Winnipeg, Manitoba,R3T 2N2, Canada.* E-mails: elhadrami@cc.umanitoba.ca; daayff@cc.umanitoba.caUsing two highly- and weakly-aggressive isolates of V. dahliae, elicited invitro with potato root extracts derived from a susceptible (Kennebec) or a moderatelyresistant cultivar (Ranger Russet), we investigated the differential accumulation ofinduced transcripts in this pathogen. A total of 573 differentially accumulatedtranscripts were detected in the two tested isolates in response to the elicitation usinga technique that combines subtractive hybridization and AFLP, along with 16primers’ combinations representing EcoRI/MseI AFLP adaptors +A/T/C/G to ensure afull coverage of the subtractive hybridization products. The number of differentiallyexpressed genes in response to the elicitation was slightly higher (5%) in the highlyaggressiveisolate as compared to the weakly-aggressive one. The recovery of 185selected transcripts from the SDS-PAGE followed by PCR re-amplification andsequencing revealed 41 matches with known sequences of assigned functions in V.dahliae pathogenicity (i.e., polygalacuronases) or with conserved hypotheticalproteins from NCBI or MIT GenBanks. The remaining sequences were unique anddid not match with any sequence currently available in these databases.67