|Norelli, John (jay) - Jay|
|Farrell, Jr., Robert|
Submitted to: Plant and Animal Genome Conference
Publication Type: Abstract Only
Publication Acceptance Date: 10/20/2006
Publication Date: 1/13/2007
Citation: Norelli, J.L., Bassett, C.L., Farrell, Jr., R.E., Baldo, A.M., Malnoy, M., Borejsza-Wysocka, E., Lalli, D., Korban, S.S., Gasic, K., Wisniewski, M.E., Aldwinckle, H.S. 2007. Functional genomic response of apple to fire blight. Plant and Animal Genome XV Conference. Final Abstract Guide, P 465, pg 217. Interpretive Summary:
Technical Abstract: The goal of this project is to use a functional genomic analysis to characterize the response of apple (Malus x domestica) to fire blight disease and in doing so, identify new opportunities for improving fire blight resistance. cDNA suppression subtractive hybridization and cDNA-AFLP analysis were used to identify genes expressed in apple in response to Erwinia amylovora infection. Additionally, bioinformatic approaches were used to identify publicly available apple ESTs uniquely associated with E. amylovora infected apple or similar to Arabidopsis ESTs associated with Pseudomonas syringae pv. tomato infection. Gene silencing is being used to elucidate the role of candidate genes in resistance and susceptibility. A multi-vector transformation approach is being evaluated for the high-throughput generation of RNAi mutants in apple. M.26 apple tissue was transformed with three single pHellsgate8-derived vectors and a mixture of the three vectors. Transformation frequency was not adversely affected by the use of multiple silencing vectors. Universal PCR primers were developed for pHellsgate8-derived vectors that can: 1) detect the presence of single or multiple EST silencing insertions in the RNAi transgenic and 2) provide sequencing template to determine EST contained in silencing insertion. A resistance assay was developed to rapidly screen apple-RNAi transgenics for changes in resistance and susceptibility to fire blight. pHellsgate8-derived silencing vectors are being constructed for 19 Malus ESTs identified as having a high-probability of being associated with response to infection. This project is supported by a National Research Initiative Competitive Grant 2005-35300-15462 from the USDA Cooperative State Research, Education, and Extension Service.