Journal Title
Title of Journal: Tree Genetics Genomes
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Abbravation: Tree Genetics & Genomes
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Publisher
Springer-Verlag
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Authors: John L Norelli Robert E Farrell Carole L Bassett Angela M Baldo Donna A Lalli Herb S Aldwinckle Michael E Wisniewski
Publish Date: 2008/06/27
Volume: 5, Issue: 1, Pages: 27-40
Abstract
Fire blight caused by the bacterium Erwinia amylovora is a destructive disease of many tree and shrub species of the Rosaceae Suppression subtractive cDNA hybridization SSH was used to identify genes that are differentially up and downregulated in apple Malus x domestica in response to challenge with E amylovora cDNA libraries were constructed from E amylovora and mockchallenged apple leaf tissue at various time intervals after challenge treatment ranging from 025 to 72 h postinoculation hpi and utilized in SSH Gel electrophoresis of PCRamplified SSH cDNAs indicated a greater quantity and size diversity in the downregulated EST population at early times after challenge 1 and 2 hpi compared to early upregulated sequences and to sequences downregulated at later 24 and 48 hpi times after challenge A total of 468 nonredundant Malus ESTs isolated by SSH in response to E amylovora challenge were characterized by bioinformatic analysis Many of ESTs identified following E amylovora challenge of apple were similar to genes previously reported to respond to bacterial challenge in Arabidopsis thaliana The results indicate that there was a substantial early 1 and 2 hpi transcriptional response in apple to fire blight disease involving both the down and upregulation of host genes Additionally genes identified responding to fire blight challenge early 1 and 2 hpi differed from those identified later 25 48 and 72 hpi in the infection processWe gratefully acknowledge Wilbur Hershberger USDA ARS Kearneysville WV USA for valuable assistance with conducting biological challenge experiments isolating RNA from challenge tissues cloning PCRamplified SSHcDNAs and editing of SSHEST sequences and for bioinformatic analyses Dr David Needleman USDA ARS Wyndmoor PA USA of the Eastern Regional Research Center’s Nucleic Acid Facility for sequencing the SSHESTs Greg Richart The Pennsylvania State University York USA for assistance with SSH Jing Ma USDA ARS Kearneysville USA for assistance with semiquantitative RTPCR John McGraw USDA ARS Kearneysville USA for assistance with sequence editing and bioinformatics and Dr Zuping Yang USDAARS Kearneysville USA for assistance with cloning PCRamplified SSHcDNAs The project was supported by the National Research Initiative of the USDA Cooperative State Research Education and Extension Service grant number 20053530015462 Additional support was provided by NutriCore NE
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