FUNCTIONAL GENOMIC RESPONSE OF APPLE TO FIRE BLIGHT

1
FUNCTIONAL GENOMIC RESPONSE OF APPLE TO FIRE
BLIGHT

• PD John (Jay) Norelli, USDA-ARS
• Co-PD Herb Aldwinckle, Cornell
• Co-PD Carole Bassett, USDA-ARS
• Co-PD Bob Farrell, Jr., Penn State U.
• A project supported by the National Research
  Initiative of USDA-CSREES

2
Project Plan

• Goal characterize the response of apple to fire
  blight disease using functional genomics.
• Objectives
• Characterize ESTs associated with apples
  response to fire blight challenge.
• Develop high-throughput system for obtaining RNAi
  mutants from pools of ESTs associated with fire
  blight.

3
Project Plan

• Objective 1 characterize ESTs
• Host resistance resistant vs. susceptible
  cultivar
• Non-host resistance compatible (E. amylovora)
  vs. incompatible (P. s. tabaci)
• Response to effector proteins hrp strain vs.
  hrp- strain
• Response to DspA/E dspE vs. dspE-

4
Project Plan

• Objective 2 high-throughput RNAi mutants
• 3 sources of ESTs
• Korban/Aldwinckle NSF Project
• McNellis NSF Project
• USDA-ARS-AFRS CRIS Project
• GATEWAY technology to facilitate rapid generation
  of RNAi constructs
• Rapid cell necrosis and resistance assay to
  evaluate mutants

5
Project Deliverables

• Apple ESTs associated with fire blight
• low abundance messages
• associated with specifics aspects of fire blight
  response
• 540-5400 RNAi mutants

6
Project Impact

• identify new opportunities for improving fire
  blight resistance
• facilitate large scale reverse genetic analysis
  in apple

7
Project Participants Director

• Jay Norelli
• USDA, ARS, Kearneysville, WV
• cloning and analysis of ESTs, generate RNAi-EST
  vectors, analyze RNAi-EST apple mutants
• Wil Hershberger ESTs
• Vacant Post Doc

8
Project Participants Co-Director

• Herb Aldwinckle
• Cornell Univ., Geneva
• Generate and analyze RNAi-EST apple mutants
• Ewa Borejza-Wysocka apple transformation
• Mickael Malnoy cDNA-AFLP

9
Project Participants Co-Director

• Carole Bassett
• USDA, ARS Kearneysville, WV
• Analysis of gene expression, bioinformatics

10
Project Participants Co-Director

• Bob Farrell
• Penn State Univ., York
• cDNA subtractions

11
Project Participants Collaborators

• Tim McNellis, Penn State Univ., Univ. Park
• Provide ESTs from current NSF project
• Schuyler Korban, Univ. Illinois, Urbana
• Provide ESTs from completed NSF project
• Angela Baldo, USDA-ARS, Geneva, NY
• Bioinformatics