Location: Plant Gene Expression Center
Project Number: 2030-22000-008-00-D
Project Type: In-House Appropriated
Start Date: Apr 23, 2012
End Date: Apr 22, 2017
The overall aim of this project is to use genomic and molecular-genetic approaches to identify and characterize mechanisms that fine tune R-gene expression for informed deployment of resistance traits in crop protection strategies, Objective 1: Generate and use genome and transcriptome analyses tools for identification of candidate sequences involved in posttranscriptional regulation of R-gene expression. Objective 2: Use functional genomics to verify crop genome and transcriptome sequences that regulate R-gene expression post-transcriptionally. Objective 3: Use characterized properties of confirmed R-gene regulatory sequences to develop genomic resources for regulation of multiple R-genes in crops of the Solanaceae.
Objective 1: We will complete analysis of the ten MIR families we previously identified, update sequence databases and use updated databases and our established bioinformatic pipeline to identify potential additional MIR families. Objective 2: We will use 5’ RACE assays to verify miRNA sequence-specific targeting and cleavage of R and R-homologue transcripts using methods we established for validation of the activity of nta-miR1 and nta-miR2 that cleave the N transcripts. Objective 3: We will use three approaches to understand the potential impact of sRNA regulation on R-gene expression and resistance. The first approach will evaluate pathogen regulation of MIR precursor expression. The second approach will use R-gene miRNA and secondary siRNA sensors to determine the impact of different levels or activity of miRNAs on siRNA production and R-transcript levels. The third aim will determine if R-gene specific small RNAs impact pathogen resistance responses.