Location: Temperate Tree Fruit and Vegetable Research
Project Number: 2092-21220-002-012-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Jul 15, 2016
End Date: Jul 14, 2020
Objective:
Characterization of virulent (Roza Pathotype) and avirulent (Race 1) Columbia Root Knot Nematode penetration during root and tuber development in S. bulbocastanum SB 22 and resistance introgressed S. tuberosum PA99N82-4
Gene expression profiling for resistance and susceptibility reactions during various stages of CRKN infection.
Approach:
Clones SB22 and PA99N82-4 will be inoculated with virulent and avirulent pathotypes of M. chitwoodi at 15 and 45 days after planting (DAP). Two reps per treatment will be carried out in the greenhouse. All the plants will be generated through tissue culture to avoid differences in the germination. Phenotyping on the response of nematode penetration and infection will be carried out at before nematode inoculation and 24hr, 5 and 10 days post inoculation. For first inoculation at 15 DAP, phenotype profiling will be carried out on roots and for second inoculation at 45DAP, phenotype profiling will be carried out on tubers .
Gene expression profiling for resistance and susceptibility will be carried out by extraction of RNA at the above mentioned time periods from roots and tubers. RNA-Seq using Illumina HiSeq 2000 will be carried out at the Center for Genome Research and Biocomputing, Oregon State University. A bioinformatics pipeline will be developed to study differential expression of genes associated with resistance and susceptibility during various stages of root and tuber development. We are currently in the process of whole genome sequencing of S. bulbocastanum SB 22. The genome information from our study will be used to identify candidates genes responsible for resistance and hypersensitive responses.
