Location: Temperate Tree Fruit and Vegetable Research
Project Number: 2092-22000-022-044-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Jul 15, 2025
End Date: Sep 30, 2026
Objective:
Identify viral pathogens that could be developed or propagated as biological control agents for potato psyllid.
Approach:
Potato psyllid adults from various non-crop hosts of psyllids including matrimony vine, bittersweet nightshade, buffalo burr, hairy nightshade, and field bindweed. Psyllids will be collected from weed stands located in the major potato growing regions of the Columbia and Snake River basins. The psyllids will be examined for presence of morphological abnormalities and will be tested for the presence of all for viruses using RT-PCR. Haplotype and Lso infection status will be determined using standard molecular techniques. Results from Objective 1 will be used to determine if psyllid virus infection rates vary among regions, psyllid haplotypes, weedy host use, and Lso infection status.
Field collected psyllids will be subjected to the HTS analysis, using 10-20 pooled psyllids per collection site. Total RNA will be extracted using a TRIzol reagent-based protocol. DNAsetreated, purified RNA will then be depleted of ribosomal RNA using the RiboMinus kit for RNA-Seq. Before proceeding further, quality of resulting ribo-depleted RNA will be checked by fragment analysis. The ribo-depleted RNA will then be submitted to Psomagen (Frederick, MD) for library preparation, multiplexing, and sequencing on the Illumina NovaSeqX platform producing 150-bp paired end reads. Bioinformatics analysis of the high-throughput sequencing data will be performed using established methods. Any matches to the custom-prepared virus sequence database entries will be confirmed through the RT-PCR or PCR verification using specific primers, with subsequent Sanger-based sequencing of amplified PCR fragments.