Salivary proteins are the initial contact between sedentary insect pests and their host plants. It is expected that one or more salivary proteins mediate the interaction between Hessian fly and wheat, in which a feeding site is established to the benefit of the fly. A survey of 52 loci annotated as insect secreted salivary proteins was conducted in 384 individuals evenly distributed among eight biotypes of Hessian fly (B, C, D, E, GP, L, O, and vH9). Amplicons were sequenced with Illumina, and sequence reads were aligned to the reference sequences from which primers had been designed. Positions of consistent base variation (998 in all) were identified and tabulated by biotype. No varying position was associated with biotype-wide virulence to any one of wheat resistance genes H3, H5, H6, H7/H8, H9, H11, H13, and H26. The multiplate pooling strategy utilized in this study is an effective, affordable way to reveal the genotype of hundreds of individuals at tens of genetic loci.
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