Characterization and silencing of the fatty acid- and retinol-binding Pp-far-1 gene in Pratylenchus penetrans

Fatty acid-and retinol-binding proteins (FARs) are unique to nematodes, and are implicated in a wide range of metabolic and parasitic related functions. Along with the in silico analyses performed in this study, three different FAR members of this family were identified in Pratylenchus penetrans. The cDNA corresponding to the fatty acid-and retinol-binding Pp-far-1 gene was cloned and herein characterized at the molecular level for the first time for this genus. The translated 185 amino acid sequence of P. penetrans FAR-1 sequence, with a predicted molecular weight of 20.82 kDa and a pI of 5.49, shares highest sequence identity to FARs of other migratory nematodes of the Pratylenchidae family (90% to P. vulnus and 80% to Radopholus similis). In situ hybridization localizes Pp-far-1 transcripts in the hypodermis of the nematode. RT-qPCR detected Pp-far-1 transcripts for all nematode developmental stages, with highest expression levels found in juveniles, adult females and adult males, respectively. Pp-far-1 is also highly expressed during infection and establishment of P. penetrans in roots of different host plants, such as corn, lily and soybean. The importance of Pp-far-1 was studied by in planta RNA interference (RNAi) assays using stable soybean hairy root lines. Targeting Pp-far-1 decreased expression of the nematode Pp-far-1 and significantly reduced the reproduction of nematodes, with a total of between 44% and 70% fewer nematodes in comparison to the average number of nematodes counted for control lines. The results indicate that suppressing the expression levels of Pp-far-1 can act as an effective target gene to control P. penetrans.