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► P. thornei and P. zeae are amenable to double stranded RNA-induced gene silencing via soaking. ► Using spermidine phosphate salt hexahydrate in soaking media results in more effective gene silencing. ► Silencing pat-10 and unc-87 of P. thornei reduces reproduction by 77–81% in carrot mini discs. ► Double stranded RNA from either nematode species silenced the corresponding gene in both species. ► RNA interference is demonstrated to be a potential control strategy for root lesion nematodes.
Root lesion nematodes (RLNs, Pratylenchus species) are a group of economically important migratory endoparasitic plant pathogens that attack host roots of major crops such as wheat and sugarcane, and can reduce crop yields by 7–15%. Pratylenchus thornei and Pratylenchus zeae were treated with double stranded RNA (dsRNA) to study gene silencing, (RNA interference, RNAi), as a potential strategy for their control. Mixed stages of nematodes of both species ingested dsRNA when incubated in a basic soaking solution in the presence of the neurostimulant octopamine. Incubation for up to 16h in soaking solutions containing 10–50mM octopamine, 0.1–1.0mg/mL FITC, and 0.5–6mM spermidine did not affect vitality. Spermidine phosphate salt hexahydrate rather than spermidine or spermidine trihydrochloride increased uptake of FITC by nematodes, and this resulted in more effective gene silencing. Silencing pat-10 and unc-87 genes of P. thornei and P. zeae resulted in paralysis and uncoordinated movements in both species, although to a higher degree in P. thornei. There was also a greater reduction in transcript of both genes in P. thornei indicating that it may be more susceptible to RNAi. For P. thornei treated with dsRNA of pat-10 and unc-87 there was a significant reduction (77–81%) in nematode reproduction on carrot mini discs over a 5week period. The results show that RLNs are clearly amenable to gene silencing, and that in planta delivery of dsRNA to target genes in these nematodes should confer host resistance. Moreover, for the two genes, dsRNA derived from either nematode species silenced the corresponding gene in both species. This implies cross-species control of nematodes via RNAi is possible.