To begin, turn on the glass needle puller, then set the heat to 280, the velocity to 170, the delay to 250, and the pole to 30. Bevel the tip of the capillary glass needle. Then using a micro injection pump, load approximately two microliters of ferritin heavy chain homology double-stranded RNA solution into the injection needle.
Transfer the agar plate containing Trichogramma dendrolimi pupae onto the compound microscope stage. Carefully insert the injection needle into the abdomen of a pupa at an approximately 30 degree angle and gradually inject five nanoliters of ferritin heavy chain homology double-stranded RNA solution. Incubate approximately 700 pupae per treatment at 25 degrees Celsius for 24 or 48 hours.
Extract RNA content from 100 pupae per replicate. From approximately 50 pupae per treatment, observe the wasps emergence. Finally, record the wasps emergence rate and deformity rate.
T.dendrolimi pupae injected with double-stranded ferritin heavy chain homology exhibited a significantly lower emergence rate compared to those injected with double-stranded GFP or without injection. In emerged wasps, 51.85%of T.dendrolimi wasps subjected to double-stranded ferritin heavy chain homology developed deformed small wings. This deformation was absent in wasps injected with double-stranded GFP or those without any injection.
Additionally, T.dendrolimi pupae injected with double-stranded ferritin heavy chain homology showed melanism, indicative of abnormal iron metabolism. The melanin was not observed in wasps injected with double-stranded GFP or those without any injection.
