Manipulation of Vacuole-Mediated Redox Homeostasis by Cyst Nematode Effectors Promotes Feeding Site Formation and Parasitism
Manipulation of Vacuole-Mediated Redox Homeostasis by Cyst Nematode Effectors Promotes Feeding Site Formation and Parasitism
Cereal cyst nematode H. avenae (Treub) Chitwood were collected from an infested wheat field in Qingdao, China. The roots of Wild type and overexpressed wheat were inoculated with Ha nematodes, and the roots were collected 3 days later, with no inoculation as a negative control. The wheat cultivar Fielder was used for transgenic studies, plant-nematode interaction analyses, and parasitism assays.Both plant species were maintained in growth chambers under controlled conditions (23°C, 16-h light/8-h dark photoperiod). First, collect cyst nematodes (Heterodera avenae) from an infested field in Qingdao, China. Then, inoculate wild-type and overexpression wheat roots with these nematodes, while maintaining a non-inoculated set of plants as a negative control. Finally, collect all root samples three days after inoculation. Total RNA was extracted using TRIzol reagent (Invitrogen, catalogue number 15596018). First-strand cDNA was synthesized from 1 μg of total RNA using the PrimeScriptTM RT reagent Kit with gDNA Eraser (Takara).  First, fragmented DNA undergoes end repair and A-tailing to create compatible ends. Then, sequencing adapters are ligated to the fragments, followed by purification and PCR amplification to generate the final library.