2.3 Plant treatment with BPH
Rice seedlings were placed in a 600 mL plastic pot, and cotton pads were placed at the base of each rice stem to prevent BPH from burrowing into the nutrient soil (Supporting Information: Figure S1). The differentially expressed genes (DEGs) were significantly enriched in the rice hormone signaling pathway after 8 h of sustained BPH attack, compared with other time intervals (Xu et al., 2021). This was also reflected in the hierarchical cluster analysis of DEGs between BPH-infested (Nl-Nip_1, Nl-Nip_2) and un-infested control (Nip_1, Nip_2) groups (Supporting Information: Figure S2). Therefore, samples used for library construction were collected from Nip or Nl-Nip rice leaf sheaths after 8 h of continuous feeding by 20 BPH female adults (Zha et al., 2023). Then, the BPHs were removed and stem samples were rapidly collected within the feeding region. The fresh weight of each sample containing four rice stems was not less than 2 grams, which were flash-frozen in liquid nitrogen and stored at -80℃ in a freezer. Un-infested rice leaf sheaths were collected as control, each treatment included two biological replicates. The rice samples infested by BPHs were named Nl-Nip_1 and Nl-Nip_2, while the control group was named Nip_1 and Nip_2.