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.