RNA Isolation and Quality Assessment

TransZol Up (Transgen, Beijing, China) was used to isolate total RNA from each sample (IR-CK, no BPH; IR56-BPH infested, IR-IR56-BPH; TN1-BPH infested, IR-TN1-BPH) (Figure 9). The isolation of total RNA was performed, as described by Nanda et al. [23 (link)]. Subsequently, the purity and concentration of total RNAs were detected using a NanoPhotometer spectrophotometer (Thermo-Fischer Scientific, Waltham, MA, USA). Then, the integrity of isolated RNA was evaluated using a Qubit RNA Assay Kit coupled with Qubit 2.0 Flurometer (Thermo-Fischer Scientific, Waltham, MA, USA) from Agilent 2100 (Thermo-Fischer Scientific, Waltham, USA). To ensure the application of qualified samples for sequencing, electrophoresis of 1% (w/v) agarose gels was used to monitor degradation and contamination of RNA.