Transcriptional Profiling of Rice Immune Response

Total RNA was extracted with an RNA Prep Pure Plant kit (Zomanbio, Beijing, China) following the manufacturer’s instructions. Each RNA sample (1 μg) was reverse-transcribed using the QuantiTect reverse transcription kit (Vazyme, Nanjing, China). The real-time quantitative PCR (RT-qPCR) was performed using an ABI 7500 system (Thermo Fisher Scientific, Waltham, MA, USA) with the SYBR Premix Ex Taq (Genestar, Beijing, China). Leaves of five-leaf-stage WT and mld1 plants were used to detect the expression of OsMLD1 and several defense marker genes. Two-leaf-stage WT seedling leaves, stems, and roots, as well as the adult flag leaves (1 L), top three leaves (2–4 L), young panicles, and stems, were used for tissue-specific expression analysis of OsMLD1, respectively. All of the experiments were performed with three technical replicates and three biological replicates. The OsACTIN gene in rice was used as an internal control. The sequences of all of the primers used in the study are listed in Table S7. Additionally, the primer information of some genes, including senescence-associated genes and PR genes, was obtained from previous studies [19 (link),22 (link)].