RNA-seq of Dendrobium nobile
For transcriptome sequencing, three replicates were set up for each set of samples, numbered DNL-A-1~ DNL-A-3, DNL-M-1~ DNL-M-3, and total RNA with a total amount of 1ug was selected, and the total RNA was used to build the transcriptome of the samples using the NEBNext Ultra II RNA Library Prep Kit for Illumina kit (New England Biolabs Inc; Ipswich, Massachusetts, USA) (Strand-Specific Library Building Kit NEBNext Ultra Directional RNA Library Prep Kit for Illumina) to prepare strand-specific libraries, which were Oligo(dT)-enriched and ion-interrupted, cDNA synthesis, double-end repair, junction ligation, AMPure XP beads screening, PCR amplification, and finally, the library was analyzed by Agilent 2100 Bioanalyzer (Agilent Technologies Inc, California, USA), Agilent High Sensitivity DNA Kit (Agilent Technologies Inc, California, USA, 5067-4626) for library quality and Pico green for total library concentration (Quantifluor-ST fluorometer, Promega, Madison, Wisconsin, USA, E6090; Quanti-Fluorescent Technology Inc. USA, E6090. Quant-iT PicoGreen dsDNA Assay Kit, Invitrogen, California, USA, P7589), qPCR quantitative detection of effective library concentration (StepOnePlus Real-Time PCR Systems, Thermo Scientific, Waltham, Massachusetts, USA) were performed before sequencing on an Illumina sequencer (Illumina Inc. , San Diego, CA, USA) for PE150 sequencing.