2.5 RNA sequencing
RNA was extracted from eighteen tissues (bark from stems, axillary buds,
immature female flowers, leaves (not fully expanded), mature leaves,
immature male inflorescence, mature male inflorescence, new shoots, leaf
buds, mature female flowers, receptive female flowers, immature fruit,
mature fruit, fruit epidermis, kernel, seed coat (testa), root, root
bark) (Table S1) sampled from one plant. An RNA-Seq library was produced
for each tissue using an NEBNext Ultra RNA Library Prep Kit (NEB,
Beverly, MA, USA). Paired end sequencing was performed on Illumina HiSeq
X Ten platform (Illumina, USA). After RNA quantification, we also pooled
equivalent amounts of RNA from each of the eighteen tissues for
full-length transcriptome sequencing. Using the purified mRNA as the
starting material, a full-length cDNA library (10-15kb) was constructed
for the PacBio Sequel platform (NEB, USA). Bioanalyzer 2100 software
(Panaro et al., 2000) was used to test the Library quality.