leaf-3
Total RNA was separately extracted from young leaves of each individual using TRIzol Reagent (Plant RNA Purification Reagent for plant tissue). The integrity and purity of the total RNA were assessed using a 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA, USA); total RNA was quantified using a NanoDrop ND-2000 (Thermo Scientific, Wilmington, DE, USA). 150-bp PE libraries were constructed following the manufacturers instructions for the Illumina TruSeqTM RNA sample preparation kit. The generated cDNA libraries were amplified using the Phusion DNA polymerase (NEB) and sequenced on the Illumina HiSeq xten.