Three mg of the total RNAwas reverse transcribed to first-strand cDNA using an iScript cDNA synthesis kit (Bio-Rad, Hercules, CA) in a final reaction volume of 20 ml including 1 ml RNase Hþ Moloney murine leukemia virus-derived reverse transcriptase pre-blended
with RNase inhibitor, and 4 ml of 5 reaction mix containing an optimized blend of oligo (dT) and random primers. The reaction was conducted as follows: one cycle at 25 C for 5 min, followed by 42 C for 30 min, and 85 C for 5 min. The first-strand cDNAs were
diluted 10-fold with nuclease-free water and stored at 80 C for further use. locules. All the samples were quick-frozen in liquid nitrogen and
stored at 80 C for RNA isolation. Three bolls each from two
different plants (biological replicates) were used for each
treatment.