Plant rna extraction kit

Maize stalk tissues were collected and frozen immediately in liquid nitrogen, and total RNA samples were isolated using the Plant RNA extraction kit (TIANDZ, Inc., Beijing, China). One microgram of total RNA was converted to cDNA using HiScript II RT SuperMix for qPCR (Vazyme, Nanjing, China) according to the manufacturer’s instructions. qRT-PCR reactions were performed with the Bio-Rad CFX96™ real-time PCR system using ChamQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China). Maize Ubc9 was used as an internal control (forward primer, 5′-acgaaggtcttccatccaaacatc-3′; reverse primer, 5′-gtttcatgggcaacaccacaatcg-3′), and the relative expression levels for target genes were calculated using the 2^–ΔΔCt method. qRT-PCRs were all performed with three biological replicates. The primers used for ZmVPS37A are as follows: ZmVPS37A-F 5′-gagctggagatttcctctcttc-3′ and ZmVPS37A-R 5′-gaagactgcggactatctgaagc-3′.