ZmHSFA2B self-regulatory loop is critical for heat tolerance in maize.

The growth and development of maize (Zea mays L.) are significantly impeded by prolonged exposure to high temperatures. Heat stress transcription factors (HSFs) play crucial roles in enabling plants to detect and respond to elevated temperatures. However, the genetic mechanisms underlying the responses of HSFs to heat stress in maize remain unclear. Thus, we aimed to investigate the role of ZmHSFA2B in regulating heat tolerance in maize. Here, we report that ZmHSFA2B has two splicing variants, ZmHSFA2B-I and ZmHSFA2B-II. ZmHSFA2B-I encodes full-length ZmHSFA2B (ZmHSFA2B-I), whereas ZmHSFA2B-II encodes a truncated ZmHSFA2B (ZmHSFA2B-II). Overexpression of ZmHSFA2B-I improved heat tolerance in maize and Arabidopsis thaliana, but it also resulted in growth retardation as a side effect. RNA-sequencing and CUT&Tag analyses identified ZmMBR1 as a putative target of ZmHSFA2B-I. Overexpression of ZmMBR1 also enhanced heat tolerance in Arabidopsis. ZmHSFA2B-II was primarily synthesized in response to heat stress and competitively interacted with ZmHSFA2B-I. This interaction consequently reduced the DNA-binding activities of ZmHSFA2B-I homodimers to the promoter of ZmMBR1. Subsequent investigations indicate that ZmHSFA2B-II limits the transactivation and tempers the function of ZmHSFA2B-I, thereby reducing the adverse effects of excessive ZmHSFA2B-I accumulation. Based on these observations, we propose that the alternative splicing of ZmHSFA2B generates a self-regulatory loop that fine-tunes heat stress response in maize.