Maize Genomics and Genetics 2025, Vol.16, No.4, 202-218 http://cropscipublisher.com/index.php/mgg 207 the future, by constructing a non-coding RNA regulatory network at the whole-genome level of corn, it will help to comprehensively understand the transcriptional regulatory system of corn in response to high-temperature stress. Figure 1 Heat tolerance is modulated by ZmHsf20 (Adopted from Li et al. 2024) Image caption: A) Construction of CRISPR/Cas9-mediated Zmhsf20 knockout lines. Two sgRNAs that specifically target ZmHsf20 were designed, leading to the identification of two mutants, Zmhsf20-1 and Zmhsf20-2. black rectangles exons, white rectangles Un-Translated Regions (UTRs) and horizontal black lines rectangles introns. B) Relative ZmHsf20 transcript levels in the leaves of V2 stage seedlings of ZmHsf20-OE and wild type (WT, KN5585). ACTIN was used as the internal control. The error bars are based on three independent experiments. The values are means ±Sd (n = 3 independent experiments). ***P < 0.001, one-way ANOVA. C) Representative photographs of V2 stage seedlings of WT, Zmhsf20-1, and Zmhsf20-2 grown at 28 °C/22 °C (top) or exposed to 45 °C HS treatment for 3 d followed by a 3-d recovery (bottom). Scale bar = 2 cm. D) Representative photographs of V2 stage seedlings of WT, ZmHsf20-OE #1, and ZmHsf20-OE #1 grown at 28 °C/22 °C (top) or exposed to 45 °C for 2 d followed by a 3-d recovery (bottom). Scale bar = 2 cm. E, F) Survival rate of seedlings after recovery at 28 °C/22 °C for 3 d in (C, D). The error bars are based on three independent experiments. The values are means ± Sd (n = 3 independent experiments). *P < 0.05, **P < 0.01, ***P < 0.001, one-way ANOVA. G, H) Ion leakage rate of V2 stage seedlings grown at 28 °C/22 °C exposed to 45 °C for 1d. The error bars are based on three independent experiments. The values are means ±Sd (n = 3 independent experiments). *P < 0.05, ***P < 0.001, one-way ANOVA. I, J) Representative photographs of leaves from V2 stage seedlings grown at 28 °C/22 °C, exposed to 45 °C for 1 d, and stained with DAB, scale bar = 1.5mm (Adopted from Li et al. 2024)
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