Heat shock factor ZmHsf17 positively regulates phosphatidic acid phosphohydrolase ZmPAH1 and enhances maize thermotolerance

Heat stress adversely impacts plant growth, development, and grain yield. Heat shock factors (Hsf), especially the HsfA2 subclass, play a pivotal role in the transcriptional regulation of genes in response to heat stress. In this study, the coding sequence of maize ZmHsf17 was cloned. ZmHsf17 contained conserved domains including a DNA binding domain, oligomerization domain, and transcriptional activation domain. The protein was nuclear localized and had transcription activation activity. Yeast two-hybrid and split luciferase complementation assays confirmed the interaction of ZmHsf17 with members of the maize HsfA2 subclass. Overexpression of ZmHsf17 in maize significantly increased chlorophyll content and net photosynthetic rate, and enhanced the stability of cellular membranes. Through integrative analysis of ChIP-seq and RNA-seq datasets, ZmPAH1, encoding phosphatidic acid phosphohydrolase of lipid metabolic pathways, was identified as a target gene of ZmHsf17. The promoter fragment..., Plant materials and growth conditions
Stress treatments
Isolation and preparation of CDS
Cloning of ZmHsf17 CDS
The instructions for using the HEATSTER platform
Subcellular localization
Transcription activity and yeast two-hybrid assay
Split-luciferase complementation
Real-time quantitative reverse transcription PCR (qRT-PCR)
SDS-PAGE and Immunoblot analysis
ChIP-seq analysis
RNA-seq analysis
ChIP assay followed by qPCR (ChIP-qPCR)
Yeast one-hybrid assay (Y1H)
Electrophoretic mobility shift assay (EMSA)
Dual luciferase reporter assay
Lipidomics analysis by LC-MS/MS, , # Data from: Heat shock factor ZmHsf17 positively regulates phosphatidic acid phosphohydrolase ZmPAH1 and enhances maize thermotolerance

The Supplementary data of JEXBOT/2024/313002.

## Description of the Data and file structure

The supplementary data include four tables named Table S1/S2 and one figure named Fig. S1 and the raw data of all experiments.

Table S1. The list of all the primers in this paper. 

In Table S1, the constructed plasmids are listed in the leftmost column, the names of the genes involved are in the middle column, and the last two columns are the forward and reverse primer sequences.

Table S2. Data analysis of ChIP-seq in *ZmHsf17* overexpression lines.

In Table S2, the 2,154 and 1,499 genes that are directly bound by ZmHsf17 at the standard temperature and 42 °C HS are listed, respectively. And 1,159 target genes that exhibit positive regulation in response to ZmHsf17 under HS conditions and the GO and KEGG enrichment annotation are listed in this Table.

...