DOI: 10.5061/dryad.r4xgxd2h6
title: Data from: Heat shock factor ZmHsf17 positively regulates phosphatidic acid phosphohydrolase ZmPAH1 and enhances maize thermotolerance

description: 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 of ZmPAH1 was bound by ZmHsf17 in protein–DNA
 interaction experiments in vivo and in vitro. Lipidomic data also
 indicated that the overexpression of ZmHsf17 increased levels of some
 critical membrane lipid components of maize leaves under heat stress. This
 research provides new insights into the role of the ZmHsf17–ZmPAH1 module
 in regulating thermotolerance in maize.
description: Plant materials and growth conditions Stress
 treatments Isolation and preparation of CDS
 Cloning of <em>ZmHsf17 </em>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
description: # 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. Fig. S1. The SDS-PAGE
 analysis of the isolation of ZmHsf17 and 6×His fusion protein. In Fig. S1,
 ZmHsf17 was prokaryotic expressed and purified in vitro, and the SDS-PAGE
 analysis of ZmHsf17 were shown. Each lane from left to right represents
 proteins marker, supernatant fluid, precipitate and MCAC eluent containing
 different concentrations of imidazole. Raw data of all charts. The raw
 data in Fig4, 5, 7, 8 and 9 of the manuscript JEXBOT/2024/313002. In the
 raw data file, Fig.4 shows the data of qPCR assays. Fig.5 includes the
 data of chlorophll content, Pn and REC of WT and overexpressing lines of
 ZmHsf17.  Fig.7 shows the data of qPCR of ZmPAH1 in ZmHsf17 overexpressing
 lines. Fig.8 shows the data of ChIP-qPCR assays of ZmPAH1 in
 overexpressing lines of ZmHsf17. Fig.9 shows raw data of lipid omics
 include the total content of all lipids and six lipid classes.