Genome‐Wide Identification of PHD Transcription Factor Family in Carya illinoinensis and Functional Characterization of CiPHD14 in Drought Stress

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Carya illinoinensis is a forestry species with significant economic and ecological importance. However, climate change may increase the frequency and severity of drought stress and natural disturbances, which could influence C. illinoinensis yields. Enhancing its drought resistance is therefore crucial for sustainable cultivation. The plant homeodomain (PHD)-finger gene family, widely identified across multiple species, plays crucial regulatory roles in stress responses. Notably, the Alfin-like (AL) subfamily has been demonstrated to participate in plant stress resistance. In this study, 62 CiPHDs were identified, of which 8 belonged to the AL subfamily. Comprehensive bioinformatics analyses were performed, and integrative analyses of transcriptome data and qRT-PCR confirmed that CiPHD14 and CiPHD61 exhibited upregulation in response to drought. Subcellular localization confirmed the nuclear positioning of CiPHD14 and CiPHD61, and yeast assays revealed no independent transcriptional activation. Dual-luciferase assay (Dual-LUC) and electrophoretic mobility shift assay (EMSA) revealed the binding of CiPHD14 and CiPHD61 to the G-rich element (5'-GTGGAG-3'). Transient overexpression of CiPHD14 in Nicotiana benthamiana significantly upregulated drought-responsive genes compared to controls. In addition, transgenic Arabidopsis thaliana lines with CiPHD14 overexpression showed enhanced drought resistance compared with wild-type (WT). This work elucidates CiPHD structures and functions while providing gene resources for breeding drought-tolerant C. illinoinensis.

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