Jina Chi

Growth Regulating Factors (GRFs) are plant-specific transcription factors that play crucial roles in regulating growth and development throughout the plant life cycle. A total of 34 Gossypium hirsutum GRF family genes were identified at the genome-wide level, which were unevenly distributed on 19 chromosomes, and were predicted to be mainly localized in the nucleus and plasma membrane. The number of GRF family genes varied greatly among different species, and they were categorized into four subfamilies (I–IV) according to their phylogenetic relationships. The G. hirsutum GRF genes possessed specific highly conserved structural domains, Trp-Arg-Cys motif (WRC) and Gln, Leu, Gln motif (QLQ), and structural analysis of the genes revealed that they contained 1–23 exons, and most of them contained UTRs. Intraspecies covariance analysis revealed that the GRF genes expanded in G. hirsutum by segmental duplication. The promoter region of the G. hirsutum GRF gene contained a large number of adversity stress response elements, as well as a small number of hormone response elements and growth and development-related response elements. Transcriptome data showed that the expression of G. hirsutum GRF genes was significantly higher in leaves than in other tissues, and some GRF genes responded to a variety of abiotic stresses. Additionally, transcriptomic sequencing revealed significantly higher expression levels of GhGRFs (e.g., GhGRF13/14/18) in embryonic callus (EC) compared to non-embryonic callus (NEC). This differential expression was validated by RT-qPCR, which confirmed that GhGRF13/14/16/20 were significantly upregulated in EC relative to NEC. These findings provide valuable candidate genes and molecular insights for improving G. hirsutum regeneration efficiency and yield-related traits through genetic manipulation, thereby accelerating the molecular breeding of elite G. hirsutum varieties.

Vascular Plant One-Zinc finger (VOZ) transcription factors are pivotal regulators of plant growth and stress adaptation, yet their functional roles in Gossypium hirsutum, a key fiber crop, remain poorly characterized. In this study, we systematically identified six VOZ genes in G. hirsutum and conducted a comprehensive analysis of their phylogenetic relationships, genomic distribution, promoter architecture, and expression profiles. Phylogenetic classification placed the GhVOZ proteins into three distinct clades, and chromosomal localization revealed that family expansion was likely driven by segmental duplication events. Promoter analysis uncovered an abundance of stress-related cis-regulatory elements, suggesting a potential role in abiotic stress signaling. Consistent with this, expression profiling demonstrated that GhVOZ1/3, GhVOZ2/4/5, and GhVOZ6 were specifically induced under drought, salt, and cold stress, respectively, with qRT-PCR further confirming their tissue-specific dynamic regulation under salt treatment. Furthermore, the GhVOZ family exhibited stage-specific expression patterns during somatic embryogenesis. GhVOZ1, GhVOZ3, and GhVOZ4 were upregulated at the early induction phase, implicating them in the initiation of cell reprogramming. In contrast, GhVOZ2 and GhVOZ4 showed sustained expression in embryogenic callus at later stages, suggesting a role in maintaining embryogenic competence, whereas GhVOZ5—preferentially expressed in non-embryogenic callus—may act as a repressor of embryogenesis. Synteny analysis further highlighted evolutionary conservation and subgenomic divergence of VOZ genes in G. hirsutum. Collectively, these findings establish GhVOZs as key regulators integrating abiotic stress response and somatic embryogenesis, providing a genetic framework for future functional studies and crop improvement.