Proline, a critical osmoregulatory compound, is integral to various plant stress responses. The P5CS gene, which encodes the rate-limiting enzyme in proline biosynthesis, known as ∆1-pyrroline-5-carboxylate synthetase, is fundamental to these stress response pathways. While the functions of P5CS genes in plants have been extensively documented, their specific roles in cotton remain inadequately characterized. In this study, we identified 40 P5CS genes across four cotton species with diverse sequence lengths and molecular weights. Phylogenetic analysis of 100 P5CS genes from nine species revealed three subgroups, with Gossypium hirsutum closely related to Gossypium barbadense. Collinearity analysis highlighted significant differences in collinear gene pairs, indicating evolutionary divergence among P5CS genes in tetraploid and diploid cotton. Exon–intron structures and conserved motifs correlated with phylogenetic relationships, suggesting functional differentiation. Stress-responsive elements in P5CS promoters suggest involvement in abiotic stress. Expression analysis under salt stress revealed differential expressions of GhP5CS genes, with GhP5CS1 emerging as a potential key regulator. Virus-induced gene silencing confirmed the pivotal role of GhP5CS1 in cotton’s salt stress response, as evidenced by increased salt sensitivity in the silenced plants. This study enhances our understanding of the functional diversity and roles of P5CS genes in cotton under stress conditions.

β-ketoacyl-CoA synthase (KCS) enzymes play a pivotal role in plants by catalyzing the first step of very long-chain fatty acid (VLCFA) biosynthesis. This process is crucial for plant development and stress responses. However, the understanding of KCS genes in maize remains limited. In this study, we present a comprehensive analysis of ZmKCS genes, identifying 29 KCS genes that are unevenly distributed across nine maize chromosomes through bioinformatics approaches. These ZmKCS proteins varied in length and molecular weight, suggesting functional diversity. Phylogenetic analysis categorized 182 KCS proteins from seven species into six subgroups, with maize showing a closer evolutionary relationship to other monocots. Collinearity analysis revealed 102 gene pairs between maize and three other monocots, whereas only five gene pairs were identified between maize and three dicots, underscoring the evolutionary divergence of KCS genes between monocotyledonous and dicotyledonous plants. Structural analysis revealed that 20 out of 29 ZmKCS genes are intronless. Subcellular localization prediction and experimental validation suggest that most ZmKCS proteins are likely localized at the plasma membrane, with some also present in mitochondria and chloroplasts. Analysis of the cis-acting elements within the ZmKCS promoters suggested their potential involvement in abiotic stress responses. Notably, expression analysis under abiotic stresses highlighted ZmKCS17 as a potential key gene in the stress response of maize, which presented an over 10-fold decrease in expression under salt and drought stresses within 48 h. This study provides a fundamental understanding of ZmKCS genes, paving the way for further functional characterization and their potential application in maize breeding for enhanced stress tolerance.

Proline, a critical osmoregulatory compound, is integral to various plant stress responses. The P5CS gene, which encodes the rate-limiting enzyme in proline biosynthesis, known as ∆1-pyrroline-5-carboxylate synthetase, is fundamental to these stress response pathways. While the functions of P5CS genes in plants have been extensively documented, their specific roles in cotton remain inadequately characterized. In this study, we identified 40 P5CS genes across four cotton species with diverse sequence lengths and molecular weights. Phylogenetic analysis of 100 P5CS genes from nine species revealed three subgroups, with Gossypium hirsutum closely related to Gossypium barbadense. Collinearity analysis highlighted significant differences in collinear gene pairs, indicating evolutionary divergence among P5CS genes in tetraploid and diploid cotton. Exon-intron structures and conserved motifs correlated with phylogenetic relationships, suggesting functional differentiation. Stress-responsive elements in P5CS promoters suggest involvement in abiotic stress. Expression analysis under salt stress revealed differential expression of GhP5CS genes, with GhP5CS1 emerging as a potential key regulator. Virus-induced gene silencing confirmed the pivotal role of GhP5CS1 in cotton’s salt stress response, as evidenced by increased salt sensitivity in the silenced plants. This study enhances our understanding of the functional diversity and stress response mechanisms of P5CS genes in cotton.