Identification and characterization of genes in response to DBP and 2,4-DTBP stress in Glehnia littoralis seedlings

Mining genes responsive to allelochemicals dibutyl phthalate (DBP) and 2,4-di-tert-butylphenol (2,4-DTBP) stress in Glehnia littoralis to provide a molecular-level scientific reference for addressing continuous cropping obstacles. This study utilized the Illumina MiSeq high-throughput sequencing platform to treat G. littoralis seedlings with exogenous allelochemicals (DBP and 2,4-DTBP). Key genes responsive to allelopathic stress were screened, and their structures and functions were analyzed. The expression of these key genes in different parts of G. littoralis was examined via qPCR, and codon usage bias in G. littoralis was evaluated. The key genes GLERF1 , GLPYL , and GLMAPKKK17-18 in G. littoralis responsive to allelopathic stress were successfully cloned. Bioinformatic predictions indicated that the proteins encoded by these three genes are hydrophilic and lack transmembrane structures. Spatial expression analysis revealed that GLERF1 and GLPYL exhibit similar expression patterns across different tissues, with the highest expression levels observed in leaves and the lowest in roots. In contrast, GLMAPKKK17_18 showed the opposite trend, displaying minimal expression in leaves but high expression in roots, flowers, and other organs. Codon usage bias analysis revealed that G. littoralis prefers codons ending with A/U. Compared with mutational pressure, natural selection exerts a greater influence on the codon usage preference of most G. littoralis genes. Subcellular localization prediction indicated that the proteins encoded by these three genes are all expressed in the nucleus. This study provides a scientific basis for subsequent continuous cropping, genetic breeding, and resource conservation of G. littoralis .