Genome‑wide analysis of the GT8 gene family in apple and functional identification of MhGolS2 in saline-alkali tolerance.

Members of the glycosyltransferase 8 (GT8) family play an important role in regulating gene expression in response to many kinds of biotic and abiotic stress. In this study, 56 members of the apple GT8 family were identified, and their gene structure, phylogenetic relationships, chromosomal localization, and promoter cis-acting elements were comprehensively analyzed. Subsequently, 20 genes were randomly selected from the evolutionary tree for qRT-PCR detection, and it was found that MhGolS2 was significantly overexpressed under stress conditions. MhGolS2 was isolated from M.halliana and transgenic Arabidopsis thaliana, tobacco and apple callus tissues were successfully obtained. The transgenic plants grew better under stress conditions with higher polysaccharide, chlorophyll and proline content, lower conductivity and MDA content, significant increase in antioxidant enzyme activities (SOD, POD, CAT) and maintenance of low Na⁺/K⁺ as compared to the wild type. Meanwhile, the expression levels of reactive oxygen species-related genes (AtSOD, AtPOD, and AtCAT), Na⁺ transporter genes (AtCAX5, AtSOS1, and AtHKT1), H⁺-ATPase genes (AtAHA2 and AtAHA8), and raffinose synthesis-related genes (AtSTS, AtRFS1, and AtMIPS) were significantly up-regulated, while the expression levels of K⁺ transporter genes (AtSKOR, AtHAK5) were reduced. Finally, the Y2H experiment confirmed the interaction between MhGolS2 and MhbZIP23, MhMYB1R1, MhbHLH60, and MhNAC1 proteins. The above results indicate that MhGolS2 can improve plant saline-alkali tolerance by promoting polysaccharide synthesis, scavenging reactive oxygen species, and increasing the activity of antioxidant enzymes. This provides excellent stress resistance genes for the stress response regulatory network in apple.