Genome-wide identification of APX genes in flax (Linum usitatissimum) and functional characterization of LuAPX12 in osmotic and salinity stress responses

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Ascorbate peroxidase (APX) is a crucial antioxidant enzyme involved in regulating hydrogen peroxide levels and signal transduction pathways associated with development and stress responses. However, information about the APX gene family and its evolution and gene function in flax has not been reported. This study identified 12 APX genes in flax and analyzed their evolutionary relationships, motif compositions, gene structures, and cis -acting elements, while also predicting potential protein-protein interactions with 226 members of the AP2 family. DREB2C, DREB1A, TOE1, and RAP2.4 likely interact with LuAPX2, LuAPX3, LuAPX7, and LuAPX8. RT-qPCR analysis of the flax APX genes revealed significant differential expression in response to mannitol-induced drought and salinity stresses in both leaf and root tissues. Moreover, the integration of GWAS and RNA-seq data from salt stress revealed that LuAPX12 may be a potential candidate gene for salt tolerance. Compared with the wild type, the heterologous transformation of this gene into Arabidopsis resulted in overexpression lines with better germination rates, APX activity, and growth status, as well as lower H 2 O 2 and MDA levels under salt and osmotic stresses. The gene was further transiently transformed into flax, and the growth status, as well as the APX and CAT activities of the transgenic lines under salt and drought stress, were greater than those of the control. A comprehensive genome-wide characterization of the flax APX gene family was conducted, and the function of the APX-R gene LuAPX12 under osmotic and salt stress was preliminarily validated. These results broaden our understanding of the core components of antioxidant defense in flax and provide a theoretical basis for molecular research and breeding for resistance to adverse stress in flax.

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