College of Horticulture Science and Technology, Hebei Normal University of Science and Technology/Hebei Key Laboratory of Horticultural Germplasm Excavation Innovative Utilization/Hebei Higher Institute Application Technology Research and Development Center of Horticultural Plant Biological Breeding, Qinhuangdao 066600, China
Abstract：The 14-3-3 protein family, constitutes a class of highly conserved regulatory proteins in plants, also known as G?box regulatory factors (GRFs), are extensively involved in various physiological processes such as cellular metabolism, signal transduction, and responses to biotic and abiotic stresses. Tamarix chinensis is a typical recretohalophyte, but the function and regulatory mechanism of its 14-3-3 family genes in response to salt stress remains unclear. To investigate the characteristics of the 14-3-3 gene family in T. chinensis and their roles in salt tolerance, the study cloned, identified, and functionally analyzed 14-3-3 family genes. The results showed that 5 14-3-3 genes, named TcGRF1 to TcGRF5, respectively, were successfully cloned, and all TcGRF proteins contained the typical conserved 14-3-3 domain. qPCR analysis revealed that the expression levels of all 5 TcGRFs genes were up-regulated to varying degrees under 0.2 and 0.4 mol/L NaCl stress, with TcGRF3 showing the most obvious difference. To further clarify the function of TcGRF3 under salt stress, transient transformation technology was used to overexpress and suppress the expression of TcGRF3 in Tamarix chinensis, respectively. Overexpression of TcGRF3 significantly increased chlorophyll content, the activities of superoxide dismutase (SOD) and peroxidase (POD)(P<0.05), and proline accumulation, while decreasing relative electrical conductivity and malondialdehyde (MDA) content (P<0.05). In contrast, TcGRF3-silenced plants showed the opposite trends. Comprehensive analysis indicated that TcGRF3 positively regulated salt tolerance in T. chinensis by enhancing reactive oxygen species (ROS) scavenging capacity, thereby mitigating membrane lipid peroxidation damage. This study provides a theoretical foundation for further elucidating the functional mechanisms of the 14-3-3 protein family in the salt stress response of T. chinensis.
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