Genome-Wide Identification of DREB Gene Family in Kiwifruit and Functional Characterization of Exogenous 5-ALA-Mediated Cold Tolerance via ROS Scavenging and Hormonal Signaling

Dehydration response element binding proteins (DREBs) have been identified as major regulators of cold acclimatization in many angiosperms. Cold stress is one of the primary abiotic stresses affecting kiwifruit growth and development. However, kiwifruit is currently one of the most widely consumed fruits worldwide because of its high nutritional value. 5-Aminolevulinic acid (5-ALA) is a nonprotein amino acid known for its distinct promotional effects on plant resistance, growth, and development. However, studies on the function of the kiwifruit DREB gene in alleviating low-temperature stress in its seedlings via exogenous 5-ALA have not been reported. Therefore, in this study, we performed a genome-wide identification of DREB gene family members in kiwifruit and analyzed the regulatory effects of exogenous 5-ALA on kiwifruit DREB genes under low-temperature stress. A total of 193 DREB genes were identified on 29 chromosomes. Phylogenetic analysis classified these genes into six subfamilies. Although there were some differences in cis-elements among subfamilies, all of them contained more biotic or abiotic stresses and hormone-related cis-acting elements. GO and KEGG enrichment analyses revealed that AcDREB plays an essential role in hormone signaling, metabolic processes, and the response to adverse stress. Under low-temperature stress, the application of exogenous 5-ALA inhibited the accumulation of APX and DHAR, promoted an increase in chlorophyll, and increased the accumulation of enzymes and substances such as 5-ALA, MDHAR, GR, ASA, GAH, and GSSH, thereby accelerating ROS scavenging and increasing the cold hardiness of kiwifruits. Functional analysis revealed that 46 differentially expressed DREB genes, especially those encoding <i>AcDREB69</i>, <i>AcDREB92</i>, and <i>AcDREB148</i>, which are involved in ethylene signaling and defense signaling, and, after the transcription of downstream target genes is activated, are involved in the regulation of low-temperature-stressed kiwifruits by exogenous 5-ALA, thus improving the cold tolerance of kiwifruits. Notably, <i>AcDREB69</i>, <i>AcDREB92</i>, and <i>AcDREB148</i> could serve as key genes for cold tolerance. This study is the first to investigate the function of AcDREB genes involved in the role of exogenous 5-ALA in regulating low-temperature stress, revealing the regulatory mechanism by which DREB is involved in the ability of exogenous 5-ALA to alleviate low-temperature stress.