Abstract
<title>Abstract</title> <p> Tubby-like proteins (TLPs) transcription factors are found in single-celled to multi-cellular eukaryotes in the form of large multigene families, and play important roles in growth development, stress responses in plants. In the current study, a total of 13 putative <italic>TLPs</italic> were identified in the grapevine genome, and unevenly distributed across 10 chromosomes. Phylogenetic analysis demonstrated that the predicted VviTLP proteins were divided into three groups (I-III). Gene structure, protein motifs, and conserved domains were analyzed to identify differences and common features among the VviTLPs. Only segmental duplication events might play key roles in the expansion of VviTLP gene family in grapevine. Analysis of publicly available gene expression data showed that VviTLP genes were differentially expressed in tissue, fruit developmental stages, and in response to abiotic stresses including drought, salt and cold stress treatment. Subcellular localization analysis showed that VamTLP3e and VamTLP7 were located in the nucleus, while VamTLP1, VamTLP3c and VamTLP10 were in plasma membranes and nucleus. Furthermore, VamTLP1, VamTLP7 and VamTLP10 had transcriptional activity, while VamTLP3c and VamTLP3e had no such activity. Overexpression of VamTLP1 enhanced cold tolerance in transient transformation grapevine and stable transformed calli by the scavenging of excessive reactive oxygen species (ROS) and promoting the upregulation of cold-responsive genes. Collectively, the results of this study provide valuable insights for future functional analysis of TLP family genes, and contribute to a better understanding of the molecular mechanisms underlying grape cold stress response, which in turn supports grapevine molecular breeding. </p>