NAC transcription factors are one of the largest families of transcriptional regulators in plants, and members of the gene family play vital roles in regulating plant growth and development processes including biotic/abiotic stress responses. However, little information is available about the NAC family in pitaya. In this study, we conducted a genome-wide analysis and a total of 64 NACs (named HuNAC1-HuNAC64) were identified in pitaya (Hylocereus). These genes were grouped into fifteen subgroups with diversities in gene proportions, exon–intron structures, and conserved motifs. Genome mapping analysis revealed that HuNAC genes were unevenly scattered on all eleven chromosomes. Synteny analysis indicated that the segmental duplication events played key roles in the expansion of the pitaya NAC gene family. Expression levels of these HuNAC genes were analyzed under cold treatments using qRT-PCR. Four HuNAC genes, i.e., HuNAC7, HuNAC20, HuNAC25, and HuNAC30, were highly induced by cold stress. HuNAC7, HuNAC20, HuNAC25, and HuNAC30 were localized exclusively in the nucleus. HuNAC20, HuNAC25, and HuNAC30 were transcriptional activators while HuNAC7 was a transcriptional repressor. Overexpression of HuNAC20 and HuNAC25 in Arabidopsis thaliana significantly enhanced tolerance to cold stress through decreasing ion leakage, malondialdehyde (MDA), and H₂O₂ and O₂⁻ accumulation, accompanied by upregulating the expression of cold-responsive genes (AtRD29A, AtCOR15A, AtCOR47, and AtKIN1). This study presents comprehensive information on the understanding of the NAC gene family and provides candidate genes to breed new pitaya cultivars with tolerance to cold conditions through genetic transformation.

Dormancy is a vital adaptive strategy in temperate and boreal plants, particularly fruit trees, enabling them to withstand harsh winter conditions and ensure survival and synchronized growth resumption in spring. This review comprehensively examines dormancy, focusing on its physiological, environmental, and molecular mechanisms. Dormancy is characterized by two distinct phases: endodormancy, which is regulated by internal plant signals and requires cold temperatures for release, and ecodormancy, which is influenced by external environmental factors. These stages are intricately linked to seasonal temperature fluctuations and the plant’s ability to synchronize growth cycles, ensuring survival through harsh winters and optimal growth in warmer seasons. The review delves into the role of chilling requirements, temperature thresholds, and hormonal regulation in the dormancy process, highlighting how these factors influence critical growth events such as budbreak, flowering, and fruiting. Plant hormones, including abscisic acid, gibberellins, and cytokinins, regulate dormancy by modulating gene expression and growth activity. Additionally, we explore the historical development of dormancy research, from early observations of chilling requirements to the formulation of the chilling hours model. Considering ongoing climate change, the review examines how rising winter temperatures may disrupt dormancy cycles, potentially affecting the timing of flowering, fruiting, and overall crop productivity. This shift necessitates new strategies for managing dormancy, particularly in regions experiencing inconsistent or insufficient chilling. The review concludes by discussing practical approaches to enhance dormancy release and mitigate the impact of environmental stress on deciduous fruit tree growth, offering insights into improving agricultural practices amidst a changing climate.

Real-time quantitative polymerase chain reaction (qRT-PCR) has been widely used in gene expression analyses due to its advantages of sensitivity, accuracy and high throughput. The stability of internal reference genes has progressively emerged as a major factor affecting the precision of qRT-PCR results. However, the stability of the expression of the reference genes needs to be determined further in different cells or organs, physiological and experimental conditions. Methods for evaluating these candidate internal reference genes have also evolved from simple single software evaluation to more reliable and accurate internal reference gene evaluation by combining different software tools in a comprehensive analysis. This study intends to provide a definitive reference for upcoming research that will be conducted on fruit trees. The primary focus of this review is to summarize the research progress in recent years regarding the selection and stability analysis of candidate reference genes for different fruit trees.