Longan (Dimocarpus longan) is a typical southern subtropical fruit tree species that is sensitive to cold stress. C-repeat binding factors (CBFs), as transcription factors, are crucial components involved in the molecular regulation of the plant response to cold stress. However, the role of CBF homologs in the cold response regulation of longan remains largely unknown. Here, three novel CBF genes, DlCBF1, DlCBF2, and DlCBF3, were cloned from longan. DlCBF1 and DlCBF2 contain an AP2 domain and PKKPAGR and DSAWR CBF signature motifs, while DlCBF3 has mutations within these conserved signature motifs. DlCBF1/2/3 were mainly localized in the nucleus and specifically bound to CRT/DRE cis-elements, resulting in strong transcriptional activation. DlCBF1/2 exhibited tissue expression specificity, and their expression was induced by low temperature, while DlCBF3 had no tissue specificity and barely responded to low temperature. DlCBF1, DlCBF2, and DlCBF3 overexpression in Arabidopsis-enhanced cold tolerance by increasing proline accumulation and reducing reactive oxygen species (ROS) content, accompanied by upregulated expression of cold-responsive genes (AtRD29A, AtCOR15A, AtCOR47, and AtKIN1) in the CBF cold stress response signaling pathway. In conclusion, the biological functions of DlCBF1/2/3 were somewhat conserved, but slow expression of DlCBF1/2 and low expression of DlCBF3 may partly cause the cold sensitivity of longan. Collectively, these results indicated that differences exist in the expression and function of CBF orthologs in the cold-sensitive plant species longan, and these findings may help to improve the understanding of the cold response regulation mechanism and provide important theoretical support for cold-tolerant breeding of longan.

Longan (Dimocarpus longan Lour.) is of great economic significance in South China for its unique taste and nutritional properties. However, longan breeding is mainly based on seedling selection, which generally results in small fruits, low flesh recovery, and few seedless germplasm. Triploid breeding is a central way to improve these problems. In this study, microspore chromosomes were doubled by colchicine and high-temperature treatment to create triploids in longans. The relationship between the development process of male gametophyte of longans and the morphological changes of male flower buds was established. Cytological observation showed that when the male flower buds were in stage I (when the diameter of the flower bud is 1.4–2.0 mm), most of the microspores were at the pachytene to diakinesis stage of meiosis, and the chromosome doubling induction effect was the best at this stage. The results showed that the 2n pollen rate induced by a high temperature of about 38 °C was higher than that induced by colchicine treatment. The highest 2n pollen rate was 5.7% and 5.5% based on the microscopic measurement method and the abnormal separation in tetrad stage estimation method, respectively. Four triploids were successfully obtained from artificial pollination with 2n pollen, with a triploid induction rate of 0.6%. This study will promote ploidy breeding in longan.

Dwarfism is a significant and desirable trait in the cultivation of horticultural crops. However, the regulatory mechanisms underlying dwarfism in perennial fruit trees remain poorly understood. In this study, we investigated the role of SRS genes in regulating plant height in Sapindaceae fruit trees, specifically litchi and longan. A total of five litchi SHORT INTERNODES (SHI)-related sequence genes (LcSRS) and four longan DlSRS genes were identified. Expression profile analysis in different tissues demonstrated that LcSRS1-LcSRS5 and DlSRS4 genes exhibited distinct expression patterns, with some genes showing tissue-specific expression. qRT-PCR analysis revealed that LcSRS5 and DlSRS4 genes had higher expression levels in the dwarf litchi ('Ziniangxi') and longan ('Shizi19') cultivars, respectively. Overexpression of the litchi LcSRS5 and the longan DlSRS4 in tobacco resulted in transgenic plants displaying a dwarf phenotype. Subcellular localization studies indicated that both LcSRS5 and DlSRS4 are localized in the nucleus of the cell. Hormone testing analysis indicated that the contents of auxin, cytokinin, and gibberellin varied among different cultivars of litchi and longan. GA3 treatment can promote the elongation of shoots in both litchi and longan. Additionally, silencing the LcSRS5 in litchi buds effectively promotes shoot growth, increases internode length, and significantly suppresses the expression of LcGA2ox1, LcGA2ox2, and LcGA2ox3 genes. Further studies confirmed that the LcSRS5 binds to the promoter of LcGA2ox3 to activate its expression. This study provides a foundation for further analysis of the functions of SRS genes in tropical fruit trees.

Longan (Dimocarpus longan Lour.) is highly sensitive to low temperature, which severely restricts its cultivation and industrial development. MYB transcription factors serve as key regulators in plant responses to cold stress. In this study, an R2R3-MYB gene DlMYB108 was cloned from ‘Shixia’ longan. Sequence analysis showed that DlMYB108 contains two typical MYB repeats and shares high homology with cold-responsive MYB108 proteins from other plants. Expression pattern analysis revealed that DlMYB108 is highly expressed in young leaves, which are more sensitive to cold stress, and is significantly induced by low-temperature treatment. Subcellular localization and transcriptional activation assays confirmed that DlMYB108 is a nuclear-localized transcriptional activator. Yeast one-hybrid and dual-luciferase assays demonstrated that DlMYB108 specifically binds to the promoters of DlCBF2 and DlCBF3 and activates their transcription. Heterologous expression of DlMYB108 in Arabidopsis significantly enhanced cold tolerance, accompanied by reduced ion leakage, malondialdehyde (MDA) content and reactive oxygen species (ROS) accumulation, as well as upregulated expression of CBF and cold-responsive genes. Collectively, DlMYB108 positively regulates longan cold tolerance through activating DlCBF2 and DlCBF3 expression, providing a valuable candidate gene for cold-tolerant longan breeding.