The seeds of Camellia oleifera accumulate a rich amount of triterpenoid saponins, which possess various biological activities and have many encouraging prospects for application. But their biosynthesis and regulatory mechanisms is still poorly understood. Here, we identified and functionally characterized five new 2,3-oxidosqualene cyclases (OSCs) from C. oleifera. CobAS1 and CobAS2 were found to act as beta-amyrin synthases, while CobAS3 showed no significant catalytic activity. CoDDS1 and CoDDS2 were identified as multifunctional OSCs, producing alpha-amyrin and beta-amyrin at a ratio of 4.5:1. We further confirmed that residue #270 is a key active site for CoOSCs, as determined by site-directed mutagenesis. Moreover, we conducted a phylogenetic analysis of the OSC family across 216 angiosperm species. Furthermore, CoERF-1 and CobHLH-2 were identified as transcription factors regulating triterpenoid saponin biosynthesis in C. oleifera by binding to the promoters of CobAS1 and CoDDS1, respectively, and enhancing their transcription. These findings elucidated the types of triterpenoid saponins present in various tissues of C. oleifera at the molecular level, establishing a foundation for their biosynthetic and regulatory mechanisms. It also supplied important candidate genes for molecular breeding of higher-quality C. oleifera and enhanced our knowledge of the active sites and evolution of the OSC in plants.

Brassinolide (BL), the sixth major plant hormone, is synthesized from phytosterols. At present, research on the relationship among BL, phytosterols, and C. oleifera fruit development remains limited. In this study, we found that 0.5 mg/L of BL promoted the growth of seed kernel cells and increased the growth of 'Huashuo' seeds. BL treatment further promoted seed development and increased phytosterol content. 24-Ethylsterol, particularly stigmasterol, demonstrated a strong correlation with seed development. Furthermore, CoSMT2 was identified as a regulator of 24-ethylsterol synthesis. In addition, the CoMYB86 gene was found to be associated with CoSMT2. The seeds of Arabidopsis thaliana overexpressing CoSMT2 were fuller than those of the wild type, with BL and phytosterol contents significantly increased. The ratio of 24-ethylsterol to 24-methylsterol also increased significantly, from 1.49 to 2.93. These results provide new insights into understanding the effects of BL on C. oleifera fruit development and offer a new perspective on the balance between phytosterols and brassinolide.

The tung tree, a crucial woody oil plant, serves as a premium raw material for eco-friendly coating production, yet its short lifespan (typically under 20 years) and lack of asexual reproduction have led to resource losses. This study surveyed wild tung trees in the Hunan–Guizhou region, focusing on older and high-fruit-yielding specimens. After two years of investigation, selected individuals were conserved in the Wugang Tung Tree Germplasm Resource Bank to provide high-quality materials for breeding superior varieties. Comparative analysis of fruit yield and commercial traits from 60 wild trees identified 12 superior individuals for secondary selection, with notable trait variations observed. Using the entropy weight-TOPSIS method, superior individual FT01 exhibited the highest relative closeness (C = 0.6836), indicating optimal overall traits, while FT01, XY12, JX01, WG25, and WG31 (all with C > 0.50) demonstrated good overall performance. Genetic diversity analysis of these 12 individuals, employing 14 SSR primers, revealed 33 alleles (average 2.2142 per primer), Shannon’s information index values ranging from 0.1973 to 0.9723 (average 0.5325), and polymorphism information content between 0.1486 and 0.5833 (average 0.3981), indicating high genetic diversity. UPGMA clustering divided the superior trees into five groups, with FT01, WG25, JX01, and XY12 in separate groups, all exhibiting high yield and large fruit size, consistent with TOPSIS results. Consequently, FT01, XY12, JX01, WG25, and WG31, with the highest comprehensive evaluation scores and richest genetic diversity, are prioritized as candidate materials for new variety selection and breeding.

BACKGROUND:Camellia oleifera Abel. is one of the four major woody oil species whose seeds produce high-grade edible oil. In recent years, the planting area of Camellia oleifera is increasing in China. However, in the process of cultivation, due to the high fruit load, the Camellia oleifera tree has small fruit and poor quality. In previous studies, we explored the optimal leaf-fruit ratio. In this study, the changes of molecular regulation mechanism of Camellia oleifera under different leaf-fruit ratios were revealed by combining physiological indexes with transcriptome data. RESULT:The physiological results showed that the content of MDA and starch in leaves increased significantly with the decrease in the leaf-to-fruit ratio. The results of transcriptome showed that there was a close relationship between leaf-fruit ratio and phenylpropanoid biosynthesis pathway. With the decrease of leaf-fruit ratio, the expression of genes related to lignin and flavonoid biosynthesis increased significantly, which promoted the synthesis of lignin and flavonoid. CONCLUSIONS:Combining physiological indicators and transcriptomics, we demonstrated that leaf-fruit ratio can significantly affect the normal growth of plants. When the fruit load is too high, the fruit as a 'sink' will consume a large amount of nutrients in the plant body and promote the biosynthesis of lignin and flavonoids in the body. The results provide a more reliable scientific basis for the study of cultivation and management of Camellia oleifera.

Camellia-oil trees are economically valuable, oil-rich species within the genus Camellia, family Theaceae. Among these species, C. oleifera, a member of Section Oleifera in the genus, is the most extensively cultivated in China. In this study, we assembled the mitochondrial genomes (mitogenomes) of two Camellia species, namely C. oleifera and C. lanceoleosa. These two species are closely related and belong to the same genus and section, with C. oleifera being hexaploid and C. lanceoleosa being diploid. The mitogenome of C. oleifera is comprised of 1,039,838 base pairs (bp), and C. lanceoleosa is comprised of 934,155 bp. Both genomes exhibit a multipartite genome structure, which is supported by our PCR experiments. We conducted codon usage and RNA editing site analysis on these two mitogenomes, which showed highly consistent results. However, analysis of repetitive sequences and mitochondrial plastid sequences (MTPTs) revealed differences between the two mitogenomes. Phylogenetic analysis indicated that these two species clustered together, suggesting a close evolutionary relationship. The collinearity analysis results showed extensive genome rearrangements in the mitogenomes of Camellia species. We successfully assembled the mitogenomes of C. oleifera and C. lanceoleosa, marking a significant advancement in understanding camellia-oil tree mitogenomes. Unlike circular mitogenomes reported before, our research confirms multiple-branched configurations in these two species. This sheds light on mitogenome structural complexities and contributes to our understanding of evolutionary processes. Additionally, these results enrich Camellia genetic resources and expand our knowledge of mitogenome variation.

Rapid alkalinization factors (RALFs) play meaningful roles in the pollination and fertilization. However, the identification and study of Camellia RALFs is lacking, particularly the function of the RALF family in regulating self-incompatibility pollen tube growth. Herein, we identified 50 RALF genes from Camellia oleifera genome, and classified them into three groups: Clades Ⅰ, Ⅱ, and Ⅲ, with 11, 12, and 27 members being included, respectively. CoRALFs were unevenly distributed in chromosomes, and the segmental duplication events mainly facilitated their expansion. Gene structure and conserved motif analyses indicated that they were highly conserved. Cis-element analysis revealed that many light responsive elements, stress responsive elements and phytohormone responsive elements were found in CoRALFs promoters. Moreover, the expression analysis showed that pollen-specific CoRALF50 responded to the elongation and stagnation of self-incompatibility pollen tube in C. oleifera. Further experiments suggested that CoRALF50 could significantly down-regulate carbohydrate metabolism pathways in pollen tubes cultured in vitro, thereby causing abnormalities in ROS level, plasma membrane biosynthesis, and cell wall biosynthesis, which demonstrating the importance of CoRALF50 in regulating pollen tube growth. Altogether, this study extends information for exploring new functions of RALFs and provides new insights into the role of RALFs in self-incompatibility pollen tube growth.

Seasonal drought has hindered the sustainable growth of the Camellia oil tree industry. While brassinolide (BL) can mitigate drought stress in plants to some extent, the regulatory mechanisms underlying BL’s effects in Camellia oil tree remain unclear. To investigate the mechanisms by which BL alleviates drought stress in Camellia oil tree, three-year-old ‘Huashuo’ cutting seedling was exposed to three experimental treatments: CK (normal watering), UW (no watering), and BL (no watering, sprayed with 1 mg∙L− 1 BL). Leaf anatomical observation, hormone quantification, and transcriptomic profiling were performed on leaf samples collected at days 2, 4, and 6 following treatments. The results demonstrated that (i) BL treatment significantly increased leaf and spongy tissue thickness; (ii) BL reduced ABA and IAA levels in leaves under drought stress; (iii) WGCNA identified three modules linked to ABA, IAA, and their corresponding hormone levels, with subsequent analysis infer NAC transcription factors (TFs) as the primary regulators in these modules. This study contributes to a deeper understanding of the role of BL in alleviating drought stress in tree crop and provides a fundamental reference for molecular breeding and genetic improvement of Camellia oil tree.

Camellia oleifera Abel. (C. oleifera) represents a significant woody edible oil species predominantly distributed in southern China. Timely flowering is essential for the growth, development and tea oil production of C. oleifera. However, the mechanisms underpinning this process remain insufficiently understood. In this study, it was demonstrated through time-course transcriptome analysis that we revealed that CoFKF1-like1 (CoFKF1) serves as a central regulatory gene in the flowering process of C. oleifera. The ectopic expression of CoFKF1 resulted in the induction of early flowering. Furthermore, it was observed that CoFKF1 interacts with the transcription factor CoMYB4 in a blue-light-dependent manner, facilitating its ubiquitination and subsequent degradation. Genetically, CoMYB4 was identified as functioning downstream of CoFKF1 by directly binding to the promoter of CoFT1 and repressing its promoter activity. In conclusion, these findings elucidate that CoFKF1 promotes flowering by reducing the stability of the CoMYB4 protein, thereby enhancing CoFT1 promoter activity. Collectively, the results provide critical insights into the flowering mechanisms of C. oleifera and present a promising avenue to optimise its flowering period via the CoFKF1-CoMYB4-CoFT1 module.

The industrial cultivation of Camellia oil tree, a key woody oil species, has faced substantial challenges due to escalating extreme weather events in recent years. To investigate rhizosphere adaptations to irrigation regimes, three-year-old saplings were subjected to four evapotranspiration (ET)-based treatments: 50 % ET (I50), 75 % ET (I75), 100 % ET (I100), and natural rainfall (Control). Soil available nutrients (dissolved organic nitrogen, ammonium nitrogen, available phosphorus) and enzyme activities (urease, phytase, acid phosphatase, beta-glucosidase, leucine aminopeptidase) were significantly lower under I100 than under I50. Metagenomic and untargeted metabolomic analyses revealed rhizosphere responses to these treatments. Shannon diversity index at phylum and genus levels were significantly higher in I50 and I75 than in I100. Relative abundances of Pseudomonadota, Acidobacteriota, Actinomycetota, and Bacteroidota increased in I50 and I75 but decreased in I100. Reduced irrigation elevated soil nitrogen (N) and phosphorus (P) cycling gene abundance. Differentially abundant metabolites were predominantly enriched in amino acid biosynthesis pathways. Integrated analysis demonstrated a positive correlation between L-glutamate e and N/P cycling genes, alongside significant negative correlations with soil and leaf water content. In summary, under irrigation deficit, Camellia oil tree roots exhibited increased L-glutamate levels alongside enriched Acidobacteriota, Actinomycetota, Chloroflexota, and Planctomycetota, collectively enhancing soil N and P cycling. These findings advance irrigation strategy optimization for drought resilience and deepen understanding of plant-environment interactions in perennial crops.

Plant abiotic stress refers to the unfavorable effects on plants caused by any abiotic factors in a specific environment, such as drought, high temperature, low temperature, etc., which cause disruption of plant physiology and metabolism, and seriously affect the growth and yield of plants. Mounting evidence demonstrates that WRKY transcription factors modulate plant abiotic stress responses by regulating sugar metabolic pathways. Sugar metabolism pathway plays an essential role in plant stress resistance, and WRKY transcription factors, as an important class of regulatory factors, have attracted wide attention for their mechanism of action in abiotic stress. Therefore, this review primarily aims to analyze the structure and classification of WRKY transcription factors, summarize the research progress on how WRKY transcription factors themselves respond to stress, and how they participate in regulating plant stress responses through sugar metabolism pathways. Through in-depth investigation of the relationship between WRKY transcription factors and sugar metabolic pathways we uncovered novel abiotic stress-related gene regulatory networks providing theoretical basis and practical guidance for genetic improvement of plants under abiotic stress.

Squalene is an extremely valuable medicinal substance. In addition to being valued for its high content of unsaturated fatty acids, Camellia oleifera is also highly regarded for its rich squalene content in the seed kernels. Comparing the squalene contents within Camellia species, it was found that the content in the seed oil of C. oleifera was higher than that of any other species. The squalene content of C. oleifera "Huashuo" (ColHS) reached 0.410 mg g-1, which was the highest. However, the squalene content in the seed kernels of ColHS did not increase gradually with the continuous maturation of the fruit. The squalene content reached its peak at 329DAP, about 0.854 mg g-1, and then decreased. With the differentially expressed genes and metabolites in the seed kernels at 329DAP and its surrounding periods, it was found that squalene content was accompanied by variations in secondary metabolites, terpenoids and flavonoids. The mevalonate (MVA) pathway played a significant role in squalene synthesis of ColHS seed kernels. In the MVA pathway, the expression patterns of four CoHMGR genes were consistent with the squalene content level. Among them, CoHMGR2 exhibited a strong correlation with squalene content. The CoHMGR2 was also found co-expressed with genes that had calcium ion-binding functions, playing a role in plant signal transduction. This study offers valuable insights into the relationship between squalene content and C. oleifera seed maturity. It also advances our understanding of the regulatory network of squalene synthesis in C. oleifera.

Introduction:Seasonal drought associated with the subtropical monsoon climate significantly impairs the growth and development of Camellia oil tree seedlings. While previous studies have established that drought stress elevates glutamate content in the rhizosphere of Camellia oil tree, the mechanisms through which glutamate modulates rhizosphere microbial community assembly remain unresolved. Methods:To investigate the effects of glutamate on the rhizosphere environment under drought stress, we conducted an experiment using three-year-old potted seedlings subjected to moderate drought. These seedlings were irrigated with 50 mL of glutamate solutions at varying concentrations (0, 1, 2, 5, and 10 mmol/L; labeled G0, G1, G2, G5, and G10, respectively). Through analysis of rhizosphere soil nutrients, enzyme activity, and bacterial community abundance (relative and absolute). Results:The study revealed the following: Concentrations of available nitrogen forms (DON, NH4 +-N, NO3 --N) increased proportionally with glutamate concentration, whereas soil pH and urease activity exhibited inverse trends. Alpha and beta diversity analyses demonstrated significant divergence in bacterial community composition across treatments. Kruskal-Wallis, ANOVA, and LEfSe analyses identified 24 bacterial phyla significantly associated with treatment differences, with their abundance patterns corresponding to nitrogen cycling gene dynamics-generally peaking at G5 before declining. Discussion:These findings collectively suggest that 5 mmol/L Glu represents a pivotal concentration influencing rhizosphere bacterial community dynamics in Camellia oil tree under drought stress.

The degree of lignification in the spring shoots of Camellia oleifera significantly affects the quality of grafted scions. However, the regulatory mechanisms of lignification are poorly understood. This study explored the effects of exogenous gibberellin A3 (GA3) on lignification in the spring shoots of C. oleifera. Five-year-old C. oleifera (cultivar, Huaxin) were subjected to treatments with different concentrations of GA3 and uniconazole. The GA effect was assessed in terms of the morphology, physiology, and molecular levels. The results indicate that exogenous GA3 markedly decreased lignin content. Transcriptomic and metabolomic analyses revealed that exogenous GA3 inhibited the expression of genes associated with the cytokinin signaling pathway, specifically downregulating histidine kinases (HK3, HK4, and HK2) and response regulators (RR23, RR26, and RR4), as well as genes involved in lignin synthesis, including hydroxycinnamoyltransferase (HCT), ferulate 5-hydroxylase (F5H), and peroxidase (PER). The concentration of intermediates involved in monolignol synthesis, such as sinapaldehyde, was altered in response to GA treatment. These findings revealed that the application of GA3 may be an efficient strategy to regulate the spring shoot growth of C. oleifera, hence enhancing scion quality.

Camellia oleifera is a kind of high-quality oil supply species. Its seeds contain rich unsaturated fatty acids and antioxidant active ingredients, which is a kind of high-quality edible oil. In this study, we used bioinformatics methods to decipher a hexaploid Camellia oil tree’s mitochondrial (mt) genome based on second-generation sequencing data. A 709,596 bp circular map of C. oleifera mt genome was found for the first time. And 74 genes were annotated in the whole genome. Mt genomes of C. oleifera and three Theaceae species had regions with high similarity, including gene composition and gene sequence. At the same time, five conserved gene pairs were found in 20 species. In all of the mt genomes, most of nad genes existed in tandem pairs. In addition, the species classification result, which, according to the gene differences in tandem with nad5 genes, was consistent with the phylogenetic tree. These initial results provide a valuable basis for the further researches of Camellia oleifera and a reference for the systematic evolution of plant mt genomes.

Background Oil-tea tree (Camellia oleifera Abel) is an important high-quality edible oil tree species in China and is also a pioneer afforestation tree species in hilly red soil areas in southern China. It can grow and bear fruit on relatively barren mountains. Although C. oleifera has strong adaptability, long-term drought still affects its normal growth and even causes plant death, which has a serious impact on the afforestation industry. Results Under drought stress, the leaf edge of the C. oleifera curled, and the leaf yellowed, drooped, and even withered and died. Moreover, the accumulation of catalase (CAT), soluble sugar (SS) and abscisic acid (ABA) gradually increased, and the CAT activity and SS content increased more in the bareroot-stage seedlings than in the container-stage seedlings. We observed that, compared with the CK treatment, the drought treatment (10 DAT) significantly reduced the net photosynthetic rate (Pn), transpiration rate (E), maximum photochemical efficiency (Fv/Fm), and actual photochemical and quantum efficiency (FPSII). The Pn in the ‘HS-CS’, ‘HS-BS’, ‘HJ-CS’ and ‘HJ-BS’ treatments decreased by 126.68%, 112.33%, 126.08% and 117.22%. We also found that the drought resistance of bareroot-stage plants was greater than that of container-stage plants. After rewatering under drought stress, multiple indices of C. oleifera were greater than normal, and physiological characteristics and anatomical structure were positively related to compensation or overcompensation. At the same time, we also found that the recovery ability of C. oleifera ‘Huashuo’ was greater than ' C. oleifera ‘Huajin’. Conclusion The results showed that drought led to yellowing and shrinkage of C. oleifera leaves, reduces photosynthetic efficiency, and ultimately affected the normal growth of C. oleifera materials. In order to prevent this situation, C. oleiferashould reduce drought stress or timely rehydration to maintain the stability of leaf structure, morphology and function. In addition, we found that the drought resistance of bareroot seedlings was greater than that of container seedlings, and the recovery ability of C. oleifera ‘Huashuo’ was stronger than that of C. oleifera ‘Huajin’.

Abstract WRKY-like transcriptional regulators are widely involved in physiological processes such as growth and development, metabolic regulation and environmental response. In this study, we obtained six CoWRKY transcription factors by yeast one-hybrid screening library with reference to the Camellia oleifera genome sequence, using squalene synthase gene (CoSQS) as bait. AOS (Antibody Optimization System) analysis showed that CoWRKY15 had the highest interactions with a confidence level of 0.9026. Bioinformatics analysis showed that CoWRKY15 encodes 346 amino acid residues, was a basic hydrophilic protein, did not contain a transmembrane region, contained one WRKY conserved structural domain and one C2H2 zinc finger structural domain. and belonged to class 2 of the WRKY gene family, and had the closest genetic distance of 0.5564 to the homologous protein of Panax quinquefolius PqWRKY1. The results of prokaryotic expression showed that the CoWRK15 protein with a size of 38.3 kD was successfully induced by adding a final concentration of 0.5 mM ITPG for 4 h at 37℃. The results of subcellular localization showed that CoWRKY15 functioned in the nucleus. The results of CoWRKY15 promoter analysis showed that 8 out of 14 cis-elements with annotatable functions were related to the light response, indicating that the expression of CoWRKY15 was strongly affected by light. The correlation analysis of CoWRKY15 expression and squalene content in Camellia oleifera seed kernels treated under different light quality conditions showed a significant positive correlation.

Light is one of the most important environmental factors for plant growth. In the production process of tung oil tree cultivation, due to the inappropriate growth of shading conditions, the lower branches are often dry and dead, which seriously affects the yield of tung oil trees. However, little is known about the key factors of light-induced tree photomorphogenesis. In this study, a total of 22 VfBBX family members were identified to provide a reference for candidate genes in tung tree seedlings. All members of the VfBBX family have different numbers of highly conserved B-box domains or CCT domains. Phylogenetic evolution clustered the VfBBX genes into four categories, and the highest density of members was on chromosome 6. Interspecific collinearity analysis suggested that there were six pairs of duplicate genes in VfBBX members, but the expression levels of all family members in different growth and development stages of the tung tree were significantly divergent. After different degrees of shading treatment and physiological data determination of tung tree seedlings, the differential expression level and chlorophyll synthesis genes correlation analysis revealed that VfBBX9 was a typical candidate nuclear localization transcription factor that was significantly differentially expressed in light response. This study systematically identified the VfBBX gene family and provided a reference for studying its molecular function, enhanced the theoretical basis for tung tree breeding, and identified excellent varieties.

Squalene synthase (SQS) is the most direct key enzyme regulating squalene synthesis. To better understand the regulatory mechanisms of squalene biosynthesis, a 1423-bp long promoter region of the CoSQS gene was isolated from Camellia oleifera. Plant CARE and PLACE analysis affirmed the existence of the core promoter elements such as TATA and CAAT boxes and transcription factor binding sites like W-box and MYB in the isolated sequence. Exogenous factors regulating the CoSQS promoter were obtained by using Yeast one-hybrid screening, and the key transcription factor CoWRKY15 was found. AOS (Antibody Optimization System) analysis showed that CoWRKY15 had the highest interactions with a confidence level of 0.9026. Bioinformatics analysis showed that CoWRKY15 belonged to class 2 of the WRKY gene family. The results of subcellular localization showed that CoWRKY15 functioned in the nucleus. The results of CoWRKY15 promoter analysis showed that 8 out of 14 cis-elements with annotatable functions were related to the light response. The region of the CoSQS promoter that interacts with CoWRKY15 is −186 bp~−536 bp. The histochemical assay and squalene content suggested that the CoSQS promoter could drive the expression of GUS gene and specific promotion of CoSQS expression. It was found that CoWRKY15 could act on the −186 bp~−536 bp CoSQS promoter to regulate the expression of CoSQS and the content of squalene in C. oleifera seed kernels.

Cytoarchitectural staining is of great importance in disease diagnosis and cell biology research. This study developed user-friendly multifunctional red-emissive carbon dots (R-CDs) for rapid cell nucleus staining via targeting nuclear proteins. R-CDs, simply prepared by electrochemical treatment of 1,2,4-benzenetriamine, exhibit strong emission at 635 nm when excited at 507 nm. The R-CDs can rapidly stain the nucleus of human SH-SY5Y, HepG2, and HUH-7 cells with a high signal-to-noise ratio owing to fluorescence enhancement after entering the nucleus. Compared to conventional cytosolic dyes such as Hoechst and DAPI, R-CDs are cheaper, more highly dispersed in water, and more stable (requiring no stringent storage conditions). The R-CDs show stable optical properties with insignificant photobleaching over 7 days and salt resistance up to 2 M of NaCl. More importantly, R-CDs, possessing a positive charge, allow rapid staining of live cells (3 min) and dead cells (10 s) in saline. According to kinetic variation, R-CDs can distinguish live cells from dead cells. Staining exhibits high efficiency in onion epidermal cells, Aspergillus niger, Caenorhabditis elegans, and human spermatozoa. The mechanism for efficient staining is based on their fast accumulation in the nucleus due to their small size and positive charge and strong interaction with nuclear proteins at amino acid residues of histidine and arginine, resulting in fluorescence enhancement by dozens of times. The developed R-CDs do not bind to DNA and would not cause genetic damage and will find various safe applications in biological and medical fields.

Tung tree (Vernicia fordii) as an important industrial raw material used to produce tung oil in China. Genetic transformation system has not been established for the tung tree, hindering research on the functions of tung tree genes and the cultivation of new stress-resistant varieties. In this study, we optimized the tissue culture regeneration system of tung tree hypocotyls, screened out a callus state suitable for Agrobacterium tumefaciens infection, and improved the efficiency of genetic transformation. RNA interference (RNAi) vector was constructed using FAD2 as the target gene. Adventitious shoots were obtained by infecting the hypocotyl calli cultured for 25 days through Agrobacterium-mediated infection. The positive adventitious shoot transformation rate of the pBI121 vector was 22.86%, and the adventitious shoot transformation rate of the RNAi vector was 7.84%. We established a complete genetic transformation system using two vectors, and the system was verified via GUS staining and real-time fluorescence quantitative analyses to ensure its stability. This system solves the technical problem of a genetic transformation system for the tung tree, and will provide technical support for studying the functions of tung tree genes and molecular breeding.