Use of d6-mooncins

Abstract

This invention relates to the field of traditional Chinese medicine cultivation technology, specifically the application of D6-abelacterol, and particularly its application in promoting the growth of Panax notoginseng, inducing resistance, and improving its quality. Experiments have shown that D6-abelacterol significantly enhances the resistance of Panax notoginseng to pathogens such as black spot disease, improving its disease resistance; simultaneously, it promotes plant growth, significantly increasing the yield and quality of Panax notoginseng, exhibiting a significant growth-promoting effect. The D6-abelacterol provided by this invention is a natural product, possessing environmentally friendly, low-toxicity, and green properties, laying the foundation for the development of the Panax notoginseng industry.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine cultivation technology, and to the application of D6-abelacterol, particularly its application in the preparation of products that promote the growth of Panax notoginseng and induce resistance. Background Technology

[0002] Panax notoginseng, a precious traditional Chinese medicine unique to my country, is rich in triterpenoid saponins, flavonoids, and other active ingredients. It possesses various medicinal properties, including hemostasis, swelling reduction, pain relief, anti-tumor effects, antioxidant effects, antidepressant effects, and lipid-lowering effects. In my country, specific regions in Yunnan and Guangxi are designated as the main growing areas for Panax notoginseng, accounting for over 90% of the national planting area and yield. However, the Panax notoginseng industry currently faces a series of severe challenges, such as serious pests and diseases, high yield but low quality, pesticide residues and heavy metal contamination, and continuous cropping obstacles, severely restricting its sustainable development. Firstly, as a perennial herbaceous plant, Panax notoginseng prefers a cool, moist growing environment and has extremely strict environmental requirements. Unsuitable growing conditions (such as high temperature, high humidity, strong light, drought, and flooding) exacerbate pests and diseases in the above-ground parts and cause continuous cropping obstacles. Black spot disease is one of the important diseases affecting Panax notoginseng in Yunnan, seriously impacting its growth. Due to the transmissible nature of the pathogenic spores, control is difficult, resulting in an annual incidence rate of 10% to 20% for Panax notoginseng, which can even exceed 80% in severe cases. Therefore, effective control measures for Panax notoginseng black spot disease urgently need to be researched and promoted to ensure the healthy growth and yield of Panax notoginseng. Secondly, over-reliance on chemical fertilizers and pesticides leads to severe fertilizer overflow, with annual nitrogen application reaching as high as 270-560 kg / hm². 2 Unlike crops, the medicinal components of Panax notoginseng rely on secondary metabolism. High fertilizer and water environments inhibit saponin accumulation, leading to root cracking, nutrient imbalance, and harmful substance residues, ultimately resulting in a significant increase in the quality and safety risks of the medicinal material. Furthermore, intensive agricultural production methods focused on yield disrupt the balance between primary and secondary metabolism in Panax notoginseng, further exacerbating the obstacles caused by continuous planting, thus leading to a decline in the quality of the medicinal material. Excessive nitrogen application (e.g., 450 kg / hm²) under high-yield models further contributes to this problem. 2 The overuse of pesticides further inhibits saponin synthesis, enriches pathogens, exacerbates continuous cropping obstacles, and leads to an industry crisis of declining quality and unsustainable cultivation. At the same time, competition for land between Panax notoginseng and traditional food crops is becoming increasingly significant. Due to the trend of converting arable land to non-grain crops, medicinal herbs are becoming increasingly scarce. This forces planting areas to continuously reclaim new forest land or slopes for cultivation, further intensifying ecological pressure and quality risks, and severely restricting the sustainable and healthy development of Panax notoginseng.

[0003] Currently, plant allelopathic effects have significant application potential in the field of green pest and disease control in agriculture, especially in stimulating induced plant resistance and inhibiting pathogen invasion. Studies have shown that plants can activate their intrinsic defense mechanisms when infected by pathogens, triggering a series of physiological and biochemical reactions to enhance disease resistance. Allelopathic substances released by plants can participate in interspecific interactions, exhibiting inhibitory effects on the growth of various pathogens and weeds. Therefore, screening compounds with plant allelopathic effects and developing environmentally friendly pesticides with high-efficiency antibacterial or anti-disease activity through structural modification and activity optimization provides a technological foundation for the innovation of green pesticides.

[0004] Existing research indicates that α-pinene can directly regulate the expression of growth and resistance-related genes in Panax notoginseng, thereby synergistically promoting its growth and enhancing its disease resistance. However, as a small-molecule monoterpene, pinene suffers from drawbacks in practical agricultural applications, such as high volatility, chemical instability, and short foliar residual effect, which limits its widespread use in field settings.

[0005] D6-Ambrettolide, chemically known as oxadiocetene, is a macrocyclic musky compound extracted from Abelmoschus mandshuriensis. Compared to α-pinene, its physicochemical properties differ significantly. D6-Ambrettolide exhibits extremely low volatility and a fragrance retention time (lasting effect) exceeding 48 hours. As an excellent fixative, it demonstrates stronger adhesion to plant leaf surfaces, providing more persistent physiological regulatory signals. Its structural stability, particularly its macrocyclic lactone structure, makes it more stable than monoterpenes under light and air exposure, facilitating the development of efficient agricultural formulations. It is commonly used as a fixative in high-end daily chemical fragrances, soap fragrances, and skincare product formulations.

[0006] A search revealed that existing technologies do not yet include the application of D6-adenosine in promoting the growth of Panax notoginseng, inducing its resistance to black spot disease, or improving its quality. Summary of the Invention

[0007] To address the shortcomings of existing technologies, this invention provides the application of D6-abelacterol.

[0008] To solve the above-mentioned technical problems, the technical solution of the present invention is as follows: This invention protects the application of D6-adenosine lactone in the preparation of products that promote the growth of Panax notoginseng, induce disease resistance in Panax notoginseng, and improve the quality of Panax notoginseng.

[0009] Furthermore, the concentration of the D6-okratin aqueous solution is 1-10 μL / L.

[0010] Furthermore, the concentration of the D6-okratin solution is 5-10 μL / L.

[0011] Furthermore, the disease mentioned is Panax notoginseng black spot disease.

[0012] Preferably, it is applied by foliar spraying or root irrigation.

[0013] Furthermore, the improvement of Panax notoginseng quality refers to increasing the content of its effective components.

[0014] Compared with the prior art, the present invention has the following beneficial effects: This invention demonstrates through experiments that D6-adenophylloidin can effectively inhibit the activity of black spot pathogens and promote the growth of Panax notoginseng, while increasing the content of Panax notoginseng saponins. This significantly reduces the amount of chemical pesticides used, overcomes the problems of continuous cropping obstacles, declining quality of medicinal materials, and serious ecological pollution in traditional farmland planting, and achieves high-quality, ecological, and sustainable production of Panax notoginseng. It also provides a new breakthrough for controlling plant diseases by utilizing allelopathic interactions of biodiversity and provides technical and theoretical support for the organic planting model of Panax notoginseng under forests.

[0015] Experiments showed that under field cultivation conditions, using surviving Panax notoginseng as the experimental material, four D6-okratin concentrations (1, 5, 10, and 20 μL / L) were applied, with four replicates per group. A blank control (CK) was also included. Plant dry and fresh weight, chlorophyll content, and disease incidence were measured. Results showed that all concentrations of D6-okratin increased the dry and fresh weight of individual Panax notoginseng plants (both aboveground and underground parts) to varying degrees. The 20 μL / L treatment showed the best effect, significantly increasing the fresh weight of underground plants by 26.70% and the fresh weight of aboveground plants by 24.77% compared to the CK.

[0016] Spraying with D6-abelacterol can effectively reduce the incidence of diseases in Panax notoginseng, and the disease index of each treatment is lower than that of the control; the 1 μL / L treatment has the best disease suppression effect, with the disease index decreasing by 18.52% compared with the CK.

[0017] Based on the results of experiments on inducing disease resistance in Panax notoginseng using three compounds, D6-okratin at a concentration of 20 μL / L was selected for resistance induction treatment. One day after the induction treatment, diseased leaves were collected and lesions were scanned. The results showed that D6-okratin reduced the lesion area by 31.84%.

[0018] D6-Abelacterol showed peak effect in promoting saponin accumulation at a concentration of 10 μL / L, with a significantly higher increase than that of the citric acid treatment groups and similar to that of the high-concentration lactulose treatment. This indicates that the regulation of notoginseng saponin synthesis by D6-Abelacterol is significantly concentration-dependent; both excessively high and low concentrations are detrimental to the efficient accumulation of saponins.

[0019] This invention, through transcriptome sequencing and analysis of treated samples, confirms that D6-adenosine can effectively enhance the stress resistance of Panax notoginseng and activate its immune response by regulating multiple metabolic and signal transduction pathways. After sample testing and analysis, the RNA was found to be qualified for transcriptome analysis. Principal component analysis (PCA) revealed that after treatment with 20 μL / D6-adenosine for one day, the transcriptional level of Panax notoginseng samples was not completely separated, but a total of 648 differentially expressed genes (DEGs) were identified, of which 326 were upregulated and 322 were downregulated. GO functional analysis showed that the upregulated DEGs were significantly enriched in functional categories related to resistance and growth, such as molecular transduction, ATP-dependent activity, catalytic activity, and stress response. KEGG enrichment analysis showed that the upregulated genes tended to cluster in pathways such as pentose-gluconate tautomerism, diterpenoid and phenylpropane biosynthesis. At the same time, enhanced expression of disease resistance genes related to plant hormone signaling, MAPK pathway, WRKY transcription factor and phenylpropane metabolism was detected, indicating that D6-okratinolone can enhance the stress resistance and immune response of Panax notoginseng by regulating multiple metabolic and signaling pathways. Attached Figure Description

[0020] Figure 1 The in vitro inhibitory effects of D6-abelacterol, lactose, and citric acid on the pathogen causing black spot disease in Panax notoginseng were investigated. Figure 1 A represents the effect of D6-adenosine on the colony diameter of pathogenic bacteria; Figure 1 B represents the effect of lactose on the colony diameter of pathogenic bacteria; Figure 1 C represents the effect of citric acid on the colony diameter of pathogenic bacteria. Data are expressed as mean ± standard error, with different lowercase letters indicating significant differences between groups (p < 0.05). Figure 2 The experiment investigated the effect of D6-abelacterol-induced resistance in Panax notoginseng; among which... Figure 2 A, Figure 2 B Figure 2 C represents the effect of D6-abelrolactone, lactulose, and citric acid on resistance to Panax notoginseng black spot fungus after 1 day of induction; data are expressed as mean ± standard error, and different lowercase letters indicate significant differences between treatments (p<0.05). Figure 3 The effect of 20 μL / LD6-adenosyllactone on the resistance of Panax notoginseng to black spot fungus after induction for 1 day is presented. Data are expressed as mean ± standard error. Different lowercase letters indicate significant differences between treatments (p < 0.05). Figure 4 The effects of three compounds on the fresh and dry weight of Panax notoginseng above and below ground; among them, Figure 4 A, Figure 4 B Figure 4 C represents the effects of D6-abelacterol, lactulose, and citric acid on the fresh and dry weights of the seven samples above and below ground. Figure 5 The effects of three compounds on the height of Panax notoginseng plants, among which Figure 5 A, Figure 5 B Figure 5 C represents the effects of D6-abelacterol, lactulose, and citric acid on the plant height of Panax notoginseng; data are expressed as mean ± standard error, and different lowercase letters indicate significant differences between treatments (p < 0.05). Figure 6 The effects of three compounds on the disease index of Panax notoginseng leaves, among which Figure 6 A, Figure 6 B Figure 6 C represents the effects of D6-okratinol, lactulose, and citric acid on the leaf disease index of Panax notoginseng; data are expressed as mean ± standard error, and different lowercase letters indicate significant differences between treatments (p < 0.05); Figure 7 The effect of different concentrations of the compound on the accumulation of notoginsenosides; among which Figure 7 A, Figure 7 B Figure 7 C represents the effects of D6-adenosine, lactulose, and citric acid on the accumulation of Panax notoginseng saponins, respectively; data are expressed as mean ± standard error, and different lowercase letters indicate significant differences between treatments (p < 0.05); Figure 8 GO secondary classification of differentially expressed genes in Panax notoginseng induced by D6-adenosine; Figure 9 Bubble plot of KEGG enrichment of differentially expressed genes in Panax notoginseng treated with D6-adenosyllactone; Figure 10 Bubble plot of KEGG enrichment for differentially expressed genes in Panax notoginseng treated with D6-adenophorol; Figure 11 The differential gene expression patterns associated with disease resistance in Panax notoginseng treated with D6-abelacterol were analyzed. Detailed Implementation

[0021] The technical solutions of the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments, but the present invention is not limited to the following technical solutions.

[0022] Example 1: Effect of D6-Abelmolactone on Panax notoginseng resistance 1.1 In vitro inhibitory effect of D6-adenosyllactone on the pathogen of Panax notoginseng black spot disease This invention first evaluated its direct inhibitory effect on the pathogen of Panax notoginseng black spot disease using the plate-on-patch method. The effects of lactulose and citric acid were also investigated simultaneously. Experiments showed that lactulose also possessed certain antibacterial potential at concentrations of 1-20 mg / L. Compared to the single nutrients lactulose and citric acid, D6-abelacterol, with its unique macrocyclic lactone structure, not only exhibited direct antibacterial activity in vitro, but also demonstrated stronger leaf adhesion ability and the potential to induce disease resistance in subsequent experiments. In the antibacterial experiment, the plate-carrying method was used. 0, 0.2, 1, 2, and 4 μL of L-6-abelrolactone were added to 200 mL of sterilized PDA cooled to 60°C, resulting in final concentrations of 0, 1, 5, 10, and 20 μL / L in the culture dishes. Similarly, 0, 0.2, 1, 2, and 4 mg of lactulose and citric acid were added to 200 mL of PDA cooled to 60°C, resulting in final concentrations of 0, 1, 5, 10, and 20 mg / L in the culture dishes. After cooling, 20 mL of the solution was poured into each culture dish, and ten plates of each concentration were prepared. Holes were punched using a puncher, and the pathogen of Panax notoginseng black spot disease was inoculated into the center of each plate. After one week of incubation, the colony diameter was counted.

[0023] The results are as follows Figure 1 As shown, D6-adenophyllactone significantly inhibited the colony diameter of *Panax notoginseng* at concentrations of 1 μL / L, 5 μL / L, 10 μL / L, and 20 μL / L, with inhibition rates of 10.79%, 13.87%, 19.88%, and 23.42%, respectively. Figure 1 A). Lactulose, when inoculated onto plates containing 1 mg / L, 5 mg / L, 10 mg / L, and 20 mg / L of *Panax notoginseng*, also exhibited a significant inhibitory effect, with inhibition rates of 4.48%, 10.49%, 14.19%, and 14.58%, respectively. Figure 1 B). Citric acid at the same concentrations of 1 mg / L, 5 mg / L, 10 mg / L, and 20 mg / L also showed significant inhibitory effects on the growth of *Panax notoginseng* black spot fungus, with inhibition rates of 7.89%, 9.44%, 12.38%, and 16.41%, respectively. Figure 1 C).

[0024] 1.2 Effect of D6-Abelmolactone Induction on Panax notoginseng Resistance Experiment D6-abelacterol at concentrations of 0, 1, 5, 10, and 20 μL / L, citric acid at concentrations of 0, 1, 5, 10, and 20 mg / L, and lactulose at concentrations of 0, 1, 5, 10, and 20 mg / L were prepared for induction. The induction method was as follows: First, select uniformly growing Panax notoginseng potted plants for transplanting, remove weeds from the soil surface, water, and let them sit for one day. Then, cover the Panax notoginseng potted plants from the bottom with a plastic bag, leaving only the above-ground parts exposed. Use tape to seal the gaps in the plastic bag to prevent the sprayed substances from affecting the soil. Place the Panax notoginseng potted plants in a transparent plastic incubator, and spray the stems and leaves of Panax notoginseng with water (CK), the prepared D6-abelacterol, citric acid, and lactulose aqueous solutions, respectively. After one day of closed induction, the pathogen of Panax notoginseng black spot disease was inoculated on the leaf surface and cultured in a humid environment. After 5 days of observation, diseased leaves were collected to further study the biological characteristics of the pathogen and its effects on Panax notoginseng plants. The diseased leaves were scanned using a root scanner, and the lesion area was counted using Photoshop software and estimated using Excel software. The effect of each treatment group in inducing resistance to black spot disease was evaluated based on the lesion area.

[0025] The results showed that after one day of induction with D6-abelacterol at concentrations of 1 μL / L, 5 μL / L, 10 μL / L, and 20 μL / L, re-inoculation with black spot pathogens significantly reduced the lesion area compared to the control group, by 17.77%, 26.28%, 28.14%, and 32.85%, respectively; among them, the lesion area was smallest under the 20 μL / L D6-abelacterol treatment. Figure 2 A). Furthermore, after one day of induction with lactulose at concentrations of 1 mg / L, 5 mg / L, 10 mg / L, and 20 mg / L, the lesion area of ​​Panax notoginseng was significantly reduced compared to the control group. Figure 2 B). Similarly, after one day of induction with 1 mg / L, 5 mg / L, 10 mg / L, and 20 mg / L citric acid, the lesion area of ​​Panax notoginseng was significantly reduced compared with the control group. Figure 2 C). However, the lesion area was smallest after treatment with 20 μL / L D6-okratin.

[0026] Resistance induction treatments were initiated using 20 μL / L D6-abelacterone, 1 mg / L lactulose, and 20 mg / L citric acid. One day after induction treatment, diseased leaves were collected and lesions were scanned. Results showed that D6-abelacterone, lactulose, and citric acid reduced the lesion area by 31.81%, 28.3%, and 31.62%, respectively. Figure 3 The experimental results are consistent with previous studies, indicating that D6-okratinolactone exhibits the best effect among the three compounds, resulting in the smallest lesion area.

[0027] Example 2: Effect of D6-Abelmolactone on the Growth of Panax notoginseng One-year-old Panax notoginseng seedlings (from the Yangjie Forest Panax notoginseng Base in Xundian County) were used. D6-okratin was applied at concentrations of 0, 1, 5, 10, and 20 μL / L, and citric acid and lactulose were applied at concentrations of 0, 1, 5, 10, and 20 mg / L. Four replicates and one control were set up, for a total of 75 plots. After successful survival of the Panax notoginseng, D6-okratin was sprayed at concentrations of 0, 1, 5, 10, and 20 μL / L, citric acid at concentrations of 0, 1, 5, 10, and 20 mg / L, and lactulose at concentrations of 0, 1, 5, 10, and 20 mg / L, for a total of 20 plots with four replicates and one control. Application was done via foliar spraying or root irrigation. The experimental plots were distributed according to a field random distribution map. The first application was made two months after the Panax notoginseng seedlings had recovered from transplant shock. The pesticide was sprayed every 15 days for a total of seven applications (August 27, 2024 – December 11, 2024). Growth indicators and disease incidence were investigated using field experimental methods.

[0028] 2.1 Effect on the fresh and dried weight of Panax notoginseng Analysis of the dry and fresh weight of individual Panax notoginseng plants showed that spraying different concentrations of D6-adenosyllactone, lactulose, and citric acid could all increase the dry and fresh weight of individual Panax notoginseng plants, both above and below ground, to varying degrees.

[0029] In the D6-abelmolide treatment group, the aboveground fresh weight of single plants treated with 10 μL / L and 20 μL / L was significantly increased by 16.80% and 23.89% compared with the control (CK), respectively; the 20 μL / L treatment had the most outstanding effect, with the underground fresh weight, aboveground fresh weight, and underground dry weight of single plants significantly increased by 26.70%, 24.77%, and 26.24% compared with CK, respectively (Figure 4A).

[0030] All lactulose concentrations promoted the accumulation of Panax notoginseng biomass: the fresh weight of the underground part of the single plant in the 1, 5, 10, and 20 mg / L treatment groups was significantly increased by 10.80%, 7.74%, 8.46%, and 14.79% respectively compared with the control group; the dry weight of the aboveground part of the single plant in the 20 mg / L treatment was significantly increased by 15.12% compared with the control group; and the dry weight of the underground part of the single plant in the 10 mg / L and 20 mg / L treatments was significantly increased by 8.45% and 17.70% respectively compared with the control group (Figure 4B).

[0031] Different concentrations of citric acid spraying also significantly improved the growth indicators of Panax notoginseng: the fresh weight of the underground part of the single plant in the 5, 10, and 20 mg / L treatment groups increased significantly by 11.94%, 3.57%, and 11.22% compared with the control group, respectively; the 1-20 mg / L treatments all significantly increased the dry weight of the aboveground part of the single plant, with increases of 8.94%, 12.66%, 10.24%, and 10.08%, respectively; the dry weight of the underground part of the single plant under each concentration treatment was also significantly higher than that under the control group, with increases of 12.12%, 17.54%, 15.23%, and 21.37%, respectively (Figure 4C).

[0032] The effects of D6-adenosyllactone, lactulose, and citric acid on the fresh and dry weight of Panax notoginseng above and below ground, among which Figure 4 A, Figure 4 B Figure 4 C represents the effects of D6-okratinol, lactulose, and citric acid on the fresh weight of the aboveground and underground parts, the dry weight of the aboveground and underground parts of a single Panax notoginseng plant, respectively; (stem represents the aboveground part, and root represents the underground part). Data are expressed as mean ± standard error, and different lowercase letters indicate significant differences between treatments (p < 0.05).

[0033] The plant height of Panax notoginseng was measured using a ruler. D6-adenosyllactone had a certain promoting effect on the plant height of Panax notoginseng, and there was a concentration effect. The plant height at a concentration of 20 μL / L was significantly higher than that at a concentration of 1 μL / L, showing a certain growth-promoting trend, but the difference was not significant compared with the control group. Figure 5 A) Within the experimental concentration range, lactulose treatment did not show a significant promoting or inhibiting effect on the height of Panax notoginseng plants. Figure 5 B), D6-adenosyllactone: The plant height of the 20 μL / L treatment was significantly higher than that of the 1 μL / L treatment. Lactulose and citric acid: None of the concentration treatments had a significant effect on the plant height of Panax notoginseng, and the plant height was not significantly different from that of the control group. Figure 5 C)

[0034] In conclusion, foliar spraying of Panax notoginseng with D6-abelacterone, lactulose, and citric acid can all increase the plant height to a certain extent. Among them, D6-abelacterone showed the best effect at a concentration of 20 μL / L; lactulose showed the best effect at a concentration of 1 mg / L; and citric acid showed the best effect at a concentration of 1 mg / L.

[0035] Field disease survey results showed that spraying with the three compounds reduced the disease index of Panax notoginseng leaves to varying degrees. Among them, the disease index was lowest at a concentration of 1 μL / L for D6-abelacterone, with a relative reduction of 18.52% compared to the control (CK) (Figure 6A); the disease index was lowest at a concentration of 5 mg / L for lactulose, with a relative reduction of 12.90% compared to the control (CK). Figure 6B). The disease index was lowest when the concentration of citric acid was 20 mg / L, with a relative decrease of 21.93% in the disease index compared to the control group (CK). Figure 6 C). This study demonstrates that treatment with D6-okratin can induce resistance to black spot disease in Panax notoginseng and promote its growth.

[0036] Example 3: Effects of different concentrations of lactulose, citric acid, and D6-adenosyllactone on the quality of Panax notoginseng. The main root of Panax notoginseng was used as the test material. It was first thoroughly rinsed with running water to remove surface soil, impurities, and fine fibrous roots. After washing, it was placed in a 60 ℃ constant temperature oven and dried to constant weight. After cooling to room temperature, it was pulverized using a high-speed universal pulverizer and passed through a 100-mesh standard sieve to obtain a uniform and fine Panax notoginseng root powder. This powder was then sealed and stored in a desiccator away from light for later use. 200 mg of Panax notoginseng powder was accurately weighed and extracted with an extraction solvent using ultrasonic assisted extraction. The extract was brought to a final volume of 15 mL. After extraction, it was centrifuged at 8000 r / min for 10 min. The supernatant was then filtered through an organic phase filter membrane to prepare the test solution. High-performance liquid chromatography (HPLC) was used for detection, with acetonitrile-water as the mobile phase gradient elution, detection wavelength 203 nm, column temperature 30 ℃, flow rate 1.0 mL / min, and injection volume 10 μL. The concentrations (mg / mL) of five monomeric saponins (R1, Re, Rg1, Rb1, and Rd) were determined by external standard method and converted into mass fractions (%) of each component. The total saponin content was then calculated by summing the results.

[0037] The results showed that treatment with different concentrations of exogenous substances had a significant effect on the accumulation of Panax notoginseng saponins.

[0038] like Figure 7 As shown: The total saponin content in all D6-abelacterol treatment groups was higher than that in the control (CK): the total saponin content in the 1 μL / L treatment group was 2.62%, an increase of 11.0% compared to CK; the 5 μL / L treatment group was 2.69%, an increase of 14.0%; the total saponin content in the 10 μL / L treatment group reached the highest value of 2.89%, a significant increase of 22.5% compared to CK. At this point, the contents of major monomeric saponins such as Rg1, Rb1, and Rd also reached their peak values ​​simultaneously. It can be seen that this concentration has the best effect on promoting saponin synthesis and is the optimal application concentration for D6-abelacterol. When the concentration was increased to 20 μL / L, the total saponin content dropped back to 2.50%, with an increase of only 6.0%. The high concentration did not show a synergistic effect, but instead showed a certain inhibitory effect. Figure 7 A).

[0039] In the lactulose treatment groups, the total saponin content was 2.91% in the 1 mg / L treatment group, an increase of 23.3% compared to the control (CK); 2.41% in the 5 mg / L treatment group, an increase of 2.1%; and 2.58% in the 10 mg / L treatment group, an increase of 9.3%. The total saponin content in the 20 mg / L treatment group reached a peak of 3.16%, a significant increase of 33.5% compared to the control (CK). At this concentration, the content of various major monomeric saponins was simultaneously the highest, indicating the best saponin accumulation effect. Figure 7 B).

[0040] Citric acid treatments at all concentrations increased the total saponin content of Panax notoginseng: the total saponin contents of the 1 mg / L, 5 mg / L, 10 mg / L, and 20 mg / L treatment groups were 2.69%, 2.63%, 2.42%, and 2.62%, respectively, representing increases of 14.1%, 11.4%, 2.5%, and 11.0% compared to the control (CK). Overall, citric acid's effect on saponin accumulation was significantly weaker than that of lactulose, and high-concentration treatments did not show a synergistic effect. Figure 7 C).

[0041] A comprehensive comparison of the three exogenous substances revealed that D6-abelacterol reached its peak effect in promoting saponin accumulation at a concentration of 10 μL / L, with a significantly higher increase than the citric acid treatment groups and similar to the effect of high-concentration lactulose treatment. This indicates that the regulation of Panax notoginseng saponin synthesis by D6-abelacterol is significantly concentration-dependent; both excessively high and low concentrations are detrimental to the efficient accumulation of saponins.

[0042] Due to differences in the activity of the compounds, the concentration units of the test reagents varied in the experiment. D6-abelacterone was measured in μL / L, while lactulose and citric acid were measured in mg / L. D6-abelacterone exhibits extremely high biological activity, achieving equivalent effects at relatively low dosages. Compared to lactulose and citric acid, it has significant advantages in application cost and dosage, resulting in better overall efficacy.

[0043] Example 4: Study on the mechanism by which D6-abelacterol enhances the stress resistance and promotes growth and quality improvement of Panax notoginseng. To further demonstrate whether D6-adenosine can enhance the stress resistance of Panax notoginseng and activate its immune response, the inventors collected Panax notoginseng samples induced by D6-adenosine for RNA sequencing analysis followed by principal component analysis (PCA). PCA results showed that after 1 day of treatment with 20 μL / L D6-adenosine, the transcriptional levels of Panax notoginseng did not completely separate on PC1 and PC2. This indicates that although gene expression changed significantly after treatment, the overall transcriptional levels still showed some similarity. Specifically, the expression of 648 genes showed significant differences, with 322 genes upregulated and 326 genes downregulated.

[0044] 4.1 GO functional classification analysis of differentially expressed genes Based on molecular function (MF), cellular components (CC), and biological processes (BP), differentially expressed genes (DEGs) in Panax notoginseng treated with D6-abelacterone were annotated and classified using GO. In terms of molecular function, the differentially expressed genes were mainly enriched in binding activity, catalytic activity, transporter activity, transcriptional regulation activity, ATP-dependent activity, molecular functional regulation activity, structural molecular activity, and antioxidant activity. In terms of cellular components, the differentially expressed genes were mainly located in cellular anatomical entities and protein-containing complexes. In terms of biological processes, the differentially expressed genes were mainly involved in metabolic processes, cellular processes, stimulus responses, biological regulation, localization, developmental processes, multicellular biological processes, interspecific interactions, growth, and reproduction. Among these functional categories, both upregulated and downregulated differentially expressed genes were abundant, indicating that genes related to resistance and growth can be significantly induced by D6-abelacterone (Figure 8).

[0045] 4.2 KEGG enrichment analysis of differentially expressed genes in Panax notoginseng after D6-okratin induction KEGG pathway enrichment analysis was performed on differentially expressed genes after D6-abelacterone treatment. The results showed that differentially expressed genes in the D6xt group were significantly enriched in pathways such as pentose-glucuronic acid tautomerism, diterpene biosynthesis, flavonoid biosynthesis, plant secondary metabolite biosynthesis, plant hormone signal transduction, carotenoid biosynthesis, and nitrogen metabolism. All of these pathways are closely related to the growth, development, and disease and stress resistance of Panax notoginseng. Differentially expressed genes in the D6st group were significantly enriched in pathways such as photosynthesis, plant MAPK signaling pathway, carotenoid biosynthesis, phenylpropane biosynthesis, photosynthetic carbon fixation, starch and sucrose metabolism, and glutathione metabolism. Among these, photosynthesis was the most significantly enriched pathway, indicating that D6-abelacterone can significantly regulate the light energy utilization and carbon metabolism processes of Panax notoginseng (Figures 9 and 10).

[0046] Based on pathway enrichment analysis data, the expression changes of Panax notoginseng genes after induction were studied, and a significant increase in the expression of differentially expressed genes related to plant growth, development, and stress resistance was observed. Figure 11It is worth noting that the study on the D6-okra lactone-induced resistance to black spot disease and its promotion of Panax notoginseng growth is significant. The DEGs related to phenylpropane biosynthesis, encoding E1.11.1.7 (peroxidase), play numerous crucial roles in plants. These enzymes help plants maintain physiological stability, thus providing them with more opportunities for survival under adversity. Peroxidases not only contribute to healthy plant growth and development but also play a key role in regulating plant adaptability to the external environment, participating in the regulation of oxidative stress responses, and in chlorophyll denitrification. These functions ensure that plants can effectively resist various environmental stresses such as high temperature, weak sunlight, strong ultraviolet radiation, drought, senescence, water transport, and climate change. Plant hormones play a vital role in regulating plant growth and immune responses. By upregulating the gene encoding AUX1 (auxin influx carrier) and synergizing with its synthesis and metabolic pathways, concentration gradients and local concentration differences can be formed, thus laying the foundation for regulating plant growth and development and "tropism" in response to internal and external stimuli. The MAPK signaling pathway plays a central role in plant disease resistance, development, and stress tolerance through multi-level phosphorylation amplification and dynamic substrate regulation. Within plants, the gene encoding WRKY33 (WRKY transcription factor 33) plays multiple key roles, primarily involved in modulating the plant's immune response, enhancing its cold tolerance, and participating in strategies to cope with various stresses. PYL (abscisic acid receptor PYR / PYL family) plays a crucial biological role in plant growth, development, and defense mechanisms. It regulates physiological processes such as seed germination and stomatal closure, and responds to abiotic stresses such as drought and high temperatures. Furthermore, E3.5.1.4 (amidase), encoded in the phenylalanine metabolic pathway, is primarily responsible for catalyzing the hydrolysis of glutamine to produce glutamate and ammonia, thereby regulating ammonia and urea synthesis, promoting muscle growth, enhancing immune function, and increasing lymphocyte numbers. This pathway not only participates in multiple metabolic activities within plants but also helps remove harmful or useless chemicals, maintains the stability of the plant's internal environment, and enhances the plant's resistance to fungal, bacterial, and other harmful microorganisms. In the glycolysis / gluconeogenesis pathway, TPI (triose phosphate isomerase) plays a crucial role in plant growth and development, while GAPDH (glyceraldehyde-3-phosphate dehydrogenase) plays a central role, primarily involved in glycolysis and the Calvin cycle—core pathways for plant energy supply and carbon metabolism. In the flavonoid biosynthesis pathway, the enzyme encoded by E2.3.1.133 (shikimate O-hydroxycinnamoyltransferase) plays a key role in plants. These enzymes can catalyze the conversion of various substrates into esters or amides, thereby positively influencing the physicochemical properties and biological activity of plant secondary metabolites.By utilizing catalysis, these novel chemicals can effectively optimize plant metabolic products, thereby enhancing biological activity. Furthermore, the enzyme encoding 4CL plays a crucial role in the biosynthesis of ubiquinone and other terpene quinones, influencing plant growth and development. Multiple studies have confirmed that the genes EREBP and MYB play vital roles in plant responses to stress, determining plant adaptability under adverse conditions.

[0047] Therefore, by performing transcriptome sequencing and analysis on the treated samples, it was confirmed that D6-okratinolone can effectively enhance the stress resistance of Panax notoginseng and activate its immune response by regulating multiple metabolic and signal transduction pathways.

[0048] In summary, using the compounds of this invention for disease control not only activates the plant's own growth and metabolism, promoting its growth and development and improving the yield and quality of Panax notoginseng, but also significantly reduces the amount of chemical fertilizers and pesticides used, providing important support for promoting green and sustainable agriculture. This method effectively reduces the use of agrochemicals, significantly improves the quality of Panax notoginseng, and is environmentally friendly.

Claims

1. Application of D6-adenosine lactone in the preparation of products that promote the growth of Panax notoginseng, induce its disease resistance, and improve its quality.

2. The application as described in claim 1, characterized in that: The concentration of the D6-adenosine aqueous solution is 1-10 μL / L.

3. The application as described in claim 2, characterized in that: The concentration of the D6-okratin lactone solution is 5-10 μL / L.

4. The application as described in claim 1, characterized in that: The disease mentioned is Panax notoginseng black spot disease.

5. The application as described in claim 1, characterized in that: Apply by foliar spraying or root irrigation.

6. The application as described in claim 1, characterized in that: The improvement of Panax notoginseng quality refers to increasing the content of its effective components.

Images