Method for inhibiting generation of branch thorns of lycium ruthenicum by using trehalose-6-dipotassium phosphate

Abstract

The invention relates to the technical field of forest biology, in particular to a method for inhibiting generation of branch thorns of lycium ruthenicum by using trehalose-6-dipotassium phosphate. And dropwise adding the trehalose-6-dipotassium phosphate solution to the terminal bud of the thorn plant of the lycium ruthenicum. The trehalose-6-dipotassium phosphate significantly inhibits the occurrence of the branch thorns of the lycium ruthenicum, has concentration dependence, has the most significant treatment effect of 480 [mu] M, can almost completely inhibit the generation of the branch thorns, and significantly inhibits the occurrence of the branch thorns in 80 [mu] M and 160 [mu] M treatment groups under the condition of hardly influencing the growth of stems and leaves. 320 [mu] M and 480 [mu] M treatment groups significantly inhibit the occurrence of branch thorns and also significantly inhibit the growth of stem lengths, which results in plant dwarfing and no thorn. In large-scale cultivation of spiny economic crops, the spiny-free characteristic solves the problem of harvesting scratching, stem and leaf growth regulation and control are matched, a supporting frame does not need to be built, protection equipment can be omitted during harvesting, and the manual harvesting efficiency is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of forestry biotechnology, in particular to a method for inhibiting the occurrence of Lycium ruthenicum thorns by trehalose-6-phosphate dipotassium salt. BACKGROUND

[0002] Lycium ruthenicum is a multi-spine shrub of Solanaceae Lycium, which is a key constructive species in the desert and semi-desert ecosystems in Northwest China, and has significant ecological and economic values. Ecologically, it is drought and salt tolerant, with developed root system, and can prevent wind and sand, and improve saline-alkali soil, thus being a pioneer plant for ecological restoration in arid and semi-arid regions. Economically, the fruits are rich in physiological active ingredients such as anthocyanins, proanthocyanidins and lycium polysaccharides, and have medicinal and edible values, thus having broad industrialization prospects in the fields of functional food and medical health care.

[0003] However, the thorns of Lycium ruthenicum are hard and sharp, which has become a core bottleneck restricting its large-scale cultivation. During harvesting, the thorns are easy to scratch the operators, and thus protective equipment needs to be additionally provided, increasing the difficulty and cost. The thorns also cause mechanical damage to the fruits, aggravate the loss during harvesting and transportation, and reduce the product quality. In cultivation, the plants with thorns and high height need to be supported to prevent lodging, increasing the cultivation cost. The development of thorns consumes the nutrients of stems, which inevitably reduces the transportation of photosynthetic products to fruits and other organs, and restricts the yield of fruits and the accumulation of core ingredients such as lycium polysaccharides. In the scenario of facility agriculture, the thorny plants also limit the planting density, and cannot adapt to the mechanical harvesting equipment, resulting in low efficiency.

[0004] To solve the problem of thorns, the existing technologies have obvious defects: physical pruning needs to be operated on each plant by hand, which is time-consuming and labor-intensive, and the thorns are easy to regenerate; chemical agents mostly rely on hormone substances, and the inhibitory effect is limited, and may also cause pesticide residues and plant deformity; traditional breeding selects thornless strains through hybridization, which has a long cycle, low efficiency, and is difficult to obtain stable genetic varieties; although gene editing technology is accurate, it has high threshold and high cost, and is difficult to be popularized on a large scale.

[0005] The present application creates a new method for completely inhibiting the occurrence of Lycium ruthenicum thorns by trehalose-6-phosphate dipotassium salt solution, which lays a solid foundation for cultivating Lycium ruthenicum with no thorns and more suitable for production, and also provides a research clue for the dwarfing of Lycium ruthenicum. SUMMARY

[0006] In order to solve the above problems, the present application provides a method for inhibiting the occurrence of Lycium ruthenicum thorns by trehalose-6-phosphate dipotassium salt, which can inhibit the occurrence of Lycium ruthenicum thorns.

[0007] In order to achieve the above purpose, the present application provides the following technical scheme:

[0008] The application provides a method for inhibiting the occurrence of Lycium ruthenicum Murr. thorn by trehalose-6-phosphate dipotassium salt, comprising the following steps: dropping a trehalose-6-phosphate dipotassium salt solution on the top bud of a Lycium ruthenicum Murr. thorny plant.

[0009] Preferably, the concentration of the trehalose-6-phosphate dipotassium salt solution is above 80 muM.

[0010] Preferably, the concentration of the trehalose-6-phosphate dipotassium salt solution is above 160 muM.

[0011] Preferably, the concentration of the trehalose-6-phosphate dipotassium salt solution is above 320 muM.

[0012] Preferably, the concentration of the trehalose-6-phosphate dipotassium salt solution is above 480 muM.

[0013] Preferably, the trehalose-6-phosphate dipotassium salt solution is dropped once a day, and the dropping is repeated for 14 days.

[0014] Preferably, the amount of the trehalose-6-phosphate dipotassium salt solution dropped on each top bud of the Lycium ruthenicum Murr. thorny plant is 10 muL.

[0015] The application has the following beneficial effects:

[0016] Exogenous application of trehalose-6-phosphate dipotassium salt can significantly inhibit the occurrence of Lycium ruthenicum Murr. thorn, and the inhibitory effect has concentration dependence, wherein the treatment effect of 480 muM is the most significant, and the thorn production can be almost completely inhibited, and the treatment groups of 80 muM and 160 muM can significantly inhibit the occurrence of thorn without affecting the growth of stems and leaves. The treatment groups of 320 muM and 480 muM can significantly inhibit the occurrence of thorn while significantly inhibiting the growth of stem length, which will lead to dwarfing and thornless of the plant. In the large-scale cultivation of thorny economic crops, the thornless characteristic solves the problem of injury during harvesting, cooperates with the growth regulation of stems and leaves (the plant height is stable within 1.2 m), does not need to build a support frame, and can save protective equipment during harvesting, thereby improving the efficiency of artificial harvesting; at the same time, the nutrient consumption of stems is reduced, which can promote the directional transport of photosynthetic products to fruits, and provides a physiological basis for fruit yield and accumulation of core functional components (such as Lycium barbarum polysaccharide). In the scenario of facility agriculture / high-density cultivation, the compact plant type of stems and leaves can improve the planting density per unit area (referring to the dwarfing regulation effect of similar crops, the planting density can be doubled), and the thornless and dwarfing compound traits can directly adapt to the mechanical harvesting equipment such as vibration type, avoid the entanglement of thorn body with mechanical parts, and reduce the falling and extrusion loss of fruits during harvesting of high plant type, thereby significantly reducing the damage rate of fruits during harvesting. BRIEF DESCRIPTION OF DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the drawings needed in the embodiments will be briefly introduced below.

[0018] Figure 1 To compare the phenotypes of new stems in different treatment groups of black goji berries, A, blank control; B, water control; C, 80 μM group; D, 160 μM group; E, 320 μM group; F, 480 μM group; white circles mark the position of the first stem node below the terminal bud before treatment. Detailed Implementation

[0019] This invention provides a method for inhibiting the occurrence of thorns on branches of black goji berries using trehalose-6-phosphate dipotassium salt, comprising the following steps: adding trehalose-6-phosphate dipotassium salt solution dropwise onto the terminal buds of thorny black goji berry plants.

[0020] In this invention, the concentration of the trehalose-6-dipotassium phosphate solution is preferably 80 μM or higher. In this invention, the concentration of the trehalose-6-dipotassium phosphate solution is preferably 160 μM or higher. In this invention, the concentration of the trehalose-6-dipotassium phosphate solution is preferably 320 μM or higher. In this invention, the concentration of the trehalose-6-dipotassium phosphate solution is preferably 480 μM or higher. In this invention, the trehalose-6-dipotassium phosphate solution is preferably added once daily for 14 days. In this invention, the amount of trehalose-6-dipotassium phosphate solution added to the terminal bud of each thorny black goji berry plant is preferably 10 μL.

[0021] To further illustrate the present invention, the following detailed description is provided in conjunction with embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0022] Example 1

[0023] 1. Experimental Methods

[0024] 1.1 Preparation of Trehalose-6-phosphate dipotassium salt solution

[0025] 1.1.1 Weighing the reagents

[0026] In a light-protected area, weigh 10.0 mg of trehalose-6-phosphate dipotassium salt powder using an electronic balance with an accuracy of 0.1 mg. Quickly transfer the powder to a clean, dry 50 mL beaker to prevent the purity of the powder from being affected by light or air exposure. Add 3-5 mL of ultrapure water to the beaker and stir slowly and evenly with a glass rod until the powder is completely dissolved. If the powder adheres to the beaker wall, gently rinse it with a small amount of ultrapure water.

[0027] 1.1.2 Dissolving and adjusting to volume

[0028] After the powder dissolves, slowly transfer the solution to a 10mL volumetric flask along a glass rod (the glass rod should be close to the inner wall of the beaker and below the graduation mark). Then, wash the inner wall of the beaker and the glass rod 2-3 times with 1mL of ultrapure water, ensuring all washings are transferred to the volumetric flask to avoid solute loss. Add ultrapure water to the volumetric flask until the liquid level is 1-2mm from the graduation mark, then use a dropper to add water dropwise (keeping the line of sight horizontal to the graduation mark) until the lowest point of the meniscus is tangent to the graduation mark. Tighten the stopper, hold the stopper down, and support the bottom of the flask. Invert and shake 5-8 times (standing upright after each inversion) to ensure the solution is thoroughly mixed. This yields a 1000mg / L stock solution [corresponding to 2006μmol / L (μM), calculated based on the molar mass of trehalose-6-dipotassium phosphate 498.46g / mol].

[0029] 1.1.3 Diluting the mother liquor to prepare the working solution

[0030] Based on a prepared 1000 mg / L (2006 μM) trehalose-6-dipotassium phosphate stock solution, four target solutions were prepared using a dilution method: The stock solution was measured using a pipette or pipette with the corresponding volume range, and then injected into a clean 10 mL volumetric flask. Following the same method, ultrapure water was added to the volumetric flask until the liquid level was 1-2 mm from the graduation mark. Dropping was then done with a dropper until the meniscus was tangent to the graduation mark. The flask was then tightly stopped, inverted, and shaken 5-8 times to mix thoroughly. The required stock solution volumes and their corresponding relationships for each target concentration were as follows: 0.4 mL for 80 μM (39.88 mg / L), 0.8 mL for 160 μM (79.75 mg / L), 1.6 mL for 320 μM (159.51 mg / L), and 2.4 mL for 480 μM (239.3 mg / L). All solutions were then diluted to 10 mL.

[0031] 1.1.4 Storage and Precautions

[0032] The trehalose-6-phosphate dipotassium salt powder of this invention was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., and the ultrapure water was obtained from the Assets and Laboratory Management Department of Shenyang Agricultural University. The prepared solution was transferred to a brown, low-temperature resistant reagent bottle, immediately sealed, and labeled with the solution name, concentration, and preparation date. It was then stored in a -20°C freezer, protected from light. The entire solution preparation process must be carried out under light-protected conditions. This can be achieved by using a light-protected operating cabinet, or by using a light-shielding cloth to completely cover the operating table and all equipment, including volumetric flasks and beakers, to ensure that the solution is not directly exposed to light during preparation and transfer.

[0033] 1.2 Obtaining Plant Materials

[0034] The black goji berry material used in the experiment was grown at the College of Forestry, Shenyang Agricultural University (Liaoning Provincial Key Laboratory of Forest Tree Genetics, Breeding and Cultivation). The specific acquisition and cultivation process is as follows:

[0035] 1.2.1 Culture medium preparation

[0036] First, weigh out MS dry powder, sucrose, and agar, place them in a pot, and add ultrapure water to make up 80% of the final total volume. Heat to a boil using an induction cooker, stirring continuously until all components are completely dissolved. Adjust the volume of the dissolved mixture to the preset mark, then add 4% (m / v) NaOH solution dropwise using a dropper, stirring constantly with a glass rod. Simultaneously monitor the pH using a pH meter, adjusting the initial pH to 6.0-6.3 (because the pH will naturally decrease by 0.1-0.3 after autoclaving at 121℃ for 20 minutes, eventually stabilizing within the target range of 5.7-6.2). Transfer the pH-adjusted culture medium to an autoclave and sterilize at 121℃ for 20 minutes. Cool before use.

[0037] 1.2.2 Propagation of Tissue Culture Seedlings

[0038] Using a sterile tissue culture seedling line of black goji berries in the laboratory tissue culture room as the subject, when the seedlings reached 3-5 cm in length, the upper half of the stem nodes was cut in a clean bench and inoculated into freshly prepared 1 / 2 MS medium. After rooting, the first generation of micro-cutting propagation was completed. This process was repeated to complete at least 10 generations of micro-cutting culture. This process not only provides sufficient experimental material for subsequent transplanting, but more importantly, through stable propagation over multiple generations, it ensures that the genetic background of all tested plants is highly consistent, avoiding interference with subsequent experimental results due to genetic differences in the materials. To improve the environmental adaptability of tissue culture seedlings after transplanting, hardening-off is carried out one week before transplanting: when the stem nodes of the tissue culture seedlings reach 5-7cm in length, the bottles along with the seedlings are transferred to a well-ventilated indoor environment with sunlight and cultured at room temperature for 3-4 days (temperature 25±2℃). After the hardening-off is completed, the caps of the culture bottles are opened and the seedlings are placed in the same environment for another 3-4 days to further enhance the seedlings' tolerance to external humidity and light.

[0039] 1.2.3 Preparations before transplanting

[0040] Select nutrient soil that meets the following parameters (pH 6.5-6.8, N / P / K ≥ 12 g / kg, moisture content ≤ 40%, organic matter ≥ 40%, silicon ≥ 0.3 g / kg), fill high-temperature inoculum bags (9.5 cm in diameter, 30 cm in length, and 0.055 mm thick) to 3 / 4 of their volume, and sterilize them in an autoclave at 121℃ for 40 minutes. After sterilization, pour out the nutrient soil and allow it to air dry indoors to remove excess moisture. Remove the hardened tissue culture seedlings and thoroughly rinse the roots with tap water to remove any residual culture medium (avoid damaging the roots during this process). Then, soak them in a pre-prepared carbendazim solution (1 g of carbendazim powder dissolved in 2 L of tap water) for approximately 30 minutes.

[0041] 1.2.4 Transplanting Operation

[0042] Fill the flowerpot with potting soil to 3 / 4 of its volume and gently compact it. Dig a hole in the soil to a depth of about 2 / 3 of the root system length. Place the soaked plant roots into the hole in a spiral shape, cover with potting soil, and water thoroughly with a fungicide solution. If the soil level drops significantly after covering, add more potting soil and water thoroughly again. After transplanting, cover the plant with a light-transmitting and breathable plastic cup to create a moisture-retaining environment for the seedlings. Then place the flowerpot in a laboratory light cultivation chamber (16 hours of light / 8 hours of darkness, temperature 25±2℃).

[0043] 1.2.5 Post-maintenance management

[0044] Every 2-3 days, observe the soil moisture in the morning. Water each pot with 130-150mL of water each time, ensuring the water is evenly distributed on the soil surface and fully penetrates to the roots. Hang yellow sticky insect traps in the cultivation room to prevent pests such as aphids and spider mites. If mycelium appears on the soil surface, remove it promptly and water with a fungicide solution. If pests occur, prepare an imidacloprid or high-efficiency cypermethrin solution according to the scale of the infestation and spray it on the plant surface every 2 days until the pests are eliminated. 30-40 days after transplanting, when the plant is growing vigorously (2-3 new stems and 4-7 cm of stem elongation), remove the plastic cup cover and allow the plant to grow naturally. After continuing cultivation for 3-4 weeks, the number of new stems will increase to 3-6 and the elongation will be 9-12 cm. More than half of the stems will be fully thorny (thorns grow at the base of the stem nodes, with 3-6 leaves clustered at the base, each cluster containing one main leaf and several leaflets). Finally, prune any thornless stems to ensure that the plant meets the experimental requirement of "thorns on the entire stem".

[0045] 1.3 Treatment method for terminal buds of black wolfberry

[0046] This invention investigates the effects of trehalose-6-phosphate dipotassium salt on the phenotypic characteristics of thorns and stems and leaves by applying different treatments to the terminal buds of thorny black goji berry plants. The specific operating procedures and control conditions are as follows:

[0047] 1.3.1 Screening and Grouping of Experimental Materials

[0048] Thorny black goji berry plants with consistent growth environment, similar growth vigor, and no pests or diseases were selected as experimental materials. Following the principle of "equal quantity and uniformity," the plants were divided into 6 groups, with 25 branches treated in each group. All branches were derived from 5 independent pots (5 branches per pot) to ensure sample independence and randomization of grouping. The specific settings for each group are as follows: 80μM trehalose-6-dipotassium phosphate treatment group (80μM group), 160μM trehalose-6-dipotassium phosphate treatment group (160μM group), 320μM trehalose-6-dipotassium phosphate treatment group (320μM group), 480μM trehalose-6-dipotassium phosphate treatment group (480μM group), water control group (terminal buds treated with an equal volume of solvent water), and blank control group (terminal buds not treated).

[0049] 1.3.2 Standardized labeling of observation stem segments

[0050] To eliminate experimental errors caused by differences in observation locations, the observation stem segments of all plants need to be uniformly marked before the experiment: use a black marker to mark the "first stem node below the terminal bud" on each observation stem. This mark serves as the reference position for subsequent measurements of indicators such as "new stem growth length and number of new stem nodes".

[0051] 1.3.3 Apical bud treatment procedure

[0052] The experiment was conducted in the light cultivation room of the College of Forestry, Shenyang Agricultural University. At 9:00 AM daily, the terminal buds of each group were treated using the following methods: For the 80μM group, 10μL of a pre-prepared 80μM trehalose-6-phosphate dipotassium solution was precisely pipetted and slowly added to the surface of the terminal buds of the spiny stems, ensuring complete adhesion (no dripping or leakage). The treatment for the 160μM, 320μM, and 480μM groups was the same as for the 80μM group, except that the solution was replaced with the corresponding concentration of trehalose-6-phosphate dipotassium solution. For the water control group, 10μL of ultrapure water (solvent) was pipetted and added to the terminal buds in the same manner to eliminate interference from the solvent itself on the growth of the terminal buds. For the blank control group, no treatment was applied to the terminal buds; only routine watering was performed.

[0053] 1.3.4 Control of test conditions

[0054] Each treatment group was repeated once daily for 14 consecutive days to ensure consistent treatment duration. During the experiment, the indoor environmental conditions for light cultivation were strictly maintained at a constant temperature of 25±2℃, with the light source provided by six 18W LED T8 plant growth lamps, and a light cycle of 16 hours of light / 8 hours of darkness per day. Simultaneously, the watering frequency and amount were standardized for each group of plants to minimize the interference of non-treatment factors on the experimental results.

[0055] 1.4 Statistical Analysis of Phenotypic Data

[0056] Phenotypic data statistics include the growth length of new stems, average number of new stem nodes, average number of new leaves, average number of leaves per cluster, average length of leaves on the fifth stem node below the terminal bud, average number of new thorns, average thorn emergence rate, and percentage of completely thornless new stems during the treatment period. The methods for measuring each indicator are as follows: the growth length of new stems was measured using vernier calipers, with the measurement being the distance from the black marker point before the experiment to the location of the terminal bud after the experiment; the average number of new stem nodes refers to the number of new stem nodes above the black marker point; the average number of new leaves is the number of new leaves above the black marker point; the average number of leaves per cluster is the total number of new stem leaves divided by the number of new stem nodes; the number of new thorns is the number of new stem nodes with thorns; the average thorn emergence rate is the ratio of the number of new thorns to the number of new stem nodes; and the percentage of completely thornless new stems is the ratio of completely thornless new stems to the total number of stems in the treatment. Microsoft Office Excel was used for data organization and preliminary analysis. SPSS 23.0 was used for significant difference analysis. One-way ANOVA combined with LSD method was used for multiple comparisons, where P < 0.05 indicated a significant difference and P < 0.01 indicated a highly significant difference.

[0057] 2 Experimental Results

[0058] 2.1 Trehalose-6-phosphate dipotassium salt inhibits thorn formation in black wolfberry branches.

[0059] Trehalose-6-phosphate dipotassium salt significantly inhibited the formation of thorns on new stem nodes of black wolfberry in a concentration-dependent manner, specifically by increasing the proportion of thornless stems, reducing the number of thorns on new branches, and decreasing the thorn emergence rate. Statistical analysis showed that the blank control group had no newly formed completely thornless stems (…). Figure 1 (A); In the water control group, 11.09% of newly sprouted stems were completely thornless, while the overall population still consisted mainly of thorny stems (A). Figure 1 The majority of the trehalose-6-phosphate groups were treated with trehalose (B); while the 80μM, 160μM, 320μM, and 480μM trehalose-6-phosphate dipotassium salt treatment groups all showed a distinct thornless phenotype. Figure 1 (C-F), of which the 480μM treatment group produced completely thornless stems ( Figure 1 The highest proportion of thornless plants was 91.18%, which almost completely eliminated thorns and had the most significant effect.

[0060] There were no significant differences in thorn size (length and width) between the treatment groups and the control group; the thorn emergence rate of all trehalose-6-phosphate dipotassium salt treatment groups was significantly lower than that of the control group (P < 0.0001, Table 1). The blank control group had the most new thorns (15.62) and the highest thorn emergence rate (71.03%) (Table 1); the water control group had 12.40 new thorns and a thorn emergence rate of 60.14%, which was not significantly different from the blank control group (Table 1), indicating that water treatment had no significant regulatory effect on the occurrence of thorns in black wolfberry, and the interference of water can be excluded. The number of new thorns in the 80 μM trehalose-6-phosphate dipotassium salt treatment group was 5.61, and the thorn emergence rate was 28.19%, which was significantly lower than that in the blank control group (P < 0.0001, Table 1). There was no significant difference between the 160 μM treatment group and the 80 μM group (P > 0.05), but both were significantly lower than the control group. The number of new thorns in the 320 μM treatment group decreased to 2.52, and the thorn emergence rate decreased to 12.04%. The 480 μM treatment group had almost no new thorns, and the thorn emergence rate was only 0.04%. The number of thorns and the thorn emergence rate in the 320 μM and 480 μM groups were significantly lower than those in the 80 μM and 160 μM groups (Table 1). The stepwise decreasing trend of thorn emergence rate and number of new branch thorns in the 80μM→160μM→320μM→480μM groups (Table 1) confirms the technical characteristic that the higher the concentration of trehalose-6-phosphate dipotassium salt, the stronger the inhibitory effect on the occurrence of thorns in black wolfberry branches.

[0061] Table 1. Effects of exogenous application of trehalose-6-phosphate dipotassium salt solution on the phenotype of potted black goji berry seedlings.

[0062]

[0063]

[0064] Note: The data in the table are the mean ± standard error of five repeated trials. The differences between data in the same row marked with different uppercase letters are extremely significant (P < 0.01, LSD), and the differences between data in the same row marked with different lowercase letters are significant (P < 0.05, LSD).

[0065] In summary, the number of new shoot thorns and the thorn emergence rate in the 80–480 μM trehalose-6-phosphate dipotassium salt treatment groups were significantly lower than those in the blank control group and the water control group (P<0.0001), confirming that trehalose-6-phosphate dipotassium salt has a strong inhibitory effect on the thorn development of black wolfberry. There was no significant difference in the number of new shoot thorns and the thorn emergence rate between the water control and the blank control (P>0.05). This result is consistent with the experimental design objective of including a water control in Section 1.3.3 to eliminate the interference of water, verifying that the water control can effectively eliminate the non-treatment interference of water on thorn development, and clarifying that the thorn inhibition effect is caused by trehalose-6-phosphate dipotassium salt.

[0066] 2.2 Effects of Trehalose-6-phosphate dipotassium salt on the phenotypic characteristics of stems and leaves of black wolfberry

[0067] Trehalose-6-phosphate dipotassium salt had a slight regulatory effect on the phenotype of new stems and leaves of black goji berries, with an overall effect weaker than its inhibitory effect on thorns. There were no significant differences in the average elongation of new stems among the blank control, water control, 80 μM treatment group, and 160 μM treatment group, except for significant reductions in the 320 μM and 480 μM treatment groups (Table 1). There were no significant differences in the number of new stem nodes and the thickness of the stem midpoint among the groups (Table 1).

[0068] The blank control group had the most new leaves, while the water control and the 80–480 μM groups showed a significant decrease. There was no significant difference in the number of leaves per cluster among the groups. In terms of leaf size, only the 480 μM treatment group showed a significant difference from the blank control group, while there were no significant differences among the other groups (Table 1).

[0069] In summary, exogenous application of trehalose-6-phosphate dipotassium salt significantly inhibited thorn formation in black wolfberry, and this inhibitory effect was concentration-dependent. The 480 μM treatment was the most effective, almost completely inhibiting thorn formation. The 80 μM and 160 μM treatments significantly inhibited thorn formation with almost no impact on stem and leaf growth. While the 320 μM and 480 μM treatments significantly inhibited thorn formation, they also significantly inhibited stem growth, leading to dwarfing and thornlessness. In the large-scale cultivation of thorny economic crops, thornlessness solves the problem of harvesting scratches. Combined with stem and leaf growth regulation (stabilizing plant height within 1.2 m), it eliminates the need for support frames, reducing the need for protective equipment during harvesting and improving manual harvesting efficiency. Simultaneously, reduced stem nutrient consumption promotes the directional transport of photosynthetic products to the fruit, providing a physiological basis for fruit yield and the accumulation of core functional components (such as wolfberry polysaccharides). In facility agriculture / high-density cultivation scenarios, the compact stem and leaf plant type can increase the planting density per unit area (referring to the dwarfing control effect of similar crops, the planting density can be doubled), and the thornless and dwarf composite traits can be directly adapted to mechanized harvesting equipment such as vibration type, avoiding the thorns from getting entangled in mechanical parts, while reducing the fruit drop and crushing loss when harvesting tall plants, thus significantly reducing the fruit harvesting damage rate.

[0070] This invention not only provides crucial clues and suitable experimental materials for elucidating the physiological mechanism of thorn formation in black goji berries, but also offers feasible regulators and technical solutions for the industrial application of thornless cultivation of black goji berries. Thornless or low-thorn content reduces branch damage and labor costs during harvesting, laying an important foundation for optimizing subsequent thorn control strategies.

[0071] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for inhibiting the occurrence of Lycium ruthenicum thorns using trehalose-6-phosphate dipotassium salt, characterized by, The method comprises the following steps: A trehalose-6-phosphate dipotassium salt solution is added dropwise to the top bud of the Lycium ruthenicum plant.

2. The method of claim 1, wherein, The concentration of the trehalose-6-phosphate dipotassium salt solution is above 80 μM.

3. The method of claim 1, wherein, The concentration of the trehalose-6-phosphate dipotassium salt solution is above 160 μM.

4. The method of claim 1, wherein, The concentration of the trehalose-6-phosphate dipotassium salt solution is above 320 μM.

5. The method of claim 1, wherein, The concentration of the trehalose-6-phosphate dipotassium salt solution is above 480 μM.

6. The method of claim 1, wherein, The trehalose-6-phosphate dipotassium salt solution is added dropwise once a day, and the process is repeated for 14 days.

7. The method of claim 1, wherein, The amount of the trehalose-6-phosphate dipotassium salt solution added dropwise to the top bud of each Lycium ruthenicum plant is 10 μL.

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