Fruit tree grafting growth promoter

Abstract

The invention discloses a fruit tree grafting growth promoter, and relates to the technical field of fruit tree cultivation. The growth promoter is prepared from the following raw materials in parts by weight: 0.01 to 0.05 part of methyl jasmonate, 0.1 to 0.3 part of salicylic acid nano microcapsules, 2 to 5 parts of bacillus velezensis, 1 to 3 parts of trichoderma harzianum, 5 to 10 parts of mesoporous silica nanoparticles, 3 to 6 parts of zinc alginate compound, 5 to 8 parts of herba portulacae extract, 2 to 4 parts of ginkgo leaf nano extract, 1 to 2 parts of chitosan oligosaccharide, 2 to 4 parts of pullulan and 100 to 200 parts of plasma activated water. The growth promoter disclosed by the invention realizes innovation of an action mechanism, prolongs the action lasting period, breaks through the adaptability limitation of a grafting technology, successfully utilizes and optimizes the micro-ecology of a grafting interface, and is environment-friendly and high in safety.

Description

Technical Field

[0001] This invention relates to the field of fruit tree cultivation technology, specifically to a fruit tree grafting growth agent. Background Technology

[0002] Fruit tree grafting, as a core technology of asexual reproduction, is a key means to ensure the breeding of superior varieties, dwarfing cultivation, improvement of stress resistance, and renewal of old orchards. The key to successful grafting lies in the rapid formation of callus tissue and the effective connection of the vascular bundle system between the cuts of the rootstock and scion. This process is subject to complex regulation by multiple factors, including plant endogenous hormones, nutritional status, environmental factors, and microbial communities.

[0003] Currently, most commercially available fruit tree grafting growth regulators are based on traditional plant hormone theory, with formulations primarily relying on simple combinations of auxins (such as indoleacetic acid (IAA), naphthaleneacetic acid (NAA), and indolebutyric acid (IBA)) and cytokinins (such as 6-benzylaminopurine (6-BA) and kinetin (KT). While these growth regulators can promote cell division to some extent, their practical applications have gradually revealed significant limitations. First, the mechanisms of action of these products are relatively simple, mainly focusing on stimulating cell division while generally neglecting the importance of the plant's systemic defense response triggered by grafting wounds. They fail to effectively activate key damage signaling pathways such as jasmonic acid, resulting in poor callus formation quality and susceptibility to pathogen infection. Second, exogenous chemical hormones are easily degraded within the plant and in the natural environment, with a short duration of action, making it difficult to maintain a stable and effective stimulating signal throughout the critical healing period. Furthermore, existing formulas are mostly "general-purpose" and lack specific consideration for the physiological characteristics of different tree species and grafting methods. Especially in distant grafting where there are significant differences in xylem structure, the survival rate is often less than 50% due to the difficulty in overcoming physiological barriers between the rootstock and scion. More importantly, existing technologies completely ignore the positive or negative impact of the microbial community in the grafting interface microecological environment on the healing process and fail to utilize the potential of beneficial endophytes to promote healing.

[0004] Therefore, developing a novel growth regulator that can synergistically regulate plant signal transduction, nutrient supply, and microecological balance has become an urgent need in the industry. Summary of the Invention

[0005] To address the above-mentioned problems, this invention provides a fruit tree grafting growth agent.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: This invention provides a fruit tree grafting growth agent, the raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.01~0.05 parts, salicylic acid nanocapsules 0.1~0.3 parts, Bacillus belye 2~5 parts, Trichoderma harzianum 1~3 parts, mesoporous silica nanoparticles 5~10 parts, zinc alginate complex 3~6 parts, purslane extract 5~8 parts, ginkgo leaf nano extract 2~4 parts, chitosan oligosaccharide 1~2 parts, pullulan polysaccharide 2~4 parts, and plasma activated water 100~200 parts.

[0007] Furthermore, the raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.03 parts, salicylic acid nanocapsules 0.2 parts, Bacillus belye 3 parts, Trichoderma harzianum 2 parts, mesoporous nano silica 8 parts, zinc alginate complex 4 parts, purslane extract 6 parts, ginkgo leaf nano extract 3 parts, chitosan oligosaccharide 1.5 parts, pullulan polysaccharide 3 parts, and plasma-activated water 150 parts.

[0008] In this formulation, methyl jasmonate and salicylic acid nanocapsules together constitute the signaling component; Bacillus belyssus and Trichoderma harzianum together constitute the microbial component; mesoporous silica nanoparticles and zinc alginate complex together constitute the nanocarrier nutrient component; purslane extract and ginkgo leaf nano-extract together constitute the plant active extract component; and chitosan oligosaccharide and pullulan polysaccharide together constitute the adjuvant component. The functions of each ingredient in the formulation are as follows: Among the signaling components, methyl jasmonic acid, as a core damage signaling molecule, activates the defense and healing responses of rootstocks and scions in advance, initiating systemic resistance; salicylic acid nanocapsules, as immune signaling molecules, enhance the innate immune response of plants through slow release via nanocapsules, forming cross-protection with the jasmonic acid pathway.

[0009] Among the microbial components, Bacillus belyssus is a beneficial endophyte that can colonize at the grafting interface, producing growth-promoting metabolites (such as IAA), extracellular polysaccharides (promoting cell adhesion), and antibacterial substances, forming a beneficial microecology; Trichoderma harzianum is a beneficial fungus that can competitively inhibit pathogens, and its hyphal network can resemble plant vascular bundles, temporarily assisting in the transport of water and nutrients, alleviating water stress in the early stages of grafting.

[0010] In the nanocarrier nutrient components, mesoporous silica nanoparticles are high-performance carriers. Their huge specific surface area and regular nanoscale pores can load and slowly release signaling molecules and nutrient elements, achieving targeted and long-lasting delivery. Zinc alginate complex has film-forming, water-retaining and nutritional functions (slow-release zinc ions). Zinc is a key cofactor for many synthetic enzymes and is essential for healing.

[0011] Among the plant active extract components, purslane extract is rich in natural plant stimulants (such as norepinephrine), providing a mild and continuous stimulation of cell division and reducing dependence on single synthetic hormones; ginkgo leaf nano-extract is rich in antioxidants such as flavonoids, and is more easily absorbed after nano-sizing, effectively clearing reactive oxygen species accumulated at the grafting wound and protecting cell membrane integrity.

[0012] Among the adjuvant components, chitosan oligosaccharide is a natural inducer that can further enhance the plant's defense response and has a certain antibacterial effect. Pullulan polysaccharide is a biodegradable film-forming agent that forms a physical protective film at the interface, reducing water evaporation and pathogen invasion.

[0013] Plasma-activated water has high permeability and activity, and as a solvent, it can enhance the solubility and bioavailability of various components.

[0014] Building upon this foundation, a positive interaction exists between signaling and microbial components. Signaling molecules create a favorable microenvironment for microbial colonization and functional expression, while microorganisms enhance signal transduction, amplifying defense and healing signals, thus forming a signal-microbe synergistic system. Simultaneously, the nanocarrier nutrient components act as a "performance multiplier" for the entire system. On one hand, they provide beneficial microorganisms with an ideal colonization "home" and nutrient source, significantly improving their survival rate and colonization density. On the other hand, the targeted delivery and sustained-release properties of the nanocarrier enable the directed release of limited signaling molecules and nutrients, maximizing efficiency. Furthermore, the composite membrane structure formed by zinc alginate and pullulan provides a physically stable and humid microenvironment for the above reactions. Based on this, the interaction of each component achieves a synergistic effect of "signal activation - microbial interaction - carrier delivery / nutrient support."

[0015] Further, the preparation method of the salicylic acid nanocapsules is as follows: accurately weigh 0.2 parts of salicylic acid and 1.0 parts of polylactic acid-glycolic acid copolymer, dissolve them together in 10 parts of dichloromethane, and slowly add them dropwise to 20 parts of an aqueous solution containing 1% (w / v) PVA under magnetic stirring at 500 rpm. Emulsify at 10000 rpm for 3 min using a high-speed disperser to form a primary emulsion; quickly pour the primary emulsion into 50 parts of an aqueous solution containing 2% (w / v) PVA, and stir at 400 rpm to form a secondary emulsion; continue stirring at 400 rpm for 4-6 h to allow the dichloromethane to completely evaporate and the microcapsules to solidify; then collect the microcapsules by centrifugation at 10000 rpm for 10 min, wash them 3 times with deionized water to remove PVA, and finally freeze-dry to obtain powdered salicylic acid nanocapsules.

[0016] Further, the preparation method of the zinc alginate complex is as follows: sodium alginate is slowly added to deionized water at 60°C and magnetically stirred at 600 rpm for 2 hours to fully dissolve it, thus preparing a 5% sodium alginate solution; under continuous stirring, the sodium alginate solution is slowly added dropwise to a 10% zinc sulfate solution, and after the addition is complete, the reaction continues for 2 hours; after the reaction is completed, the precipitate is allowed to stand and precipitate is collected, the supernatant is discarded, and the gel-like precipitate is repeatedly washed with deionized water until no sulfate ions are detected; the washed precipitate is dried in a 60°C forced-air drying oven to constant weight, and finally pulverized through a 200-mesh sieve to obtain a light yellow powdered zinc alginate complex.

[0017] Furthermore, the molar ratio of sodium alginate to zinc sulfate is 1:1.2.

[0018] Furthermore, the growth agent is prepared by the following method: S1: Weigh all raw materials according to the formula weight parts and set aside; S2: Place 2 / 3 of the total amount of plasma-activated water in a sterile preparation tank, maintain the temperature at 25±2℃, and slowly add mesoporous silica nanoparticles and zinc alginate complex while stirring at a low speed of 200rpm. Then increase the stirring speed to 800rpm and supplement with ultrasonic dispersion treatment until a uniform and stable suspension is formed, which is the base liquid. S3: Take a small amount of the base solution and mix Bacillus belye and Trichoderma harzianum into a paste for later use; S4: Adjust the stirring speed of the base solution to 400 rpm, and add methyl jasmonate, purslane extract, ginkgo leaf nano extract and chitosan oligosaccharide in sequence. After each component is added, continue stirring for 10 minutes to ensure that it is completely dissolved or evenly dispersed. Then add salicylic acid nanocapsules and gently stir at 300 rpm for 20 minutes to avoid damaging the microcapsule structure, and obtain an active mixture. S5: Reduce the stirring speed of the active mixture to 150 rpm, slowly add the paste from step S3, and gently stir for 30 minutes to ensure uniform dispersion of the bacterial agent and obtain a microbial suspension. S6: Dissolve pullulan in the remaining plasma-activated water, add it to the microbial suspension, and stir at 150 rpm for 15 min to obtain the final product.

[0019] Furthermore, in step S2, the ultrasonic dispersion power is 400W, the frequency is 40kHz, and the time is 30min.

[0020] Further, the method of using the growth agent is as follows: (1) Scion treatment: Soak the base of the scion in a 300-fold diluted solution of the growth agent for 15 minutes, take it out and dry it before grafting; (2) Rootstock treatment: Apply a small amount of the original solution of the growth agent to the cut of the rootstock before grafting to form a protective film; (3) Post-grafting management: After grafting, spray the area around the interface with a 500-fold diluted solution of the growth agent once every 7 to 10 days for 2 to 3 consecutive times.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. A fundamental shift in the mechanism of action: By introducing signaling molecules such as methyl jasmonate, the present invention changes the target of action from "supplementing exogenous hormones" to "activating endogenous plant defense and healing", enabling plants to shift from passively receiving stimuli to actively repairing themselves, resulting in a more fundamental and efficient effect.

[0022] 2. Extended duration of action: The present invention uses salicylic acid nanocapsules, mesoporous nano silica and other slow-release ingredients to achieve controllable and sustained release of active ingredients, ensuring stable support throughout the grafting healing process.

[0023] 3. Overcoming the limitations of grafting technology: The formula of this invention takes into account the needs of signal, nutrition, microorganisms and other aspects, so that it shows significant effects on both conventional grafting and distant grafting (significantly improving the grafting survival rate, promoting the rapid and large-scale generation of callus tissue and accelerating the lignification process of the interface, etc.), greatly expanding the application boundaries of grafting technology.

[0024] 4. Successfully utilized and optimized the grafting interface micro-ecology: This invention scientifically combines beneficial bacteria with signal molecules and nanocarriers to create a superior colonization environment for beneficial bacteria, maximizing their growth-promoting and antibacterial potential, and realizing the mutualistic symbiosis between plants and microorganisms. Through the three-in-one system design of "signal-microorganism-carrier nutrition", a significant synergistic effect was generated.

[0025] 5. Environmentally friendly and highly safe: It makes extensive use of natural extracts, biodegradable materials and beneficial microorganisms, reducing the amount and residue of chemically synthesized hormones, and is safe for the environment and non-target organisms.

[0026] Based on this, the present invention provides a novel, efficient, safe, and universally applicable fruit tree grafting growth agent through multi-component compounding, which is of great significance to the development of the fruit tree industry. Detailed Implementation

[0027] To make the objectives and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention.

[0028] Unless otherwise specified, the instruments, reagents, and materials used in the following embodiments are all conventional instruments, reagents, and materials already available in the prior art and can be obtained through legitimate commercial channels. Unless otherwise specified, the experimental methods and detection methods used in the following embodiments are all conventional experimental methods and detection methods already available in the prior art.

[0029] Methyl jasmonate (chemically pure, ≥95%), Bacillus vesiculosus (wettable powder, viable count ≥1.0×10⁻⁶) 10 CFU / g), Trichoderma harzianum (wettable powder, viable count ≥5.0×10⁻⁶). 9 CFU / g), mesoporous silica nanoparticles (pore size 2-10nm, specific surface area 500m²). 2 / g), chitosan oligosaccharide (molecular weight ≤3000Da).

[0030] Preparation Example 1 Preparation of salicylic acid nanocapsules: S1: Preparation of colostrum: Accurately weigh 0.2 parts of salicylic acid and 1.0 parts of polylactic acid-glycolic acid copolymer (PLGA), dissolve them together in 10 parts of dichloromethane (organic phase), and slowly add them dropwise to 20 parts of an aqueous solution containing 1% (w / v) polyvinyl alcohol (PVA) under magnetic stirring at 500 rpm. Emulsify at 10000 rpm for 3 min using a high-speed disperser to form a water-in-oil (W / O) colostrum; S2: Preparation of double emulsion: Quickly pour the colostrum into 50 parts of an aqueous solution containing 2% (w / v) PVA and stir at 400 rpm to form a water-in-oil-in-water (W / O / W) double emulsion; S3: Solvent evaporation and curing: Stir continuously at 400 rpm for 4-6 hours to completely evaporate the dichloromethane and solidify the microcapsules; then, collect the microcapsules by centrifugation at 10000 rpm for 10 min, wash them three times with deionized water to remove PVA, and finally freeze-dry them to obtain powdered salicylic acid nanocapsules (particle size 300-500 nm), which are stored at 4℃ in the dark.

[0031] Preparation Example 2 Preparation of zinc alginate complex: S1: Slowly add sodium alginate to deionized water at 60°C and stir magnetically at 600 rpm for 2 hours to fully dissolve it, thus preparing a 5% sodium alginate solution. S2: While stirring continuously, slowly add sodium alginate solution dropwise to 10% zinc sulfate solution (the molar ratio of sodium alginate to zinc sulfate is 1:1.2). After the addition is complete, continue the reaction for 2 hours. S3: After the reaction is complete, let the precipitate stand, discard the supernatant, collect the gel-like precipitate, and wash it repeatedly with deionized water until no sulfate ions are detected (using barium chloride solution for detection). S4: The washed precipitate was dried in a 60℃ forced-air drying oven to constant weight, and finally pulverized through a 200-mesh sieve to obtain a light yellow powdered zinc alginate complex.

[0032] Preparation Example 3 Preparation of purslane extract: S1: Pretreatment: The dried whole purslane herb was pulverized and passed through a 40-mesh sieve, placed in a Soxhlet extractor, and an appropriate amount of petroleum ether was added (the material-liquid ratio was 1:8). The extract was heated in a water bath at 75°C and refluxed for 5 hours to remove fat. S2: Extraction and Concentration: After drying the defatted medicinal residue at a low temperature of 35℃, add 70% ethanol solution (solid-liquid ratio 1:15) and reflux extract twice at 75℃, 1.5h each time; combine the two extracts and concentrate under reduced pressure until there is no alcohol odor to obtain the extract. S3: Purification: The extract was suspended in water and purified by passing it through an AB-8 macroporous adsorption resin column. It was eluted with water and 30% ethanol, and the target fraction (norepinephrine) was collected. S4: Drying: After concentrating the target fraction, spray drying is performed (inlet temperature 180℃, outlet temperature 85℃) to obtain a brownish-yellow powder.

[0033] Preparation Example 4 Preparation of Ginkgo Leaf Nanoparticle Extract: S1: Pretreatment: The dried ginkgo leaves are crushed and enzymatically hydrolyzed for 2 hours at 50℃ and pH 5.0 using a compound enzyme (cellulase and pectinase mixed in a 1:1 ratio) aqueous solution to destroy the cell wall. S2: Extraction: After enzymatic hydrolysis, add an appropriate amount of surfactant (Tween-80) and a certain concentration of ethanol aqueous solution, and perform ultrasonic-assisted extraction (ultrasonic power 400W, time 30min). S3: Purification: After the extract is filtered through a ceramic membrane, it is purified by column chromatography using a combination of polyamide resin and macroporous resin, and the fraction with high content of flavonoid glycosides and lactones is collected. S4: Nano-sizing: The purified extract is subjected to nano-sizing using high-pressure homogenization technology, with the pressure controlled at 800-1000 bar and cyclicated 10-15 times to reduce the particle size to 100-500 nm. S5: Drying: Finally, vacuum freeze-drying (freezing at -45℃ for 3 hours, then freezing at -20℃ for 15 hours at 10 Pa, and then drying at 30℃ for 6 hours) was performed to obtain loose nanoparticles.

[0034] Preparation Example 5 Preparation of plasma-activated water: An atmospheric pressure plasma jet device was used, with high-purity argon (99.99% purity) as the working gas and a flow rate of 10 L / min. Deionized water was placed in the treatment tank, with the plasma jet nozzle 1 cm above the water surface. The treatment power was 150 W, and the treatment time was 15 min. The pH and ORP values ​​were measured immediately after treatment. Once the requirements were met (pH = 6.5-7.0, ORP ≤ -600 mV), the water was sealed and stored, and used within 24 hours.

[0035] Example 1 This embodiment provides a fruit tree grafting growth agent. The raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.01 parts, salicylic acid nanocapsules 0.1 parts, Bacillus vesicles 2 parts, Trichoderma harzianum 1 part, mesoporous silica nanoparticles 5 parts, zinc alginate complex 3 parts, purslane extract 5 parts, ginkgo leaf nano extract 2 parts, chitosan oligosaccharide 1 part, pullulan polysaccharide 2 parts, and plasma activated water 100 parts.

[0036] This growth agent was prepared by the following method: S1: Weigh all raw materials according to the formula weight parts and set aside; S2: Place 2 / 3 of the total amount of plasma-activated water in a sterile preparation tank, maintain the temperature at 25±2℃, and slowly add mesoporous silica nanoparticles and zinc alginate complex while stirring at a low speed of 200rpm. Then increase the stirring speed to 800rpm and supplement with ultrasonic dispersion (power 400W, frequency 40kHz) for 30min until a uniform and stable suspension is formed, which is the base liquid. S3: Take a small amount of the base solution (twice the amount of microorganisms) and mix Bacillus belye and Trichoderma harzianum into a paste for later use; S4: Adjust the stirring speed of the base solution to 400 rpm, and add methyl jasmonate, purslane extract, ginkgo leaf nano extract and chitosan oligosaccharide in sequence. After each component is added, continue stirring for 10 minutes to ensure that it is completely dissolved or evenly dispersed. Then add salicylic acid nanocapsules and gently stir at 300 rpm for 20 minutes to obtain an active mixture. S5: Reduce the stirring speed of the active mixture to 150 rpm, slowly add the paste from step S3, and gently stir for 30 minutes to ensure uniform dispersion of the bacterial agent and obtain a microbial suspension. S6: Dissolve pullulan in the remaining plasma-activated water, add it to the microbial suspension, and stir at 150 rpm for 15 min to obtain the final product.

[0037] Example 2 The difference from Example 1 is that the raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.03 parts, salicylic acid nanocapsules 0.2 parts, Bacillus vesicles 3 parts, Trichoderma harzianum 2 parts, mesoporous nano silica 8 parts, zinc alginate complex 4 parts, purslane extract 6 parts, ginkgo leaf nano extract 3 parts, chitosan oligosaccharide 1.5 parts, pullulan polysaccharide 3 parts, and plasma activated water 150 parts.

[0038] Example 3 The difference from Example 1 is as follows: 0.05 parts methyl jasmonate, 0.3 parts salicylic acid nanocapsules, 5 parts Bacillus belye, 3 parts Trichoderma harzianum, 10 parts mesoporous silica nanoparticles, 6 parts zinc alginate complex, 8 parts purslane extract, 4 parts ginkgo leaf nano extract, 2 parts chitosan oligosaccharide, 4 parts pullulan polysaccharide, and 200 parts plasma-activated water.

[0039] Comparative Example 1 The difference from Example 2 is that the raw material formula and weight parts of the growth agent are: 0.03 parts of methyl jasmonate, 0.2 parts of salicylic acid microcapsules, and 150 parts of plasma-activated water.

[0040] Comparative Example 2 The difference from Example 2 is that the raw material formula and weight parts of the growth agent are: 3 parts of Bacillus vesiculosus, 2 parts of Trichoderma harzianum, and 150 parts of plasma-activated water.

[0041] Comparative Example 3 The difference from Example 2 is that the raw material formula and weight parts of the growth agent are: 8 parts of mesoporous nano silica, 4 parts of zinc alginate complex, and 150 parts of plasma activated water.

[0042] Comparative Example 4 The difference from Example 2 is that the raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.03 parts, salicylic acid microcapsules 0.2 parts, Bacillus vesiculosus 3 parts, Trichoderma harzianum 2 parts, and plasma-activated water 150 parts.

[0043] Comparative Example 5 The difference from Example 2 is that the raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.03 parts, salicylic acid microcapsules 0.2 parts, mesoporous nano silica 8 parts, zinc alginate complex 4 parts, and plasma activated water 150 parts.

[0044] Comparative Example 6 The difference from Example 2 is that the raw material formula and weight parts of the growth agent are: 3 parts of Bacillus vesiculosus, 2 parts of Trichoderma harzianum, 8 parts of mesoporous nano silica, 4 parts of zinc alginate complex, and 150 parts of plasma activated water.

[0045] Comparative Example 7 The difference from Example 2 is that the raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.03 parts, salicylic acid microcapsules 0.2 parts, Bacillus vesiculosus 3 parts, Trichoderma harzianum 2 parts, mesoporous nano silica 8 parts, zinc alginate complex 4 parts, and plasma activated water 150 parts.

[0046] Comparative Example 8 Commercially available grafting growth agent (purchased from Shandong Yunhai Dahua Agricultural Technology Co., Ltd., the main components of which are 2.5% naphthaleneacetic acid and 2.5% indolebutyric acid).

[0047] Comparative Example 9 Blank control (water).

[0048] Experimental Example 1 During the 2024 growing season, a grafting experiment was conducted in Jingning County, Pingliang City, Gansu Province (typical continental climate, spring drought, challenging experimental conditions, which can effectively test the product's stress resistance and stability).

[0049] 1. Experimental Materials and Design: (1) Grafting combinations: Three grafting combinations representing different grafting compatibility were carefully selected to test their wide adaptability: Combination A (high affinity): Apple "Red Fuji" (scion) / Xinjiang wild apple (rootstock). Both are grafted and have strong affinity, serving as a basic verification method.

[0050] Combination B (medium affinity): Pear 'Zaosu Pear' (scion) / Pear 'Du' (rootstock). These are different genera within the same family, and their healing is easily affected under drought conditions. This combination was used to verify their stress-resistant healing abilities.

[0051] Combination C (low affinity): Pear "Korla Fragrant Pear" (scion) / Papaya (rootstock). This interfamily distant grafting is extremely difficult and is used to verify the limits and breakthroughs of the technology.

[0052] (2) Experimental treatments: 12 treatment groups were set up for each combination (corresponding to the growth agents of Examples 1-3 and Comparative Examples 1-9, respectively). The grafting method was uniformly the cleft grafting method operated by local skilled technicians. The specific treatment method was as follows: Scion treatment: Soak the base of the scion (about 3-5cm) in a 300-fold diluted growth agent solution for 15 minutes, take it out and let it dry before grafting; Rootstock treatment: Before grafting, apply a small amount of undiluted growth promoter to the cut of the rootstock to form a protective film. Post-grafting management: After grafting, spray the area around the graft union with a 500-fold diluted growth agent every 7-10 days for 2-3 consecutive times. For special cases such as distant grafting or grafting under adverse conditions such as high and low temperatures or drought, the spraying concentration can be increased to 200-300 times, and the spraying frequency can be increased to once every 5-7 days to continuously enhance the resistance to adverse conditions.

[0053] (3) Evaluation indicators and measurement methods: Survival rate (%): Statistical results were obtained 30 days after grafting.

[0054] Callus index (0-5): Score obtained by sampling 10 days after grafting.

[0055] Lignification degree of the graft union (%): The percentage of the graft union that is completely lignified is determined by dissection 45 days after grafting.

[0056] Physiological and biochemical indicators: β-1,3-glucanase activity (U / g) and endogenous jasmonic acid content (ng / g): measured 7 days post-grafting. On the 7th day post-grafting (peak period of injury response and defense response), a tissue sample (approximately 1-2 grams) containing the entire healing surface was taken by making incisions approximately 5 mm above (scion) and below (rootstock) of the graft union using a scalpel for measurement.

[0057] Chlorophyll fluorescence parameter (Fv / Fm): Measured 15 days after grafting to reflect the early photosynthetic health of the grafted seedling. On the 15th day after grafting, select leaves from the upper and middle parts of the scion that are in good growth condition and fully expanded, avoiding the main vein, and measure the value in the middle of the leaf.

[0058] 2. Test Results Table 1. Effects of different treatments on survival and healing of three grafting combinations. ,

[0059] Note: Different letters after the same column of data in the same group indicate that... P The difference was statistically significant at the <0.05 level, and the same applies below.

[0060] Table 2 Effects of different treatments on physiological and biochemical indicators of grafts ,

[0061] As shown in Table 1, in terms of grafting effect, groups 1-3 of this embodiment are significantly better than groups 1-9 of the comparative examples in terms of grafting survival rate, callus formation index, and lignification degree of the graft union. This result confirms that the growth agent of this embodiment has broad-spectrum adaptability and effectiveness, and in particular solves the technical bottleneck of poor adaptability and low survival rate of existing technologies for distant grafting.

[0062] As shown in Table 2, regarding the intrinsic mechanism, compared with comparative groups 1-9, the endogenous jasmonic acid content and β-1,3-glucanase activity in the graft union tissue of Examples 1-3 were significantly increased. This indicates that it successfully activated the plant's systemic defense and healing response centered on the jasmonic acid signaling pathway, rather than relying solely on passive stimulation from exogenous hormones as in traditional techniques, thus solving the problem of a single mechanism of action. Simultaneously, the chlorophyll fluorescence parameter (Fv / Fm) of the scion leaves in Examples 1-3 was significantly better than that of the control groups, strongly demonstrating the good recovery of the graft's physiological functions and the efficient establishment of vascular bundle connections between the rootstock and scion, reflecting its excellent persistence and stress resistance, overcoming the short persistence of traditional products.

[0063] Based on the results in Tables 1 and 2, the growth agents in Examples 1-3 significantly improved grafting survival rate, promoted rapid and abundant callus formation, and accelerated the lignification process at the graft union. Furthermore, they effectively activated β-1,3-glucanase activity at the grafting wound, which helps enhance resistance; simultaneously, they significantly increased the content of endogenous jasmonic acid, thereby actively regulating gene expression and metabolism related to wound healing; and they also improved the chlorophyll fluorescence properties of the scion leaves, effectively enhancing the photosynthetic system efficiency of the scion and providing more sufficient energy for healing and early growth.

[0064] Furthermore, by comparing the effects of Examples 1-3 (full formulation), Comparative Example 1 (signal), Comparative Example 2 (microorganisms), Comparative Example 3 (carrier nutrition), Comparative Example 4 (signal + microorganisms), Comparative Example 5 (signal + carrier nutrition), Comparative Example 6 (microorganisms + carrier nutrition), and Comparative Example 7 (signal + microorganisms + carrier nutrition), it was found that the effect of any single component among signal, microorganisms, and carrier nutrition is relatively limited. However, when combined in pairs, they show a preliminary additive effect of 1+1>2. The combined use of the three components produces a synergistic effect of 1+1+1>>3. This fully demonstrates that there is an interaction between signal, microorganisms, and carrier nutrition: the nanocarrier ensures the slow release of signal molecules and the stable colonization of microorganisms, while the life activities of microorganisms amplify the signal response of plants, and nutrient supply provides the material basis for this process. The three components work together to produce a synergistic effect of "signal-microorganism-carrier nutrition".

[0065] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A fruit tree grafting growth agent, characterized by: The raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.01~0.05 parts, salicylic acid nanocapsules 0.1~0.3 parts, Bacillus vesicularis 2~5 parts, Trichoderma harzianum 1~3 parts, mesoporous silica nanoparticles 5~10 parts, zinc alginate complex 3~6 parts, purslane extract 5~8 parts, ginkgo leaf nano extract 2~4 parts, chitosan oligosaccharide 1~2 parts, pullulan polysaccharide 2~4 parts, and plasma activated water 100~200 parts. ​ 2. The fruit tree grafting growth agent according to claim 1, characterized by: The raw material formula and weight parts of the growth agent are as follows: methyl jasmonate 0.03 parts, salicylic acid nanocapsules 0.2 parts, Bacillus vesicles 3 parts, Trichoderma harzianum 2 parts, mesoporous nano silica 8 parts, zinc alginate complex 4 parts, purslane extract 6 parts, ginkgo leaf nano extract 3 parts, chitosan oligosaccharide 1.5 parts, pullulan polysaccharide 3 parts, and plasma activated water 150 parts.

3. The fruit tree grafting growth agent according to claim 1 or 2, characterized in that: The preparation method of the salicylic acid nanocapsules is as follows: accurately weigh 0.2 parts of salicylic acid and 1.0 parts of polylactic acid-glycolic acid copolymer, dissolve them together in 10 parts of dichloromethane, and slowly add them dropwise to 20 parts of an aqueous solution containing 1% (w / v) PVA under magnetic stirring at 500 rpm. Emulsify at 10000 rpm for 3 min using a high-speed disperser to form a primary emulsion. Quickly pour the primary emulsion into 50 parts of an aqueous solution containing 2% (w / v) PVA and stir at 400 rpm to form a secondary emulsion. Continue stirring at 400 rpm for 4-6 h to allow the dichloromethane to completely evaporate and the microcapsules to solidify. Subsequently, collect the microcapsules by centrifugation at 10000 rpm for 10 min, wash them three times with deionized water to remove PVA, and finally freeze-dry to obtain powdered salicylic acid nanocapsules.

4. The fruit tree grafting growth agent according to claim 1 or 2, characterized in that: The preparation method of the zinc alginate complex is as follows: sodium alginate is slowly added to deionized water at 60°C and magnetically stirred at 600 rpm for 2 hours to fully dissolve it, thus preparing a 5% sodium alginate solution; under continuous stirring, the sodium alginate solution is slowly added dropwise to a 10% zinc sulfate solution, and the reaction continues for 2 hours after the addition is complete; after the reaction is completed, the precipitate is allowed to stand and the supernatant is discarded, the gel-like precipitate is collected, and it is repeatedly washed with deionized water until no sulfate ions are detected; the washed precipitate is dried in a 60°C forced-air drying oven to constant weight, and finally pulverized through a 200-mesh sieve to obtain a light yellow powdered zinc alginate complex.

5. The fruit tree grafting growth agent according to claim 4, characterized in that: The molar ratio of sodium alginate to zinc sulfate is 1:1.

2.

6. A fruit tree grafting growth agent according to any one of claims 1-5, characterized in that: The growth agent is prepared by the following method: S1: Weigh all raw materials according to the formula weight parts and set aside; S2: Place 2 / 3 of the total amount of plasma-activated water in a sterile preparation tank, maintain the temperature at 25±2℃, and slowly add mesoporous silica nanoparticles and zinc alginate complex while stirring at a low speed of 200rpm. Then increase the stirring speed to 800rpm and supplement with ultrasonic dispersion treatment until a uniform and stable suspension is formed, which is the base liquid. S3: Take a small amount of the base solution and mix Bacillus belye and Trichoderma harzianum into a paste for later use; S4: Adjust the stirring speed of the base solution to 400 rpm, and add methyl jasmonate, purslane extract, ginkgo leaf nano extract and chitosan oligosaccharide in sequence. After each component is added, continue stirring for 10 minutes to ensure that it is completely dissolved or evenly dispersed. Then add salicylic acid nanocapsules and gently stir at 300 rpm for 20 minutes to obtain an active mixture. S5: Reduce the stirring speed of the active mixture to 150 rpm, slowly add the paste from step S3, and gently stir for 30 minutes to ensure uniform dispersion of the bacterial agent and obtain a microbial suspension. S6: Dissolve pullulan in the remaining plasma-activated water, add it to the microbial suspension, and stir at 150 rpm for 15 min to obtain the final product.

7. The fruit tree grafting growth agent according to claim 6, characterized in that: In step S2, the ultrasonic dispersion power is 400W, the frequency is 40kHz, and the time is 30min.

8. A fruit tree grafting growth agent according to any one of claims 1-7, characterized in that: The method of using the growth agent is as follows: (1) Scion treatment: Soak the base of the scion in a 300-fold diluted solution of the growth agent for 15 minutes, take it out and dry it before grafting; (2) Rootstock treatment: Apply a small amount of the original solution of the growth agent to the cut of the rootstock before grafting to form a protective film; (3) Post-grafting management: After grafting, spray the area around the interface with a 500-fold diluted solution of the growth agent once every 7 to 10 days for 2 to 3 consecutive times.

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