A double-effect time sequence nano healing agent for cross-family distant grafting and a matching grafting method thereof

Abstract

The application provides a double-effect time sequence nano healing agent for cross-species distant grafting and a grafting method thereof, and belongs to the technical field of plant grafting. The application initiates a double-component time sequence cooperation + nano targeted anchoring slow release + stock-scion homologous signal adaptation integrated technology system, which comprises a pre-antioxidant dredging agent and a main-effect healing promoting film forming agent; and is used for a grafting incision in a process and in stages. The application first applies an amino modified nano silicon slow release carrier, a composite enzymatic dredging system and a homologous leaf signal extraction liquid to a distant grafting scene, and solves the problems of grafting incision oxidation browning, callose deposition, poor vascular bundle connection, high cyst rate, slow healing and false activity in cross-species distant grafting through six-effect cooperation of antioxidant barrier, enzymatic dredging, nano targeted delivery, biomimetic skeleton film forming, homologous signal adaptation and bacteriostatic stress resistance, and is suitable for grafting of the same family but different species, such as Ebenaceae, Rutaceae, Bignoniaceae and Myrtaceae, and has a high survival rate.

Description

Technical Field

[0001] This invention belongs to the field of plant grafting technology, specifically relating to a dual-effect time-series nano-healing agent and its exclusive grafting method for high-difficulty distant grafting involving transploidy, transphenology, and different genera within the same family. Background Technology

[0002] Distant grafting is a key horticultural technique for integrating superior plant traits, enhancing stress resistance, and expanding suitable growing ranges, and it is widely used in agricultural production. Grafting between different genera within the same family can combine the superior economic traits of the scion with the strong stress resistance of the rootstock, effectively overcoming the growth limitations of a single variety, and is an important technical means in current horticultural breeding and cultivation.

[0003] However, grafting between distant relatives of the same family but different genera faces common technical bottlenecks: (1) There are significant differences in genetic background and ploidy between the scion and rootstock. Abnormal deposition of callose during the healing process can hinder the transport of plasmodesmata, leading to vascular bundle connectivity problems, limiting the normal transport of photosynthetic products from the scion to the rootstock, thus restricting root development and ultimately affecting the normal growth of the grafted seedling; (2) After the grafting cut is exposed to air, the phenolic substances released by cell breakage are oxidized to form a brown isolation layer, while a large amount of reactive oxygen species are produced, which not only damages the integrity of the cambium cells, but also directly inhibits the effective growth of callus tissue. (3) When grafting across phenological periods, the imbalance between the supply and demand of nutrients in the rootstock and scion is prominent. In addition, the callus formation is slow and the incidence of cysts is high, which leads to the frequent occurrence of "false survival" phenomenon. Although the scion can sprout and shoot for a short time by relying on its own nutrients, it dies quickly due to the interruption of subsequent water and nutrient supply. (4) The functional design of commercially available grafting healing agents is not targeted. They do not have comprehensive functions such as interface directional protection, slow release of active ingredients to retain water, and breathable film support. The active ingredients are easily lost, making it difficult to efficiently solve the healing problem in distant grafting, resulting in a low survival rate and seriously restricting the application of large-scale seedling cultivation.

[0004] Currently available grafting healing agents, including those with existing patents, are mostly single-formula, single-application designs that can only achieve simple antibacterial, moisturizing, and bud-promoting effects. They have three major drawbacks: First, they cannot address the sequential pain points of distant grafting, such as "early-stage oxidative necrosis and later-stage healing voids," and a single agent cannot simultaneously address both early-stage unblocking and later-stage shaping. Second, the active ingredients are highly water-soluble and easily lost, resulting in a short efficacy period that cannot cover the callus differentiation cycle of more than 20 days in distant grafting. Third, there are no targeted technical means to alleviate the rejection between rootstock and scion species, leading to extremely low survival rates for cross-ploidy and cross-phenological grafting.

[0005] Meanwhile, existing technologies are merely simple superpositions of conventional raw materials, lacking systematic synergistic design, dedicated process adaptation, and targeted functional modification, thus failing to overcome the technical barriers of highly challenging distant grafting. Based on the gaps and deficiencies in existing technologies, this invention innovatively establishes a three-in-one technical system of dual-component time-sequential synergy, nano-targeted sustained release, and homologous signal adaptation. Through extensive screening experiments, the optimal ratio and process parameters were determined, solving the long-standing industry problems of low survival rate and poor stability in distant grafting.

[0006] Therefore, developing a highly efficient and adaptable nano-healing agent specifically for distant grafting has become a core technical problem that urgently needs to be solved in the field of plant grafting. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of existing technologies, such as poor adaptability, lack of timing regulation, short efficacy, inability to alleviate distant grafting rejection, and insufficient creativity, and to provide a dual-effect timing nano-healing agent for distant grafting and its adapted grafting method. This invention breaks through the limitations of conventional healing agents' single function and single-use, achieving precise healing throughout the entire cycle of complex distant grafting through the timing division of labor between two agents, specific modification for nanomaterial grafting, adaptation to homologous signals, and synergistic effect of multiple components.

[0008] To achieve the above objectives, the present invention adopts the following original technical solution: The core innovation of this invention, which distinguishes it from all existing technologies, lies in its four unique and non-obvious features, without any prior art combinations to suggest otherwise: First, it features a unique dual-component, time-sequential action system with two agents acting at different times: a pre-treatment antioxidant and unblocking agent (early-stage protection) + a main-effect healing and film-forming agent (late-stage shaping). This system precisely addresses the core pain points of different healing stages in distant grafting, completing antioxidant, callus removal, and vascular unblocking functions in the pre-treatment stage, and completing cell differentiation, film-forming shaping, and stress resistance and survival in the later stage. This fills the industry gap where a single agent cannot meet the healing needs of both the pre- and post-treatment cycles. Second, it pioneers a customized application for grafting pesticide nano-slow-release materials, enabling nano-targeted delivery: amino-modified nano-silicon carriers improve adhesion and slow-release properties, achieving in-situ anchoring at the interface, long-term slow-release of active ingredients, and enhanced adsorption capacity of the cambium layer; Third, a unique rootstock-scion homologous signal extraction and matching technology, homologous signal matching: By inducing species recognition through specific signaling molecules in the leaves of the rootstock and scion themselves, the immune rejection response to distant grafting is specifically reduced. The plant extracts used in existing technologies are all single exogenous plant extracts, which can only play a general antioxidant or antibacterial role. The present invention uses a mixed low-temperature extract of healthy leaves from both the rootstock and the scion in the current year, which contains specific signaling molecules from both the rootstock and the scion (such as systemin, jasmonic acid, salicylic acid, etc.). These signaling molecules can induce the rootstock and scion cells to recognize each other, reduce immune rejection response, and promote the fusion of rootstock and scion cells and vascular bundle connectivity. This adaptation mechanism is an original design that has not been disclosed in the prior art and is absolutely not a substitute for conventional additives.

[0009] Fourth, it features a unique multi-component synergistic system that deeply couples five functions: compound enzymatic hydrolysis, nano-slow release, homologous adaptation, biomimetic membrane formation, and multi-dimensional regulation. The compound antioxidants scavenge reactive oxygen species and inhibit browning; the targeted cell wall degrading enzyme group degrades callosity and unblocks vascular bundles; the quaternary growth regulators promote callus formation, vascular bundle differentiation, and bud germination; the biopolysaccharide complex forms a membrane for water retention, stress resistance, and antibacterial properties; the nanocellulose acts as a biomimetic scaffold, guiding orderly healing and reducing cysts; and the homologous signal leaf extract alleviates distant rejection and improves affinity. This results in superior technical effects that cannot be achieved by single components or existing technology combinations, and cannot be easily screened by conventional experiments in this field.

[0010] This invention provides a dual-effect time-series nano-healing agent for distant grafting, comprising a pre-antioxidant unblocking agent and a main-effect healing film-forming agent; The pre-antioxidant unclogging agent comprises the following components by weight percentage: 0.15%~0.6% compound antioxidant, 0.12%~0.6% directed cell wall degrading enzyme group, 0.25%~1.1% amino-modified nano-silicon carrier, 4%~9% flavonoid vitamin C plant-derived liquid, 4%~9% rootstock and scion homologous signal leaf extract, and the balance being water; the pH value of the pre-antioxidant unclogging agent is 5.2~6.6; The main active ingredient for promoting healing and film formation comprises the following components by weight percentage: 0.015%~0.09% quaternary compound growth regulator, 0.06%~0.25% biopolysaccharide complex, 0.25%~1.6% nanofiber framework material, 0.0015%~0.009% cytokinin synergist, 0.06%~0.22% antibacterial agent, 0.012%~0.06% compound calcium and magnesium nutrients, 4%~9% flavonoid and vitamin C plant-derived liquid, 4%~9% rootstock and scion homologous signal leaf extract, and the remainder being water; The extract of the homologous signal leaves of the scion and rootstock is a mixed aqueous extract of fresh scion leaves and fresh rootstock leaves of the corresponding grafting combination.

[0011] Preferably, the composite antioxidant comprises N-acetylcysteine, glutathione, and ascorbate phosphate, wherein the mass ratio of N-acetylcysteine, glutathione, and ascorbate phosphate is (2.5~3.5):1:1.

[0012] Preferably, the directed cell wall degrading enzyme group includes β-1,3-glucanase, pectin lyase and cellulose hemicellulase; the mass ratio of β-1,3-glucanase, pectin lyase and cellulose hemicellulase is (1.5~2.5):(0.9~1.1):(0.45~0.55).

[0013] Preferably, the amino-modified nano-silicon carrier is nano-silica modified with KH550+KH560 composite coupling agent, with a particle size of 60~90nm and a Zeta potential of -30~-40mV.

[0014] Preferably, the quaternary compound growth regulator includes naphthaleneacetic acid, indolebutyric acid, 24-epibrassinolide, and bud-promoting agent; the mass ratio of naphthaleneacetic acid, indolebutyric acid, 24-epibrassinolide, and 6-BA is (2~4):(0.5~1.5):(0.5~1.5):(0.4~0.6).

[0015] Preferably, the biopolysaccharide complex comprises chitosan oligosaccharide, sodium alginate, and konjac glucomannan, wherein the mass ratio of chitosan oligosaccharide, sodium alginate, and konjac glucomannan is (3~5):(0.5~1.5):(0.4~0.6). The nanofiber framework material is plant-derived nanocellulose.

[0016] Preferably, the cytokinin enhancer is zeatin nucleoside.

[0017] Preferably, the flavonoid vitamin C plant-derived liquid is a fragrant lemon infusion.

[0018] Preferably, the pre-antioxidant unclogging agent is prepared and used immediately or prepared as a lyophilized powder for compound use; the main healing film-forming agent is stored in a sealed container at 2℃~8℃; The aforementioned dual-effect time-series nano-healing agent for distant grafting is suitable for distant grafting of different genera within the same family of Ebenaceae, Rutaceae, Bignoniaceae, and Myrtaceae.

[0019] This invention provides a method for using the dual-effect sequential nano-healing agent for distant grafting of different genera within the same family. The main healing film-forming agent is applied to the cambium area of ​​the rootstock and scion, left to stand for 10-15 seconds, and then inserted and aligned with the cambium layer. A pre-prepared antioxidant and unblocking agent is then applied to the rootstock-scion joint and the surrounding 0.5cm area, and the grafting film is used to seal it.

[0020] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a dual-effect time-sequential nanocomposite system specifically for distant grafting of different genera within the same family. It consists of a pre-antioxidant and a main-effect healing-promoting film-forming agent. The pre-antioxidant and the main-effect healing-promoting film-forming agent form a time-sequential synergistic system, covering the entire process from early interface protection to later healing and shaping. It integrates multiple functions such as antioxidant and browning prevention, cell wall-assisted degradation, nanocarrier slow release, biomimetic film-forming support, plant-derived synergistic antibacterial and growth regulation. It systematically solves the common interface healing problems in distant grafting of different genera within the same family, such as oxidative browning of the grafting interface, callus deposition, poor vascular bundle connectivity, high cyst incidence, slow healing, and cross-phenological nutrient supply and demand imbalance, thereby achieving broad-spectrum compatibility, efficient healing, and stable survival.

[0021] The dual-effect time-series nanocomposite system for distant grafting of different genera within the same family provided by this invention is highly versatile and widely adaptable. It can be stably applied to the cross-phenological and cross-ploidy distant grafting of fruit trees and economic forest trees from multiple families and genera, such as Ebenaceae, Rutaceae, Bignoniaceae, and Myrtaceae. The key survival and healing indicators are significantly better than the blank control. At the same time, it is suitable for various climatic environments, such as tropical and subtropical high temperature and humidity, and temperate winter low temperature. The raw materials are readily available, the preparation is simple, the use is safe, and there are no pesticide residues. It is suitable for large-scale seedling production and industrial promotion, and has outstanding practical value and broad application prospects. Attached Figure Description

[0022] Figure 1 A diagram illustrating the scientific application method for the pre-antioxidant and the main healing film-forming agent is shown below: 1- scion incision, 2- rootstock, 3- pre-antioxidant and dredging agent, 4- main healing film-forming agent.

[0023] Figure 2 Schematic diagram for applying the grafting agent; 1- Scion incision, 2- Rootstock, 3- Pre-antioxidant and unblocking agent, 4- Main active healing and film-forming agent.

[0024] Figure 3 This is a photograph of a grafted hexaploid temperate persimmon (scion) × diploid tropical chocolate pudding fruit Diospyros digynaJacq (rootstock).

[0025] Figure 4 A photograph of a grafted citrus fruit (Ehime 38, scion of Rutaceae, Citrus genus) × wampee (rootstock of Rutaceae, Wampee genus).

[0026] Figure 5 A photo of a grafted specimen of different genera in the Rutaceae family: Chun Jian Ba ​​Ba Mandarin (scion of Rutaceae, Citrus genus) × Fragrant Lemon (rootstock of Rutaceae, Lemon genus).

[0027] Figure 6 Different genera within the Bignoniaceae family: *Tabebuia purpurea* (scion of *Tabebuia* genus, Bignoniaceae family) HandroanthusGenus) × Magenta Trumpet Tree (Rootstock Bignoniaceae, Quercus genus) Tabebuia (A picture of a grafted specimen)

[0028] Figure 7 Different genera within the Bignoniaceae family: *Tecoma stans* (scion, Bignoniaceae family, *Tecoma stans* genus) × *Tecoma stans* (rootstock, Bignoniaceae family, *Tecoma stans* genus) Tabebuia (Appendix) Actual object image.

[0029] Figure 8 This is a picture of a grafted specimen of Corymbia ficifolia (scion, Corymbia genus, Myrtaceae) × Eucalyptus urophylla (rootstock, Eucalyptus genus, Myrtaceae). Detailed Implementation

[0030] This invention provides a dual-effect time-series nano-healing agent for distant grafting of different genera within the same family, comprising a pre-antioxidant unblocking agent and a main-effect healing film-forming agent.

[0031] The pre-treatment antioxidant unblocking agent comprises the following components by weight percentage: 0.15%~0.6% compound antioxidant, 0.12%~0.6% directed cell wall degrading enzyme group, 0.25%~1.1% amino-modified nano-silicon carrier, 4%~9% flavonoid vitamin C plant-derived liquid, 4%~9% rootstock and scion homologous signal leaf extract, and the balance being water; preferably, it comprises 0.3%~0.5% compound antioxidant, 0.3%~0.5% directed cell wall degrading enzyme group, 0.5%~0.9% amino-modified nano-silicon carrier, 5%~8% flavonoid vitamin C plant-derived liquid, 5%~8% rootstock and scion homologous signal leaf extract, and the balance being water.

[0032] In this invention, the pH value of the pre-antioxidant unclogging agent is preferably 5.2 to 6.6.

[0033] In this invention, the composite antioxidant includes N-acetylcysteine, glutathione, and ascorbate phosphate. The preferred mass ratio of N-acetylcysteine, glutathione, and ascorbate phosphate is (2.5~3.5):1:1, more preferably (2.8~3.2):1:1, and most preferably 3:1:1.

[0034] In this invention, the directed cell wall degrading enzyme group includes β-1,3-glucanase, pectin lyase, and cellulose hemicellulase; the preferred mass ratio of β-1,3-glucanase, pectin lyase, and cellulose hemicellulase is (1.5~2.5):(0.9~1.1):(0.45~0.55), more preferably (1.8~2.2):1:0.5, and most preferably 2:1:0.5.

[0035] In this invention, the amino-modified nano-silicon carrier is preferably nano-silica modified with KH550+KH560 composite coupling agent, with a particle size of 60~90nm and a Zeta potential of -30~-40mV; purchased from Jiangsu Xianfeng Nano XFNANO store on Taobao.

[0036] In this invention, the functions of the pre-antioxidant unblocking agent are: to remove reactive oxygen species, inhibit browning, precisely degrade callosity, unblock vascular bundles, improve adhesion, and slowly release active ingredients.

[0037] In this invention, the main active healing film-forming agent comprises the following components by weight percentage: 0.015%~0.09% quaternary composite growth regulator, 0.06%~0.25% biopolysaccharide complex, 0.25%~1.6% nanofiber framework material, 0.0015%~0.009% cytokinin synergist, 0.06%~0.22% antibacterial agent, 0.012%~0.06% compound calcium and magnesium nutrients, 4%~9% flavonoid vitamin C plant-derived liquid, and rootstock-scion homologous signaling leaf. The extract comprises 4%–9% of the total volume, with the remainder being water. Preferred components include: 0.03%–0.07% of a quaternary compound growth regulator, 0.10%–0.20% of a biological polysaccharide complex, 0.5%–1.5% of a nanofiber framework material, 0.002%–0.007% of a cytokinin synergist, 0.1%–0.2% of an antibacterial agent, 0.02%–0.05% of a compound calcium-magnesium nutrient solution, 5%–8% of a flavonoid-vitamin C plant-derived extract, 5%–8% of a rootstock-scion homologous signal leaf extract, with the remainder being water.

[0038] In this invention, the quaternary composite growth regulator preferably includes naphthaleneacetic acid, indolebutyric acid, 24-epibrassinolide, and 6-BA; the mass ratio of naphthaleneacetic acid, indolebutyric acid, 24-epibrassinolide, and 6-BA is preferably (2~4):(0.5~1.5):(0.5~1.5):(0.4~0.6), more preferably (2.5~3.5):(0.8~1.2):(0.8~1.3):(0.45~0.55), and even more preferably 3:1:1:0.5.

[0039] In this invention, the biopolysaccharide complex includes chitosan oligosaccharide, sodium alginate, and konjac glucomannan. The preferred mass ratio of chitosan oligosaccharide, sodium alginate, and konjac glucomannan is (3~5):(0.5~1.5):(0.4~0.6), more preferably (3.5~4.5):(0.7~1.3):(0.45~0.56), and most preferably 4:1:0.5.

[0040] In this invention, the nanofiber skeleton material is nanocellulose, and its content in the main active healing film-forming agent is 0.25%~1.6%, preferably 0.3%~0.8%, more preferably 0.5% (5.0g); the nanocellulose was purchased from the Taobao store Sailuna Technology Store.

[0041] In this invention, the cytokinin enhancer is preferably zeatin nucleoside.

[0042] In this invention, the antibacterial agent includes a mixture of carbendazim and pyraclostrobin; the preferred mass ratio of the carbendazim and pyraclostrobin mixture is 1:1.

[0043] In this invention, the compound calcium-magnesium nutrient comprises calcium nitrate and magnesium sulfate; the mass ratio of calcium nitrate to magnesium sulfate is (1.5~2.5):1, more preferably (1.8~2.2):1, and most preferably 2:1.

[0044] In this invention, the flavonoid vitamin C plant-derived liquid is preferably a fragrant lemon infusion.

[0045] In this invention, the homologous signal leaf extract of the scion and rootstock is a mixed aqueous extract of fresh scion leaves and fresh rootstock leaves of the corresponding grafting combination. Specifically, fresh scion leaves and fresh rootstock leaves of the corresponding grafting combination are mixed, purified water is added, and the mixture is extracted in a constant temperature water bath at 40-50°C for 2-3 hours. After filtration to remove impurities and sterilization clarification, the extract is obtained. In this invention, the preferred mass ratio of the mixed fresh scion leaves to fresh rootstock leaves is (1-2):(2-1), more preferably 1:1; the preferred mass of the purified water is 8-12 times the total mass of the fresh scion leaves and fresh rootstock leaves, more preferably 9-11 times, and most preferably 10 times.

[0046] In this invention, the main active healing film-forming agent has the following functions: promoting callus differentiation, guiding vascular bundle directional connectivity, inhibiting bacteria and preventing cysts, strengthening interface firmness, retaining water and forming film, and providing long-lasting sustained release.

[0047] The dual-effect sequential nano-healing agent for distant grafting of plants from different genera within the same family described in this invention is suitable for distant grafting of plants from different genera within the Ebenaceae, Rutaceae, Bignoniaceae, and Myrtaceae families.

[0048] This invention also provides a dual-effect time-sequential nano-healing agent for distant grafting of different genera within the same family. The main healing film-forming agent is applied to the cambium area of ​​the rootstock and scion, left to stand for 10-15 seconds, and then inserted and aligned with the cambium. A pre-prepared antioxidant and unblocking agent is then applied to the rootstock-scion joint and the surrounding 0.5cm area, and sealed with a grafting film.

[0049] In practice, the pre-antioxidant and unblocking agent should be prepared and used immediately or mixed with freeze-dried powder; the main healing-promoting film-forming agent should be sealed and refrigerated. Application should follow the principles of applying to the rootstock first, then the scion, applying a thin, even layer, and focusing on the cambium layer, avoiding contact with the bud. Remove the grafting film 40-50 days after grafting, when the bud is 4-6 cm long and the leaves have unfolded. For winter grafting, it is preferable to preheat the main healing-promoting film-forming agent to 16-22℃. For summer grafting, apply 0.25-0.45 mL / plant of the pre-antioxidant and unblocking agent 6-8 days after grafting. In this invention, the main healing-promoting film-forming agent and the pre-antioxidant and unblocking agent can also be mixed as a single component for use, depending on the application requirements.

[0050] In this invention, during the early grafting stage (0–3 days): a pre-applied antioxidant and unblocking agent is used to prevent browning and open channels, creating conditions for healing; during the middle and late grafting stages (3–22 days): a main-effect film-forming agent is used to promote healing, build up the graft, and enhance resistance to adverse conditions, achieving efficient and stable healing.

[0051] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0052] In the following embodiments: Quaternary compound growth regulator (mass ratio): Naphthaleneacetic acid (NAA): Indolebutyric acid (IBA): 24-epibrassinolide (EBR): 6-benzyladenine (6-BA) = 3:1:1:0.5.

[0053] The nanofiber skeleton material is nanocellulose, which was purchased from the Taobao store "Sailuna Technology".

[0054] Fragrant lemon extract: Fresh fragrant lemon slices are selected, extracted in a constant temperature water bath at 45℃ for 2.5 hours, and then filtered to remove impurities, sterilized and clarified.

[0055] Preparation method of 40℃ warm water extract of leaves from rootstock and scion of the same origin: Select fresh scion leaves and fresh rootstock leaves of the corresponding grafting combination and mix them at a mass ratio of 1:1. Add purified water at a material-to-liquid mass ratio of 1:10 and extract in a constant temperature water bath at 45℃ for 2.5h. After filtration to remove impurities and sterilization and clarification, the extract is obtained.

[0056] Example 1

[0057] Pre-treatment antioxidant unclogging agent (1000 g): N-acetylcysteine ​​2.0 g, glutathione 0.66 g, ascorbate phosphate 0.66 g, β-1,3-glucanase 1.5 g, pectin lyase 0.75 g, cellulose hemicellulase 0.4 g, nano-silica modified with KH550+KH560 composite coupling agent 4.0 g, fragrant lemon extract 50 g, rootstock and scion leaf extract at 40℃, balance sterile water, pH 5.8, freshly prepared and used immediately.

[0058] Main active ingredients for promoting healing and film formation (1000 g): 0.25 g of quaternary compound growth regulator, 0.4 g of chitosan oligosaccharide, 0.3 g of sodium alginate, 0.315 g of konjac glucomannan, 5.0 g of nanofiber skeleton material, 0.02 g of zeatin nucleoside (ZR), 1.5 g of compound antibacterial agent (including carbendazim and pyraclostrobin 1:1); 0.225 g of compound calcium and magnesium nutrients (0.15 g of calcium nitrate and 0.075 g of magnesium sulfate), 55 g of fragrant lemon extract, 55 g of 40°C warm water extract of rootstock and scion leaves, with the remainder being sterile water.

[0059] Application method: Apply the main active healing film-forming agent to the cambium layer, with a thickness of 0.1~0.2 mm and a dosage of 0.2~0.4 mL / plant. Let it stand for 10~15 s before grafting. Then apply the pre-antioxidant and unblocking agent to the 0.5 cm area of ​​the joint, with a dosage of 0.6~0.8 mL / plant. Seal with grafting film.

[0060] Control group setup

[0061] Control group (CK): No healing-inducing agent.

[0062] Example 2 (Pre-antioxidant and unclogging agent: free of glutathione, ascorbate phosphate, and cellulase)

[0063] Pre-treatment antioxidant unclogging agent (1000 g): N-acetylcysteine ​​2.0 g, β-1,3-glucanase 1.5 g, pectin lyase 0.75 g, nano-silica modified with KH550+KH560 composite coupling agent 4.0 g, fragrant lemon extract 50 g, rootstock and scion homologous leaf extract at 40℃ warm water 50 g, balance is sterile water, pH 5.8, prepare and use immediately.

[0064] Main active ingredients for promoting healing and film formation (1000 g): 0.25 g of quaternary compound growth regulator, 0.4 g of chitosan oligosaccharide, 0.3 g of sodium alginate, 0.315 g of konjac glucomannan, 5.0 g of nanofiber skeleton material, 0.02 g of zeatin nucleoside (ZR), 1.5 g of compound antibacterial agent (including carbendazim and pyraclostrobin 1:1); 0.225 g of compound calcium and magnesium nutrients (0.15 g of calcium nitrate and 0.075 g of magnesium sulfate), 55 g of fragrant lemon extract, 55 g of 40°C warm water extract of rootstock and scion leaves, with the remainder being sterile water.

[0065] Example 3 (Main active healing and film-forming agent: perfume-free lemon extract and rootstock- and scion-derived leaf extract at 40°C)

[0066] Pre-treatment antioxidant unclogging agent (1000 g): N-acetylcysteine ​​2.0 g, N-acetylcysteine ​​glutathione 0.66 g, ascorbate phosphate 0.66 g, β-1,3-glucanase 1.5 g, pectin lyase 0.75 g, cellulose hemicellulase 0.4 g, nano-silica modified with KH550+KH560 composite coupling agent 4.0 g, fragrant lemon extract 50 g, 40℃ warm water extract of rootstock and scion leaves, balance sterile water, pH 5.8, prepare and use immediately.

[0067] Main active ingredients for promoting healing and film formation (1000 g): 0.25 g of quaternary compound growth regulator, 0.4 g of chitosan oligosaccharide, 0.3 g of sodium alginate, 0.315 g of konjac glucomannan, 5.0 g of nanofiber skeleton material, 0.02 g of zeatin nucleoside (ZR), 1.5 g of compound antibacterial agent (including carbendazim and pyraclostrobin 1:1); 0.225 g of compound calcium and magnesium nutrients (0.15 g of calcium nitrate and 0.075 g of magnesium sulfate), with the remainder being sterile water.

[0068] Experimental Example

[0069] General Instructions for Experimental Design

[0070] The experimental subjects included distant grafting combinations of different genera within the same family such as Ebenaceae, Rutaceae, Bignoniaceae, and Myrtaceae, with 50 plants in each group and consistent management conditions.

[0071] Test grafting combination

[0072] Ebenaceae: Hexaploid temperate Diospyros digyna Jacq (scion) × diploid tropical chocolate pudding fruit (rootstock)

[0073] Different genera within the Rutaceae family: Ehime 38 citrus (scion, Rutaceae, Citrus genus) × Wampee (rootstock, Rutaceae, Wampee genus)

[0074] Different genera within the Rutaceae family: Chun Jian Ba ​​Ba Mandarin (scion of Rutaceae, Citrus genus) × Fragrant Lemon (rootstock of Rutaceae, Lemon genus)

[0075] Different genera within the Bignoniaceae family: Tabebuia purpurea (scion of Tabebuia genus in Bignoniaceae family) Handroanthus Genus) × Magenta Trumpet Tree (Rootstock Bignoniaceae, Quercus genus) Tabebuia (genus)

[0076] Different genera within the Bignoniaceae family: *Tecoma stans* (scion, Bignoniaceae family, *Tecoma stans* genus) × *Tabebuia chrysantha* (rootstock, Bignoniaceae family, *Tabebuia* genus) Handroanthus (genus)

[0077] Myrtaceae: Corymbia ficifolia (scion, Myrtaceae, Corymbia genus) × Eucalyptus urophylla (rootstock, Myrtaceae, Eucalyptus genus)

[0078] Each grafting combination included Example 1 (the optimal formula of this invention) and a control group CK (without pre-prepared antioxidant and main healing film-forming agent), with 50 plants in each group. The soil pH of the experimental plots was 5.5-6.0, and the fertility, light, and humidity were consistent. Conventional grafting operations (cleft grafting) were used uniformly. The observed indicators included: budding rate, average budding time, grafting survival rate, browning rate of grafting interface, and secondary branching germination rate.

[0079] Table 1. Control experiment on budding rate of distant grafting of multiple genera and families.

[0080] Table 2. Survival rate control experiment of distant grafting of multiple genera and families at 60 days.

[0081] Browning is considered to have occurred if, upon visual inspection 60 days after grafting, the grafting point turns dark brown, the browning area exceeds 50%, or the entire circumference turns black.

[0082] Table 3. Grafting Interface Antioxidant Browning Control Experiment

[0083] After the grafted new shoots grow to 30 cm in length, pinch off the tips to promote the growth of secondary branches. Observe the growth rate of secondary branches uniformly after 90 days.

[0084] Table 4. Control Experiment on Secondary Branch Sprouting Rate

[0085] As can be seen from the above embodiments, the dual-effect time-series nanocomposite system for distant grafting of different genera and species provided by the present invention has strong versatility and wide applicability. Its key survival and healing indicators are significantly better than the blank control. It is suitable for various climatic environments such as tropical and subtropical high temperature and humidity, and temperate winter low temperature, and has outstanding practical value and broad application prospects.

[0086] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A double effect time sequence nano-healing agent for cross-species distant hybrid grafting, characterized in that, It consists of a pre-treatment targeted unblocking solution and a main-effect healing film-forming agent. The pre-antioxidant unclogging agent comprises the following components by weight percentage: 0.15%~0.6% compound antioxidant, 0.12%~0.6% directed cell wall degrading enzyme group, 0.25%~1.1% amino-modified nano-silicon carrier, 4%~9% flavonoid vitamin C plant-derived liquid, 4%~9% rootstock and scion homologous signal leaf extract, and the balance being water; the pH value of the pre-antioxidant unclogging agent is 5.2~6.6; The main active ingredient for promoting healing and film formation comprises the following components by weight percentage: 0.015%~0.09% quaternary compound growth regulator, 0.06%~0.25% biopolysaccharide complex, 0.25%~1.6% nanofiber framework material, 0.0015%~0.009% cytokinin synergist, 0.06%~0.22% compound antibacterial agent, 0.012%~0.06% compound calcium and magnesium nutrients, 4%~9% flavonoid and vitamin C plant-derived liquid, 4%~9% rootstock and scion homologous signal leaf extract, and the remainder being water; The homologous signal leaf extract of the scion and rootstock is a mixed aqueous extract of fresh scion leaves and fresh rootstock leaves of the corresponding grafting combination. The composite antioxidant comprises N-acetylcysteine, glutathione, and ascorbate phosphate, wherein the mass ratio of N-acetylcysteine, glutathione, and ascorbate phosphate is (2.5~3.5):1:1; The targeted cell wall degrading enzyme group includes β-1,3-glucanase, pectin lyase, and cellulose hemicellulase; the mass ratio of β-1,3-glucanase, pectin lyase, and cellulose hemicellulase is (1.5~2.5):(0.9~1.1):(0.45~0.55). The amino-modified nano-silicon carrier is nano-silica modified with KH550+KH560 composite coupling agent, with a particle size of 60~90nm and a zeta potential of -30~-40mV.

2. The dual-effect time-sequential nano-healing agent for distant grafting of different genera within the same family as described in claim 1, characterized in that, The quaternary composite growth regulator includes naphthaleneacetic acid, indolebutyric acid, 24-epibrassinolide, and 6-BA; the mass ratio of naphthaleneacetic acid, indolebutyric acid, 24-epibrassinolide, and 6-BA is (2~4):(0.5~1.5):(0.5~1.5):(0.4~0.6).

3. The dual-effect time-sequential nano-healing agent for distant grafting of different genera within the same family as described in claim 2, characterized in that, The biopolysaccharide complex includes chitosan oligosaccharide, sodium alginate, and konjac glucomannan, wherein the mass ratio of chitosan oligosaccharide, sodium alginate, and konjac glucomannan is (3~5):(0.5~1.5):(0.4~0.6). The nanofiber framework material is plant-derived nanocellulose, and the particle size of the nanofiber framework material is 70~120nm.

4. The dual-effect time-sequential nano-healing agent for distant grafting of different genera within the same family as described in claim 3 or 4, characterized in that, The cytokinin enhancer is zeatin nucleoside.

5. The dual-effect time-sequential nano-healing agent for distant grafting of different genera within the same family as described in claim 1, characterized in that, The flavonoid vitamin C plant-derived liquid is a compound extract of fragrant lemon.

6. The dual-effect time-sequential nano-healing agent for distant grafting of different genera within the same family as described in claim 1, characterized in that, The pre-antioxidant unblocking agent is prepared and used immediately or made into a freeze-dried powder for compound use; the main healing film-forming agent is stored in a sealed container at 2℃~8℃; The dual-effect sequential nano-healing agent for distant grafting of plants from different genera within the same family is suitable for distant grafting of plants from different genera within the Ebenaceae, Rutaceae, Bignoniaceae, and Myrtaceae families.

7. The method of using the dual-effect time-sequential nano-healing agent for distant grafting of different genera within the same family as described in any one of claims 1 to 6, characterized in that, Apply the pre-antioxidant and unblocking agent to the cambium area of ​​the rootstock and scion, let it stand for 10-15 seconds, then insert and align the cambium layers; then apply the main healing and film-forming agent to the rootstock-scion joint and the surrounding 0.5cm area, and seal with grafting film.

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