Method for 3D printing of grafting auxiliary sleeve and application thereof

Abstract

The invention discloses a preparation method of a grafting sleeve. The preparation method comprises the following steps: 1) acquiring stock diameter data R1, scion diameter data R2 and scion length L; (2) data of the wide diameter and the narrow diameter of a grafting casing pipe are obtained, and the grafting casing pipe comprises a stock casing pipe and a scion casing pipe; 3) inputting the wide diameter data and the narrow diameter data of the stock sleeve and the scion sleeve into a 3D printer; and 4) taking a thermoplastic elastic material as an ink material, and obtaining the grafting sleeve through a 3D printer. The invention also provides the grafting sleeve prepared by the preparation method and a grafting method.

Description

A method and application of 3D printing grafting auxiliary sleeve

[0001] Priority information

[0002] none. Technical Field

[0003] The present invention relates to the field of bioinformatics, and in particular to a method for 3D printing a grafting auxiliary cannula and applications thereof. Background Art

[0004] Grafting is a method of asexual plant propagation in which the branch or bud (scion) of one plant is connected to the stem or root (stock) of another plant to grow into a complete plant. It is an economical and efficient method used in the breeding and production of horticultural crops. Its main purpose is to preserve the excellent characteristics of both the scion and the stock. It is widely used in common fruits and vegetables such as apples, pears, cherries, walnuts, chestnuts, dates, persimmons, grapes, citrus oranges, lychees, kiwis, tomatoes, and eggplants. Currently, the most commonly used grafting method is cleft grafting, while approach grafting and plug grafting are also widely used. Grafting in crop production is mainly done manually, which is labor-intensive and inefficient. At the same time, the efficiency and survival rate of grafting are highly dependent on the skills of the grafting technicians.

[0005] During the manual grafting process, cambium alignment and binding are the most time-consuming and uncertain steps, making grafting efficiency and success highly dependent on the skilled worker. Inexperienced binding can cause the scion and rootstock to shift during grafting, resulting in cambium misalignment. Prolonged grafting can lead to repeated re-applying of the graft, increasing the risk of infection. Loose binding can cause cambium misalignment during wound healing. Scion wrapping is time-consuming, and pre-wrapping is sometimes required to improve efficiency. During periods of high grafting workload, orchards hire large numbers of temporary workers and devote significant time and effort to training them. Even with this effort, grafting efficiency and survival rates cannot be guaranteed. Low survival rates lead to significant losses in seedling costs. Furthermore, current methods require extensive labor to remove the sealing tape after the graft and rootstock wounds have healed, preventing it from hindering the growth of the successfully grafted plants, further adding to costs. Therefore, there is a need for further development of grafting devices that are easy to use and can guarantee grafting efficiency and survival rates.

[0006] Summary of the Invention

[0007] The purpose of the present invention is to solve at least one of the above technical defects, especially the technical defects of the prior art such as high labor consumption, low grafting efficiency and low survival efficiency.

[0008] The present invention provides a method for preparing a grafting sleeve. According to an embodiment of the present invention, the method includes the following steps: 1) obtaining rootstock diameter data R1, scion diameter data R2, and scion length L; 2) obtaining wide diameter and narrow diameter data of the grafting sleeve, wherein the grafting sleeve includes a rootstock sleeve and a scion sleeve; 3) inputting the wide diameter data and narrow diameter data of the rootstock sleeve and the scion sleeve into a 3D printer; 4) using a thermoplastic elastic material as an ink material to obtain the grafting sleeve through a 3D printer. According to a specific embodiment of the present invention, the wide and narrow diameter data of the grafting sleeve are obtained by a certain calculation method using the rootstock diameter data, the scion diameter data, and the length of the scion, and the wide and narrow diameter data of the grafting sleeve are combined with a 3D printer to prepare the corresponding rootstock sleeve and scion sleeve. The method for preparing the grafting sleeve described in the present application can quickly and effectively obtain a grafting sleeve adapted to any diameter range, flexibly prepare a sleeve of suitable diameter, and reduce the labor and time costs during grafting.

[0009] According to an embodiment of the present invention, the method for preparing the grafting sleeve may further include at least one of the following additional technical features:

[0010] According to an embodiment of the present invention, the stock diameter data and the scion diameter data are obtained by scanning the diameters of the stock and the scion by a scanning device, and the length L of the scion is obtained by scanning the full length of the scion by a scanning device. The scanning device is not particularly limited, and any device that can scan the diameter size of the stock or scion can be used, such as a laser scanner, a radar scanner, or a smartphone with a scanning function. The full length of the scion is the longest distance obtained by connecting the two ends of the scion by a point.

[0011] According to an embodiment of the present invention, the diameter data of the rootstock is obtained by scanning the diameter of the rootstock within 0-5cm from the incision cross section by a scanning device, and the diameter data of the scion is obtained by scanning the diameter of the scion at 0.1-1.5cm from the incision cross section by a scanning device. Those skilled in the art will understand that the incision cross section of the rootstock refers to the incision cross section of the rootstock that needs to be pretreated, matched and positioned with the scion, and the incision cross section of the scion refers to the incision cross section of the scion that needs to be pretreated, matched and positioned with the rootstock. Depending on the diameter range of the plant, the length of the scion and the rootstock when positioned is different. Therefore, when the stock diameter data is obtained, the length range of the scanning distance from the incision cross section can be adjusted. Similarly, when the scion diameter data is obtained, the length range of the scanning distance from the incision cross section can also be adjusted within an appropriate range.

[0012] According to an embodiment of the present invention, the diameter data of the rootstock is obtained by scanning the diameter of the rootstock within 0-3 cm from the incision cross section by a scanning device, and the diameter data of the scion is obtained by scanning the diameter of the scion at 0.1-1 cm from the incision cross section by a scanning device. According to some specific embodiments of the present invention, the inventors perform diameter scanning measurements on the rootstock along its length direction, and the scanning measurement range is the rootstock diameter within a length of 0-3 cm from the incision cross section of the rootstock, for example, the rootstock diameter is scanned at 0.1 cm, 0.5 cm, 0.7 cm, 0.8 cm, 0.9 cm, 1.0 cm, 1.5 cm, 2.0 cm, 2.5 cm and 3.0 cm of the incision cross section of the rootstock to obtain diameter data of a plurality of the rootstocks. The inventors performed a diameter scanning measurement on the scion along its length direction, and the scanning measurement range was the scion diameter at a length of 0-3 cm from the cut cross section of the stock. For example, the stock diameter was scanned at 0.1 cm, 0.5 cm, 0.7 cm, 0.8 cm, 0.9 cm or 1.0 cm of the scion cut cross section to obtain the diameter data of the scion.

[0013] According to an embodiment of the present invention, the wide diameter data of the stock sleeve is the maximum value of R1 minus 1-3mm. The inventors found that when the wide diameter data of the stock sleeve is too large, it is difficult to ensure the sealing of the grafting sleeve after it is fixed. When the wide diameter data of the stock sleeve is too small, the stock is difficult to install with the sleeve during the grafting operation. When the wide diameter data of the stock sleeve is the maximum value of R1 minus 1-3mm, it can ensure the sealing of the rear interface of the sleeve and facilitate the grafting operation.

[0014] According to an embodiment of the present invention, the wide diameter data of the stock sleeve is the value of the maximum value of R1 minus 1mm. According to a specific embodiment of the present invention, the inventor performs a diameter scanning measurement on the stock along its length direction, and the range of the scanning measurement is the stock diameter within a length of 0-3cm from the incision cross section of the stock, for example, the stock diameter is scanned at 0.1cm, 0.5cm, 0.7cm, 0.8cm, 0.9cm, 1.0cm, 1.5cm, 2.0cm, 2.5cm and 3.0cm from the incision cross section of the stock to obtain diameter data of multiple stock, then the wide diameter data of the stock sleeve is the value obtained by subtracting 1mm from the maximum value of the diameter data of multiple stock obtained at 0.1cm, 0.5cm, 0.7cm, 0.8cm, 0.9cm, 1.0cm, 1.5cm, 2.0cm, 2.5cm and 3.0cm from the incision cross section of the stock.

[0015] According to an embodiment of the present invention, the narrow diameter data of the rootstock sleeve is 100%-130% of the minimum value of R1.

[0016] According to an embodiment of the present invention, the narrow diameter data of the rootstock sleeve is smaller than the wide diameter data of the rootstock sleeve.

[0017] According to an embodiment of the present invention, the narrow diameter data of the stock sleeve is the minimum value of R1. According to a specific embodiment of the present invention, the inventor performs a diameter scanning measurement on the stock along its length direction, and the scanning measurement range is the stock diameter within a length of 0-3 cm from the incision cross section of the stock. For example, the stock diameter is scanned at 0.1 cm, 0.5 cm, 0.7 cm, 0.8 cm, 0.9 cm, 1.0 cm, 1.5 cm, 2.0 cm, 2.5 cm and 3.0 cm from the incision cross section of the stock to obtain diameter data of multiple stock, then the narrow diameter data of the stock sleeve is the minimum value of the diameter data of multiple stock obtained at 0.1 cm, 0.5 cm, 0.7 cm, 0.8 cm, 0.9 cm, 1.0 cm, 1.5 cm, 2.0 cm, 2.5 cm and 3.0 cm from the incision cross section of the stock. The inventors found that when the diameter data of the stock sleeve is too large, it is difficult to ensure the sealing of the grafting sleeve after it is fixed. When the diameter data of the stock sleeve is too small, the stock is difficult to install with the sleeve during the grafting operation. Therefore, the grafting sleeve needs to select a suitable diameter size to ensure the sealing of the rear interface of the sleeve and the simplicity of the grafting operation, so as to moisturize and resist infection of the grafting wound.

[0018] According to an embodiment of the present invention, the wide diameter data of the scion sleeve is the diameter value of the scion at a distance of 1-2 cm from the incision cross section thereof. According to a specific embodiment of the present invention, the inventors perform a diameter scanning measurement on the scion along its length direction, scanning and measuring the diameter of the stock at a distance of 1-2 cm from the incision cross section of the scion, such as scanning the scion diameter at 1 mm, 1.5 mm, 2.0 mm, 2.5 mm, 3.0 mm, 5.0 mm, 1.0 cm, 1.5 cm, and 2.0 cm from the incision cross section of the stock to obtain the diameter data of the scion.

[0019] According to an embodiment of the present invention, the wide diameter data of the scion sleeve is the diameter value of the scion at a distance of 1-3 mm from the cut cross section thereof.

[0020] According to an embodiment of the present invention, the wide diameter data of the scion sleeve is the diameter value of the scion at a distance of 2 mm from the cut cross section thereof.

[0021] According to an embodiment of the present invention, the narrow diameter data of the scion sleeve is the diameter value of the scion at a distance of (0.65-0.75) L from its incision cross section, preferably the diameter value of the scion at a distance of 0.7 L from its incision cross section. According to some specific embodiments of the present invention, the inventors took a diameter scanning measurement of the length direction of the scion incision cross section, and the scanning measurement distance from the incision cross section of the scion is 0.65-0.75 times the length of the full length of the scion. For example, when the length L (full length) of the scion is 5 cm, the narrow diameter data of the scion sleeve is any scion diameter value at a distance of (5*0.65) to (5*0.75) cm from the incision cross section of the scion. When the narrow diameter data of the scion sleeve is the diameter value of the scion at a distance of 0.65-0.75 L from its incision cross section, it can ensure that the grafting sleeve is close to the scion, has good airtightness and is convenient for installation operation, and has a moisturizing and anti-infection effect on the grafting wound.

[0022] According to an embodiment of the present invention, the thermoplastic elastic material includes: styrene block copolymer, thermoplastic polyolefin elastomer, thermoplastic vulcanizate, thermoplastic polyurethane, thermoplastic copolyester or thermoplastic polyamide.

[0023] According to an embodiment of the present invention, the stock sleeve and the scion sleeve are hollow frustum-shaped transparent sleeves.

[0024] According to an embodiment of the present invention, the stock sleeve and the scion sleeve include a sleeve body, a pre-opening is provided on the sleeve body along its length, a transparent covering layer is provided on the pre-opening, and the material of the covering layer is the thermoplastic elastic material. The tube body and the pre-opening are both covered with a covering layer, and the material of the covering layer is the thermoplastic elastic material. The sleeve body and the pre-opening covering layer can be the same thermoplastic elastic material or different thermoplastic materials.

[0025] According to an embodiment of the present invention, the width of the pre-opening of the stock sleeve is 0.5-1.5 mm, and the thickness is 0.1-0.3 mm. The thickness of the pre-opening is smaller than the thickness of the sleeve body, so that the pre-opening can be opened after the grafted plant survives, allowing the grafting sleeve to fall off.

[0026] According to an embodiment of the present invention, the thickness of the sleeve body is 0.5-1.5 mm.

[0027] According to an embodiment of the present invention, a clamping portion is provided on the outer circumference of one of the stock sleeve and the scion sleeve at the wide diameter, and a buckling portion is provided on the outer circumference of the other, and the clamping portion and the buckling portion can be engaged and connected.

[0028] In a second aspect, the present invention provides a grafting sleeve. According to an embodiment of the present invention, the grafting sleeve is prepared using the method described in the first aspect. The grafting sleeve according to an embodiment of the present invention can be effectively used for plant grafting, reducing grafting time and labor costs, and significantly improving grafting efficiency and the survival rate of grafted plants.

[0029] In a third aspect of the present invention, the present invention provides a grafting method. According to an embodiment of the present invention, the method comprises the following steps:

[0030] The grafting sleeve described in the second aspect is respectively installed on the stock and the scion; the lower end of the scion is embedded in the upper end of the stock, wherein the stock and the scion are pretreated and installed with the grafting sleeve; the grafting sleeve is fixed by the clamping portion and the buckling portion of the grafting sleeve. Those skilled in the art will understand that when grafting is performed using this method, the stock and the scion can be installed with the grafting sleeve first and then pretreated, or they can be pretreated first and the pretreated stock and scion are installed with the grafting sleeve. The method according to the embodiment of the present invention can effectively reduce the labor cost and time cost during grafting, and improve the grafting efficiency and the survival rate of the grafted plants.

[0031] According to an embodiment of the present invention, the grafting method may further include at least one of the following additional technical features:

[0032] According to an embodiment of the present invention, the length of the rootstock and the scion is 4-7 cm.

[0033] According to an embodiment of the present invention, the diameter of the scion is 1-2 cm.

[0034] According to an embodiment of the present invention, the diameter of the rootstock is greater than or equal to the diameter of the scion.

[0035] According to an embodiment of the present invention, the pretreatment includes: exposing the cambium of the scion, the exposure position is 2-4 cm extending from the maximum diameter of the scion to the bottom end of the scion, and the exposed surface is more than one.

[0036] According to an embodiment of the present invention, after the pretreatment, the lower end of the scion forms a wedge-shaped structure. The lower end of the scion is the part where the scion and the stock are positioned. It will be understood by those skilled in the art that the shape of the lower end of the scion can be adjusted as needed, as long as it is a shape that is convenient for the survival of the grafted plant.

[0037] According to an embodiment of the present invention, the pretreatment further comprises: cutting downward perpendicularly to the cut cross section of the rootstock, wherein the depth of the cutting is longer than the length of the exposed surface of the scion. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0039] FIG1 is a schematic diagram of the operation of a traditional grafting method provided by an embodiment of the present invention;

[0040] FIG2 is a schematic diagram of a process of printing a grafting auxiliary sleeve using a 3D printer and a structural diagram of the grafting auxiliary sleeve according to an embodiment of the present invention. DETAILED DESCRIPTION

[0041] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0042] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of the technical features being referred to. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "plurality" means at least two, such as two, three, etc., unless otherwise specifically defined.

[0043] To facilitate understanding of the present invention, a brief introduction is given here:

[0044] The present invention designs a kind of 3D printing rapid grafting auxiliary sleeve, which can replace the adhesive tape binding operation in the grafting process, play the role of moisturizing and anti-infective of the grafting wound, and make the scion and the rootstock cambium automatically aligned and tightly fitted (no loosening during the wound healing process), greatly improving the grafting efficiency, reducing the difficulty of personnel training, and reducing the number of technicians. The traditional method also has the situation of using the sleeve method to assist in improving efficiency, provided that the diameter of the plant material scion to be grafted is basically the same as that of the rootstock and can be adapted to the sleeve diameter, so the scope of application is narrow.

[0045] The present invention first uses a laser radar scanning distance measurement method to confirm the basic shape and size of the stock and scion, and converts it into the corresponding grafting sleeve size through an independently designed calculation method, and then uses a 3D printing method to respectively produce two sections of sleeves suitable for the scion and the stock for rapid grafting. The sleeve can be adapted to both manual grafting operations and fully automatic grafting machines. In the selection of two types of sleeve 3D materials, since the stock sleeve needs to have both moisturizing and binding functions, it is necessary to select a printing material with both thermoplasticity and elasticity. At this stage, thermoplastic elastomers (TPEs) are preferred, including styrene block copolymers (TPS), thermoplastic polyolefin elastomers (TPO), thermoplastic vulcanizates (TPV), thermoplastic polyurethanes (TPU), thermoplastic copolyesters (TPC), thermoplastic polyamides (TPA), etc.; and the main function of the scion sleeve is moisturizing, and the elasticity requirements are reduced, and there are more materials to choose from. However, considering the convenience of operation, the scion sleeve should be made of the same material as the stock sleeve. The stock sleeve is a transparent round tube with an inner diameter that matches the stock diameter, a pre-engraved structure at the upper end (for the scion section), and a pre-opening on the side (not completely disconnected); the scion sleeve is a truncated cone-shaped transparent round tube with a wide lower end (for the stock end), a narrow upper end (in the direction of plant growth), a pre-engraved structure at the lower end, and a pre-opening on the side. During the grafting operation, the stock sleeve is first placed on the stock, and then the stock is processed, that is, cut from the middle, the scion is placed in the scion sleeve and the bottom is exposed for cutting (a wedge-shaped structure is cut out with a blade), and finally the scion wedge-shaped structure is inserted into the stock incision and the two sections of the conduit interface are engaged. The wound of the grafted plant gradually heals and continues to grow. The grown plant bursts through the sleeve, and the sleeve falls off naturally. The material can be 100% recycled and reused.

[0046] When dealing with small-scale grafting tasks, a smartphone (or other portable scanner) lidar + miniaturized 3D printer solution can be used. It is portable and movable. The scions and rootstocks are manually scanned in batches and then the sleeves are printed in batches. After printing is completed, the sleeve operations are carried out in an orderly manner.

[0047] When dealing with large-scale grafting tasks, the production process can be modularized and integrated with automated rootstock processing and automated joining operations to form a fully automated solution. After the rootstock and scion are scanned with the lidar module, the information is transmitted to the sleeve 3D printing module to make the sleeve. At the same time, the rootstock is transmitted to the incision module for disinfection and incision processing. Then the scion, rootstock and sleeve are transmitted to the joining module for combination to realize mass production of grafted plants.

[0048] As shown in Figure 2, the grafting sleeve (stock sleeve and scion sleeve) obtained by the method described in the present application includes a sleeve body, and the sleeve body is provided with one or more pre-openings along its length direction. The pre-opening is provided with a covering layer. The material of the covering layer is the same as that of the sleeve body, but the thickness of the pre-opening covering layer is smaller than that of the sleeve body, so as to facilitate the expansion of the grafting sleeve after the plant grows. The stock sleeve and the scion sleeve are in the shape of a truncated cone, wherein, in the stock sleeve, the narrow diameter end (narrow end) of the stock sleeve wraps the cut end (upper end) of the stock, the wide diameter end (wide end) includes the lower end of the stock, and the narrow diameter end of the stock sleeve is provided with a snap-fit ​​portion. In some cases, the diameters of the upper and lower ends of the stock are basically the same; in the scion sleeve, the wide diameter end (wide end) of the scion sleeve wraps the cut end (lower end) of the scion, the narrow diameter end (narrow end) includes the upper end of the scion, and the wide diameter end of the scion sleeve is provided with a snap-fit ​​portion. The snap-fit ​​portion and the snap-fit ​​portion can be engaged and connected, and the stock sleeve and the scion sleeve can be fixed during the grafting operation, so that the grafting sleeve has good sealing properties.

[0049] In the present invention, the "cambium" refers to a meristematic tissue located between the xylem and phloem in the roots and stems of gymnosperms and dicotyledons.

[0050] Example 1

[0051] In this example, a grafting auxiliary cannula was obtained by 3D printing. Experiments were conducted at the State Key Laboratory of Agrigenomics, Shenzhen BGI Life Sciences Institute, Longgang District, Shenzhen. The grafting efficiency and survival rate of the traditional grafting method and the grafting method using the 3D grafting auxiliary cannula were compared. The specific experimental procedures are as follows:

[0052] 1.1 Preparation of long purple eggplant scion

[0053] Before sowing eggplant, soak the seeds in warm water at about 50 degrees for about 2 hours, and then soak them in a gibberellin solution for 22 hours to accelerate germination. After the eggplant seeds sprout, sow them in loose, breathable and well-drained potting soil. After the eggplant seedlings grow stably, transplant them into land with deep, soft, breathable and well-drained soil. They will grow into plants that can be used for grafting in about 60 days. The scion comes from the side branches of the plant body. The part where the side branches are connected to the stem is cut off with grafting shears. The selected side branches are about 20 cm long and the stem is about 1.2 cm thick. After selecting a suitable scion, the scion needs to be pre-treated before the grafting operation: all the leaves of the scion are cut off, leaving one terminal bud and several axillary buds.

[0054] 1.2 Preparation of eggplant rootstock

[0055] Interspecific hybridization was performed using wild Solanum wrightii as the female parent and wild Solanum torvum as the male parent. Immature hybrid fruits were obtained 22 days after cross-pollination and surface sterilized with NaClO. The embryos in the fruits were cut and separated with a sterile blade and transferred to an induction proliferation medium (MS + 2 mg / L ZR) and cultured at 25 ± 1 °C. The induced embryoids were transferred to a regeneration medium (MS + 2 mg / L 6-BA + 0.5 mg / L IAA) to obtain regenerated buds, which were then transferred to a rooting medium (MS + 0.4 mg / L After 10 days, the in vitro cultured seedlings were taken out, and the adhering culture medium was gently removed with distilled water. The seedlings were transplanted into a hardening medium composed of coconut coir, vermiculite and perlite in a volume ratio of 1:1:1. They were watered with 1 / 2MS nutrient solution every 2 weeks. The seedlings were covered with plastic wrap to keep them moist and maintained under strong light. After 6 days, the film was gradually removed. The hardened eggplant plantlets quickly grew into large seedlings in the medium and were then transplanted into soil or large seedling bags for growth. After about four months, they grew into plants suitable for grafting.

[0056] 1.3 Traditional grafting method

[0057] The 50 scions and 50 rootstocks from Sections 1.1 and 1.2 were grafted using the traditional grafting method. The traditional method was performed by technicians with many years of grafting experience. The specific steps are as follows:

[0058] 1) Pre-treat the scion: Use plastic wrap to tie the scion clockwise or counterclockwise and obliquely upward to the terminal bud, leaving the terminal bud exposed to the air and leaving 5 cm of bare branches at the bottom;

[0059] 2) Select a suitable rootstock stem side branch or stem trunk: the cross-sectional diameter of the selected side branch should be larger than the cross-sectional diameter of the scion (stem thickness). Use the grafting scissors to cut perpendicular to the epidermis of the side branch. Generally, the side branch incision surface is perpendicular to the branch trunk. The incision is kept smooth and neat. In this experiment, the rootstock is treated by cleft grafting. The blade of the utility knife is perpendicular to the widest side of the incision cross section and cut vertically downward. The depth is greater than the length of the scion cut surface, generally 5 cm is appropriate.

[0060] 3) When grafting, first leave 5 cm of branches at the root of the pretreated scion, use a tool knife with the blade facing the widest cross section of the branch, and cut out the cambium part of the scion at a certain angle. The same operation is performed on the reverse side of the cut surface, along a certain angle. The two cut surfaces are kept parallel and the length of the cut surface is kept at 3 cm. Finally, a wedge-shaped structure is formed at the bottom of the scion; then the scion can be inserted into the treated stock branch incision, and the cambium side of the scion must be firmly attached to the stock. The cambium on the same side of the incision is aligned, and a part of the cut surface of the scion can be retained above the incision. After the cambium of the scion and the cambium of the rootstock are aligned, they need to be tightly combined; then, the grafting wound is sealed and tied with a professional self-adhesive grafting film; after the tying is completed, the date of grafting and the variety of scion are written on a label and marked on the grafted rootstock. After about 14 days, it can be judged whether the plant has survived. After the top bud of the scion is stretched out, it can be judged that the grafted scion has survived. If it survives, the plastic wrap of the scion part can be untied depending on the situation.

[0061] 1.4 3D Printing Grafting Assisted Sleeve Method

[0062] Use a 3D printing device to quickly print 3D grafting auxiliary sleeves to graft the 50 scions and 50 rootstocks prepared in parts 1.1 and 1.2. The specific steps are as follows:

[0063] 1) First, 50 prepared scions and rootstocks were scanned using an iReal 2E handheld scanner. The diameters of the scions and rootstocks at approximately 5 cm from their respective cut cross sections were obtained. This data was then automatically entered into the Prusa Slicer software to adjust the pre-set rootstock and scion sleeve combination model. The basic logic for converting the LiDAR-scanned scion and rootstock dimensions to the 3D-printed sleeve dimensions was as follows: both the rootstock and the scion sleeve were hollow, frustum-shaped sleeves. The diameter of the rootstock within 3 cm of the cut cross section was measured. The narrowest diameter of the rootstock within 3 cm of the cut cross section was used as the inner diameter of the stock sleeve at the narrow end, r'2. The widest diameter of the rootstock within 3 cm of the cut cross section, minus 1 mm, was used as the inner diameter of the stock sleeve at the wide end, r'1. The diameter of the scion at a distance of 2 mm from the incision cross section is measured as the inner diameter r1 of the wide end of the scion sleeve; the length of the scion is measured as L, and the value of 0.7 times L is calculated to take the diameter at a distance of 0.7L from the incision cross section as the inner diameter r2 of the narrow end of the scion sleeve. Use Prusa i3 MK3S+ preloaded with PolyFlex TPU95 filament to print 50 scions and stock sleeves at one time, wherein the process of printing to obtain the scion or stock sleeve is shown in Figure 2, and the obtained stock sleeve is a truncated cone-shaped transparent round tube with an inner diameter adapted to the stock diameter, a reserved interlocking structure at the upper end (scion section), and a reserved thin transparent material (not completely disconnected) on the side (as shown in Figure 2); the scion sleeve is a truncated cone-shaped transparent round tube with a wide lower end (stock end), a narrow upper end (plant growth direction), a reserved interlocking structure at the lower end, and a reserved thin transparent material on the side (as shown in Figure 2).

[0064] 2) At the same time as step 1), select a suitable rootstock stem side branch or stem trunk. In this experiment, the cross-sectional diameter of the selected side branch rootstock must be larger than the scion stem. First, put the 3D printed rootstock sleeve on the rootstock, and then process the rootstock. The grafting scissors are perpendicular to the epidermis of the side branch. Generally, the side branch incision surface is perpendicular to the branch trunk. The incision is kept smooth and neat. The rootstock is processed by cleft grafting. The blade of the utility knife is perpendicular to the widest side of the incision cross section. Cut vertically downward. The depth is greater than the length of the scion cut surface, generally 5 cm. It is appropriate to leave 5 cm of branches at the root of the scion, place the scion in the above-mentioned 3D printed scion sleeve and expose the bottom for cutting, use the blade of the utility knife to face the widest cross-section of the branch, and cut out the cambium part of the scion at a certain inclination angle. The same operation is performed on the opposite side of the cut surface, and the two cut surfaces are kept parallel at a certain inclination angle. The length of the cut surface should be kept at 3 cm. Finally, a wedge-shaped structure is formed at the bottom of the scion. Finally, the scion wedge structure is inserted into the incision of the rootstock and the two sections of the catheter interface are embedded.

[0065] 3) After connecting the printed sleeve to the cut scion and rootstock using the above steps, write the grafting date and scion variety on a label and mark it on the grafted rootstock. After about 14 days, determine whether the plant has survived. When the terminal bud of the scion stretches out, it can be determined that the grafted scion has survived. As the surviving plant gradually grows, the sleeve will automatically crack and fall off.

[0066] 1.5 Experimental Results Analysis

[0067] Experimental results show that when grafting using traditional methods, each grafting process takes approximately 8 minutes, from pre-wrapping the scion with plastic wrap to completing the final self-adhesive grafting film binding, and a total of 400 minutes to complete 50 grafts. However, when grafting using the 3D-printed rapid grafting-assisted sleeve method, preparing the scion and stock incisions takes approximately 2 minutes, for a total of 100 minutes. During this time, the scion and stock sleeves can be printed simultaneously, so the entire process only takes 100 minutes, which is only 1 / 4 of the time consumed by traditional grafting. When grafting using traditional methods, the number of surviving plants was 47, with a survival rate of 94%. When grafting using the 3D-printed sleeve-assisted grafting method, the number of surviving plants was 49, with a survival rate of 98%, which is higher than the traditional grafting method performed by experienced grafting technicians.

[0068] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0069] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A method for preparing a grafting sleeve, characterized in that: The following steps are involved: 1) Obtaining stock diameter data R1, scion diameter data R2 and the length L of the scion; 2) obtaining data on the wide diameter and narrow diameter of a grafting sleeve, wherein the grafting sleeve includes a stock sleeve and a scion sleeve; 3) inputting the wide diameter data and narrow diameter data of the stock sleeve and the scion sleeve into a 3D printer; 4) Using thermoplastic elastic material as ink material, the grafting sleeve is obtained by a 3D printer.

2. The method according to claim 1, characterized in that The stock diameter data and the scion diameter data are obtained by scanning the diameters of the stock and the scion with a scanning device, and the length L of the scion is obtained by scanning the full length of the scion with a scanning device; Optionally, the diameter data of the rootstock is obtained by scanning the diameter of the rootstock at 0-5 cm from its cut cross section with a scanning device, and the diameter data of the scion is obtained by scanning the diameter of the scion at 0.1-1.5 cm from its cut cross section with a scanning device; Optionally, the diameter data of the rootstock is obtained by scanning the diameter of the rootstock at 0-3 cm from its cut cross section with a scanning device, and the diameter data of the scion is obtained by scanning the diameter of the scion at 0.1-1 cm from its cut cross section with a scanning device.

3. The method according to claim 1, characterized in that The wide diameter data of the stock sleeve is the value of the maximum value of R1 minus 1-3 mm; Optionally, the wide diameter data of the stock sleeve is the value of the maximum value of R1 minus 1 mm; Optionally, the narrow diameter data of the rootstock sleeve is 100%-130% of the minimum value of R1; Optionally, the narrow diameter data of the stock sleeve is smaller than the wide diameter data of the stock sleeve; Optionally, the narrow diameter data of the rootstock sleeve is the minimum value of R1.

4. The method according to claim 1, wherein The wide diameter data of the scion sleeve is the diameter value of the scion at a distance of 1-2 cm from the cut cross section; Optionally, the wide diameter data of the scion sleeve is the diameter value of the scion at a distance of 1-3 mm from the cut cross section thereof; Optionally, the wide diameter data of the scion sleeve is the diameter value of the scion at a distance of 2 mm from the cut cross section thereof; Optionally, the narrow diameter data of the scion sleeve is the diameter value of the scion at a distance of 0.65-0.75 times the L from the cut cross section thereof, preferably the diameter value of the scion at a distance of 0.7 times the L from the cut cross section thereof.

5. The method according to claim 1, wherein The thermoplastic elastic material includes: styrene block copolymer, thermoplastic polyolefin elastomer, thermoplastic vulcanizate, thermoplastic polyurethane, thermoplastic copolyester or thermoplastic polyamide.

6. The method according to claim 1, characterized in that The stock sleeve and the scion sleeve are hollow frustum-shaped transparent sleeves; Optionally, the stock sleeve and the scion sleeve comprise a sleeve body, the sleeve body is provided with one or more pre-openings along its length, the pre-openings are provided with a transparent covering layer, and the covering layer is made of the thermoplastic elastic material; Optionally, the pre-opening of the stock sleeve has a width of 0.5-1.5 mm and a thickness of 0.1-0.3 mm; Optionally, the thickness of the sleeve body is 0.5-1.5 mm; Optionally, one of the stock sleeve and the scion sleeve has a clamping portion on its outer circumference at its wide diameter, and the other has a buckling portion on its outer circumference, and the clamping portion and the buckling portion can be engaged and connected.

7. A grafting sleeve, characterized in that: The grafting sleeve is prepared by the method according to any one of claims 1 to 6.

8. A grafting method, characterized in that: The following steps are involved: Installing the grafting sleeve according to claim 7 on the rootstock and the scion respectively; Embedding the lower end of the scion into the upper end of the rootstock, wherein the rootstock and scion are pre-treated and installed with the grafting sleeve; The grafting sleeve is fixed by the clamping portion and the buckling portion of the grafting sleeve.

9. The method according to claim 8, characterized in that The length of the stock and scion is 4-7 cm; Optionally, the diameter of the scion is 1-2 cm; Optionally, the diameter of the rootstock is 1-6 cm, and the diameter of the rootstock is greater than or equal to the diameter of the embedded scion.

10. The method according to claim 8, characterized in that The pretreatment comprises: exposing the cambium of the scion, wherein the exposure position is 2-4 cm extending from the maximum diameter of the scion to the bottom end of the scion, and the exposed surface of the scion is more than one; Optionally, after the pretreatment, the lower end of the scion forms a wedge-shaped structure; Optionally, the pre-treating further comprises cutting downwardly perpendicular to the cut cross-section of the rootstock.

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