Tissue culture device and culture method for sugarcane tissue culture seedlings

Abstract

The invention discloses a tissue culture device and method for sugarcane tissue culture plants.The tissue culture device comprises explant filling equipment, culture medium filling equipment, screw cap equipment and a tissue culture frame, and the inlet end and the outlet end of the explant filling equipment are connected with a first conveying belt and a second conveying belt respectively; the inlet end of the culture medium filling device is connected with the tail end of the second conveying belt, the outlet end of the culture medium filling device is connected with a third conveying belt, the culture medium filling device is communicated with at least three culture solution temporary storage tanks, the inlet end of the cap screwing device is connected with the tail end of the third conveying belt, and the outlet end of the cap screwing device is connected with a fourth conveying belt. The inlet end of the tissue culture frame is connected with the tail end of the fourth conveying belt, and the outlet end of the tissue culture frame is connected with a fifth conveying belt which is connected with the first conveying belt. Through cooperative use of the explant filling equipment, the culture medium filling equipment, the screw cap equipment and the tissue culture frame, industrial, automatic and large-scale production of sugarcane tissue culture seedlings is realized, and the working efficiency is effectively improved.

Description

Technical Field

[0001] The present invention relates to the technical field of sugarcane tissue culture, and specifically relates to a tissue culture device and a cultivation method for sugarcane tissue culture seedlings. Background Art

[0002] Sugarcane is a temperate and tropical crop and is the raw material for making sucrose. It can be refined into ethanol as an energy alternative. China has a long history of sugarcane cultivation, and the planting areas are extensive, covering many provinces such as Yunnan, Guangdong, and Hainan. However, at present, the sugarcane seed stems in most sugarcane-growing areas in China are still in the stage of self-use. After years of continuous asexual seed retention and planting, various diseases have accumulated, causing the degeneration of variety characteristics, which directly affects the quality and yield of sugarcane. Obtaining healthy seedlings through tissue culture technology can improve the quality of sugarcane. However, at present, the production and cultivation efficiency of healthy sugarcane seedlings is low and far from meeting the planting needs. During the cultivation process of sugarcane tissue culture seedlings, long-term reliance on manual operations is required. The placement of explants, the filling of culture media, the replacement of culture media at different culture stages, and the placement of culture bottles all require manual operations, which easily lead to contamination, low automation, and affect the survival rate and production efficiency of tissue culture seedlings.

[0003] The patent application with the Chinese patent publication number CN117923404 A discloses a fully automated production line for cleaning and filling culture media for sugarcane virus-free tissue culture, including a first conveyor and a second conveyor. The conveying surface of the first conveyor and the conveying surface of the second conveyor are on the same plane; a flipping type ultrasonic cleaning machine, which is fixedly installed between one end of the first conveyor and one end of the second conveyor; an automatic quantitative filling machine, which is fixedly installed on the second conveyor; a capping machine body, which is arranged on the second conveyor. Although this patent realizes the automation of culture bottle cleaning and culture medium filling, after filling, it still needs to be manually carried to the tissue culture rack and placed manually one by one for tissue culture. Moreover, operations such as the placement of explants and the replacement of tissue culture media still need to be manually carried out, the process connection efficiency is low, and the possibility of being contaminated due to a large amount of manual contact affects the survival rate and production efficiency of tissue culture seedlings.

[0004] The patent application with the Chinese patent publication number CN 111512966 A discloses a tissue culture rack with a vertically circulating movement for tomato tissue culture baskets, including a main frame body. The main frame body is slidably arranged on the bottom plate. Two runway discs are installed on the main frame body, and two driving discs are arranged on the main frame body. The driving discs are coaxially connected to chain discs. This patent is convenient for adjusting to a suitable operation position for tomato tissue culture by the operator. The tissue culture bottle is placed above the bottle holder in the tissue culture frame at the bottom, effectively reducing the space occupied by the tissue culture frame and facilitating the taking and placing of tissue culture bottles. However, the culture bottles still need to be manually carried and placed one by one, and the production efficiency is also low. Summary of the Invention

[0005] The main object of the present invention is to overcome the defects existing in the above-mentioned background technology, and to provide a tissue culture device for sugarcane tissue culture seedlings and a cultivation method for sugarcane tissue culture seedlings.

[0006] To achieve the above object, the tissue culture device for sugarcane tissue culture seedlings proposed by the present invention includes an explant filling device, a culture medium filling device, a capping device and a tissue culture rack. The inlet end and the outlet end of the explant filling device are respectively connected with a first conveyor belt and a second conveyor belt. The culture bottle is conveyed into the explant filling device by the first conveyor belt. After the explant is put into the culture bottle, it is conveyed to the next process by the second conveyor belt. The inlet end of the culture medium filling device is connected to the end of the second conveyor belt, and the outlet end of the culture medium filling device is connected with a third conveyor belt. The culture medium filling device is communicated with at least three culture solution storage tanks. The culture bottle conveyed out from the second conveyor belt enters the culture medium filling device, and the prepared culture medium stored in the culture solution storage tank is quantitatively filled into the culture bottle and then conveyed to the next process by the third conveyor belt. The inlet end of the capping device is connected to the end of the third conveyor belt, and the outlet end of the capping device is connected with a fourth conveyor belt. The capping device includes a cap sorting component and a capping component. A lower cap guide rail is connected between the cap sorting component and the capping component. The culture bottle conveyed from the third conveyor belt and the bottle cap sliding down from the lower cap guide rail enter the capping component to be screwed and tightened, and then are conveyed to the next process by the fourth conveyor belt. The inlet end of the tissue culture rack is connected to the end of the fourth conveyor belt, and the outlet end of the tissue culture rack is connected with a fifth conveyor belt. The fifth conveyor belt is connected with the first conveyor belt. After the culture bottle is screwed with the cap, it enters the tissue culture rack through the fourth conveyor belt for cultivation. By the combined use of the explant filling device, the culture medium filling device, the capping device and the tissue culture rack, the industrialized, automated and large-scale production of sugarcane tissue culture seedlings is realized, and the production efficiency is effectively improved.

[0007] Further, the explant filling device includes a first bottle inlet star wheel, a dispenser, an explant storage hopper and a first filling machine body. The first bottle inlet star wheel is rotatably arranged on the first filling machine body. The explant storage hopper is fixedly arranged on the first filling machine body. The bottom of the explant storage hopper is communicated with a plurality of feeding pipes. The dispenser is communicated with one end of the feeding pipe and corresponds to the conveying track of the first bottle inlet star wheel. By putting the processed explants into the explant storage hopper, when the culture bottle rotates to the lower part of the dispenser, the dispenser drops into the culture bottle to automatically fill the explants.

[0008] Further, the dispenser includes a hollow housing, a dispensing wheel rotatably disposed inside the housing, a plurality of grooves formed on the outer peripheral surface of the dispensing wheel, and a dispensing motor connected to the wheel shaft of the dispensing wheel. After the explants fall into the grooves, the dispensing wheel is driven to rotate by the dispensing motor. When the grooves rotate to the bottom surface of the housing, the explants in the grooves automatically fall into the culture bottles below.

[0009] Further, the culture medium filling device includes a second filling body, a second bottle inlet star wheel, a bottle supporting plate, a central rotating shaft, and a plurality of filling valve assemblies. The central rotating shaft is rotatably and vertically disposed on the second filling body. The second bottle inlet star wheel, the bottle supporting plate, and the filling valve assemblies are all fixed on the central rotating shaft. A star wheel guard plate is fixed around the second bottle inlet star wheel. The bottle supporting plate is disposed below the second bottle inlet star wheel. A plurality of the filling valve assemblies are disposed above the second bottle inlet star wheel and correspond to the conveying track of the second bottle inlet star wheel. The central rotating shaft has a hollow structure. A rotary joint is provided at the top end of the central rotating shaft. The culture medium temporary storage tank is communicated with the rotary joint through a pipeline. A central distribution ring is communicated on the central rotating shaft. A plurality of the filling valve assemblies are all communicated with the central distribution ring. By adopting a rotary filling method, the prepared culture medium is quantitatively filled into the culture bottles. A plurality of filling valve assemblies can fill a plurality of culture bottles simultaneously, realizing a continuous process of "bottle inlet - filling - bottle outlet" without intermittent pauses, and the production efficiency is much higher than that of a linear filling device.

[0010] Further, the filling valve assembly includes a mounting plate, a valve body, a valve needle, and a positioning plate. The mounting plate is fixedly sleeved on the central rotating shaft. The valve body is circumferentially mounted on the mounting plate and can slide up and down. The positioning plate is fixedly mounted above the mounting plate through a support rod. A first spring is sleeved on the valve body. The bottom end of the first spring abuts against the top surface of the mounting plate. A first guide wheel is provided at the top end of the valve body. A first guide rail section corresponding to the first guide wheel is provided on the bottom surface of the positioning plate. A positioning block is provided at the bottom end of the valve body. The valve needle is inserted through the valve body. A second guide wheel is provided at the top end of the valve needle. A second guide rail section corresponding to the second guide wheel is provided on the outer peripheral surface of the positioning plate. A second spring is sleeved on the valve needle. A valve cavity is provided inside the valve body. The valve cavity is communicated with the central distribution ring through a liquid distribution pipe. The valve needle has a hollow tubular structure. A plurality of liquid inlet holes are formed on the side wall of a section of the valve needle body located in the valve cavity. Through the cooperation of the first guide wheel and the first guide rail section, the culture bottle is clamped between the positioning block and the bottle supporting plate to prevent the culture bottle from shaking during filling. Through the cooperation of the second guide wheel and the second guide rail section, the valve needle is communicated with the valve cavity on the valve body to carry out filling or stop filling.

[0011] Further, a number of top holes are provided on the bottle supporting plate, a top rod is provided in each top hole, a third guide wheel is provided at the bottom of the top rod, a support ring is provided on the support rod, a third guide rail section corresponding to the third guide wheel is provided on the support ring, a liquid discharge hole is provided on the bottom surface of the culture bottle, a first one-way blocking piece is provided at the liquid discharge hole, one end of the first one-way blocking piece is hinged to the inner bottom surface of the culture bottle, the top end of the top rod corresponds to the liquid discharge hole, a liquid injection hole is provided on the bottle cap, a second one-way blocking piece is provided at the liquid injection hole, one end of the second one-way blocking piece is hinged to the inner bottom surface of the bottle cap, and a partition net is provided in the culture bottle. Through the cooperation of the third guide wheel and the third guide rail section, when the culture bottle needs to discharge the original culture medium, the top rod rises to push open the first one-way blocking piece, and when new culture medium needs to be filled, the valve needle pushes down to push open the second one-way blocking piece and extends into the culture bottle.

[0012] Further, the tissue culture rack includes a rack body, runway discs are symmetrically provided on the rack body, a number of tissue culture baskets are provided on the runway discs, and the number of tissue culture baskets can rotate cyclically around the runway discs on both sides. The tissue culture basket includes a connecting frame, a carrying conveyor belt and a bottle arranging assembly. The carrying conveyor belt is laid on the connecting frame, the bottle arranging assembly is arranged on the carrying conveyor belt, a transverse moving guide rod is provided on the connecting frame, and both ends of the bottle arranging assembly are slidably connected to the transverse moving guide rod. A first lead screw is provided on the bottle arranging assembly, and one end of the first lead screw is connected with a first driving motor. After the culture bottle enters the tissue culture basket, the bottle arranging assembly arranges and stores the culture bottles on the carrying conveyor belt, avoiding the culture bottles being squeezed together and affecting the light.

[0013] Further, the bottle arranging assembly includes a moving frame, a bottle separating screw and a screw guard plate. The bottle separating screw is rotatably arranged on the moving frame, one end of the bottle separating screw is connected with a second driving motor, the screw guard plate is slidably arranged on the moving frame, a second lead screw is provided on the screw guard plate, and a third driving motor is provided at one end of the second lead screw. After the culture bottles are separated by a certain distance by the bottle separating screw, they are then arranged and placed, so as to reserve sufficient light spacing between the culture bottles.

[0014] Further, a buffer bed is connected to the fifth conveyor belt. The buffer bed is used for buffer storage of the culture bottles unloaded from the tissue culture rack, avoiding affecting the bottle unloading efficiency of the tissue culture bottles after a failure occurs in the subsequent process.

[0015] The present invention also provides a cultivation method for sugarcane tissue culture seedlings, which includes the following steps: S1. Preparation: Select tender sugarcane stems as explants. After the explants are disinfected, they are placed in the explant storage hopper of the explant filling device. At the same time, the induction medium, proliferation medium, and rooting medium are prepared and stored in the culture solution temporary storage tank respectively. The culture bottles and bottle caps are disinfected for standby; S2. Explant filling: Start the explant filling device. Place the disinfected culture bottles on the first conveyor belt and convey them into the first bottle inlet star wheel of the explant filling device. The explants enter the grooves of the delivery wheel from the explant storage hopper along the delivery pipe. When the culture bottle rotates to the position directly below the delivery device, the delivery motor drives the delivery wheel to rotate, and the explants in the grooves fall into the culture bottle; S3. Medium filling: Start the medium filling device. The culture bottles enter the second bottle inlet star wheel of the medium filling device from the second conveyor belt. During the rotation of the culture bottles, the first guide wheel rotates into the first guide rail section, and the valve body of the filling valve assembly is pressed down. The positioning block presses the mouth of the culture bottle, so that the culture bottle is clamped between the bottle supporting plate and the positioning block. The culture bottle continues to rotate with the second bottle inlet star wheel. When the second guide wheel rotates into the second guide rail section, the valve needle is pressed down. After the valve needle is pressed down, the liquid inlet hole on it communicates with the valve cavity, and the medium in the corresponding culture solution temporary storage tank enters the valve needle and is injected into the culture bottle. After the second guide wheel exits the second guide rail section, the valve needle moves up under the drive of the second spring, and the liquid inlet hole is misaligned with the valve cavity to stop filling. After the first guide wheel exits the first guide rail section, the valve body moves up to release the culture bottle; S4. Sealing the culture bottle cap: Start the capping device. The culture bottles enter the capping assembly from the third conveyor belt. The bottle caps stored in the cap sorting assembly slide down along the lower cap guide rail to the mouths of the culture bottles and enter the capping assembly together, and are tightened in the capping assembly; S5. Placing on the shelf for cultivation: Start the tissue culture rack. The culture bottles are conveyed into the tissue culture baskets of the tissue culture rack through the fourth conveyor belt. The culture bottles enter the carrying conveyor belt of the tissue culture basket and enter the bottle distribution assembly. After being separated by a certain distance by the bottle separating screw, the first driving motor drives the first lead screw to rotate, driving the bottle distribution assembly to drive the culture bottles to move horizontally. After moving in place, the third driving motor drives the second lead screw to rotate, driving the screw guard plate to slide up. The bottle distribution assembly retreats back to the starting point. Repeat until the tissue culture basket is full, and then the tissue culture basket rotates along the runway disc to load the next empty tissue culture basket. All tissue culture baskets are loaded with tissue culture bottles and then tissue culture is carried out; S6. Replacing the tissue culture medium: When replacing the culture medium during the tissue culture process, start the carrying conveyor belt. The culture bottles are conveyed along the fifth conveyor belt, pass through the first conveyor belt and the second conveyor belt, and re-enter the second bottle inlet star wheel of the medium filling device. During the rotation of the culture bottles, the third guide wheel rotates into the third guide rail section, and the ejector rod rises into the drain hole to lift the first one-way blocking piece up and turn it over. The original culture medium in the culture bottle is discharged from the drain hole. After discharging, the third guide wheel exits the third guide rail section, and the first one-way blocking piece covers the drain hole again for sealing. The second guide wheel rotates into the second guide rail section, and the valve needle is pressed down into the injection hole to push open the second one-way blocking piece, and the required culture medium is poured in.

[0016] The beneficial effects of the present invention include: through the coordinated use of the explant filling device, the culture medium filling device, the capping device, and the tissue culture rack, the placement of explants, the filling of the culture medium, the replacement of the culture medium at different culture stages, the placement of culture bottles, etc. do not require manual operation, realizing the industrialized, automated, and large-scale production of sugarcane tissue culture seedlings, breaking the situation that the traditional tissue culture method seriously relies on manual operation. Compared with the traditional tissue culture method, the operation efficiency is increased by more than 10 times, and the possibility of the culture medium being contaminated caused by manual contact can be avoided, improving the survival rate of tissue culture seedlings while improving the operation efficiency. Brief Description of the Drawings

[0017] Figure 1 It is the overall schematic diagram of the tissue culture device in the embodiment of the present invention.

[0018] Figure 2 It is the schematic diagram of the capping device in the embodiment of the present invention.

[0019] Figure 3 It is the schematic diagram of the explant filling device in the embodiment of the present invention.

[0020] Figure 4 It is the installation schematic diagram of the dispenser in the embodiment of the present invention.

[0021] Figure 5 It is the schematic diagram of the culture medium filling device in the embodiment of the present invention.

[0022] Figure 6 It is the installation schematic diagram of the filling valve assembly in the embodiment of the present invention.

[0023] Figure 7 It is the installation schematic diagram of the first guide rail section and the second guide rail section in the embodiment of the present invention.

[0024] Figure 8 It is the assembly sectional view of the valve body and the valve needle in the embodiment of the present invention.

[0025] Figure 9 It is the installation schematic diagram of the ejector rod in the embodiment of the present invention.

[0026] Figure 10 It is the assembly schematic diagram of the culture bottle and the bottle cap in the embodiment of the present invention.

[0027] Figure 11 It is the schematic diagram of the tissue culture basket in the embodiment of the present invention.

[0028] Figure 12 It is the schematic diagram of the bottle arranging assembly in the embodiment of the present invention.

[0029] Figure numerals: 1 explant filling device; 101 first bottle inlet star wheel; 102 dispenser; 1021 housing; 1022 dispensing wheel; 1023 groove; 1024 dispensing motor; 103 explant storage bucket; 104 first filling body; 105 dispensing tube; 2 culture medium filling device; 201 second filling body; 202 second bottle inlet star wheel; 203 bottle supporting plate; 2031 top hole; 2032 push rod; 2033 third guide wheel; 2034 support ring; 2035 third guide rail segment; 204 central rotating shaft; 205 filling valve assembly; 2051 mounting plate; 2052 valve body; 2053 valve needle; 2054 positioning plate; 2055 first spring; 2056 first guide wheel; 2057 first guide rail segment; 2058 positioning block; 2059 second guide wheel; 2060 second guide rail segment; 2061 second spring; 2 062 valve chamber; 2063 liquid inlet hole; 2064 liquid dispensing tube; 206 star wheel guard plate; 207 rotary joint; 208 pipeline; 209 support rod; 3 capping equipment; 301 capping assembly; 302 capping assembly; 303 lower cover guide rail; 304 capping head; 4 tissue culture rack; 401 rack body; 402 runway plate; 403 tissue culture basket; 404 connection frame; 405 carrying conveyor belt; 406 bottle distribution assembly; 406 1 moving rack; 4062 bottle separation screw; 4063 screw guard plate; 4064 second drive motor; 4065 second screw rod; 4066 third drive motor; 407 transverse guide rod; 408 first screw rod; 409 first drive motor; 5 first conveyor belt; 6 second conveyor belt; 7 culture bottle; 8 third conveyor belt; 9 fourth conveyor belt; 10 fifth conveyor belt; 11 culture solution temporary storage tank; 12 buffer bed; 13 bottle cap. DETAILED DESCRIPTION

[0030] In order to make the technical problems, technical solutions and beneficial effects to be solved by the embodiments of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0031] It should be noted that when a component is referred to as being "fixed to" or "disposed on" another component, it can be directly on the other component or indirectly on the other component. When an element is referred to as being "connected to" another element, it can be directly connected to the other element or indirectly connected to the other element. In addition, connection can be used for both fixing and circuit connection.

[0032] It should be understood that the terms "length", "width", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating the orientation or position relationship, are based on the orientation or position relationship shown in the accompanying drawings, and are only for the convenience of describing the embodiments of the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention.

[0033] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of the features. In the description of the embodiments of the present invention, the meaning of "plurality" is two or more, unless otherwise clearly and specifically defined.

[0034] See also Figures 1 to 12In one embodiment, a tissue culture device for sugarcane tissue culture seedlings disclosed in the present invention is installed in a sterile culture room with a purification level of 100,000, and the tissue culture device includes an explant filling device 1, a culture medium filling device 2, a capping device 3 and a tissue culture rack 4; a first conveyor belt 5 and a second conveyor belt 6 are respectively connected to the inlet and outlet ends of the explant filling device 1, and a culture bottle 7 is conveyed into the explant filling device 1 by the first conveyor belt 5, and the explant after the disinfection treatment is put into the culture bottle 7 and then conveyed to the next process via the second conveyor belt 6, the inlet end of the culture medium filling device 2 is connected to the end of the second conveyor belt 6, and the outlet end of the culture medium filling device 2 is connected to the third conveyor belt 8, and the culture medium filling device 2 is connected to the outlet end of the culture medium filling device 2. 2 is connected with at least three culture solution temporary storage tanks 11, which are used to store prepared culture media used in different tissue culture stages. The culture bottles 7 conveyed from the second conveyor belt 6 enter the culture medium filling device 2, and the prepared culture media stored in the culture solution temporary storage tank 11 are quantitatively filled into the culture bottles 7 and then conveyed to the next process via the third conveyor belt 8; the inlet end of the capping device 3 is connected to the end of the third conveyor belt 8, and the outlet end of the capping device 3 is connected to the fourth conveyor belt 9. The capping device 3 includes a capping assembly 301 and a capping assembly 302. A lower cover guide rail 303 is connected between the capping assembly 301 and the capping assembly 302, and a lower cover guide rail 303 is stored on the capping assembly 301. There is a bottle cap 13 for sealing the culture bottle 7. After the bottle cap 13 is sorted by the cap arrangement assembly 301, the cover opening of the bottle cap 13 is uniformly facing downward and enters the lower cover guide rail 303, and the bottle cap 13 slides down onto the culture bottle 7. The culture bottle 7 conveyed from the third conveyor belt 8 and the bottle cap 13 sliding down from the lower cover guide rail 303 enter the cap screwing assembly 302, and are screwed and tightened by the cap screwing head 304, and then conveyed to the next process via the fourth conveyor belt 9; the inlet end of the tissue culture rack 4 is connected to the end of the fourth conveyor belt 9, and the outlet end of the tissue culture rack 4 is connected to the fifth conveyor belt 10, and the fifth conveyor belt 10 is connected to the first conveyor belt 5. After the culture bottle 7 is screwed with the cap, it enters the tissue culture rack 4 via the fourth conveyor belt 9. Arrange and place for cultivation. When one stage of cultivation is completed, the culture bottle 7 is conveyed from the tissue culture rack 4 via the fifth conveyor belt 10 and returned to the first conveyor belt 5, and finally enters the culture medium filling device 2 to replace the culture medium. The culture medium used in the present invention is a liquid culture medium, which is convenient for quantitative filling and replacement by the culture medium filling device 2. In this embodiment, through the coordinated use of the explant filling device 1, the culture medium filling device 2, the capping device 3 and the tissue culture rack 4, the industrialized, automated and large-scale production of sugarcane tissue culture seedlings is realized, breaking the situation that the traditional tissue culture method is heavily dependent on manual work, and the operation efficiency is improved by more than 10 times compared with the traditional tissue culture method.

[0035] In a specific example, the explant filling device 1 includes a first bottle-input star wheel 101, a dispenser 102, an explant storage bucket 103 and a first filling body 104. The first bottle-input star wheel 101 is rotatably set on the first filling body 104, and the explant storage bucket 103 is fixedly set on the first filling body 104. The bottom of the explant storage bucket 103 is connected to a plurality of delivery tubes 105. The dispenser 102 corresponding to the number of delivery tubes 105 is connected to one end of the delivery tube 105 and corresponds to the conveying track of the first bottle-input star wheel 101. After the explant is disinfected, it is placed in the explant storage bucket 103 for use. The dispenser 102 includes a hollow shell 1021, and the bottom end of the shell 1021 is an open structure. A delivery wheel 1022 is rotatably provided inside the shell 1021. A plurality of grooves 1023 are provided on the outer peripheral surface of the wheel 1022, and a delivery motor 1024 is connected to the axle of the delivery wheel 1022; in the initial stage of tissue culture, the sterilized culture bottle 7 is placed on the first conveyor belt 5 and conveyed into the first bottle-entering star wheel 101, and the explant enters the groove 1023 of the delivery wheel 1022 from the explant storage bucket 103 along the delivery tube 105. When the culture bottle 7 rotates to enter directly below the shell 1021 of the dispenser 102, the delivery motor 1024 drives the delivery wheel 1022 to rotate. When the groove 1023 rotates to align with the opening at the bottom of the shell 1021, the explant in the groove 1023 falls into the culture bottle 7 under the action of its own gravity, completing the automatic canning of the implant, without the need for manual transfer of the explant for bottling, effectively improving production work efficiency.

[0036] In a specific example, the culture medium filling equipment 2 includes a second filling body 201, a second bottle-inlet star wheel 202, a bottle-supporting plate 203, a central rotating shaft 204 and a plurality of filling valve assemblies 205. The central rotating shaft 204 is rotatably arranged vertically on the second filling body 201. The second bottle-inlet star wheel 202, the bottle-supporting plate 203 and the filling valve assembly 205 are all fixed on the central rotating shaft 204. A star wheel guard plate 206 is fixed to the periphery of the second bottle-inlet star wheel 202. The bottle-supporting plate 203 is arranged below the second bottle-inlet star wheel 202. The plurality of filling valve assemblies 205 are arranged above the second bottle-inlet star wheel 202 and correspond to the conveying track of the second bottle-inlet star wheel 202. The central rotating shaft 204 is a hollow structure. 4 is provided with a rotary joint 207 at the top, the culture solution temporary storage tank 11 is connected to the rotary joint 207 through a pipeline 208, the central rotating shaft 204 is connected to a central distribution ring 2041, and a plurality of filling valve assemblies 205 are all connected to the central distribution ring 2041. Specifically, the second bottle inlet star wheel 202, the bottle supporting plate 203 and the filling valve assembly 205 are driven by the central rotating shaft 204 to rotate synchronously; the culture medium filling equipment 2 adopts a rotary filling method to quantitatively fill the prepared culture medium into the culture bottle 7, and a plurality of filling valve assemblies 205 can fill multiple culture bottles 7 at the same time, realizing a continuous process of "bottle inlet-filling-bottle outlet" without intermittent pauses, and the production efficiency is much higher than that of a linear filling equipment.

[0037] In a specific example, the filling valve assembly 205 includes a mounting plate 2051, a valve body 2052, a valve needle 2053 and a positioning plate 2054. The mounting plate 2051 is fixedly sleeved on the central rotating shaft 204. The valve body 2052 is mounted around the mounting plate 2051 and can slide up and down. The positioning plate 2054 is fixedly mounted above the mounting plate 2051 through a support rod 209. The positioning plate 2054 does not rotate with the central rotating shaft 204. A first spring 2055 is sleeved on the valve body 2052. The bottom end of the first spring 2055 abuts against the top surface of the mounting plate 2051. A first guide wheel 2056 is provided on the top of the valve body 2052. The bottom surface of the positioning plate 2054 is provided with a first guide wheel abutting against the first guide wheel. The valve body 2052 is provided with a first guide rail section 2057 corresponding to the wheel 2056, a positioning block 2058 is provided at the bottom end of the valve body 2052, a valve needle 2053 is penetrated on the valve body 2052, a second guide wheel 2059 is provided on the top of the valve needle 2053, a second guide rail section 2060 corresponding to the second guide wheel 2059 is provided on the outer peripheral surface of the positioning plate 2054, a second spring 2061 is sleeved on the valve needle 2053, a valve cavity 2062 is provided inside the valve body 2052, the valve cavity 2062 is communicated with the central distribution ring 2041 through a liquid distribution pipe 2064, the valve needle 2053 is a hollow tubular structure, and a tube body of the valve needle 2053 is located in the valve cavity 2062. The side wall is provided with a plurality of liquid inlet holes 2061. 63. After the culture bottle 7 enters the second bottle-inlet star wheel 202, the first guide wheel 2056 drives into the first guide rail section 2057, driving the valve body 2052 to overcome the elastic force of the first spring 2055 and move the positioning block 2058 downward to the bottle mouth of the culture bottle 7, clamping the culture bottle 7 between the positioning block 2058 and the bottle supporting plate 203 to prevent the culture bottle 7 from shaking during rotation, thereby ensuring the stability of the filling process; after the filling is completed, the first guide wheel 2056 drives out of the first guide rail section 2057, and under the action of the first spring 2055, the valve body 2052 moves upward to release the culture bottle 7, and the culture bottle 7 can be conveyed from the second bottle-inlet star wheel 202 to the third conveyor belt 8; when the culture bottle 7 rotates to the filling When the second guide wheel 2059 drives into the second guide rail section 2060, the valve needle 2053 is pressed downward, and the liquid inlet hole 2063 on the tube body of the valve needle 2053 enters the valve cavity 2062 during the downward movement of the valve needle 2053. The culture medium in the valve cavity 2062 enters the interior of the valve needle 2053 along the liquid inlet hole 2063 and flows downward to be injected into the clamped culture bottle 7 below for automatic filling. When the second guide wheel 2059 drives out of the second guide rail section 2060, under the action of the second spring 2061, the valve needle 2053 moves upward, and the liquid inlet hole 2063 and the valve cavity 2062 are misaligned to stop filling, thereby realizing quantitative filling of the culture medium. There is no need for manual quantitative filling, which effectively improves production efficiency.

[0038] In a specific example, the tissue culture rack 4 includes a frame 401, on which a runway disk 402 is symmetrically provided, and a plurality of tissue culture baskets 403 are provided on the runway disk 402. The plurality of tissue culture baskets 403 can circulate around the runway disks 402 on both sides, and the tissue culture baskets 403 include a connecting frame 404, a carrying conveyor belt 405 and a cloth bottle assembly 406. The carrying conveyor belt 405 is laid on the connecting frame 404, and the cloth bottle assembly 406 is arranged on the carrying conveyor belt 405. A transverse guide rod 407 is provided on the connecting frame 404, and both ends of the cloth bottle assembly 406 are slidably connected to the transverse guide rod 407. A first screw rod 408 is provided on the cloth bottle assembly 406, and one end of the first screw rod 408 is connected to a first driving motor 409. Through the above structural arrangement, the tissue culture rack 4 can simultaneously accommodate multiple tissue culture baskets 403, which can improve the space occupancy rate and facilitate the placement of the culture bottle 7. When the tissue culture basket 403 rotates to the bottom, it docks with the fourth conveyor belt 9 and the starting end of the carrier conveyor belt 405, and the end of the carrier conveyor belt 405 docks with the starting end of the fifth conveyor belt 10. The filled and capped culture bottle 7 enters the bottle cloth assembly 406 on the carrier conveyor belt 405, and the first drive motor 409 drives the first screw rod 408. The first screw rod 408 drives the bottle distribution assembly 406 to push the culture bottles 7 for placement, and there is no need for manual placement, which further improves work efficiency. Until a group of culture baskets 403 is full of culture bottles 7, the tissue culture baskets 403 rotate around the runway disk 402, and the empty tissue culture baskets 403 rotate to the lowest position and dock with the fourth conveyor belt 9. This cycle continues until all tissue culture baskets 403 are filled with culture bottles 7. After the cultivation is completed, the carrying conveyor belt 405 is started to transport the culture bottles 7 on the carrying conveyor belt 405 to the fifth conveyor belt 10 to exit the tissue culture rack 4.

[0039] In a preferred embodiment, the bottle distribution assembly 406 includes a movable frame 4061, a bottle separation screw 4062 and a screw guard plate 4063. The bottle separation screw 4062 can be rotatably set on the movable frame 4061, one end of the bottle separation screw 4062 is connected to a second drive motor 4064, the screw guard plate 4063 is slidably set on the movable frame 4061, a second screw 4065 is provided on the screw guard plate 4063, and a third drive motor 4066 is provided at one end of the second screw 4065. When the tissue culture bottle 7 enters the bottle distribution assembly 406, the second drive motor 4064 drives the bottle distribution screw 4062 to rotate. Under the action of the bottle distribution screw 4062, the tissue culture bottle 7 is pulled apart between the bottle distribution screw 4062 and the screw guard plate 4063, so that there is at least 4 cm between two adjacent tissue culture bottles 7. After the movable frame 4061 is driven by the first screw rod 408 to carry the tissue culture bottle 7 horizontally to the placement position, the third drive motor 4066 drives the second screw rod 4065 to rotate, the screw guard plate 4063 slides upward, and the movable frame 4061 returns to the starting point to complete the bottle distribution. The bottle body spacing of each row of tissue culture bottles 7 is controlled by the bottle distribution screw 4062, and the bottle body spacing of each column of tissue culture bottles 7 is controlled by the position of the movable frame 4061 driven by the first screw rod 408, which effectively avoids the tissue culture bottles 7 from being squeezed together to affect the light and avoids developmental abnormalities caused by insufficient light.

[0040] In a preferred embodiment, a buffer bed 12 is connected to the fifth conveyor belt 10. When the tissue culture bottles 7 are conveyed from the tissue culture rack 4, they are buffered and stored on the buffer bed 12 to avoid blockage of the fifth conveyor belt 10 and affect the efficiency of unloading the tissue culture bottles 7 after a malfunction occurs in the subsequent process.

[0041] In a preferred embodiment, a plurality of top holes 2031 are provided on the bottle supporting plate 203, a top rod 2032 is provided in the top hole 2031, the top rod 2032 can move up and down in the top hole 2031, a third guide wheel 2033 is provided at the bottom of the top rod 2032, a support ring 2034 is provided on the support rod 209, a third guide rail section 2035 corresponding to the third guide wheel 2033 is provided on the support ring 2034, a drainage hole 701 is provided on the bottom surface of the culture bottle 7, a first one-way plugging piece 702 is provided at the drainage hole 701, one end of the first one-way plugging piece 702 is hinged to the inner bottom surface of the culture bottle 7, and the top end of the top rod 2032 is hinged to the drainage hole Corresponding to 701, a liquid injection hole 1301 is provided on the bottle cap 13, and a second one-way plugging piece 1302 is provided at the liquid injection hole 1301. One end of the second one-way plugging piece 1302 is hinged to the inner bottom surface of the bottle cap 13. A partition net 703 is provided in the culture bottle 7. After the explant is put into the culture bottle 7, it falls on the partition net 703 to prevent the explant from falling directly on the bottom surface of the culture bottle 7. When the first one-way plugging piece 702 turns upward, it hits the tissue culture seedlings that have been cultured for a period of time, causing their root system to break and affecting their growth. Specifically, a torsion spring (not shown) is sleeved on the hinge axis of the first one-way plugging piece 702 and the second one-way plugging piece 1302. The first one-way plugging piece 702 or the second one-way plugging piece 1302 is hinged to the inner bottom surface of the bottle cap 13. When the second one-way plugging piece 1302 is not acted upon by an external force, the first one-way plugging piece 702 and the second one-way plugging piece 1302 can respectively block the drainage hole 701 and the injection hole 1301 under the action of the torsion spring; in the present embodiment, when the tissue culture medium needs to be replaced, the culture bottle 7 covered with the bottle cap 13 is conveyed out from the tissue culture rack 4 and re-enters the culture medium filling device 2, and after the culture bottle 7 is clamped between the positioning block 2058 and the bottle supporting plate 203, the third guide wheel 2033 drives into the third guide rail section 2035, and the push rod 2032 moves upward to push the first one-way plugging piece 702 upward to flip and open the drainage hole 701, and the original culture medium in the culture bottle 7 is filled with The culture medium is discharged from the drainage hole 701 (the diameter of the top rod 2032 is smaller than the inner diameter of the drainage hole 701). After the original culture medium is discharged, the three guide wheels 2033 move out of the third guide rail section 2035, the top rod 2032 moves down, and the first one-way plugging piece 702 re-blocks the drainage hole 701. At the same time, the second guide wheel 2059 moves into the second guide rail section 2060, and the valve needle 2053 moves down to flip the second one-way plugging piece 1302 to open the injection hole 1301. The bottom end of the valve needle 2053 enters the culture bottle 7, and the new culture medium is re-injected for filling, thereby realizing the automatic replacement of the culture medium. Compared with the traditional method of replacing the culture medium, the work efficiency is greatly improved.

[0042] Please refer again Figures 1 to 12 In one embodiment, a method for cultivating sugarcane tissue culture seedlings disclosed herein utilizes the above-mentioned tissue culture device to cultivate sugarcane tissue culture seedlings, comprising the following steps:

[0043] S1. Preparation: Select axillary buds, stem tip leaf sheaths, leaf tissues, root tip meristems or tender stems of sugarcane as explants. After disinfection, the explants are placed in the explant storage bucket 103 of the explant filling device 1 for standby use. At the same time, prepare the induction culture medium, proliferation culture medium and rooting culture medium, and store them in the culture solution temporary storage tank 11 for standby use. Disinfect the culture bottle 7 and the bottle cap 13 for standby use. The induction culture medium includes MS+6-BA1.5-2.0mg / L+NAA0.2-0.6mg / L+2,4-EBR1-2mg / L+medamba 0.02-0.05mg / L+sucrose 40g / L+antibacterial agent; the proliferation culture medium includes MS+6-BA2.0-4.0mg / L g / L+NAA0.1-0.3mg / L+sucrose 40g / L+antibacterial agent, the rooting culture medium includes 1 / 2MS+NAA6mg / L+sucrose 60g / L+antibacterial agent; S2, explant filling: start the explant filling equipment 1, put the sterilized culture bottle 7 empty bottle into the first conveyor belt 5 and convey it into the first bottle inlet star wheel 101 of the explant filling equipment 1, the explant enters the groove 1023 of the delivery wheel 1022 from the explant storage bucket 103 along the delivery tube 105, when the culture bottle 7 rotates to enter the bottom of the delivery device 102, the delivery motor 1024 drives the delivery wheel 1022 to rotate, and the explant in the groove 1023 falls into the culture bottle 7 under the action of gravity for explant filling. S3, culture medium filling: start the culture medium filling device 2, the culture bottle 7 loaded with explants enters the second bottle inlet star wheel 202 of the culture medium filling device 2 from the second conveyor belt 6, the culture bottle 7 is driven to rotate, the first guide wheel 2056 rotates and drives into the first guide rail section 2057, the valve body 2052 of the filling valve assembly 205 is pressed down, and the positioning block 2058 is pressed down synchronously to press the bottle mouth of the culture bottle 7, so that the culture bottle 7 is clamped between the bottle support plate 203 and the positioning block 2058, the culture bottle 7 continues to rotate with the second bottle inlet star wheel 202, when the second guide wheel 2059 rotates and drives into the second guide rail section 2060, the valve needle 205 3 is pressed down, after the valve needle 2053 is pressed down, the liquid inlet hole 2063 on the valve needle 2053 enters the valve cavity 2062 and is connected with the valve cavity 2062, and the culture medium in the corresponding culture medium temporary storage tank 11 enters the valve needle 2053 and is injected into the culture bottle 7 for filling. After the second guide wheel 2059 drives out of the second guide rail section 2060, the valve needle 2053 moves up driven by the second spring 2061, and the liquid inlet hole 2063 rises and is misaligned with the valve cavity 2062 to stop filling. After the first guide wheel 2056 drives out of the first guide rail section 2057, the valve body 2052 moves up to release the culture bottle 7; in the initial stage, the induction culture medium in step S1 is first filled into the culture bottle 7.S4, capping of culture bottle 7: start the capping device 3, the culture bottle 7 filled with culture medium enters the capping assembly 302 from the third conveyor belt 8, and the bottle caps 13 stored in the capping assembly 301 slide down along the lower cover guide rail 303 to the bottle mouth of the culture bottle 7 and enter the capping assembly 302 together, and are tightened in the capping assembly 302. S5, shelf cultivation: start the tissue culture rack 4, the culture bottle 7 after tightening the bottle cap 13 is conveyed to the tissue culture basket 403 of the tissue culture rack 4 through the fourth conveyor belt 9, the culture bottle 7 enters the carrying conveyor belt 405 of the tissue culture basket 403 and enters the bottle distribution assembly 406, and after being pulled apart by the bottle separation screw 4062 for a certain distance, the first drive motor 409 drives the first screw 408 to rotate, and drives the bottle distribution assembly 406 to drive the culture bottle 7 to move horizontally, and after moving into place, the third drive motor 4066 drives the second screw 4065 to rotate , driving the screw guard plate 4063 to slide upward, the bottle assembly 406 retreats and returns to the starting point, and repeats until the tissue culture basket 403 is full. The tissue culture basket 403 rotates along the runway plate 402, and the empty tissue culture basket 403 rotates to the lowest position and then repeats the above basket loading operation until all tissue culture baskets 403 are loaded with culture bottles 7 and then opened for tissue culture, to avoid the tissue culture bottles 7 being squeezed together to affect the light, and to avoid developmental abnormalities caused by insufficient light. The induction culture time is 10-15 days, and the culture temperature is 25-28°C. S6, replacing tissue culture medium: after the induction culture is completed, the culture medium is replaced to enter the proliferation culture stage, the carrying conveyor belt 405 is started, the culture bottle 7 is conveyed along the fifth conveyor belt 10 through the first conveyor belt 5 and the second conveyor belt 6 to re-enter the second bottle inlet star wheel 202 of the culture medium filling device 2, and during the process of the culture bottle 7 being driven to rotate, the third guide wheel 2033 rotates and enters the third guide rail section 2035, the push rod 2032 rises and enters the drainage hole 701 to lift up the first one-way plugging piece 702, and the original induction medium in the culture bottle 7 is discharged from the drainage hole 701. After the drainage, the third guide wheel 2033 drives out of the third guide rail section 2035, the first one-way plugging piece 702 covers the drainage hole 701 again to seal it, the second guide wheel 2059 rotates and drives into the second guide rail section 2060, and the valve needle 2 053 is pressed down, the injection hole 1301 enters the valve cavity 2062, the bottom end of the valve needle 2053 pushes open the second one-way plugging piece 1302, and the proliferation culture medium is poured in. Then the culture bottle 7 returns to the tissue culture basket 403 for proliferation culture. The proliferation culture time is 8-12 days, and the proliferation culture temperature is 25-30°C; after the proliferation culture is completed, the above method is repeated to replace the rooting culture medium for rooting culture. The rooting culture time is 10-15 days, and the culture temperature is 25-28°C. The entire cultivation process can not only automatically fill the culture medium but also automatically replace the culture medium. The culture bottle 7 can automatically enter the tissue culture rack 4 for discharge, avoiding the probability of contamination caused by manual contact, improving the operation efficiency and survival rate, and improving the operation efficiency by more than 10 times compared with the traditional tissue culture method.

[0044] The above content is a further detailed description of the present invention in combination with specific / preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For ordinary technicians in the technical field to which the present invention belongs, without departing from the concept of the present invention, they can also make several substitutions or modifications to these described embodiments, and these substitutions or modifications should be regarded as belonging to the protection scope of the present invention. In the description of this specification, the description of reference terms "an embodiment", "some embodiments", "preferred embodiments", "examples", "specific examples" or "some examples" means that the specific features, structures, materials or characteristics described in combination with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily target 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 the absence of mutual contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples. Although the embodiments of the present invention and their advantages have been described in detail, it should be understood that various changes, substitutions and modifications can be made herein without departing from the scope of protection of the patent application.

Claims

1. A tissue culture device for sugarcane tissue culture seedlings, characterized in that: include An explant filling device, wherein the inlet and outlet ends of the explant filling device are respectively connected to a first conveyor belt and a second conveyor belt, the culture bottle is conveyed into the explant filling device by the first conveyor belt, and the explant is put into the culture bottle and then conveyed to the next process via the second conveyor belt; A culture medium filling device, wherein the inlet end of the culture medium filling device is connected to the end of the second conveyor belt, the outlet end of the culture medium filling device is connected to the third conveyor belt, the culture medium filling device is connected to at least three culture solution temporary storage tanks, the culture bottles conveyed from the second conveyor belt enter the culture medium filling device, the prepared culture medium stored in the culture solution temporary storage tank is quantitatively filled into the culture bottle, and then conveyed to the next process via the third conveyor belt; A capping device, wherein the inlet end of the capping device is connected to the end of the third conveyor belt, the outlet end of the capping device is connected to the fourth conveyor belt, the capping device comprises a cap sorting assembly and a capping assembly, a lower cap guide rail is connected between the cap sorting assembly and the capping assembly, the culture bottles conveyed from the third conveyor belt and the bottle caps sliding down from the lower cap guide rail enter the capping assembly for capping and tightening, and then are conveyed to the next process via the fourth conveyor belt; A tissue culture rack, wherein the inlet end of the tissue culture rack is connected to the end of the fourth conveyor belt, the outlet end of the tissue culture rack is connected to the fifth conveyor belt, the fifth conveyor belt is connected to the first conveyor belt, and the culture bottles enter the tissue culture rack for cultivation via the fourth conveyor belt after the caps are screwed on.

2. The tissue culture device for sugarcane tissue culture seedlings according to claim 1, characterized in that: The explant filling equipment includes a first bottle-input star wheel, a dispenser, an explant storage bucket and a first filling body, wherein the first bottle-input star wheel is rotatably arranged on the first filling body, the explant storage bucket is fixedly arranged on the first filling body, a plurality of dispensing tubes are connected to the bottom of the explant storage bucket, the dispenser is connected to one end of the dispensing tubes and corresponds to the transmission track of the first bottle-input star wheel.

3. The tissue culture device for sugarcane tissue culture seedlings according to claim 2, characterized in that: The dispenser comprises a hollow shell, a dispensing wheel is rotatably arranged inside the shell, a plurality of grooves are arranged on the outer peripheral surface of the dispensing wheel, and a dispensing motor is connected to the wheel axle of the dispensing wheel.

4. The tissue culture device for sugarcane tissue culture seedlings according to any one of claims 1 to 3, characterized in that: The culture medium filling equipment includes a second filling body, a second bottle-inlet star wheel, a bottle-supporting plate, a central rotating shaft and a plurality of filling valve assemblies. The central rotating shaft is rotatably arranged vertically on the second filling body. The second bottle-inlet star wheel, the bottle-supporting plate and the filling valve assembly are all fixed on the central rotating shaft. A star wheel guard plate is fixed to the periphery of the second bottle-inlet star wheel. The bottle-supporting plate is arranged below the second bottle-inlet star wheel. A plurality of the filling valve assemblies are arranged above the second bottle-inlet star wheel and correspond to the transmission track of the second bottle-inlet star wheel. The central rotating shaft is a hollow structure. A rotating joint is arranged at the top of the central rotating shaft. The culture medium temporary storage tank is connected to the rotating joint through a pipeline. A central distribution ring is connected to the central rotating shaft. A plurality of the filling valve assemblies are all connected to the central distribution ring.

5. The tissue culture device for sugarcane tissue culture seedlings according to claim 4, characterized in that: The cam is an airtight container, and the cam is secured to the interior of the valve body with a spring, and the cam is secured to the interior of the valve body with a spring, and the cam is secured to the interior of the valve body with a spring.

6. The tissue culture device for sugarcane tissue culture seedlings according to claim 5, characterized in that: The bottle supporting plate is provided with a plurality of top holes, a top rod is provided in the top hole, a third guide wheel is provided at the bottom of the top rod, a support ring is provided on the support rod, a third guide rail section corresponding to the third guide wheel is provided on the support ring, a drainage hole is provided on the bottom surface of the culture bottle, a first one-way plugging piece is provided at the drainage hole, one end of the first one-way plugging piece is hinged to the inner bottom surface of the culture bottle, the top end of the top rod corresponds to the drainage hole, an injection hole is provided on the bottle cap, a second one-way plugging piece is provided at the injection hole, one end of the second one-way plugging piece is hinged to the inner bottom surface of the bottle cap, and a partition net is provided in the culture bottle.

7. The tissue culture device for sugarcane tissue culture seedlings according to claim 6, characterized in that: The tissue culture rack includes a frame body, on which runway disks are symmetrically arranged, on which a plurality of tissue culture baskets are arranged, and the plurality of tissue culture baskets can circulate and rotate around the runway disks on both sides, and the tissue culture basket includes a connecting frame, a carrying conveyor belt and a bottle cloth assembly, the carrying conveyor belt is laid on the connecting frame, the bottle cloth assembly is arranged on the carrying conveyor belt, a transverse guide rod is provided on the connecting frame, and both ends of the bottle cloth assembly are slidably connected to the transverse guide rod, and a first screw rod is provided on the bottle cloth assembly, and one end of the first screw rod is connected to a first driving motor.

8. The tissue culture device for sugarcane tissue culture seedlings according to claim 7, characterized in that: The bottle distribution assembly includes a moving frame, a bottle separation screw and a screw guard plate. The bottle separation screw is rotatably arranged on the moving frame, one end of the bottle separation screw is connected to a second driving motor, the screw guard plate is slidably arranged on the moving frame, a second screw is arranged on the screw guard plate, and a third driving motor is arranged at one end of the second screw.

9. The tissue culture device for sugarcane tissue culture seedlings according to claim 8, characterized in that: The fifth conveyor belt is connected with a buffer bed.

10. A method for cultivating sugarcane tissue culture seedlings, characterized in that: Cultivating sugarcane tissue culture seedlings using the tissue culture device according to any one of claims 1 to 9 comprises the following steps: S1. Preparation: Select sugarcane tender stems as explants, and place the explants into the explant storage bucket of the explant filling equipment after disinfection. At the same time, prepare the induction medium, proliferation medium and rooting medium, and store them in the culture solution temporary storage tank respectively. Disinfect the culture bottle and the bottle cap for standby use. S2, explant filling: start the explant filling equipment, put the sterilized culture bottle into the first bottle feeding star wheel of the explant filling equipment through the first conveyor belt, and the explant enters the groove of the delivery wheel from the explant storage bucket along the delivery tube. When the culture bottle rotates and enters directly under the delivery device, the delivery motor drives the delivery wheel to rotate, and the explant in the groove falls into the culture bottle; S3, culture medium filling: start the culture medium filling equipment, the culture bottle enters the second bottle inlet star wheel of the culture medium filling equipment from the second conveyor belt, the culture bottle is driven to rotate, the first guide wheel rotates into the first guide rail section, the valve body of the filling valve assembly is pressed down, the positioning block presses the bottle mouth of the culture bottle, so that the culture bottle is clamped between the bottle support plate and the positioning block, the culture bottle continues to rotate with the second bottle inlet star wheel, when the second guide wheel rotates into the second guide rail section, the valve needle is pressed down, and the liquid inlet hole on the valve needle is connected with the valve cavity after the valve needle is pressed down, the culture medium in the corresponding culture liquid temporary storage tank enters the valve needle and is injected into the culture bottle, after the second guide wheel drives out of the second guide rail section, the valve needle moves up driven by the second spring, the liquid inlet hole and the valve cavity are misaligned to stop filling, after the first guide wheel drives out of the first guide rail section, the valve body moves up to release the culture bottle; S4, culture bottle capping: start the capping equipment, the culture bottles enter the capping assembly from the third conveyor belt, and the bottle caps stored in the cap sorting assembly slide along the lower cover guide rail to the bottle mouth of the culture bottle and enter the capping assembly together, and are tightened in the capping assembly; S5, shelf cultivation: start the tissue culture rack, the culture bottle is conveyed into the tissue culture basket of the tissue culture rack through the fourth conveyor belt, the culture bottle enters the carrying conveyor belt of the tissue culture basket and enters the bottle cloth assembly, and after being pulled apart by the bottle separation screw by a certain distance, the first drive motor drives the first screw rod to rotate, drives the bottle cloth assembly to drive the culture bottle to move horizontally, and after moving into place, the third drive motor drives the second screw rod to rotate, drives the screw guard plate to slide upward, and the bottle cloth assembly retreats and returns to the starting point, repeats until the tissue culture basket is full, and the tissue culture basket rotates along the runway disk to load an empty tissue culture basket, and all tissue culture baskets are loaded with tissue culture bottles and opened for tissue culture; S6. Replacement of tissue culture medium: When replacing the culture medium during the tissue culture process, start the carrying conveyor belt, and the culture bottle is conveyed along the fifth conveyor belt, through the first conveyor belt and the second conveyor belt, and re-enters the second bottle inlet star wheel of the culture medium filling equipment. During the process of the culture bottle being driven to rotate, the third guide wheel rotates and enters the third guide rail section, and the push rod rises and enters the drainage hole to lift up the first one-way plugging piece, and the original culture medium in the culture bottle is discharged from the drainage hole. After the drainage, the third guide wheel drives out of the third guide rail section, and the first one-way plugging piece covers the drainage hole again to seal it. The second guide wheel rotates and enters the second guide rail section, and the valve needle is pressed down and enters the injection hole to push open the second one-way plugging piece, and the required culture medium is poured in.

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