Efficient breeding method for healthy sugarcane seedlings

Abstract

The invention relates to the technical field of improved variety breeding of crops, in particular to an efficient breeding method for healthy sugarcane seedlings, which comprises the following steps: step 1, taking materials and inducing; step 2, callus differentiation; step 3, multiplication culture; step 4, root promoting culture; 5, heeling in and transplanting: transplanting rooted tissue culture seedlings outdoors, carrying out single-plant field planting to obtain healthy sugarcane original seedlings, and transplanting the healthy sugarcane original seedlings into a field; step 6, tillering pruning and cuttage: after the sugarcane original seedlings are tillered, cutting off main stems, and performing cuttage breeding through a cuttage device after soaking treatment. According to the method, a large number of healthy sugarcane seedlings are obtained through the tissue culture technology, the seedlings have the advantages of being high in growth speed, good in quality and the like, meanwhile, an oblique cutting mode in the cutting technology is combined, tillering stems are reused, the breeding efficiency is improved, and then efficient breeding of the healthy sugarcane seedlings is achieved.

Description

Technical Field

[0001] The invention relates to the technical field of crop improved variety breeding, and in particular to a method for efficiently breeding healthy sugarcane seedlings. Background Art

[0002] As a temperate and tropical crop, sugarcane is not only the main raw material for making sucrose, but also can be used to extract ethanol as an energy substitute. It is rich in sugar, water, and a variety of vitamins, fats, proteins, organic acids, calcium, iron and other substances that are beneficial to human metabolism. In recent years, with the increasing demand for healthy diets and renewable energy, the sugarcane planting and processing industry has shown a booming trend. Especially in China, sugarcane is widely planted in Guangxi, Yunnan, Guangdong, Hainan and other places, and is gradually expanding to wider areas. However, sugarcane planting also faces many challenges, such as frequent pests and diseases, variety degeneration, and rising planting costs, which urgently need to be solved through efficient breeding technology.

[0003] Although the existing technology for breeding healthy sugarcane seedlings has improved the quality and yield of seedlings to a certain extent, there are still some obvious defects. In particular, sugarcane seedlings detoxified by tissue culture will have multiple tillers during the growth process, and these tillers will squeeze the main stem and affect the growth and development of the main stem. The traditional treatment method is often to directly discard the stem segments of these tillers, which not only wastes resources but also increases production costs. In order to solve this problem, the prior art attempts to re-cut the stem segments of the tillers to achieve reuse and thus achieve the purpose of rapid breeding. However, this method has also encountered many difficulties in the implementation process. First, due to the relatively low growth points and nutrient content of the tillering stem segments, direct cuttings are often difficult to ensure their survival rate. Secondly, during the cutting process, since the main stem is young and fragile, if it is all buried in the soil, it is easy to be invaded by microorganisms and pathogens in the soil, resulting in the occurrence of rot. In order to solve the problems of secondary utilization of tillering main stems and the influence of cutting methods on the main stems, a method for efficient breeding of healthy sugarcane seedlings is proposed. Summary of the invention

[0004] In order to solve the above problems, the present invention provides a method for efficiently breeding healthy sugarcane seedlings. A large number of healthy sugarcane seedlings are obtained through tissue culture technology. These seedlings have the characteristics of fast growth rate and excellent quality. At the same time, combined with the oblique insertion method in the cutting technology, not only the tillering stem segments are reused, but also the breeding efficiency is improved, thereby achieving efficient breeding of healthy sugarcane seedlings.

[0005] In order to achieve the above object, the technical solution of the present invention is as follows: A method for efficiently breeding healthy sugarcane seedlings comprises the following steps:

[0006] Step 1, material collection and induction: select disease-free and insect-pest-free, healthy sugarcane stem tips as explants, peel off the outer leaf sheaths of the sugarcane stem tips, disinfect them and move them under sterile conditions, peel off the tissues outside the stem tip growth point with an inoculation needle and a scalpel under sterile conditions until the growth point is exposed, cut off 1-2 mm of stem tip tissue, soak it in sterile water and inoculate it on MS stem tip tissue culture medium, and after several weeks of culture, the stem tip tissue sprouts several terminal buds to form clustered seedlings;

[0007] Step 2, callus differentiation: continue to peel off the outer leaf sheath under sterile conditions, leaving the heart leaf with a diameter of 0.5 cm, cut it into 1-2 mm thin slices, inoculate it on the MS heart leaf tissue culture medium, add 2,4-D to the inoculated MS heart leaf tissue culture medium, culture it for several days to form white callus, transfer the callus to MS differentiation medium, select cytokinin and auxin to culture the callus, and observe the differentiation of green seedlings after 10 days of culture;

[0008] Step 3, proliferation culture: the clustered seedlings induced in step 1 are transferred to MS proliferation medium, gibberellin, 6-benzyladenine and naphthaleneacetic acid are added, 7 days is taken as one cycle, each time the clustered seedlings are cut into several clusters, transferred to MS proliferation medium, and proliferated for 1 to 2 generations, and as the number of proliferation generations increases, the concentrations of gibberellin, 6-benzyladenine and naphthaleneacetic acid added are gradually reduced; the green seedlings differentiated from the callus in step 2 are transferred to MS strong seedling medium, 6-benzyladenine and naphthaleneacetic acid are added, and proliferated for 1 to 2 generations, and the concentrations of 6-benzyladenine and naphthaleneacetic acid added are gradually reduced;

[0009] Step 4: Root promotion culture: transfer the clustered bud seedlings and green seedlings differentiated from callus tissue after proliferation culture to MS rooting medium. After 7 days of culture, wait for white roots to differentiate from the basal part. Count the rooting rate of the medium inoculation bottle, and transplant the tissue culture seedlings with a rooting rate of more than 60% and strong growth;

[0010] Step 5, transplanting and transplanting: transplant the rooted tissue culture seedlings outdoors, open the cap of the inoculation bottle, harden the seedlings for 2-3 days, then take out the plants with intact root systems from the inoculation bottle, wash away the culture medium attached to the root system, keep the clumps of 5-6 green seedlings / clump during the root promotion culture in step 4, cut off about 1 / 3 of the leaves on the upper part of the pseudostem, disinfect the plants and transplant them in the seedbed in the greenhouse, irrigate with MS rooting culture medium mother solution every 5-7 days, and plant them in the greenhouse when the seedlings grow to more than 20 cm. After 40 days, obtain healthy sugarcane seedlings and transplant them to the field;

[0011] Step 6, tillering pruning and cutting: After transplanting the sugarcane seedlings, wait for tillering to occur, cut off the main stem, cut off the leaves of the main stem, leaving 2 to 3 cm leaves close to the ear, remove the dry leaves, and then soak the main stem in the seed soaking solution for 1 to 2 hours and then send it into the cutting device. The main stem is cut into another large field through the cutting device. During cutting, the angle between the main stem and the row furrow is 20°, and the oblique insertion depth is 7 to 10 cm, and the length left on the ground is 8 to 15 cm.

[0012] Furthermore, in step 1, the disinfection treatment is to rinse the sugarcane stems with tap water, first disinfect the surface with 75% alcohol, then disinfect with 2% NaClO solution for 10 to 15 minutes, and then rinse with sterile water for 4 to 5 times.

[0013] Furthermore, in step 1, activated carbon and polyvinylpyrrolidone need to be added to the MS induction medium during the cultivation of the shoot apex tissue, and the shoot apex tissue explants are continuously transferred to fresh MS induction medium according to the browning of the shoot apex tissue explants.

[0014] Furthermore, in step 2, the concentration of 2,4D is 3.0 mg / L; the cytokinin for callus culture is 1.0 mg / L 6-benzyladenine, the auxin is 0.01 mg / L naphthaleneacetic acid, and 30 g / L sucrose and 0.6% agar powder are added.

[0015] Further, in step three, 0.5 g / L gibberellin, 2.0 mg / L 6-benzyladenine and 0.1 mg / L naphthaleneacetic acid were added to the MS proliferation medium, and 30 g / L sucrose was added; 1.0 mg / L 6-benzyladenine and 0.05 mg / L naphthaleneacetic acid were added to the MS seedling medium, and 30 g / L sucrose was added.

[0016] Further, in step 4, the concentration of MS rooting medium is 1 / 2MS medium and 15.0 mg / L naphthylacetic acid.

[0017] Further, in step 4, the rooting rate calculation formula is:

[0018]

[0019] Furthermore, in steps 2 to 4, the callus induction stage is dark culture, and the differentiation, proliferation and rooting stages are all light culture, with a temperature of 28±2° C. and a light intensity of 2000 lx.

[0020] Furthermore, in step five, the disinfection treatment uses 1 / 1000 potassium permanganate solution for 5 to 6 minutes, and the planting medium of the transplanted seedbed is all disinfected sandy soil.

[0021] Furthermore, in step six, the seed soaking solution is a mixture of 50-200 mg / L chlorantraniliprole, 50-200 mg / L clofoscar and 200-500 mg / L prochloraz.

[0022] The above scheme has the following beneficial effects:

[0023] 1. This scheme, through tissue culture technology, especially the combination of stem tip culture and heart leaf tissue culture, can quickly induce a large number of healthy sugarcane seedlings. In this process, the use of sterile operations and fine culture conditions effectively shortens the breeding cycle of seedlings, thereby greatly improving the breeding efficiency. At the same time, combined with the oblique insertion method in cutting technology, the utilization rate and breeding speed of seedlings are further improved; through the combination of tillering pruning and oblique insertion technology, not only the tillering stem segments are reused, but also the waste of seedlings is reduced. At the same time, the sterile operation and fine management used in the entire breeding process reduce the probability of disease and insect pests, thereby reducing the use of pesticides and fertilizers. These measures jointly reduce production costs and improve economic benefits

[0024] 2. This scheme selects pest-free and healthy sugarcane stems as explants to ensure the genetic quality and health of the seedlings. Through a sophisticated tissue culture process, such as adding activated carbon and polyvinylpyrrolidone to prevent browning, and optimizing the composition of the culture medium and light conditions, the growth potential and stress resistance of the seedlings are further enhanced. The sugarcane seedlings bred in this way have the characteristics of fast growth and excellent quality, providing a solid foundation for subsequent sugarcane planting.

[0025] 3. This scheme, using the above-mentioned efficient breeding method, can quickly breed a large number of healthy sugarcane seedlings to meet the demand for seedlings in the sugarcane planting industry. At the same time, by optimizing the breeding process and management measures, the yield and quality of sugarcane can be improved, promoting the sustainable development of the sugarcane planting industry.

[0026] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a schematic diagram of the method steps of an embodiment of the method for efficiently breeding healthy sugarcane seedlings of the present invention;

[0028] Figure 2 It is a side cross-sectional schematic diagram of a cutting device of an embodiment of a method for efficiently breeding healthy sugarcane seedlings of the present invention;

[0029] Figure 3 This is an axonometric schematic diagram of a cutting assembly according to an embodiment of the method for efficiently breeding healthy sugarcane seedlings of the present invention.

[0030] The figure marks in the drawings of the specification include: 1. car body; 2. motor; 3. connecting rod; 4. wheel; 5. driving sprocket; 6. belt; 7. connecting plate; 8. digging plate; 9. implantation arm; 10. slide groove; 11. rack; 12. fixed plate; 13. connecting shaft; 14. gear; 15. slider; 16. electric clamp; 17. sugarcane box; 18. box cover; 19. connecting shaft. DETAILED DESCRIPTION

[0031] The technical solution of the present invention will be described clearly and completely below 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 creative work are within the scope of protection of the present invention.

[0032] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying 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 limiting the present invention. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0033] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0034] The following is further described in detail through specific implementation methods:

[0035] Embodiment 1:

[0036] As attached Figure 1 As shown: A method for efficiently breeding healthy sugarcane seedlings, comprising the following steps:

[0037] Step 1, material collection and induction: select the sugarcane stem tip that is free of pests and diseases and grows robustly as the explant, peel off the outer leaf sheath from the sugarcane stem tip, rinse the sugarcane stem with tap water, first disinfect the surface with 75% alcohol, then disinfect with 2% NaClO solution for 10-15 minutes, then rinse with sterile water for 4-5 times for disinfection and move to sterile conditions, under sterile conditions, use an inoculation needle and a scalpel to peel off the tissue outside the stem tip growth point until the growth point is exposed, cut off 1-2 mm of stem tip tissue, soak it in sterile water and then inoculate it on MS stem tip tissue culture medium, add activated carbon and polyvinylpyrrolidone to the MS induction culture medium during the cultivation of the stem tip tissue, continuously transfer the stem tip tissue explant to fresh MS induction culture medium according to the browning of the stem tip tissue explant, and after several weeks of cultivation, the stem tip tissue sprouts several terminal buds to form clustered seedlings.

[0038] Step 2, callus differentiation: continue to peel off the outer leaf sheath under sterile conditions, leaving the heart leaf with a diameter of 0.5 cm, cut into 1-2 mm thin slices, and inoculate them into MS heart leaf tissue culture medium. Add 2,4-D with a concentration of 3.0 mg / L to the inoculated MS heart leaf tissue culture medium, and culture it in the dark for several days to form white callus. Transfer the callus to MS differentiation medium, select 1.0 mg / L 6-benzyladenine as cytokinin and 0.01 mg / L naphthaleneacetic acid as auxin, and add 30 g / L sucrose and 0.6% agar powder. Carry out light culture on the callus at a temperature of 28±2°C and a light intensity of 2000lx. Observe the differentiation of green seedlings after 10 days of culture.

[0039] Step 3, proliferation culture: the clustered seedlings induced in step 1 are transferred to MS proliferation medium, 0.5g / L gibberellin, 2.0mg / L 6-benzyladenine and 0.1mg / L naphthaleneacetic acid are added, and 30g / L sucrose is added. With 7 days as a cycle, the clustered seedlings are cut into several clusters each time, and transferred to MS proliferation medium for proliferation for 1 to 2 generations. As the proliferation generation increases, the concentrations of gibberellin, 6-benzyladenine and naphthaleneacetic acid added are gradually reduced; the green seedlings differentiated from the callus in step 2 are transferred to MS seedling medium, 1.0mg / L 6-benzyladenine and 0.05mg / L naphthaleneacetic acid are added, and 30g / L sucrose is added. They are proliferated for 1 to 2 generations, and the concentrations of 6-benzyladenine and naphthaleneacetic acid added are gradually reduced. The proliferation culture is light culture, the temperature is 28±2°C, and the light intensity is 2000lx.

[0040] Step 4, root promotion culture: transfer the clustered bud seedlings and green seedlings differentiated from callus tissue after proliferation culture to MS rooting medium with a concentration of 1 / 2MS medium and 15.0mg / L naphthaleneacetic acid for light culture, the temperature is 28±2℃, the light intensity is 2000lx, and after 7 days of culture, white roots are differentiated from the basal part. Count the rooting rate of the medium inoculation bottle, and transplant the tissue culture seedlings with a rooting rate of more than 60% and strong growth. The rooting rate calculation formula is:

[0041]

[0042] Step 5, transplanting and transplanting: transplant the rooted tissue culture seedlings outdoors, open the cap of the inoculation bottle, harden the seedlings for 2 to 3 days, then take out the plants with complete root systems from the inoculation bottle, wash away the culture medium attached to the roots, retain the clumps of 5 to 6 green seedlings / clump during the root promotion culture in step 4, cut off about 1 / 3 of the leaves on the upper part of the pseudostem, disinfect the plants with 1 / 1000 potassium permanganate solution for 5 to 6 minutes, and then transplant them in the seedbed in the greenhouse. Water them with MS rooting culture medium mother solution every 5 to 7 days. When the seedlings grow to more than 20 cm, they are transplanted in the greenhouse. After 40 days, healthy sugarcane seedlings are obtained and transplanted to the field. The planting medium of the transplanted seedbed is all disinfected sandy soil.

[0043] Step six, tillering pruning and cutting: after transplanting the sugarcane seedlings, wait for tillering to occur in the sugarcane seedlings, cut off the main stem, cut off the leaves of the main stem, leaving 2 to 3 cm leaves close to the ear, remove the dry leaves, and then soak the main stem in a seed soaking solution mixed with 50 to 200 mg / L chlorantraniliprole, 50 to 200 mg / L chlorantraniliprole and 200 to 500 mg / L prochloraz for 1 to 2 hours, then send it into the cutting device, and use the cutting device to cut the main stem into another large field. During cutting, the angle between the main stem and the row furrow is 20°, the oblique insertion depth is 7 to 10 cm, and the length left on the ground is 8 to 15 cm.

[0044] The specific implementation process is as follows:

[0045] Table 1 Survival rate and tillering number of main stem full-buried planting and cutting planting

[0046]

[0047]

[0048] The results of full burying and cutting planting of sugarcane main stems show that the survival rate of cuttings is significantly higher than that of full burying plants, and the number of effective tillers of oblique planting is also higher than that of full burying plants.

[0049] Embodiment 2:

[0050] As attached Figure 2 and Figure 3 As shown, the difference from Example 1 is that the cutting device includes a cutting machine, the cutting machine signal is connected to a controller, the cutting machine includes a car body 1 and a plurality of wheels 4 rotatably connected to both sides of the car body 1, and the bottom of the car body 1 is bottomless.

[0051] A motor 2 is fixedly connected to the inner wall of the vehicle box 1, and the motor 2 is connected to the controller signal. A cutting assembly for cutting the main stem of sugarcane is coaxially fixedly connected to the output shaft of the motor 2. A digging assembly for trenching is also coaxially fixedly connected to the output shaft of the motor 2. A sugarcane box 17 is fixedly connected to the side of the vehicle box 1 away from the motor 2. The side of the sugarcane box 17 close to the cutting assembly is without side panels, and a box cover 18 is hinged on the top of the sugarcane box 17.

[0052] The excavation assembly includes a driving sprocket 5 connected to the output shaft of the motor 2 by a coaxial key, a transmission shaft is provided at the bottom of the car body 1, symmetrically arranged connecting plates 7 are welded on the side walls of the bottom of the car body 1, the transmission shaft is rotatably connected between the symmetrically arranged connecting plates 7, a driven sprocket is welded on the transmission shaft, a belt 6 is provided for transmission between the driving sprocket 5 and the driven sprocket, and a number of digging plates 8 are also welded on the transmission shaft.

[0053] The implant assembly includes a connecting shaft 19 coaxially welded to the output shaft of the motor 2, a connecting rod 3 is welded to the end of the connecting shaft 19 away from the motor 2, an implant arm 9 is hingedly connected to the end of the connecting rod 3 away from the motor 2, a rack 11 is welded to the bottom of the implant arm 9, a connecting shaft 13 is arranged in the carriage 1, and both ends of the connecting shaft 13 are rotatably connected to symmetrically arranged fixed plates 12, and the sides of the fixed plates 12 away from the connecting shaft 13 are welded to the inner wall of the carriage 1, and the connecting shaft 13 away from the fixed plate 12 is welded to the inner wall of the carriage 1. A gear 14 is coaxially welded to the end, and the rack 11 is meshed with the gear 14. A slide groove 10 is opened on the side wall of the cutting arm 9. A rotating rod is coaxially sleeved on the connecting shaft 13. A slider 15 is welded on the rotating rod. The slider 15 is sleeved on the top of the cutting arm 9 and the slider 15 and the slide groove 10 are slidably matched. An electric clamp 16 is screwed to the end of the cutting arm 9 away from the connecting rod 3. The electric clamp 16 is connected to the controller signal, and the sugarcane box 17 is located on the movement trajectory of the electric clamp 16.

[0054] The specific implementation process is as follows: the grafting machine is placed in the field, the soaked sugarcane main stems are loaded into the sugarcane box 17, and the tender branches of the sugarcane main stems face the side close to the grafting arm 9. The grafting machine is started by the controller. During the forward movement of the grafting machine, the controller starts the motor 2, so that the driving sprocket 5 rotates with the output shaft of the motor 2, and the driven sprocket rotates under the transmission of the belt 6, and drives the transmission shaft to rotate, thereby driving the digging plate 8 to start rotating to dig the sugarcane ditch.

[0055] At the same time, driven by the motor 2, the connecting rod 3 rotates with the rotation of the output shaft of the motor 2, thereby driving the grafting arm 9 to perform reciprocating arc motion. At this time, the rack 11 on the grafting arm 9 is meshed with the gear 14, and because the slider 15 on the rotating rod limits the grafting arm 9, the grafting arm 9 can only perform reciprocating arc motion within the limiting range of the slider 15. When the grafting arm 9 slides obliquely upward, the electric clamp 16 extends into the sugarcane box 17, and at the same time, the controller controls the electric clamp 16 to clamp the main stem of the sugarcane. When the grafting arm 9 slides obliquely downward, the electric clamp 16 approaches the sugarcane ditch, and due to the pushing action of the grafting arm 9, the main stem of the sugarcane is obliquely inserted into the ground. Due to the meshing of the rack 11 of the grafting arm 9 and the gear 14 and the limiting of the slider 15, when the main stem of the sugarcane is grafted After completion, the electric clamp 16 releases the main stem of the sugarcane and moves in an arc again until it clamps the main stem of the sugarcane to be inserted in the sugarcane box 17. The cutting machine can continuously and stably perform cutting operations, greatly improving work efficiency. Through the automated cutting machine, the time and labor required for manual cutting can be significantly reduced; and because the angle between the main stem and the row furrow is 20° during cutting, and the oblique insertion depth is 7 to 10 cm, there are often subjective errors in manual cutting. Through the limiting of the cutting arm 9 by the slider 15, the depth and angle of the oblique insertion of the sugarcane main stem into the ground are fixed each time, which improves the accuracy and consistency of cutting. Through precise control of the cutting depth and position, it can ensure that the sugarcane main stem can grow better in the soil, which helps to improve the sugarcane breeding efficiency.

[0056] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A method for efficiently breeding healthy sugarcane seedlings, characterized in that: The following steps are involved: Step 1, material collection and induction: select disease-free and insect-pest-free, healthy sugarcane stem tips as explants, peel off the outer leaf sheaths of the sugarcane stem tips, disinfect them and move them under sterile conditions, peel off the tissues outside the stem tip growth point with an inoculation needle and a scalpel under sterile conditions until the growth point is exposed, cut off 1-2 mm of stem tip tissue, soak it in sterile water and inoculate it on MS stem tip tissue culture medium, and after several weeks of culture, the stem tip tissue sprouts several terminal buds to form clustered seedlings; Step 2, callus differentiation: continue to peel off the outer leaf sheath under sterile conditions, leaving the heart leaf with a diameter of 0.5 cm, cut it into 1-2 mm thin slices, inoculate it on the MS heart leaf tissue culture medium, add 2,4-D to the inoculated MS heart leaf tissue culture medium, culture it for several days to form white callus, transfer the callus to MS differentiation medium, select cytokinin and auxin to culture the callus, and observe the differentiation of green seedlings after 10 days of culture; Step 3, proliferation culture: the clustered seedlings induced in step 1 are transferred to MS proliferation medium, gibberellin, 6-benzyladenine and naphthaleneacetic acid are added, 7 days is taken as one cycle, each time the clustered seedlings are cut into several clusters, transferred to MS proliferation medium, and proliferated for 1 to 2 generations, and as the number of proliferation generations increases, the concentrations of gibberellin, 6-benzyladenine and naphthaleneacetic acid added are gradually reduced; the green seedlings differentiated from the callus in step 2 are transferred to MS strong seedling medium, 6-benzyladenine and naphthaleneacetic acid are added, and proliferated for 1 to 2 generations, and the concentrations of 6-benzyladenine and naphthaleneacetic acid added are gradually reduced; Step 4: Root promotion culture: transfer the clustered bud seedlings and green seedlings differentiated from callus tissue after proliferation culture to MS rooting medium. After 7 days of culture, wait for white roots to differentiate from the basal part. Count the rooting rate of the medium inoculation bottle, and transplant the tissue culture seedlings with a rooting rate of more than 60% and strong growth; Step 5, transplanting and transplanting: transplant the rooted tissue culture seedlings outdoors, open the cap of the inoculation bottle, harden the seedlings for 2-3 days, then take out the plants with intact root systems from the inoculation bottle, wash away the culture medium attached to the root system, keep the clumps of 5-6 green seedlings / clump during the root promotion culture in step 4, cut off about 1 / 3 of the leaves on the upper part of the pseudostem, disinfect the plants and transplant them in the seedbed in the greenhouse, irrigate with MS rooting culture medium mother solution every 5-7 days, and plant them in the greenhouse when the seedlings grow to more than 20 cm. After 40 days, obtain healthy sugarcane seedlings and transplant them to the field; Step 6, tillering pruning and cutting: After transplanting the sugarcane seedlings, wait for tillering to occur, cut off the main stem, cut off the leaves of the main stem, leaving 2 to 3 cm leaves close to the ear, remove the dry leaves, and then soak the main stem in the seed soaking solution for 1 to 2 hours and then send it into the cutting device. The main stem is cut into another large field through the cutting device. During cutting, the angle between the main stem and the row furrow is 20°, and the oblique insertion depth is 7 to 10 cm, and the length left on the ground is 8 to 15 cm.

2. The method for efficiently breeding healthy sugarcane seedlings according to claim 1, characterized in that: In step 1, the disinfection treatment is to rinse the sugarcane stems with tap water, first disinfect the surface with 75% alcohol, then disinfect with 2% NaClO solution for 10 to 15 minutes, and then rinse with sterile water for 4 to 5 times.

3. The method for efficiently breeding healthy sugarcane seedlings according to claim 2, characterized in that: In step 1, during the culture of the shoot apex tissue, activated carbon and polyvinylpyrrolidone need to be added to the MS induction medium, and the shoot apex tissue explants are continuously transferred to fresh MS induction medium according to the browning of the shoot apex tissue explants.

4. The method for efficiently breeding healthy sugarcane seedlings according to claim 3, characterized in that: In step 2, the concentration of 2,4D is 3.0 mg / L; the cytokinin for callus culture is 1.0 mg / L 6-benzyladenine, the auxin is 0.01 mg / L naphthaleneacetic acid, and 30 g / L sucrose and 0.6% agar powder are added.

5. The method for efficiently breeding healthy sugarcane seedlings according to claim 4, characterized in that: In step three, 0.5 g / L gibberellin, 2.0 mg / L 6-benzyladenine and 0.1 mg / L naphthaleneacetic acid were added to the MS proliferation medium, and 30 g / L sucrose was added; 1.0 mg / L 6-benzyladenine and 0.05 mg / L naphthaleneacetic acid were added to the MS seedling medium, and 30 g / L sucrose was added.

6. The method for efficiently breeding healthy sugarcane seedlings according to claim 5, characterized in that: In step 4, the concentration of MS rooting medium is 1 / 2MS medium and 15.0 mg / L naphthyl acetic acid.

7. The method for efficiently breeding healthy sugarcane seedlings according to claim 6, characterized in that: In step 4, the rooting rate calculation formula is:

8. The method for efficiently breeding healthy sugarcane seedlings according to claim 7, characterized in that: In step 2 to step 4, the callus induction stage is dark culture, and the differentiation, proliferation and rooting stages are all light culture, with a temperature of 28±2°C and a light intensity of 2000lx.

9. The method for efficiently breeding healthy sugarcane seedlings according to claim 8, characterized in that: In step five, the disinfection treatment is carried out using 1 / 1000 potassium permanganate solution for 5 to 6 minutes, and the planting medium of the transplanted seedbed is all disinfected sandy soil.

10. The method for efficiently breeding healthy sugarcane seedlings according to claim 9, characterized in that: In step six, the seed soaking solution is prepared by mixing 50-200 mg / L chlorantraniliprole, 50-200 mg / L clofoscar and 200-500 mg / L prochloraz.

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