Cultivation method of healthy sugarcane seedlings

Abstract

The invention discloses a method for cultivating healthy sugarcane seedlings. The method comprises the following steps: (1) detoxifying the seedlings by adopting a botanical disinfectant; (2) accelerating germination induction: adopting an antioxidant to prevent browning; (3) multiplication culture; (4) rooting culture; (5) seedling hardening; and (6) transplanting and temporary planting by using a special culture medium. According to the cultivation method, the high budding rate with high stability can be achieved, the period is shortened by 9-12 days compared with a conventional seedling cultivation period, sugarcane bud tip callus cultivation does not need to be cut, the production efficiency is improved, browning of sugarcane buds can be effectively inhibited, growth, proliferation and rooting of the seedlings are promoted, the transplanting survival rate is increased, healthy sugarcane seedlings are cultivated, and the method is suitable for large-scale popularization and application. And a high-quality seedling source is provided for the sugarcane industry.

Description

Technical Field

[0001] The invention relates to the technical field of sugarcane planting, and in particular to a method for cultivating healthy sugarcane seedlings. Background Art

[0002] Sugarcane (Saccharum officinarum), a perennial tall solid herb with a strong and well-developed rhizome and a stalk height of 3-6 meters. It is widely planted in southern tropical regions such as Taiwan, Fujian, Guangdong, Hainan, Guangxi, Sichuan, and Yunnan. Sugarcane is suitable for planting in places with fertile soil, sufficient sunshine, and a large temperature difference between winter and summer. Sugarcane is a temperate and tropical crop, a raw material for making sucrose, and can be refined into ethanol as an energy substitute. Sugarcane is a crop that uses axillary buds for asexual reproduction. Due to repeated planting for many years, it is very susceptible to repeated infection by seedling-borne pathogens, resulting in reduced yield and quality, thereby shortening the perennial root period and degeneration of the species. It is necessary to prevent seedlings from carrying bacteria (viruses) and spreading diseases and pests, such as sugarcane perennial root dwarfing disease and mosaic disease. Therefore, in order to obtain healthy sugarcane seedlings, it is necessary to detoxify the seedlings. The common seedling detoxification method is warm water detoxification treatment technology that generally does not exceed 58°C. The detoxification effect of sugarcane seedlings is closely related to the warm water treatment temperature and treatment time. Too high temperature and too long treatment time will destroy the sugarcane bud tissue, which is not conducive to the growth of the bud. Too low temperature and too short time will reduce the detoxification rate of the sugarcane buds. Therefore, the existing conventional warm water detoxification technology is difficult to achieve the goal of complete detoxification. Different varieties of sugarcane should be treated with water baths of different temperatures and durations. The parameters are difficult to define, resulting in a lack of stability in the detoxification effect and failure to ensure that the sugarcane buds grow into healthy seedlings. In addition, browning is a common problem in sugarcane tissue culture. Browning of sugarcane buds usually refers to the browning of the young buds of sugarcane. This may be due to the oxidation of phenolic substances in the cells into quinones under aerobic conditions, which in turn produce colored substances through non-enzymatic reactions, leading to tissue browning. Browning not only affects the ornamental value of sugarcane, but may also affect its quality and food safety.

[0003] Patent: A method for cultivating healthy sugarcane seedlings (application number 202211071949.1). The patent uses warm water detoxification technology. By setting a temperature higher than the conventional detoxification temperature and extending the constant temperature treatment time, the bud meristem tissue is directly taken for tissue culture without planting and germination, and sugarcane seedlings without streak mosaic virus are cultivated under the requirement of high germination rate. The patent only uses valerian as the test material, and the stability of the high germination rate cannot be seen, and the browning problem of sugarcane buds is not considered; Patent: A method for cultivating high-yield sugarcane using healthy sugarcane seeds (application number 201710024667.9), including detoxification treatment, seedling value-added, seedling coating, field disinfection, mechanical sowing and planting and harvesting, and discloses the preparation of sugarcane seed bud coating agent and green bactericidal disinfectant, but the cultivation cycle of the patent is relatively long, and the detoxification treatment temperature is too high to easily cause damage to the sugarcane seed buds. Summary of the invention

[0004] In view of the above shortcomings, the present invention provides a method for cultivating healthy sugarcane seedlings, which can achieve a high germination rate and a high seedling rate with high stability, shorten the cultivation period, effectively inhibit the browning of sugarcane buds, and ensure the high quality of sugarcane seedlings. The specific technical scheme is as follows:

[0005] A method for cultivating healthy sugarcane seedlings comprises the following steps:

[0006] (1) Seedling detoxification: Cut 2-3.5 cm stem tips or axillary buds from newly harvested sugarcane stems and place them in a plant-derived disinfectant for ultrasonic detoxification for 30-60 minutes;

[0007] (2) Germination induction: The stem tips or axillary buds after the virus-free treatment are rinsed with sterile water for 3-5 times and then placed in an induction medium for germination. After the sugarcane buds are induced, the outer old leaves are peeled off, and then the sugarcane buds are immersed in an antioxidant. After the immersion is completed, they are transferred to a proliferation medium for proliferation culture;

[0008] (3) Proliferation culture: The sugarcane buds are cultured for 12-14 days to obtain bud clusters;

[0009] (4) Rooting culture: Select individual shoots from the proliferation culture and transfer them to the rooting medium for rooting culture. Cultivate for 16-18 days to obtain rooted seedlings.

[0010] (5) Hardening: Transfer the rooted seedlings from the culture room to the hardening room for hardening;

[0011] (6) Transplanting and transplanting: transplant the seedlings after hardening treatment into a seedling tray containing a transplanting matrix, wherein the transplanting matrix is ​​flush with the upper surface of the seedling tray, and the healthy sugarcane seedlings obtained in step (5) are planted in the seedling tray filled with nutrient soil and watered thoroughly, and an arch shed is built and covered with a plastic film or a sunshade cloth. After the sugarcane seedlings grow out and have 3 leaves, the plastic film or the sunshade cloth is removed. When the seedlings grow to 4-5 true leaves, the healthy sugarcane seedlings are obtained.

[0012] In the seedling detoxification step, the plant-derived disinfectant is prepared by mixing matrine, azadirachtin, rotenone, water and a dispersant in a weight ratio of 10:10:0.5-5:5000:10-50; and the dispersant is an alkyl sulfonate.

[0013] In the germination induction step, the formula of the induction culture medium is: 1800 mg / L potassium nitrate, 1650 mg / L ammonium nitrate, 270 mg / L potassium dihydrogen phosphate, 370 mg / L magnesium sulfate, 9 mg / L zinc sulfate, 100 mg / L inositol, 0.25 mg / L sodium molybdate, 0.5 mg / L pyridoxine hydrochloride, 0.1 mg / L thiamine hydrochloride, 2 mg / L glycine, and 10 mg / L chitosan oligosaccharide.

[0014] In the germination induction step, the inoculated induction culture medium is placed in an incubator at a temperature of 30-33° C. and a relative humidity of 75-80% for cultivation, the light intensity is set to 1500-2000 lux, and the light time is 8-12 hours per day.

[0015] In the germination induction step, the sugarcane buds are immersed in the antioxidant for 10-60 minutes; the antioxidant comprises, by weight: 1-5 parts of vitamin C, 0.1-2 parts of beta-carotene, 0.1-2 parts of astaxanthin, 0.1-2 parts of silymarin, and 5-8 parts of ganoderma lucidum polysaccharide.

[0016] In the proliferation culture step, the proliferation culture medium is MS+6-BA (6-benzyladenine) 1.0 mg / L+GA (gibberellic acid) 1.0 mg / L+NAA (naphthaleneacetic acid) 2.0 mg / L+cysteine ​​5.0 mg / L; the rooting culture medium is MS+NAA (naphthaleneacetic acid) 5.0 mg / L+2,4-D (2,4-dichlorophenoxyacetic acid) 2.5 mg / +IBA (indolebutyric acid) 0.2 mg / L+GA (gibberellic acid) 0.5 mg / L+AC (abscisic acid) 0.1%+sucrose 30 g / L.

[0017] The seedling hardening step specifically includes: a. preliminary adjustment, on the first day of seedling hardening, the relative humidity of the seedling hardening room is kept at 75-78%, and the temperature is kept at 28-30°C; b. mid-term adjustment, on the second to third day, the relative humidity of the seedling hardening room is kept at 70-75%, and the temperature is kept at 26-28°C; c. late stabilization, on the fourth to fifth day, the seedlings are taken out of the seedling hardening room and placed in a cool and ventilated place outdoors to avoid rain and direct sunlight.

[0018] In the transplanting and transplanting step, the transplanting matrix is ​​formed by mixing loess, biochar, vermiculite and coconut bran in a weight ratio of 3-4:0.5:0.5:1.

[0019] In the transplanting and canopy step, water is poured once a day to keep the canopy substrate moist; carbendazim with a concentration of 0.2% is sprayed once every 2-3 days; fertilization begins one week after transplanting, and sulfur-based compound fertilizer is selected, which is dissolved in water at a mass ratio of 1:(300-400) and then sprayed or poured on the seedlings, and poured once every 4-5 days;

[0020] The antioxidant is used at a concentration of 0.2-0.7 g / L.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1. The present invention provides a method for cultivating healthy sugarcane seedlings, including seedling detoxification, germination induction, proliferation culture, rooting culture, seedling hardening and transplanting steps, which can achieve a high and stable high germination rate, shorten the cultivation period of conventional seedlings by 9-12 days, and do not need to cut the tip of the sugarcane buds for callus cultivation, thereby improving production efficiency. The method can also effectively inhibit the browning of sugarcane buds, promote seedling growth, proliferation and rooting, improve transplant survival rate, cultivate healthy sugarcane seedlings, and provide a high-quality seedling source for the sugarcane industry.

[0023] 2. Matrine, azadirachtin and rotenone are all broad-spectrum and highly effective fungicides (insecticides). The present invention uses matrine, azadirachtin and rotenone as plant-based disinfectants to detoxify seedlings. The inventors of the present invention found that the synergistic effect of matrine, azadirachtin and rotenone can reduce the stimulation to the tender sugarcane seedlings and effectively detoxify the sugarcane seedlings. The present invention is suitable for a variety of sugarcane varieties, and there is no need to explore the temperature and time of warm water detoxification according to different sugarcane varieties. The seedling rate reaches 97.5%, the germination rate reaches 96.0%, and the germination rate has high stability.

[0024] 3. In the germination induction step of the present invention, the induction culture medium formula can provide sufficient nutrients for the stem tip or axillary bud after detoxification, and promote the germination and growth of sugarcane buds. Among them, potassium nitrate, ammonium nitrate and other macroelements ensure the basic growth needs of plants; potassium dihydrogen phosphate provides phosphorus and potassium elements, which are beneficial to cell division and growth; magnesium sulfate provides magnesium elements, which participates in physiological processes such as chlorophyll synthesis; trace elements such as zinc sulfate and sodium molybdate ensure the normal physiological metabolism of plants; inositol and chitosan oligosaccharides can induce plants to improve the stress resistance of seedlings and enhance the vitality of sugarcane buds. Soaking sugarcane buds in antioxidants can effectively prevent browning of sugarcane buds. Vitamin C, β-carotene, astaxanthin, and silymarin play an antioxidant role together. Ganoderma lucidum polysaccharides can preserve sugarcane buds, protect the cell structure and function of sugarcane buds, prolong the storage time of sugarcane buds, and assist in providing nutrients to improve the quality and survival rate of sugarcane buds. Antioxidants also shorten the subsequent proliferation culture and rooting culture cycle, thereby shortening the entire seedling cultivation cycle by 9-12 days.

[0025] 4. The gradual seedling hardening method of the present invention provides a buffering process for the seedlings by gradually adjusting the temperature and relative humidity, so that the seedlings gradually adapt to the external environment, help enhance the stress resistance and adaptability of the seedlings, and improve the survival rate of the seedlings after transplanting.

[0026] 5. The proliferation medium of the present invention is MS+6-BA (6-benzyladenine) 1.0mg / L+GA (gibberellin) 1.0mg / L+NAA (naphthaleneacetic acid) 2.0mg / L+cysteine ​​5.0mg / L, and the rooting medium is MS+NAA (naphthaleneacetic acid) 5.0mg / L+2,4-D (2,4-dichlorophenoxyacetic acid) 2.5mg / +IBA (indolebutyric acid) 0.2mg / L+GA (gibberellin) 0.5mg / L+AC (abscisic acid) 0.1%+sucrose 30g / L. On the basis of MS medium, a variety of plant growth hormones are introduced to promote sugarcane bud proliferation and rooting. 6-BA and cysteine ​​in the proliferation medium can effectively prevent and control sugarcane bud browning. Cysteine ​​can inhibit phenolic substances from being oxidized to quinones under aerobic conditions, thereby inhibiting the non-enzymatic reaction from producing colored substances. At the same time, the medium formula of the present invention can also effectively shorten the proliferation and rooting cycle and promote the growth and development of sugarcane buds.

[0027] 6. The transplanting matrix is ​​made of loess, biochar, vermiculite and coconut bran in a specific proportion, which has good air permeability, water retention and fertility. Loess provides basic soil structure and nutrients, biochar can improve soil structure, provide an attachment environment for microorganisms, and increase soil water retention. Vermiculite and coconut bran improve the air permeability of the transplanting matrix and provide a suitable environment for the growth of seedlings. DETAILED DESCRIPTION

[0028] The specific embodiments of the present invention are described in detail below, but it should be understood that the protection scope of the present invention is not limited by the specific embodiments.

[0029] Example 1

[0030] The method for cultivating healthy sugarcane seedlings of this embodiment comprises the following steps:

[0031] (1) Seedling detoxification: Matrine, azadirachtin, rotenone, water, and calcium alkyl sulfonate were mixed uniformly in a weight ratio of 10:10:0.5:5000:10 to prepare a botanical disinfectant. A 3.5 cm stem tip was cut from a newly harvested sugarcane stem and placed in the botanical disinfectant for 60 min of ultrasonic detoxification at 500 W.

[0032] (2) Germination induction: The detoxified stem tips or axillary buds were rinsed with sterile water for three times and then placed in an induction medium for germination. The inoculated induction medium was placed in an incubator at a temperature of 30°C and a relative humidity of 75%. The light intensity was set to 1500 lux and the light time was 8 hours per day. By adjusting the light quality ratio and the irradiation time, cell elongation and photosynthesis were promoted, and the growth and germination of axillary buds were optimized. After the sugarcane buds were induced to grow, the outer old leaves were peeled off, and then the sugarcane buds were placed in an antioxidant and soaked for 60 minutes. After the soaking was completed, they were transferred to a proliferation medium for proliferation culture;

[0033] The antioxidants include, by weight: 1 part of vitamin C, 0.1 part of beta-carotene, 0.1 part of astaxanthin, 0.1 part of silymarin, and 5 parts of ganoderma lucidum polysaccharide, which are diluted with water to 0.2 g / L when used;

[0034] The formula of the induction medium is: potassium nitrate 1800 mg / L, ammonium nitrate 1650 mg / L, potassium dihydrogen phosphate 270 mg / L, magnesium sulfate 370 mg / L, zinc sulfate 9 mg / L, inositol 100 mg / L, sodium molybdate 0.25 mg / L, pyridoxine hydrochloride 0.5 mg / L, thiamine hydrochloride 0.1 mg / L, glycine 2 mg / L, chitosan oligosaccharide 10 mg / L.

[0035] (3) Proliferation culture: The sugarcane buds were cultured for 12 days to obtain bud clusters; the proliferation culture medium was MS + 6-BA 1.0 mg / L + GA 1.0 mg / L + NAA 2.0 mg / L + cysteine ​​5.0 mg / L;

[0036] (4) Rooting culture: Select individual bud clusters from the proliferation culture and transfer them to a rooting medium for rooting culture. Culture for 16 days to obtain rooted seedlings. The rooting medium is MS + NAA 5.0 mg / L + 2,4-D 2.5 mg / L + IBA 0.2 mg / L + GA 0.5 mg / L + AC 0.1% + sucrose 30 g / L.

[0037] (5) Hardening: The rooted seedlings are transferred from the culture room to the hardening room for hardening, which specifically includes: a. Initial adjustment: on the first day of hardening, the relative humidity in the hardening room is maintained at 75% and the temperature is maintained at 28°C; b. Mid-term adjustment: on the 2nd to 3rd day, the relative humidity in the hardening room is maintained at 70% and the temperature is maintained at 26°C; c. Late stabilization: on the 4th to 5th day, the seedlings are taken out of the hardening room and placed in a cool and ventilated place outdoors to avoid rain and direct sunlight.

[0038] (6) Transplanting and canopy planting: transplant the seedlings after hardening treatment into a seedling tray containing a canopy substrate, with the canopy substrate flush with the upper surface of the seedling tray; plant the healthy sugarcane seedlings obtained in step (5) into the seedling tray filled with nutrient soil, water thoroughly, build an arch shed, and cover with a plastic film; water once a day to keep the canopy substrate moist; spray with 0.2% carbendazim every 2 days; start fertilizing one week after transplanting, select sulfur-based compound fertilizer, dissolve it in water at a mass ratio of 1:300, and spray or water the seedlings, and then water them every 4 days; after the sugarcane seedlings grow out and have 3 leaves, remove the plastic film, and when the seedlings grow to 4 true leaves, the healthy sugarcane seedlings can be obtained;

[0039] The transplanting matrix is ​​a mixture of loess, biochar, vermiculite and coconut bran in a weight ratio of 3:0.5:0.5:1.

[0040] Example 2

[0041] The method for cultivating healthy sugarcane seedlings of this embodiment comprises the following steps:

[0042] (1) Seedling detoxification: Matrine, azadirachtin, rotenone, water, and calcium alkyl sulfonate were mixed uniformly in a weight ratio of 10:10:5:5000:50 to prepare a plant-based disinfectant. A 2 cm stem tip or axillary bud was cut from a newly harvested sugarcane stem and placed in the plant-based disinfectant for ultrasonic detoxification for 30 min.

[0043] (2) Germination induction: After the detoxified shoot tip or axillary bud is rinsed with sterile water for 5 times, it is placed in an induction medium for germination. The inoculated induction medium is placed in an incubator at a temperature of 33°C and a relative humidity of 80%. The light intensity is set to 2000 lux and the light time is 12 hours per day. By adjusting the light quality ratio and the irradiation time, cell elongation and photosynthesis are promoted, and the growth and germination of the axillary buds are optimized. After the sugarcane buds are induced to grow, the outer old leaves are peeled off, and then the sugarcane buds are placed in an antioxidant and soaked for 10 minutes. After the soaking is completed, they are transferred to a proliferation medium for proliferation culture;

[0044] The antioxidants include, by weight: 5 parts of vitamin C, 2 parts of beta-carotene, 2 parts of astaxanthin, 2 parts of silymarin, and 8 parts of ganoderma lucidum polysaccharide, which are diluted with water to 0.7 g / L when used;

[0045] The formula of the induction medium is: potassium nitrate 1800 mg / L, ammonium nitrate 1650 mg / L, potassium dihydrogen phosphate 270 mg / L, magnesium sulfate 370 mg / L, zinc sulfate 9 mg / L, inositol 100 mg / L, sodium molybdate 0.25 mg / L, pyridoxine hydrochloride 0.5 mg / L, thiamine hydrochloride 0.1 mg / L, glycine 2 mg / L, chitosan oligosaccharide 10 mg / L.

[0046] (3) Proliferation culture: The sugarcane buds were cultured for 14 days to obtain bud clusters; the proliferation culture medium was MS + 6-BA 1.0 mg / L + GA 1.0 mg / L + NAA 2.0 mg / L + cysteine ​​5.0 mg / L;

[0047] (4) Rooting culture: Select individual bud clusters from the proliferation culture and transfer them to a rooting medium for rooting culture. Culture for 18 days to obtain rooted seedlings. The rooting medium is MS + NAA 5.0 mg / L + 2,4-D 2.5 mg / L + IBA 0.2 mg / L + GA 0.5 mg / L + AC 0.1% + sucrose 30 g / L.

[0048] (5) Hardening: The rooted seedlings are transferred from the culture room to the hardening room for hardening, which specifically includes: a. Initial adjustment: on the first day of hardening, the relative humidity in the hardening room is maintained at 78% and the temperature is maintained at 30°C; b. Mid-term adjustment: on the 2nd to 3rd day, the relative humidity in the hardening room is maintained at 75% and the temperature is maintained at 28°C; c. Late stabilization: on the 4th to 5th day, the seedlings are taken out of the hardening room and placed in a cool and ventilated place outdoors to avoid rain and direct sunlight.

[0049] (6) Transplanting and canopying: transplant the seedlings after hardening treatment into a seedling tray containing a canopy substrate, with the canopy substrate flush with the upper surface of the seedling tray; plant the healthy sugarcane seedlings obtained in step (5) into the seedling tray filled with nutrient soil, water them thoroughly, build an arch shed, and cover them with a plastic film or a sunshade cloth; water them once a day to keep the canopy substrate moist; spray them with 0.5% carbendazim every 3 days; start fertilizing one week after transplanting, use a sulfur-based compound fertilizer, dissolve it in water at a mass ratio of 1:400, and then spray or water the seedlings, and then water them every 5 days; after the sugarcane seedlings grow out and have 3 leaves, remove the plastic film or sunshade cloth, and when the seedlings grow to 5 true leaves, the healthy sugarcane seedlings can be obtained;

[0050] The transplanting matrix is ​​a mixture of loess, biochar, vermiculite and coconut bran in a weight ratio of 4:0.5:0.5:1.

[0051] Example 3

[0052] The method for cultivating healthy sugarcane seedlings of this embodiment comprises the following steps:

[0053] (1) Seedling detoxification: Matrine, azadirachtin, rotenone, water, and calcium alkyl sulfonate were mixed uniformly in a weight ratio of 10:10:2:5000:25 to prepare a botanical disinfectant. A 3 cm stem tip or axillary bud was cut from a newly harvested sugarcane stem and placed in the botanical disinfectant for ultrasonic detoxification for 45 min.

[0054] (2) Germination induction: After the detoxified shoot tip or axillary bud is rinsed with sterile water for 4 times, it is placed in an induction medium for germination. The inoculated induction medium is placed in an incubator at a temperature of 32° C. and a relative humidity of 78%. The light intensity is set to 1800 lux and the light time is 10 hours per day. By adjusting the light quality ratio and the irradiation time, cell elongation and photosynthesis are promoted, and the growth and germination of the axillary buds are optimized. After the sugarcane buds are induced to grow, the outer old leaves are peeled off, and then the sugarcane buds are placed in an antioxidant and soaked for 10-60 minutes. After the soaking is completed, they are transferred to a proliferation medium for proliferation culture;

[0055] The antioxidants include, by weight: 3 parts of vitamin C, 1 part of beta-carotene, 0.8 parts of astaxanthin, 1 part of silymarin, and 6 parts of ganoderma lucidum polysaccharide, which are diluted with water to 0.5 g / L when used;

[0056] The formula of the induction medium is: potassium nitrate 1800 mg / L, ammonium nitrate 1650 mg / L, potassium dihydrogen phosphate 270 mg / L, magnesium sulfate 370 mg / L, zinc sulfate 9 mg / L, inositol 100 mg / L, sodium molybdate 0.25 mg / L, pyridoxine hydrochloride 0.5 mg / L, thiamine hydrochloride 0.1 mg / L, glycine 2 mg / L, chitosan oligosaccharide 10 mg / L.

[0057] (3) Proliferation culture: The sugarcane buds were cultured for 13 days to obtain bud clusters; the proliferation culture medium was MS + 6-BA 1.0 mg / L + GA 1.0 mg / L + NAA 2.0 mg / L + cysteine ​​5.0 mg / L;

[0058] (4) Rooting culture: Select individual shoots from the proliferation culture and transfer them to the rooting medium for rooting culture. After culturing for 17 days, rooted seedlings were obtained. The rooting medium consisted of MS + NAA 5.0 mg / L + 2,4-D 2.5 mg / L + IBA 0.2 mg / L + GA 0.5 mg / L + AC 0.1% + sucrose 30 g / L.

[0059] (5) Hardening: The rooted seedlings are transferred from the culture room to the hardening room for hardening, which specifically includes: a. Initial adjustment: on the first day of hardening, the relative humidity in the hardening room is maintained at 76% and the temperature is maintained at 29°C; b. Mid-term adjustment: on the 2nd to 3rd day, the relative humidity in the hardening room is maintained at 72% and the temperature is maintained at 27°C; c. Late stabilization: on the 4th to 5th day, the seedlings are taken out of the hardening room and placed in a cool and ventilated place outdoors to avoid rain and direct sunlight.

[0060] (6) Transplanting and canopy planting: transplant the seedlings after hardening treatment into a seedling tray containing a canopy substrate, with the canopy substrate flush with the upper surface of the seedling tray; plant the healthy sugarcane seedlings obtained in step (5) into the seedling tray filled with nutrient soil and water thoroughly; build an arch shed, and cover with a plastic film or a sunshade cloth; water once a day to keep the canopy substrate moist; spray with 0.3% carbendazim every 3 days; start fertilizing one week after transplanting, select sulfur-based compound fertilizer, dissolve it in water at a mass ratio of 1:350, and spray or water the seedlings, and then water them every 4 days; after the sugarcane seedlings grow out and have 3 leaves, remove the plastic film or sunshade cloth, and when the seedlings grow to 4 true leaves, the healthy sugarcane seedlings can be obtained;

[0061] The transplanting matrix is ​​a mixture of loess, biochar, vermiculite and coconut bran in a weight ratio of 3.5:0.5:0.5:1.

[0062] Comparative Example 1

[0063] Compared with Example 3, the seedling detoxification method is 58° C. water bath for 60 min, and the rest is the same as Example 3.

[0064] Comparative Example 2

[0065] Compared with Example 3, the seedling detoxification method is 53° C. water bath for 120 min, and the rest is the same as Example 3.

[0066] Comparative Example 3

[0067] Compared with Example 3, the seedling detoxification method is to soak the stem tip in 0.1% mercuric chloride solution for 60 minutes and then rinse it with sterile water for 3 times, and the rest is the same as Example 3.

[0068] Comparative Example 4

[0069] Compared with Example 3, the proliferation medium is MS+GA 1.0 mg / L+NAA 2.0 mg / L, the rooting medium is MS+NAA 5.0 mg / L+GA 0.5 mg / L+sucrose 30 g / L, and the rest is the same as Example 3.

[0070] Comparative Example 5

[0071] Compared with Example 3, in the germination induction step, the sugarcane buds are not immersed in antioxidants, but are directly transferred to the proliferation culture medium for proliferation culture. The rest is the same as Example 3.

[0072] Comparative Example 6

[0073] Compared with Example 3, the antioxidant is only vitamin C, and the rest is the same as Example 3.

[0074] Comparative Example 7

[0075] Compared with Example 3, the transplanting substrate is commercially available loess, and the rest is the same as Example 3.

[0076] Comparative Example 8

[0077] Compared with Example 3, the formula of the induction medium is (without inositol and chito-oligosaccharide): potassium nitrate 1800 mg / L, ammonium nitrate 1650 mg / L, potassium dihydrogen phosphate 270 mg / L, magnesium sulfate 370 mg / L, zinc sulfate 9 mg / L, sodium molybdate 0.25 mg / L, pyridoxine hydrochloride 0.5 mg / L, thiamine hydrochloride 0.1 mg / L, glycine 2 mg / L, and the rest is the same as Example 3.

[0078] Comparative Example 9

[0079] Compared with Example 3, the plant-based disinfectant is prepared by mixing matrine, water, and calcium alkyl sulfonate in a weight ratio of 10:5000:25, and the rest is the same as Example 3.

[0080] Comparative Example 10

[0081] Compared with Example 3, the plant-derived disinfectant is prepared by mixing azadirachtin, water, and calcium alkyl sulfonate in a weight ratio of 10:5000:25, and the rest is the same as Example 3.

[0082] Comparative Example 11

[0083] Compared with Example 3, the plant-derived disinfectant is prepared by mixing rotenone, water, and calcium alkyl sulfonate in a weight ratio of 2:5000:25, and the rest is the same as Example 3.

[0084] Comparative Example 12

[0085] Compared with Example 3, the plant-based disinfectant is prepared by mixing matrine, azadirachtin, water, and calcium alkyl sulfonate in a weight ratio of 10:10:5000:25, and the rest is the same as Example 3.

[0086] Comparative Example 13

[0087] Compared with Example 3, the plant-based disinfectant is prepared by mixing matrine, rotenone, water, and calcium alkyl sulfonate in a weight ratio of 10:2:5000:25, and the rest is the same as Example 3.

[0088] Comparative Example 14

[0089] Compared with Example 3, the plant-derived disinfectant is prepared by mixing azadirachtin, rotenone, water, and calcium alkyl sulfonate in a weight ratio of 10:2:5000:25, and the rest is the same as Example 3.

[0090] This application selects Guitang No. 42, New Taiwan Sugar No. 22, and Guiliu 05136 as test materials. The following are the test data of Examples 1-3 and Comparative Examples 1-14.

[0091]

[0092]

[0093]

[0094]

[0095] Budding rate = number of buds at the stem tip (axillary buds) ÷ number of cut stem tips (axillary buds) × 100%;

[0096] Seedling rate = number of seedlings / number of sprouts × 100%;

[0097] Virus-free rate = number of virus-free plants ÷ number of mature seedlings × 100%;

[0098] Browning rate = number of browning buds ÷ number of cut stem tips (axillary buds) × 100%

[0099] As can be seen from the above table, matrine, azadirachtin and rotenone are all broad-spectrum and highly effective fungicides (insecticides). The present invention uses matrine, azadirachtin and rotenone as plant-derived disinfectants to detoxify seedlings. The inventors of the present invention have found that the synergistic effect of matrine, azadirachtin and rotenone can reduce the stimulation to the tender sugarcane seedlings while effectively detoxifying the sugarcane seedlings. It is suitable for a variety of sugarcane varieties, and there is no need to explore the temperature and time of warm water detoxification according to different sugarcane varieties. The detoxification rate is as high as 99%, the emergence rate is 96%, and the emergence stability is very high in different varieties. 2 Although the use of warm water for detoxification can also achieve a high detoxification rate, the germination rate in different sugarcane varieties is not very stable. Compared with Comparative Example 3, Example 3 has better detoxification and germination effects, and avoids the phytotoxicity and pollution of chemical reagents; From the comparison of Comparative Examples 4-6 and Comparative Example 8 with Example 3, it can be seen that the induction culture medium formula of the present invention can provide sufficient nutrients for the stem tip or axillary buds after detoxification, promote the germination and growth of sugarcane buds, and the synergistic effect of inositol and chitosan oligosaccharides can induce plants to improve the stress resistance of seedlings and enhance the vitality of sugarcane buds; placing the sugarcane buds in the antioxidant medium of the present invention The sugarcane buds are soaked in an antioxidant, which can effectively prevent the browning of sugarcane buds. Vitamin C, β-carotene, astaxanthin and silymarin play an antioxidant role together. Ganoderma lucidum polysaccharide can preserve the sugarcane buds, protect the cell structure and function of the sugarcane buds, prolong the storage time of the sugarcane buds, and assist in providing nutrients to improve the quality and survival rate of the sugarcane buds. The antioxidant also shortens the subsequent proliferation culture and rooting culture cycle. It can be seen from Comparative Example 4 and Example 3 that 6-BA and cysteine ​​in the proliferation culture medium can effectively prevent and control the browning of sugarcane buds. Cysteine ​​can inhibit the oxidation of phenolic substances into quinones under aerobic conditions, thereby inhibiting The non-enzymatic reaction produces colored substances, which greatly reduces the browning rate of sugarcane buds; from Comparative Example 7 and Example 3, it can be seen that the transplanting matrix of the present invention can help improve the seedling rate and also shorten the cultivation period; from the comparison of Comparative Examples 9-14 and Example 3, it can be seen that only using a single component of matrine, azadirachtin, and rotenone or a combination of two of them as a plant-derived disinfectant, the germination rate and the detoxification rate are both lower than those of Example 3. The matrine, azadirachtin, and rotenone of the present invention can achieve a synergistic effect, can reduce the stimulation to the tender sugarcane seedlings, improve the germination rate, and can effectively detoxify the sugarcane seedlings.

[0100] The foregoing description of specific exemplary embodiments of the present invention is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present invention to the precise form disclosed, and it is clear that many changes and variations can be made based on the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present invention and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present invention and various different selections and changes. The scope of the present invention is intended to be limited by the claims and their equivalents.

Claims

1. A method for cultivating healthy sugarcane seedlings, characterized in that: The steps include: (1) Seedling detoxification: Cut 2-3.5 cm stem tips or axillary buds from newly harvested sugarcane stems and place them in a plant-derived disinfectant for ultrasonic detoxification for 30-60 minutes; (2) Germination induction: The stem tips or axillary buds after the virus-free treatment are rinsed with sterile water for 3-5 times and then placed in an induction medium for germination. After the sugarcane buds are induced, the outer old leaves are peeled off, and then the sugarcane buds are immersed in an antioxidant. After the immersion is completed, they are transferred to a proliferation medium for proliferation culture; (3) Proliferation culture: The sugarcane buds are cultured for 12-14 days to obtain bud clusters; (4) Rooting culture: Select individual shoots from the proliferation culture and transfer them to the rooting medium for rooting culture. Cultivate for 16-18 days to obtain rooted seedlings. (5) Hardening: Transfer the rooted seedlings from the culture room to the hardening room for hardening; (6) Transplanting and transplanting: transplant the seedlings after hardening treatment into a seedling tray containing a transplanting matrix, wherein the transplanting matrix is ​​flush with the upper surface of the seedling tray, and the healthy sugarcane seedlings obtained in step (5) are planted in the seedling tray filled with nutrient soil and watered thoroughly, and an arch shed is built and covered with a plastic film or a sunshade cloth. After the sugarcane seedlings grow out and have 3 leaves, the plastic film or the sunshade cloth is removed. When the seedlings grow to 4-5 true leaves, the healthy sugarcane seedlings are obtained.

2. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: In the seedling detoxification step, the plant-derived disinfectant is prepared by mixing matrine, azadirachtin, rotenone, water and a dispersant in a weight ratio of 10:10:0.5-5:5000:10-50; and the dispersant is an alkyl sulfonate.

3. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: In the germination induction step, the formula of the induction culture medium is: 1800 mg / L potassium nitrate, 1650 mg / L ammonium nitrate, 270 mg / L potassium dihydrogen phosphate, 370 mg / L magnesium sulfate, 9 mg / L zinc sulfate, 100 mg / L inositol, 0.25 mg / L sodium molybdate, 0.5 mg / L pyridoxine hydrochloride, 0.1 mg / L thiamine hydrochloride, 2 mg / L glycine, and 10 mg / L chitosan oligosaccharide.

4. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: In the germination induction step, the inoculated induction culture medium is placed in an incubator at a temperature of 30-33° C. and a relative humidity of 75-80% for cultivation, the light intensity is set to 1500-2000 lux, and the light time is 8-12 hours per day.

5. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: In the germination induction step, the sugarcane buds are immersed in the antioxidant for 10-60 minutes; the antioxidant comprises, by weight: 1-5 parts of vitamin C, 0.1-2 parts of beta-carotene, 0.1-2 parts of astaxanthin, 0.1-2 parts of silymarin, and 5-8 parts of ganoderma lucidum polysaccharide.

6. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: In the proliferation culture step, the proliferation culture medium is MS+6-BA 1.0mg / L+GA 1.0mg / L+NAA 2.0mg / L+cysteine ​​5.0mg / L; the rooting culture medium is MS+NAA 5.0mg / L+2,4-D 2.5mg / +IBA 0.2mg / L+GA 0.5mg / L+AC 0.1%+sucrose 30g / L.

7. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: The seedling hardening step specifically includes: a. preliminary adjustment, on the first day of seedling hardening, the relative humidity of the seedling hardening room is kept at 75-78%, and the temperature is kept at 28-30°C; b. mid-term adjustment, on the second to third day, the relative humidity of the seedling hardening room is kept at 70-75%, and the temperature is kept at 26-28°C; c. late stabilization, on the fourth to fifth day, the seedlings are taken out of the seedling hardening room and placed in a cool and ventilated place outdoors to avoid rain and direct sunlight.

8. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: In the transplanting and transplanting step, the transplanting matrix is ​​formed by mixing loess, biochar, vermiculite and coconut bran in a weight ratio of 3-4:0.5:0.5:

1.

9. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: In the transplanting and canopy step, water is poured once a day to keep the canopy substrate moist; carbendazim with a concentration of 0.2-0.5% is sprayed once every 2-3 days; and fertilization begins one week after transplanting, and sulfur-based compound fertilizer is selected, which is dissolved in water at a mass ratio of 1:(300-400) and then sprayed or poured on the seedlings, and poured once every 4-5 days.

10. The method for cultivating healthy sugarcane seedlings according to claim 1, characterized in that: The antioxidant is used at a concentration of 0.2-0.7 g / L.

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