CN111699972A - Construction method of breeding system of virus-free seed potatoes - Google Patents
Construction method of breeding system of virus-free seed potatoes Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G2/00—Vegetative propagation
- A01G2/10—Vegetative propagation by means of cuttings
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Botany (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Cultivation Of Plants (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a construction method of a virus-free potato seed breeding system, belonging to the technical field of potato seed breeding. The method comprises the steps of screening and culturing potato virus-free basic seedlings, strengthening screening and culturing the virus-free basic seedlings, selecting the virus-free basic seedlings, culturing the virus-free seedlings, carrying out section propagation on the virus-free seedlings, carrying out production and propagation of water culture seedlings, hardening seedlings before transplanting, producing potato virus-free breeder seeds, producing improved seeds, breeding two or high mountains of the breeder seeds and the like.
Description
Technical Field
The invention belongs to the technical field of breeding of potato seeds, and particularly relates to a construction method of a virus-free potato seed breeding system.
Background
With the development of economy, potatoes are converted from traditional grain crops into economic crops integrating grain, vegetables, feed and processing raw materials, and particularly, the proposal of the national potato staple feeding strategy plays a vital role in adjusting rural industrial structures and increasing the income of farmers. Meanwhile, the popularization of the detoxified seed potatoes restores the physiological function and the production characteristic of the potato variety, prevents the variety degeneration of the potatoes caused by the continuous infection and accumulation of the viruses, and ensures that the quality and the yield of the good variety are achieved. On the basis of the production technology of the virus-free potato seeds, the construction research of the virus-free potato seed rapid propagation system is further developed according to the rapid propagation rule of asexual propagation crops, and the popularization and the application of the virus-free potato seed production system are accelerated.
Disclosure of Invention
The invention aims to solve the defects of the prior art, and provides a construction method of a virus-free potato seed breeding system, which can timely obtain the corresponding previous seed supply quantity required by the production of various grades of potatoes according to the actual production requirement, and accurately regulate and control the production of various grades of potatoes.
The invention adopts the following technical scheme:
the invention provides a construction method of a breeding system of virus-free seed potatoes, which comprises the following steps:
s001 screening and culturing of potato virus-free basic seedlings: the method comprises the steps of screening and culturing S002 enhanced detoxification base seedlings, selecting S003 detoxification base seedlings, culturing S004 detoxification seedlings, segmenting and propagating S005 detoxification seedlings, producing and expanding propagation S006 water culture seedlings, and hardening seedlings before S007 transplanting;
s008 potato virus-free stock production: transplanting the S009 hydroponic seedlings to a greenhouse medium to culture original seeds, culturing the S010 hydroponic seedlings to culture potato by liquid nutrient solution, and culturing the original seeds by the S011 greenhouse medium for the next year;
the production method of the S012 stock comprises the following steps: the method is realized by S013 stock high-mountain breeding and S014 water culture seedling high-mountain breeding;
the production method of the improved S015 seeds comprises the following steps: the stock produced by the steps S013 and S014 is bred in the second mountain or mountain breeding way of the S016 stock.
S017 comprehensive regulation and control of the third-level propagation donor system of the potato virus-free seed potato: constructing a three-level breeding donor system model of the virus-free potato seeds by using the seedling quantity and the production seed potato quantity data of the periodic production process of the potato seeds at all levels, and timely regulating and controlling the S007-S014 key steps according to the actual production requirements to obtain the corresponding last-level seed supply quantity required by the production of the potato seeds at all levels.
Furthermore, the screening and culturing of the S002 enhanced detoxification base seedling specifically comprises the following steps: taking stem tip meristem detoxification culture and virus detection as quality assurance, using a tissue culture room to detoxify the stem tip meristem of seed potatoes, inducing the seed potatoes in a test tube to form virus-free test-tube seedlings, then carrying out virus detection one by one, expanding and propagating the virus-free test-tube seedlings qualified in virus detection into wide-mouthed bottles, and randomly extracting 1 in each 10 wide-mouthed bottles for virus detection and screening to ensure that the basic seedlings are really virus-free;
the S003 virus-free basic seedling is selected specifically as follows: preferentially selecting a robust detoxified test-tube seedling as a basic seedling for rapid propagation of the detoxified seedling;
the culture of the S004 virus-free seedlings specifically comprises the following steps: using a test-tube seedling culture container, adopting an MS culture medium, culturing the virus-free seedlings at 22-25 ℃, illuminating for 16 hours every day with illumination intensity of 2000-;
the S005 virus-free seedling cutting propagation specifically comprises the following steps: performing single-section cutting cuttage when the virus-free seedlings grow to 6-7 sections, and then inoculating the virus-free seedlings into a culture box for propagation; the strong virus-free test-tube plantlet is inserted in an MS culture medium and takes root in 7-9 days, the plantlet can grow into 2-3 leaves 11-13 days after the young bud is emerged from the leaf axilla, a stem 6-7 joints can be formed in 20-25 days, the height of the plantlet is 7-10cm, the stem thickness is 0.6-0.8mm, and the number of the leaves is 5-8 plantlets, the virus-free plantlet can grow rapidly at proper temperature and can be used as a virus-free strong plantlet to perform water culture plantlet production or cut propagation;
the S006 water culture seedling production and propagation specifically comprises the following steps: cutting detoxicated seedlings into sections, performing cutting water culture, inducing the water culture cutting seedlings to root by a rooting nutrient solution in an intelligent greenhouse, enabling the original roots to grow 3 days after cutting, enabling the original roots to root 5-7 days after cutting, adding the rooting water culture cutting seedlings into a growth nutrient solution, enabling the water culture seedlings to grow rapidly at proper temperature, enabling buds to grow into small plants with the plant height of 7-10cm, the stem thickness of 0.12cm and 6-7 nodes of stems after the buds grow from leaf axils for 10-12 days, and enabling the small plants to be used as detoxified strong seedlings to perform section propagation, water culture potato production, original stock production or stock production;
the hardening-seedling before the S007 transplantation specifically comprises the following steps: opening a greenhouse window before transplanting to the field, allowing the hydroponic seedlings to naturally grow for 2-3 days, and then transplanting to the field.
Furthermore, the S009 water-cultured seedling is transplanted to the greenhouse medium culture stock specifically as follows: adopting greenhouse or net shed isolation facilities and an insect-proof net with more than 40 meshes, removing weeds and leveling the soil of a sunlight greenhouse, a self-control greenhouse or a net shed, and uniformly spreading quicklime, an insecticide and a bactericide; dividing the prepared ground surface into cells or placing a culture tray, putting a substrate with the thickness of 6-8 cm, thoroughly watering, loosening, and scraping the substrate; transplanting the water culture seedlings which are cultured and tempered by natural illumination into vermiculite, planting the seedlings into a matrix with the row spacing of 6cm multiplied by 7cm and the depth of 2cm-2.5cm, wherein the seedlings are preferably transplanted with 2 leaves of vermiculite, and spraying little water after planting; shading with a shading net for 5-7 days after transplanting, at the temperature of 22-25 ℃ and the relative humidity of 95%; covering the transplanted water culture seedlings with a small plastic arch shed, and keeping moisture and keeping the temperature for 7-12 days; in the moisture-keeping stage, the humidity of the vermiculite is kept at 100%, the temperature is 25-28 ℃, and the humidity is not higher than 30 ℃; removing the small arched shed after the water culture seedlings take roots and survive; watering and subculturing the nutrient solution in time according to the conditions of the seedlings and the vermiculite, keeping the water content of the matrix between 50 and 70 percent in the whole growth period, and strictly managing in a greenhouse during the production period.
The water culture of the S010 water culture seedlings by the liquid nutrient solution is as follows: in the intelligent water culture greenhouse, 1 season of water-cultured potatoes are produced in the idle time of autumn and winter, wherein the time is from the first ten days of 9 months to the middle and last ten days of 2 months of the next year; the culture medium adopts strong seedling nutrient solution, and is replaced periodically for 20-25 days; the virus-free test-tube plantlet is 20-25 days old, 7-10cm high, 0.6-0.8mm thick stem, 5-8 leaves, 2400 plants/m2The density of the culture medium is inserted in a culture disc; the temperature of the greenhouse is kept at 20 +/-5 ℃ in the daytime, the illumination intensity is kept at 3500-When the current is over; the produced hydroponic potatoes can be used for producing and culturing stock seeds in a greenhouse and can also be used for planting and producing stock seeds in high-altitude fields.
Furthermore, the bactericide in the step S009 is Lesper Benth or carbendazim, the substrate comprises vermiculite and compound fertilizer, 2000kg of vermiculite is mixed with 1kg of compound fertilizer (N: P: K: 17:17:17) for mixing, the pH value of the vermiculite is required to be less than or equal to 8.2, the particle diameter is 2mm-4mm, and the vermiculite is fluffy and has less impurities.
Furthermore, the S011 next-year greenhouse medium culture stock comprises the following specific steps: the planting density is 40 plants/square meter, the net room is watered regularly to prevent and control plant diseases and insect pests, the paclobutrazol is sprayed at 10 g/mu before flowering to prevent the plants from growing excessively, and the planting density in the net room is 40 plants/m2The number of the potatoes and the ratio of the standard seed potatoes reach 100.5 grains/m respectively286.59 percent; the S013 original stock mountain breeding specifically comprises the following steps: the planting density of the original seeds is 5000 grains/mu, 60kg of potassium sulfate compound fertilizer is applied as base fertilizer before each mu of planting, 7.5kg of urea is additionally applied to each mu in the later period, the field is cultivated and hogged in due time to prevent and treat plant diseases and insect pests, and 10g of paclobutrazol is sprayed per mu before flowering to prevent the excessive growth of plants.
The alpine breeding of the S014 water culture seedlings is specifically as follows: planting virus-free water culture strong seedlings or virus-free water culture potatoes in a field with the altitude of 1200-1800m original seed base in spring, controlling the original seed production through density control, wherein the planting density of the virus-free water culture strong seedlings is 12000-15000 plants/mu, applying 15kg of potassium sulfate compound fertilizer as base fertilizer before each mu of planting, and applying 7.5kg of urea at the later stage; the planting density of the detoxified water-cultured potatoes is 5000 grains/mu, 60kg of potassium sulfate compound fertilizer is applied as base fertilizer before each mu of planting, 7.5kg of urea is additionally applied to each mu in the later period, the field is timely intertillage and weeding are carried out to prevent and control plant diseases and insect pests, and 10g of paclobutrazol is sprayed per mu before flowering to prevent the excessive growth of plants; every 667m in the field2When 1.5 ten thousand of virus-free water culture strong seedlings are planted, the ratio of the harvested seed potatoes to the standard seed potatoes (10g-20g) reaches 3.94 ten thousand seeds/667 m respectively2、87.98％。
Further, the breeding of the S016 elite II mountain or mountain is specifically as follows: after the previous crops are harvested, timely ploughing and cleaning the stubbles, and deeply ploughing the soil by over 25 cm; the fertilizing mode takes a base fertilizer as a main part and takes additional fertilizer as an auxiliary part, and the additional application of a foliar fertilizer is properly carried out, wherein the base fertilizer takes fermented and decomposed farmyard manure and biological organic fertilizer as key points, and 1500 kilograms of farmyard manure and 120 kilograms of biological organic fertilizer are applied per mu; the method is characterized in that the farmyard manure is deficient, 50-100 kg of special compound fertilizer or compound fertilizer for potatoes is applied to each mu, 100-200 kg of biological organic fertilizer is matched to be used as base fertilizer to be applied to furrows at one time, 4000-5000 holes of original seeds are planted in each mu in fine seed production, and attention is paid to direct seeding and shortage is paid to seed searching.
Compared with the prior art, the invention has the beneficial effects that:
1. the test-tube plantlet has reliable detoxification effect. The stem tip meristem detoxification culture and virus detection are used as quality assurance, a tissue culture room is used for carrying out stem tip meristem detoxification on potato varieties, then virus detection is carried out one by one, virus detection qualified detoxification test-tube seedlings are propagated into wide-mouth bottles, 1 virus is randomly extracted from every 10 wide-mouth bottles for virus detection and identification, and the true detoxification of basic seedlings is ensured.
2. Performing datamation and scale on water culture seedling propagation: 2400 water-cultured seedlings can be produced once by a self-made culture disc with 1 square meter, one period is 10-12 days, seedlings are propagated, the water-cultured seedlings can be propagated for 3 months in the last half year, and 5-6 months are used as a transplanting period. The growth period of the water culture seedlings is recorded in the production process to form a data model, the seedling emergence period of the water culture seedling culture disc is optimal for 7 times, and the seedling emergence amount in the period is 16800. One 667m2The intelligent greenhouse can be used for placing more than 400 1m2The cultivation plate can produce 672 tens of thousands of potato virus-free hydroponic seedlings in one production period, and can be used for 448 mu of field close planting cultivation.
3. Diversification of original stock production: except the traditional water culture seedling substrate culture original seed, the water culture potato can also be produced by the culture of the water culture seedling nutrient solution in autumn and winter. On one hand, the production efficiency of the original stock is obviously improved by a diversified production mode, on the other hand, the outdoor production risk of the original stock cultured by the water culture seedling substrate is obviously reduced by combining two production modes, the average number of single-plant tuber bearing is 1.5 per tuber in autumn and winter, about 3500 individual tuber bearing can be induced per square meter, and one tuber bearing is 667m2The intelligent greenhouse is provided with more than 400 1m2The culture tray can produce about 140 thousands of potato virus-free hydroponics potatoes in autumn.
4. The production cycle of the original seeds is adjustable: the method is characterized in that while a two-year detoxification potato seed planting technology is perfected, a rapid propagation technology of water-cultured potatoes is utilized, water-cultured seedlings or water-cultured potatoes are directly cultured in a high-altitude large-area field, seeding density is adjusted to control stock seed production, batch standard stock seeds are produced within one year and are directly applied in production, and the detoxification potato seed planting period is successfully shortened to one year.
5. The method comprises the following steps of optimizing production regulation of various potato seeds by using a potato virus-free seed potato three-stage propagation donor line: and constructing a three-stage propagation donor system model of the potato virus-free seed potatoes by utilizing the seedling (seed) quantity required in the periodic production process of each stage of seed potatoes and the production seed potato quantity data, timely acquiring the corresponding previous stage seed supply quantity required by the production of each stage of seed potatoes according to the actual production requirement, and accurately regulating and controlling the production of each stage of seed potatoes.
6. The nutrient solution and other reagents used by the invention are relatively common, the used cost is low, the main components of the waste liquid after the test are a large amount of or trace elements, no toxin exists, and the pollution degree to the surrounding environment after the waste liquid and the sewage are treated is small.
Drawings
FIG. 1 is a flow chart of a construction method of a breeding system of a virus-free seed potato;
FIG. 2 is a schematic structural diagram of a homemade culture tray used in the present invention;
wherein, 18, foam board; 19. a drain hole; 20. a nutrient solution injection valve; 21. a water culture tray; 22. a drain valve; 23. a waste discharge pipeline.
Detailed Description
The present invention is further illustrated by the following examples, which are intended to be purely exemplary and are not intended to limit the scope of the invention, which is defined in the appended claims, as may be amended by those skilled in the art upon reading the present invention.
The present invention will be described in further detail with reference to examples.
The water culture disc 21 used in the invention is a cuboid which is hollow inside and is made of PC (polycarbonate), the length of the cuboid is 1.3m, the width of the cuboid is 1.0m, the height of the cuboid is 0.05m, the surface of the water culture disc is provided with a plurality of foam plates 18, the length of each foam plate 18 is 20cm, the width of each foam plate is 25cm, through holes are arranged on the foam plates 18, the length of each through hole is 2cm, the middle part of the water culture disc 21 is provided with a water drainage hole 19, the upper layer of each water drainage hole 19 is provided with a; one corner of the water culture disc 21 is provided with a nutrient solution injection valve 20, the nutrient solution injection valve 20 is a water tap with the caliber of 2cm, the other end of the nutrient solution injection valve is connected with a nutrient preparation container, and the container is 2.5m away from the ground. The middle part of the lower part of the water culture disc 21 is connected with a waste discharge pipeline 23, and a liquid discharge valve 22 is arranged on the waste discharge pipeline 23.
Examples
As shown in fig. 1, the method for constructing a breeding system of a virus-free seed potato provided in this embodiment includes the following steps:
s001 screening and culturing of potato virus-free basic seedlings: comprises the steps of screening and culturing S002 enhanced detoxification base seedlings, selecting S003 detoxification base seedlings, culturing S004 detoxification seedlings, segmenting and propagating S005 detoxification seedlings, producing and expanding propagation S006 water culture seedlings and hardening seedlings before S007 transplantation.
S002 enhanced detoxification basic seedling screening and culturing: the stem tip meristem detoxification culture and virus detection are taken as quality assurance, a tissue culture room is utilized to detoxify the stem tip meristem of the seed potato, a detoxified test tube seedling is induced in a test tube to form, virus detection and screening are carried out on the detoxified test tube seedling according to the proportion of 1:10, and the basic seedling is ensured to be really detoxified.
S003 detoxification basic seedling selection: preferably, a robust detoxified test-tube seedling is selected as a basic seedling for rapid propagation of the detoxified seedling.
Culturing S004 virus-free seedlings by taking a plastic square box with the specification of 5cm × 5cm × 9cm as a test-tube seedling culture container and adopting 5 layers of culture shelves, wherein the area of each culture shelf is 1.2m × 0.48.48 m to 0.576m2Each box is stored with 14 × 5 culture boxes with × 2 layers of 140, each box is transplanted with 25 single-section stem segments of the virus-free seedlings, the culture mode of the test-tube seedlings is adopted, MS culture medium is adopted, the temperature for propagating the virus-free seedlings is 22-25 ℃, the virus-free seedlings are irradiated for 16 hours every day, the illumination intensity is 2000-3000Lx, and the virus-free seedlings are strictly disinfected and prevent the air pollution of a sterile room.
S005 cutting propagation of virus-free seedlings: and carrying out single-section cutting cuttage when the virus-free seedlings grow to 6-7 sections, and then inoculating the virus-free seedlings into a culture box for propagation. The strong virus-free test-tube plantlet is inserted in MS culture medium, and rooted in 5-7 days, the young bud can grow into 2-3 leaf plantlet after 10-12 days, the stem node 6-7 nodes can be formed in 20-25 days, the height of the plantlet is 7-10cm, the stem thickness is 0.6-0.8mm, the number of leaves is 5-8, the virus-free plantlet can grow rapidly at proper temperature, and the virus-free test-tube plantlet can be used as virus-free strong plantlet to perform water culture plantlet production or cut propagation. The breeding cycle requires 21-25 days every season and the production is annual.
S006 water culture seedling production and propagation: a1.3 m long, 1.0m wide and 0.05m high PC tray was prepared as a hydroponics culture tray (FIG. 2), and 35L of a nutrient solution for inducing formation of root progenitors (in which 9.023g of ammonium nitrate, 10.391g of potassium nitrate, 0.93g of potassium dihydrogen phosphate, 0.202g of magnesium sulfate, 7.284g of anhydrous calcium chloride, 0.61g of ferrous sulfate and 0.815g of disodium ethylenediaminetetraacetate were dissolved in 35L of ultrapure water) was placed on each tray, and a foam plastic plate having a planting plant pitch of 2cm and a row pitch of 2cm was floated on the nutrient solution. Cutting detoxicated seedling into sections, water planting, adding rooting nutrient solution 35L (comprising 54.141g of ammonium nitrate, 62.344g of potassium nitrate, 5.578g of monopotassium phosphate, 12.141g of magnesium sulfate, 10.927g of anhydrous calcium chloride, 0.914g of ferrous sulfate, 1.222g of disodium ethylenediamine tetraacetate, 27.235mg of potassium iodide, 8.203mg of sodium molybdate, 0.82mg of copper sulfate, 0.82mg of cobalt chloride, 731.73mg of manganese sulfate, 282.192mg of zinc sulfate and 203.441mg of boric acid dissolved in 35L of ultrapure water) to induce the water planting cutting seedling to root, 3 days after cutting, the original root body takes place, about 5-7 days to root, adding the rooting water planting cutting seedling into secondary propagation (strong seedling) nutrient solution 35L (comprising 90.234g of ammonium nitrate, 103.906g of potassium nitrate, 9.297g of monopotassium phosphate, 20.234g of magnesium sulfate, 18.211g of anhydrous calcium chloride, 1.523g of ferrous sulfate, 2.037g of disodium ethylenediamine tetraacetate, 45.391mg of potassium iodide, 13.672mg of sodium molybdate, 1.367mg of copper sulfate, 1.367mg of cobalt chloride, 1219.542mg of manganese sulfate, 470.317mg of zinc sulfate and 339.066mg of boric acid are dissolved in 35L of ultrapure water), the seedlings can grow rapidly under proper temperature, the sprouts can grow into small plants with the plant height of 7-10cm, the stem thickness of 0.12cm and 6-7 nodes of stem nodes after being emitted from leaf axils, and the small plants can be used as virus-free strong seedlings for segment propagation, water culture potato production, original seed production or original seed production.
Hardening seedlings before S007 transplantation: the water culture seedling production greenhouse has the advantages of large space, smooth air circulation, sufficient illumination, strong growth of potato virus-free seedlings, no seedling hardening for other greenhouse cuttage, more than 99.8 percent of cuttage seedling survival rate and strong growth. Opening a greenhouse window before transplanting to the field, allowing the hydroponic seedlings to naturally grow for 2-3 days, and then transplanting to the field.
S008 potato virus-free stock production: the method can be realized by transplanting S009 water-cultured seedlings to greenhouse medium culture stock seeds, culturing S010 water-cultured seedlings to water-cultured potatoes by liquid nutrient solution, and culturing the stock seeds by the greenhouse medium for S011 years.
S009 original strain cultured in greenhouse medium: adopts isolation facilities such as a greenhouse, a net shed and the like and is provided with an insect-proof net with more than 40 meshes. Removing weeds and leveling the soil of a sunlight greenhouse, a self-control greenhouse or a net shed, and uniformly spreading quicklime and a pesticide and a bactericide (Lespeben, carbendazim and the like). Selecting new sterile bricks on the finished ground surface, dividing the new sterile bricks into cells or manufacturing a bracket, placing a proper PC culture tray, pouring a substrate (mainly vermiculite, wherein the pH value of the vermiculite is required to be less than or equal to 8.2, the particle diameter is 2-4 mm, the substrate is fluffy and has less impurities) into the culture tray, wherein the thickness of the substrate is 6-8 cm, thoroughly watering the substrate, and then loosening and scraping the substrate. And (3) preparing sterilized clothes, caps, gloves and masks, 75% alcohol or 10% lime water in the isolation room. Transplanting the water culture seedlings cultivated and exercised by natural illumination into vermiculite, planting the seedlings with the row spacing of 6cm multiplied by 7cm into a matrix with the depth of 2cm-2.5cm, preferably exposing 2 leaves of the vermiculite during transplanting, and spraying little water after planting. After transplanting, the shading net shades the sun for 5 to 7 days at the temperature of 22 to 25 ℃ and the relative humidity of 95 percent. And (3) covering the transplanted water culture seedlings by a small plastic arched shed (preferably a white shed film), and keeping moisture and heat for 7-12 days. The humidity of the vermiculite is kept at 100 percent in the moisture-keeping stage, and the temperature is 25-28 ℃ and is not more than 30 ℃. And (5) removing the small arched shed after the water culture seedlings take roots and survive. Watering and subculturing to proliferate (strengthen seedlings) nutrient solution in time according to the conditions of seedlings and vermiculite, wherein the water content of the substrate in the whole growth period is kept between 50 and 70 percent, the greenhouse is strictly managed during production, smoking is strictly prohibited, and idle personnel are prohibited to enter the greenhouse, and the entering personnel must wear sterilized clothes, and the sterilization treatment is well carried out to prevent the invasion of disease sources.
S010 hydroponic seedling liquid nutrient solution culture hydroponic potato: in the intelligent water culture greenhouse, 1 season of water-cultured potatoes are produced in the idle time of autumn and winter, wherein the idle time is from the first ten days of 9 months to the middle and last ten days of the next 2 months. The culture dish is made of PC plate, the culture medium adopts subculture multiplication (strong seedling) nutrient solution, and the culture dish is periodically replaced for 20-25 days. The virus-free test-tube plantlet is about 20-25 days old, 7-10cm high, 0.6-0.8mm thick stem, 5-8 leaves in terms of 2400 plants/m2The density of (A) is inserted in a PC culture plate. The temperature of the greenhouse is kept at 20 +/-5 ℃ in the daytime, the illumination intensity is kept at 3500-. The produced hydroponic potatoes can be used for producing and culturing stock seeds in a greenhouse and can also be used for producing stock seeds in a field with high altitude by straight pulling.
The production method of the S012 stock comprises the following steps: the method is realized by S013 stock high-mountain breeding and S014 hydroponic seedling high-mountain breeding.
S013 original stock mountain breeding specifically comprises the following steps: the planting density of the original seeds is 5000 grains/mu, 60kg of potassium sulfate compound fertilizer is applied as base fertilizer before each mu of planting, 7.5kg of urea is additionally applied to each mu in the later period, the field is cultivated and hogged in due time to prevent and treat plant diseases and insect pests, and 10g of paclobutrazol is sprayed per mu before flowering to prevent the excessive growth of plants.
In S014 spring, planting the virus-free water-cultured strong seedlings or virus-free water-cultured potatoes in a field with the altitude of 1200-1800m original seed base, controlling the original seed production through density control, wherein the planting density of the virus-free water-cultured strong seedlings is 12000-15000 plants/mu, 15kg of potassium sulfate compound fertilizer is applied as base fertilizer before each mu of planting, and 7.5kg of urea is applied in later period per mu. The planting density of the detoxified water-cultured potatoes is 5000 grains/mu, 60kg of potassium sulfate compound fertilizer is applied as base fertilizer before each mu of planting, 7.5kg of urea is additionally applied to each mu in the later period, the field is timely intertillage and weeding are carried out to prevent and control plant diseases and insect pests, and 10g of paclobutrazol is sprayed per mu before flowering to prevent the excessive growth of plants. Every 667m in the field2When 1.5 ten thousand of virus-free water culture strong seedlings are planted, the ratio of the harvested seed potatoes to the standard seed potatoes (10g-20g) reaches 3.94 ten thousand seeds/667 m respectively2、87.98％。
The production method of the improved S015 seeds comprises the following steps: the stock produced by the steps S013 and S014 is bred in the second mountain or mountain breeding way of the S016 stock.
S016 breeding of two high mountains or two high mountains of the original variety: after the previous crops are harvested, timely ploughing and cleaning the stubbles, and deeply ploughing the soil by over 25 cm; the fertilizing mode takes base fertilizer as a main part and additional fertilizer as an auxiliary part, and is to increase the application of foliar fertilizer properly, wherein the base fertilizer takes fermented and decomposed farmyard manure and biological organic fertilizer as key points, and 1500 kilograms of farmyard manure and 100 kilograms of biological organic fertilizer are applied per mu; the special compound fertilizer or compound fertilizer for the potatoes is applied by 50-100 kg per mu of land which is lack of farmyard manure, and is matched with 200 kg of biological organic fertilizer as a base fertilizer to be applied into furrows at one time. The method is suitable for planting about 5000 holes of the original seeds in each mu of fine seed production, and the direct seeding of the seed potatoes is concerned with seedling searching and filling.
S017 comprehensive regulation and control of the third-level propagation donor system of the potato virus-free seed potato: establishing a three-stage propagation donor system model of the virus-free potato seeds by using the seedling (seed) demand and the production seed potato quantity data in the periodic production process of each stage of potato seeds, and timely regulating and controlling key steps of S007-S014 and the like according to the actual production demand to obtain the corresponding last-stage seed supply quantity required by the production of each stage of potato seeds.
TABLE 1 hydroponics of potato seedlings inducing root primordium formation nutrient solution preparation
Note: the preparation of the nutrient solution comprises two steps of preparation of stock solution and preparation of the nutrient solution; A. b, C indicates different stock solutions, which need to be prepared separately, after the stock solution is prepared, the stock solution can be used after being mixed uniformly according to the corresponding volume ratio of A, B, C stock solution dosage and ultrapure water; tables 2 and 3 are the same.
TABLE 2 preparation of rooting nutrient solution for hydroponic potato seedlings
TABLE 3 preparation of nutrient solution for subculture multiplication (strong seedling)
The embodiments of the present invention have been described in detail with reference to the above examples, but the present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the scope of the present invention, which is defined in the appended claims.
Claims (6)
1. The construction method of the breeding system of the virus-free seed potatoes is characterized by comprising the following steps:
s001 screening and culturing of potato virus-free basic seedlings: the method comprises the steps of screening and culturing S002 enhanced detoxification base seedlings, selecting S003 detoxification base seedlings, culturing S004 detoxification seedlings, segmenting and propagating S005 detoxification seedlings, producing and expanding propagation S006 water culture seedlings, and hardening seedlings before S007 transplanting;
s008 potato virus-free stock production: transplanting the S009 hydroponic seedlings to a greenhouse medium to culture original seeds, culturing the S010 hydroponic seedlings to culture potato by liquid nutrient solution, and culturing the original seeds by the S011 greenhouse medium for the next year;
the production method of the S012 stock comprises the following steps: the method is realized by S013 stock high-mountain breeding and S014 water culture seedling high-mountain breeding;
the production method of the improved S015 seeds comprises the following steps: the stock produced by the steps S013 and S014 is bred in the second mountain or mountain breeding way of the S016 stock.
S017 comprehensive regulation and control of the third-level propagation donor system of the potato virus-free seed potato: constructing a three-level breeding donor system model of the virus-free potato seeds by using the seedling quantity and the production seed potato quantity data of the periodic production process of the potato seeds at all levels, and timely regulating and controlling the S007-S014 key steps according to the actual production requirements to obtain the corresponding last-level seed supply quantity required by the production of the potato seeds at all levels.
2. The method for constructing a breeding system of potato virus-free seed potatoes as claimed in claim 1, wherein the screening and cultivation of the S002 enhanced virus-free base seedlings specifically comprises: taking stem tip meristem detoxification culture and virus detection as quality assurance, using a tissue culture room to detoxify the stem tip meristem of seed potatoes, inducing the seed potatoes in a test tube to form virus-free test-tube seedlings, then carrying out virus detection one by one, expanding and propagating the virus-free test-tube seedlings qualified in virus detection into wide-mouthed bottles, and randomly extracting 1 in each 10 wide-mouthed bottles for virus detection and screening to ensure that the basic seedlings are really virus-free;
the S003 virus-free basic seedling is selected specifically as follows: preferentially selecting a robust detoxified test-tube seedling as a basic seedling for rapid propagation of the detoxified seedling;
the culture of the S004 virus-free seedlings specifically comprises the following steps: using a test-tube seedling culture container, adopting an MS culture medium, culturing the virus-free seedlings at 22-25 ℃, illuminating for 16 hours every day with illumination intensity of 2000-;
the S005 virus-free seedling cutting propagation specifically comprises the following steps: performing single-section cutting cuttage when the virus-free seedlings grow to 6-7 sections, and then inoculating the virus-free seedlings into a culture box for propagation; the strong virus-free test-tube plantlet is inserted in an MS culture medium and takes root in 7-9 days, the plantlet can grow into 2-3 leaves 11-13 days after the young bud is emerged from the leaf axilla, a stem 6-7 joints can be formed in 20-25 days, the height of the plantlet is 7-10cm, the stem thickness is 0.6-0.8mm, and the number of the leaves is 5-8 plantlets, the virus-free plantlet can grow rapidly at proper temperature and can be used as a virus-free strong plantlet to perform water culture plantlet production or cut propagation;
the S006 water culture seedling production and propagation specifically comprises the following steps: cutting detoxicated seedlings into sections, performing cutting water culture, inducing the water culture cutting seedlings to root by a rooting nutrient solution in an intelligent greenhouse, enabling the original roots to grow 3 days after cutting, enabling the original roots to root 5-7 days after cutting, adding the rooting water culture cutting seedlings into a growth nutrient solution, enabling the water culture seedlings to grow rapidly at proper temperature, enabling buds to grow into small plants with the plant height of 7-10cm, the stem thickness of 0.12cm and 6-7 nodes of stems after the buds grow from leaf axils for 10-12 days, and enabling the small plants to be used as detoxified strong seedlings to perform section propagation, water culture potato production, original stock production or stock production;
the hardening-seedling before the S007 transplantation specifically comprises the following steps: opening a greenhouse window before transplanting to the field, allowing the hydroponic seedlings to naturally grow for 2-3 days, and then transplanting to the field.
3. The method for constructing the potato virus-free seed breeding system as claimed in claim 1, wherein the transplanting of the S009 hydroponic seedlings to the greenhouse medium culture stock comprises: adopting greenhouse or net shed isolation facilities and an insect-proof net with more than 40 meshes, removing weeds and leveling the soil of a sunlight greenhouse, a self-control greenhouse or a net shed, and uniformly spreading quicklime, an insecticide and a bactericide; dividing the prepared ground surface into cells or placing a culture tray, putting a substrate with the thickness of 6-8 cm, thoroughly watering, loosening, and scraping the substrate; transplanting the water culture seedlings which are cultured and tempered by natural illumination into vermiculite, planting the seedlings into a matrix with the row spacing of 6cm multiplied by 7cm and the depth of 2cm-2.5cm, wherein the seedlings are preferably transplanted with 2 leaves of vermiculite, and spraying little water after planting; shading with a shading net for 5-7 days after transplanting, at the temperature of 22-25 ℃ and the relative humidity of 95%; covering the transplanted water culture seedlings with a small plastic arch shed, and keeping moisture for 7-12 days; in the moisture-keeping stage, the humidity of the vermiculite is kept at 100%, the temperature is 25-28 ℃, and the humidity is not higher than 30 ℃; removing the small arched shed after the water culture seedlings take roots and survive; watering and subculturing nutrient solution in time according to the conditions of the seedlings and the vermiculite, wherein the water content of the substrate is kept between 50 and 70 percent in the whole growth period, and strict management is carried out in a greenhouse during the production period;
the water culture of the S010 water culture seedlings by the liquid nutrient solution is as follows: in the intelligent water culture greenhouse, 1 season of water-cultured potatoes are produced in the idle time of autumn and winter, wherein the time is from the first ten days of 9 months to the middle and last ten days of 2 months of the next year; the culture medium adopts strong seedling nutrient solution, and is replaced periodically for 20-25 days; the virus-free test-tube plantlet is 20-25 days old, 7-10cm high, 0.6-0.8mm thick stem, 5-8 leaves, 2400 plants/m2The density of the culture medium is inserted in a culture disc; the temperature of the greenhouse is kept at 20 +/-5 ℃ in the daytime, the illumination intensity is kept at 3500-; the produced hydroponic potatoes can be used for producing and culturing stock seeds in a greenhouse and can also be used for planting and producing stock seeds in high-altitude fields.
4. The method for constructing the breeding system of the detoxified potato seeds of claim 3, wherein the bactericide in step S009 is Lespeben or carbendazim, the matrix comprises vermiculite and compound fertilizer, the vermiculite is mixed with 1kg of compound fertilizer according to 2000kg of vermiculite, the pH value of the vermiculite is required to be less than or equal to 8.2, the particle diameter is 2mm-4mm, and the vermiculite is fluffy and has less impurities.
5. The method of claim 1 wherein said seed potato is used in a potato de-virulent breeding systemThe construction method is characterized in that the S011 next-year greenhouse medium culture stock comprises the following steps: the planting density is 40 plants/square meter, the net room is watered regularly to prevent and control plant diseases and insect pests, the paclobutrazol is sprayed at 10 g/mu before flowering to prevent the plants from growing excessively, and the planting density in the net room is 40 plants/m2The number of the potatoes and the ratio of the standard seed potatoes reach 100.5 grains/m respectively286.59 percent; the S013 original stock mountain breeding specifically comprises the following steps: the planting density of the original seeds is 5000 grains/mu, 60kg of potassium sulfate compound fertilizer is applied as base fertilizer before each mu of planting, 7.5kg of urea is additionally applied to each mu in the later period, the field is timely intertillage and weeding are carried out to prevent and control plant diseases and insect pests, and 10 g/mu of paclobutrazol is sprayed before flowering;
the alpine breeding of the S014 water culture seedlings is specifically as follows: planting virus-free water culture strong seedlings or virus-free water culture potatoes in a field with the altitude of 1200-1800m original seed base in spring, controlling the original seed production through density control, wherein the planting density of the virus-free water culture strong seedlings is 12000-15000 plants/mu, applying 15kg of potassium sulfate compound fertilizer as base fertilizer before each mu of planting, and applying 7.5kg of urea at the later stage; the planting density of the detoxified water-cultured potatoes is 5000 grains/mu, 60kg of potassium sulfate compound fertilizer is applied as base fertilizer before each mu of planting, 7.5kg of urea is additionally applied to each mu in the later period, the field is timely intertillage and weeding are carried out to prevent and control plant diseases and insect pests, and 10 g/mu of paclobutrazol is sprayed before flowering; every 667m in the field2When 1.5 ten thousand of virus-free water culture strong seedlings are planted, the ratio of the harvested seed potatoes to the standard seed potatoes reaches 3.94 ten thousand seeds/667 m respectively2、87.98％。
6. The method for constructing a breeding system of a virus-free potato seed of claim 1, wherein the breeding of S016 elite II mountain or mountain is as follows: after the previous crops are harvested, timely ploughing and cleaning the stubbles, and deeply ploughing the soil by over 25 cm; the fertilizing mode takes a base fertilizer as a main part and takes additional fertilizer as an auxiliary part, and the additional application of a foliar fertilizer is properly carried out, wherein the base fertilizer takes fermented and decomposed farmyard manure and biological organic fertilizer as key points, and 1500 kilograms of farmyard manure and 120 kilograms of biological organic fertilizer are applied per mu; the method is characterized in that the farmyard manure is deficient, 50-100 kg of special compound fertilizer or compound fertilizer for potatoes is applied to each mu, 100-200 kg of biological organic fertilizer is matched to be used as base fertilizer to be applied to furrows at one time, 4000-5000 holes of original seeds are planted in each mu in fine seed production, and attention is paid to direct seeding and shortage is paid to seed searching.
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| CN112335520A (en) * | 2020-11-03 | 2021-02-09 | 刘克锋 | Potato virus-free seedling culture substrate produced by mechanical composition method and manufacturing method thereof |
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| CN115088569A (en) * | 2022-06-13 | 2022-09-23 | 江苏宝德农业科技有限公司 | Breeding method of virus-free potato seeds |
| CN115039682A (en) * | 2022-07-07 | 2022-09-13 | 湖北恩施中国南方马铃薯研究中心 | Breeding method for quickly obtaining large number of potato seed potatoes |
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