CN111448986A - Production and preparation method of virus-free seed potatoes - Google Patents

Abstract

The invention discloses a production and preparation method of a virus-free seed potato, which comprises the following steps: selecting materials, sterilizing, peeling and inoculating stem tips, culturing basic seedlings, identifying viruses, expanding propagation, transplanting, harvesting and sorting stock seeds and storing the stock seeds. The invention has the beneficial effects that: high production efficiency, good detoxification effect, complete steps, strong operability, accordance with the growth characteristics and habits of the potatoes, and wide popularization.

Description

Production and preparation method of virus-free seed potatoes

The technical field is as follows:

the invention relates to the field of agriculture, in particular to a production and preparation method of virus-free seed potatoes.

Background art:

the potatoes are susceptible to virus infection in the planting process, and when the conditions are proper, the potatoes can proliferate, transport and accumulate in the potato blocks in the plants, and are transmitted by generations and aggravated year by year. The growth potential of the plants is obviously deteriorated, tubers are reduced, the yield is continuously reduced, the quality is gradually deteriorated, the edibility and the commodity are seriously influenced, the varieties lose the original excellent properties, and the degeneration phenomenon of the potatoes is caused. The multiplication of potato virus is very close to the normal metabolism of plants, and no medicament capable of killing the virus and not damaging the plants is found at present, so that the virus hazard is an incurable symptom of the potato at one time. The rapid development of the shoot tip tissue culture virus-free plant technology provides an effective way for solving the harm of the potato virus. The production of healthy virus-free seed potatoes by using a tissue culture technology becomes one of the most advanced technical measures for solving the potato degeneration in the current production. The yield, quality and stress resistance of the planted detoxified seed potatoes are improved to a great extent, so that the tissue culture technology becomes an important means for improving the productivity of the potatoes.

The invention content is as follows:

the invention aims to provide a production and preparation method of potato virus-free seed potatoes, which has the advantages of high production efficiency, good virus-free effect, complete steps, strong operability, accordance with the growth characteristics and habits of potatoes and large-scale popularization.

The invention is realized by the following technical scheme:

the invention relates to a production and preparation method of a virus-free seed potato, which comprises the following steps:

step 1, selecting materials, selecting a single plant with original seed characteristic characteristics in the potato growing season, soaking the selected tubers for 20min by 0.50-1mg/kg gibberellin after harvesting, and then placing the tubers on a sand bed in a greenhouse or planting the tubers in sterile pot soil for bud cultivation; or cutting the explant from the field plant, and soaking the terminal bud or lateral bud together with part of the petiole and stem segment in 2-10% sodium hypochlorite solution for 10-15 min; shearing the cultured material to obtain 2-3cm bud tip when the bud grows to 5cm, and removing the visible leaf with forceps to obtain stem tip raw material;

step 2, sterilizing, namely wrapping the stem tip raw material obtained in the step 1 by using gauze, placing the wrapped material in tap water for washing for 25-35min, soaking the wrapped material in 0.10-0.20% mercuric chloride solution for 8-10min in a sterile room, sterilizing the wrapped material by using 70% alcohol for 10-20s, finally washing the wrapped material by using sterile water for 3-5 times, and sucking water on the surface of the wrapped material by using sterilized filter paper;

step 3, stripping and inoculating the stem tip, wherein the stem tip is inserted into a culture medium in a test tube or a triangular flask after being stripped by 0.1-0.3 mm;

step 4, culturing the basic seedlings, namely placing the inoculated test tubes or triangular flasks in a culture room for culture;

step 5, virus identification, namely carrying out virus identification on the seedlings which are stripped and cultured through the stem tips and cut into segments and propagated for a plurality of bottles, and identifying virus-free seedlings without viruses through indicator plants for use in the step 6;

step 6, expanding propagation, namely, taking the virus-free seedlings without main viruses after identification into an inoculation chamber for inspection and sorting, cutting the virus-free seedlings, and then putting the cut virus-free seedlings on a culture medium into a culture chamber for culture to obtain tissue culture seedlings;

step 7, transplanting, namely, selecting robust pollution-free tissue culture seedlings with 23-27 days of seedling age, 4-5cm of plant height and more than 3 leaves, hardening the seedlings in a natural light greenhouse, cutting the seedlings into a seedling bed, and paving vermiculite;

8, harvesting and sorting original seeds, wherein after the plants grow for 80-120 days, 85% of potato blocks grow to be more than 2.0 g and then can be harvested; stopping watering and nutrient solution 10-15 days before harvesting, and harvesting after vermiculite is dried; when in harvest, the plants are pulled out, the potato blocks are shaken off, the potato blocks on the surface of vermiculite are picked up, and then the rest potato blocks are screened and collected; the harvested potato blocks are air-dried, aired, aged, sieved and classified according to sizes, and packaged into breathable packaging bags.

And 9, storing the original seeds, and after the storage chamber is disinfected, placing the seeds according to the variety, specification and quantity specification after the prestoring is finished.

Preferably: step 3, the stem tip stripping and inoculation comprises the following steps:

step a, sterilizing an inoculation chamber, cleaning the ground of the inoculation chamber, spraying 70% alcohol for dust reduction, and irradiating with an ultraviolet lamp for 20 min;

step b, disinfecting tools and a table board, and thoroughly cleaning hands, a dissecting mirror, a dissecting needle, tweezers, a blade and an ultra-clean working table board by using 70% alcohol or benzalkonium bromide;

c, dissecting and inoculating, namely taking the forceps fixing material with the left hand and taking the dissecting needle with the right hand simultaneously to peel off the young leaves layer by layer under a dissecting mirror until a growth point with two leaf primordia is exposed, cutting off the stem tip with only one small leaf primordia, and quickly inoculating the stem tip on a culture medium subjected to high-pressure sterilization, wherein 1 stem tip is inoculated in each bottle; the scalpel, the dissecting needle and the forceps inoculation tool are used once and then are soaked in 70% alcohol, and then are burned and cooled for standby;

d, packaging, namely rotating and burning the gauze cotton balls on flame for 2-8 seconds, immediately plugging the bottle openings with the gauze cotton balls after inoculation is finished, wrapping the gauze cotton balls with tinfoil paper or kraft paper, fastening the gauze cotton balls with ropes or rubber bands, marking, and noting the material names and the inoculation dates.

Preferably: the culture medium used in the stem tip stripping and inoculation in the step 3 is an MS solid culture medium; step 4, in the culture of the basic seedlings, the initial illumination intensity of the culture is set to be 1000lx, the illumination intensity is increased to 2000lx after 4 weeks, and the illumination intensity is increased to 4000lx after the stem tip grows to 1cm and the height is 5-6 weeks; the illumination time is 16-20 h; the culture temperature is 23-27 ℃; when the stem tip tissue turns green, transferring the stem tip tissue to a half-amount MS culture medium added with 0.50mg2.4-D and 1mg ZT per liter for further culture; after callus proliferation and formation of multiple bud primordia, transfer to hormone-free half-amount MS culture medium.

Preferably: in the step 5, in virus identification, after cutting and breeding a plurality of bottles of seedlings cultured by stem tip stripping, one part of offspring of each plant is reserved for preservation, and the other part is subjected to virus identification; and (3) eliminating all positive reaction patients after detection by a serological method, identifying the plants with negative reaction by using an indicator plant, and identifying virus-free seedlings by using the indicator plant for propagation expansion in the step (6).

Preferably: the expanding propagation in step 6 comprises the following steps:

e, preparing materials, eliminating the polluted virus-free seedlings, and classifying and numbering the healthy virus-free seedlings according to varieties; sterilizing a plurality of 150ml triangular bottles or wide-mouth bottles, gauze, cotton balls, tinfoil, kraft paper, rubber bands, surgical scissors, culture dishes and alcohol lamps; then adding 30ml of MS culture medium without any growth regulator into each bottle, and carrying out autoclaving treatment;

f, cutting and cuttage, namely cutting the non-toxic seedlings out of the bottles on a clean bench, cutting the non-toxic seedlings into stem sections with one leaf in a culture dish, and uniformly laying or cuttage on a culture medium, wherein 25 seedlings are put in each bottle;

step g, packaging and culturing, namely covering a bottle cover, marking date, variety and name, and then sending the bottle cover into a culture room for culturing to obtain test-tube plantlets;

step h, culturing strong seedlings in a culture room, namely reducing the culture temperature of the test-tube seedlings propagated for the last time to 18-20 ℃, increasing the illumination intensity to 3000-; and trace elements and organic components in the MS culture medium are subtracted, and chlormequat chloride is additionally added;

and step i, hardening the seedlings in a culture room, and removing the bottle stoppers of the triangular bottles or the wide-mouth bottles 7 days before planting to harden the seedlings to obtain the tissue culture seedlings.

Preferably: in the step f, in the cutting and cuttage, when the nontoxic seedlings in the bottle are cut off and taken out, a leaf is remained at the base part of the small seedling, and 1/3 culture medium is added; and g, in the packaging culture, setting the temperature of the culture chamber to be 25-28 ℃, setting the illumination condition to be continuously irradiated for 7 days under 1000-1500lx, and repeating the step 6 to expand and propagate until the required number is reached after the plantlets grow into 3-5 leaves.

Preferably: 7, in the transplanting, hardening the seedlings for 4-6 days, setting the temperature to be 15-20 ℃ and setting the distance between vessels to be 5 cm;

meanwhile, the seedbed is treated, namely phoxim and carbendazim are sprayed, 80-mesh insect-proof nets and 8cm vermiculite are paved, two bags of organic fertilizer and 40kg of compound fertilizer are added into the vermiculite in each shed, and the mixture is uniformly stirred; the watering amount reaches 95 percent, and the water is leveled for use;

then, cutting, namely soaking the tweezers in potassium permanganate, disinfecting the arms of operators by potassium permanganate, soaking the tissue culture seedlings in rooting powder for 1 minute for cutting, wherein the cutting depth is 2-2.5 cm; after cuttage, agricultural streptomycin and Kelu spraying are used, and a small arched shed is built for heat preservation and moisture preservation; meanwhile, setting up a sunshade net for 5-7 days, setting the temperature in the shed to be 22-25 ℃, the relative humidity to be 95 percent and the temperature in the small arch shed to be 25-28 ℃; removing the arched shed after rooting for 7 to 12 days;

watering and nutrient solution every 7-10 days after the arched shed is removed, so as to keep the water content of the matrix at 60-70%; when the plant is raised to 7-8 leaves, vermiculite with the thickness of 5-6cm is cultured.

Preferably: in the step 9 of stock seed storage, firstly, pre-storing harvested potato blocks in a ventilated and dried storehouse for 15-20 days; simultaneously, before storage, the storage is fumigated by formaldehyde, spread with quicklime and sprayed with a bactericide for disinfection treatment; the temperature of the storage is set to be 2-4 ℃, the humidity is 75-85%, and the interior of the storage is ventilated regularly and kept clean.

Preferably: in the cutting and cuttage in the step f, firstly, the basic seedling bottle and the culture medium bottle are opened, the bottle caps cannot be used in a mixed mode, and the cover openings are upward when the basic seedling bottle and the culture medium bottle are placed on the table top;

then, shearing basic seedlings in the basic seedling bottles by using scissors after disinfection and cooling, and placing the basic seedlings in the culture medium bottles, wherein the scissors need to disinfect once when each basic seedling is sheared, and the tweezers need to disinfect once when each tissue culture seedling is cut, and the disinfection time is 30 seconds;

during cuttage, each section is 1CM long with at least one leaf, 25 leaves are placed in each bottle of the stem tip, and 25 leaves are placed in each bottle of the stem section; after the cuttage is finished, the bottle body is marked, and the written date, variety and shearer number are sent into a culture room in a unified way.

The invention has the beneficial effects that: the method has the advantages of high production efficiency, good detoxification effect, complete steps, strong operability, accordance with the growth characteristics and habits of the potatoes, and large-scale popularization.

The specific implementation mode is as follows:

the invention will be further illustrated with reference to specific embodiments:

example (b): a production method of a virus-free seed potato of potato comprises the following steps:

step 1, selecting materials, selecting a single plant with original seed characteristic characteristics in the potato growing season, soaking the selected tubers for 20min by 0.75mg/kg gibberellin after harvesting, and then placing the tubers on a sand bed in a greenhouse or planting the tubers in sterile pot soil for bud cultivation; or cutting the explant from the field plant, and soaking the terminal bud or lateral bud together with part of petiole and stem segment in 5% sodium hypochlorite solution for 12 min; shearing 2.5cm bud tips of the cultured material when the buds grow to 5cm, and peeling off the visible leaves by using tweezers to obtain a stem tip raw material;

step 2, sterilizing, namely wrapping the stem tip raw material obtained in the step 1 by using gauze, placing the wrapped material in tap water for washing for 30min, then soaking the wrapped material in 0.15% mercuric chloride solution for 10min in a sterile room, sterilizing the wrapped material by using 70% alcohol for 15s, finally washing the wrapped material by using sterile water for 5 times, and absorbing the water on the surface of the wrapped material by using sterilized filter paper;

step 3, stripping and inoculating the stem tip, wherein the stem tip is inserted into a culture medium in a test tube or a triangular flask after being stripped by 0.1-0.3 mm; specifically, the method comprises the following steps:

step a, sterilizing an inoculation chamber, cleaning the ground of the inoculation chamber, spraying 70% alcohol for dust reduction, and irradiating with an ultraviolet lamp for 20 min;

step b, disinfecting tools and a table board, and thoroughly cleaning hands, a dissecting mirror, a dissecting needle, tweezers, a blade and an ultra-clean working table board by using 70% alcohol or benzalkonium bromide;

c, dissecting and inoculating, namely taking the forceps fixing material with the left hand and taking the dissecting needle with the right hand simultaneously to peel off the young leaves layer by layer under a dissecting mirror until a growth point with two leaf primordia is exposed, cutting off the stem tip with only one small leaf primordia, and quickly inoculating the stem tip to the MS solid culture medium subjected to high-pressure sterilization, wherein 1 stem tip is inoculated in each bottle; the scalpel, the dissecting needle and the forceps inoculation tool are used once and then are soaked in 70% alcohol, and then are burned and cooled for standby;

and d, packaging, namely rotating and burning the gauze cotton balls on flame for 6 seconds, immediately plugging the bottle openings with the gauze cotton balls after inoculation is finished, wrapping the gauze cotton balls with tinfoil paper or kraft paper, fastening the gauze cotton balls with cotton ropes or rubber bands, marking, and noting the material names and the inoculation dates.

Step 4, culturing the basic seedlings, namely placing the inoculated test tubes or triangular flasks in a culture room for culture; specifically, the initial illumination intensity of the culture was set to 1000lx, and increased to 2000lx after 4 weeks, and when the stem tip was grown to 1cm in height and 6 weeks, the illumination intensity was increased to 4000 lx; the illumination time is 18 h; the culture temperature is 25 ℃; when the stem tip tissue turns green, transferring the stem tip tissue to a half-amount MS culture medium added with 0.50mg2.4-D and 1mg ZT per liter for further culture; after callus proliferation and formation of multiple bud primordia, transfer to hormone-free half-amount MS culture medium.

Step 5, virus identification, namely carrying out virus identification on the seedlings which are stripped and cultured through the stem tips and cut into segments and propagated for a plurality of bottles, and identifying virus-free seedlings without viruses through indicator plants for use in the step 6; specifically, after cutting and propagating the seedlings which are subjected to stem tip stripping culture for several bottles, a part of offspring of each plant is reserved for preservation, and the other part is subjected to virus identification; and (3) eliminating all positive reaction patients after detection by a serological method, identifying the plants with negative reaction by using an indicator plant, and identifying virus-free seedlings by using the indicator plant for propagation expansion in the step (6).

Step 6, expanding propagation, namely, taking the virus-free seedlings without main viruses after identification into an inoculation chamber for inspection and sorting, cutting the virus-free seedlings, and then putting the cut virus-free seedlings on a culture medium into a culture chamber for culture to obtain tissue culture seedlings; specifically, the method comprises the following steps:

e, preparing materials, eliminating the polluted virus-free seedlings, and classifying and numbering the healthy virus-free seedlings according to varieties; sterilizing a plurality of 150ml triangular bottles or wide-mouth bottles, gauze, cotton balls, tinfoil, kraft paper, rubber bands, surgical scissors, culture dishes and alcohol lamps; then adding 30ml of MS culture medium without any growth regulator into each bottle, and carrying out autoclaving treatment;

f, cutting and cuttage, namely cutting the non-toxic seedlings out of the bottles on a clean bench, cutting the non-toxic seedlings into stem sections with one leaf in a culture dish, and uniformly laying or cuttage on a culture medium, wherein 25 seedlings are put in each bottle; when the nontoxic seedlings in the bottle are cut off and taken out, a leaf is remained at the base part of the small seedling, and 1/3 culture medium is added; and g, in the packaging culture, setting the temperature of the culture room to be 26 ℃, setting the illumination condition to be 1200lx, continuously irradiating for 7 days, and repeating the step 6 to expand the propagation till the required number is reached after the plantlets grow into 3-5 leaves.

Step g, packaging and culturing, namely covering a bottle cover, marking date, variety and name, and then sending the bottle cover into a culture room for culturing to obtain test-tube plantlets;

step h, culturing strong seedlings in a culture room, namely, reducing the culture temperature of the test-tube seedlings propagated at the last time to 18 ℃, improving the illumination intensity to 3500lx, and setting the illumination time to be 20h every day; and trace elements and organic components in the MS culture medium are subtracted, and chlormequat chloride is additionally added;

and step i, hardening the seedlings in a culture room, and removing the bottle stoppers of the triangular bottles or the wide-mouth bottles 7 days before planting to harden the seedlings to obtain the tissue culture seedlings.

Step 7, transplanting, namely, selecting robust pollution-free tissue culture seedlings with the seedling age of 25 days, the plant height of 4-5cm and more than 3 leaves, hardening the seedlings in a natural light greenhouse, and then cutting the seedlings into a seedling bed; the seedling exercising time is 6 days, the temperature is set to be 18 ℃, and the distance between vessels is set to be 5 cm; meanwhile, the seedbed is treated, namely phoxim and carbendazim are sprayed, 80-mesh insect-proof nets and 8cm vermiculite are paved, two bags of organic fertilizer and 40kg of compound fertilizer are added into the vermiculite in each shed, and the mixture is uniformly stirred; the watering amount reaches 95 percent, and the water is leveled for use;

then, cutting, namely soaking the tweezers in potassium permanganate, disinfecting the arms of operators by potassium permanganate, soaking the tissue culture seedlings in rooting powder for 1 minute for cutting, wherein the cutting depth is 2.5 cm; after cuttage, agricultural streptomycin and Kelu spraying are used, and a small arched shed is built for heat preservation and moisture preservation; meanwhile, a sunshade net is set up for 7 days, the temperature in the greenhouse is set to be 24 ℃, the relative humidity is 95%, and the temperature in the small-arch greenhouse is controlled to be 26 ℃; removing the arched shed after rooting for 10 days; after the arched shed is removed, watering and nutrient solution every 8 days to keep the water content of the matrix at 60-70%; when the plants are raised to 7-8 leaves, vermiculite with a thickness of 6cm is cultivated.

8, harvesting and sorting original seeds, wherein after the plants grow for 80-120 days, 85% of potato blocks grow to be more than 2.0 g and then can be harvested; stopping watering and nutrient solution 12 days before harvesting, and harvesting after vermiculite is dried; when in harvest, the plants are pulled out, the potato blocks are shaken off, the potato blocks on the surface of vermiculite are picked up, and then the rest potato blocks are screened and collected; the harvested potato blocks are air-dried, aired, aged, sieved and classified according to sizes, and packaged into breathable packaging bags.

And 9, storing the original seeds, and after the storage chamber is disinfected, placing the seeds according to the variety, specification and quantity specification after the prestoring is finished. Specifically, the harvested potato blocks are pre-stored in a ventilated and dried storehouse for 180 days; simultaneously, before storage, the storage is fumigated by formaldehyde, spread with quicklime and sprayed with a bactericide for disinfection treatment; the temperature of the storage is set to 4 ℃ and the humidity is 80%, and the storage is ventilated regularly and kept clean.

Wherein: in the cutting and cuttage in the step f, firstly, the basic seedling bottle and the culture medium bottle are opened, the bottle caps cannot be used in a mixed mode, and the cover openings are upward when the basic seedling bottle and the culture medium bottle are placed on the table top;

then, shearing basic seedlings in the basic seedling bottles by using scissors after disinfection and cooling, and placing the basic seedlings in the culture medium bottles, wherein the scissors need to disinfect once when each basic seedling is sheared, and the tweezers need to disinfect once when each tissue culture seedling is cut, and the disinfection time is 30 seconds;

during cuttage, each section is 1CM long with at least one leaf, 25 leaves are placed in each bottle of the stem tip, and 25 leaves are placed in each bottle of the stem section; after the cuttage is finished, the bottle body is marked, and the written date, variety and shearer number are sent into a culture room in a unified way.

Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments or portions thereof without departing from the spirit and scope of the invention.

Claims (9)

1. A production and preparation method of a virus-free seed potato is characterized by comprising the following steps: the method comprises the following steps:

step 1, selecting materials, soaking tubers in gibberellin of 0.50-1mg/kg for 20min after potatoes are harvested, and then placing the tubers on a sand bed in a greenhouse or planting the tubers in sterile pot soil for bud cultivation; or cutting the explant from the field plant, and soaking the terminal bud or lateral bud together with part of the petiole and stem segment in 2-10% sodium hypochlorite solution for 10-15 min;

shearing the cultured material to obtain 2-3cm bud tip when the bud grows to 5cm, and removing the visible leaf with forceps to obtain stem tip raw material;

step 2, sterilizing, namely wrapping the stem tip raw material obtained in the step 1 by using gauze, placing the wrapped material in tap water for washing for 25-35min, soaking the wrapped material in 0.10-0.20% mercuric chloride solution for 8-10min in a sterile room, sterilizing the wrapped material by using 70% alcohol for 10-20s, finally washing the wrapped material by using sterile water for 3-5 times, and sucking water on the surface of the wrapped material by using sterilized filter paper;

step 3, stripping and inoculating the stem tip, wherein the stem tip is inserted into a culture medium in a test tube or a triangular flask after being stripped by 0.1-0.3 mm;

step 4, culturing the basic seedlings, namely placing the inoculated test tubes or triangular flasks in a culture room for culture;

step 5, virus identification, namely carrying out virus identification on the seedlings which are stripped and cultured through the stem tips and cut into segments and propagated for a plurality of bottles, and identifying virus-free seedlings without viruses through indicator plants for use in the step 6;

step 6, expanding propagation, namely, taking the virus-free seedlings without main viruses after identification into an inoculation chamber for inspection and sorting, cutting the virus-free seedlings, and then putting the cut virus-free seedlings on a culture medium into a culture chamber for culture to obtain tissue culture seedlings;

step 7, transplanting, namely, selecting robust pollution-free tissue culture seedlings with 23-27 days of seedling age, 4-5cm of plant height and more than 3 leaves, hardening the seedlings in a natural light greenhouse, cutting the seedlings into a seedling bed, and paving vermiculite;

8, harvesting and sorting original seeds, wherein after the plants grow for 80-120 days, 85% of potato blocks grow to be more than 2.0 g and then can be harvested; stopping watering and nutrient solution 10-15 days before harvesting, and harvesting after vermiculite is dried; when in harvest, the plants are pulled out, the potato blocks are shaken off, the potato blocks on the surface of vermiculite are picked up, and then the rest potato blocks are screened and collected; the harvested potato blocks are air-dried, aired, aged, sieved and classified according to sizes, and packaged into breathable packaging bags.

And 9, storing the original seeds, and after the storage chamber is disinfected, placing the seeds according to the variety, specification and quantity specification after the prestoring is finished.

2. The method of producing a detoxified seed potato as set forth in claim 1, characterized in that: the step 3 of stem tip stripping and inoculation comprises the following steps:

step a, sterilizing an inoculation chamber, cleaning the ground of the inoculation chamber, spraying 70% alcohol for dust reduction, and irradiating with an ultraviolet lamp for 20 min;

step b, disinfecting tools and a table board, and thoroughly cleaning hands, a dissecting mirror, a dissecting needle, tweezers, a blade and an ultra-clean working table board by using 70% alcohol or benzalkonium bromide;

c, dissecting and inoculating, namely taking the forceps fixing material with the left hand and taking the dissecting needle with the right hand simultaneously to peel off the young leaves layer by layer under a dissecting mirror until a growth point with two leaf primordia is exposed, cutting off the stem tip with only one small leaf primordia, and quickly inoculating the stem tip on a culture medium subjected to high-pressure sterilization, wherein 1 stem tip is inoculated in each bottle; the scalpel, the dissecting needle and the forceps inoculation tool are used once and then are soaked in 70% alcohol, and then are burned and cooled for standby;

d, packaging, namely rotating and burning the gauze cotton balls on flame for 2-8 seconds, immediately plugging the bottle openings with the gauze cotton balls after inoculation is finished, wrapping the gauze cotton balls with tinfoil paper or kraft paper, fastening the gauze cotton balls with ropes or rubber bands, marking, and noting the material names and the inoculation dates.

3. The method for producing potato seeds of claim 2, wherein: the culture medium used in the stem tip stripping and inoculation in the step 3 is an MS solid culture medium; in the step 4, in the culture of the basic seedlings, the initial illumination intensity is set to be 1000lx, the illumination intensity is increased to 2000lx after 4 weeks, and the illumination intensity is increased to 4000lx after the stem tip grows to 1cm and the height is 5-6 weeks; the illumination time is 16-20 h; the culture temperature is 23-27 ℃;

when the stem tip tissue turns green, transferring the stem tip tissue to a half-amount MS culture medium added with 0.50mg2.4-D and 1mg ZT per liter for further culture; after callus proliferation and formation of multiple bud primordia, transfer to hormone-free half-amount MS culture medium.

4. The method of producing a detoxified seed potato as set forth in claim 1, characterized in that: in the step 5, in virus identification, after cutting and breeding a plurality of bottles of seedlings cultured by stem tip stripping, one part of offspring of each plant is reserved for preservation, and the other part is subjected to virus identification; and (3) eliminating all positive reaction patients after detection by a serological method, identifying the plants with negative reaction by using an indicator plant, and identifying virus-free seedlings by using the indicator plant for propagation expansion in the step (6).

5. The method of producing a detoxified seed potato as set forth in claim 1, characterized in that: the step 6 comprises the following steps:

e, preparing materials, eliminating the polluted virus-free seedlings, and classifying and numbering the healthy virus-free seedlings according to varieties; sterilizing a plurality of 150ml triangular bottles or wide-mouth bottles, gauze, cotton balls, tinfoil, kraft paper, rubber bands, surgical scissors, culture dishes and alcohol lamps; then adding 30ml of MS culture medium without any growth regulator into each bottle, and carrying out autoclaving treatment;

f, cutting and cuttage, namely cutting the non-toxic seedlings out of the bottles on a clean bench, cutting the non-toxic seedlings into stem sections with one leaf in a culture dish, and uniformly laying or cuttage on a culture medium, wherein 25 seedlings are put in each bottle;

step g, packaging and culturing, namely covering a bottle cover, marking date, variety and name, and then sending the bottle cover into a culture room for culturing to obtain test-tube plantlets;

step h, culturing strong seedlings in a culture room, namely reducing the culture temperature of the test-tube seedlings propagated for the last time to 18-20 ℃, increasing the illumination intensity to 3000-; and trace elements and organic components in the MS culture medium are subtracted, and chlormequat chloride is additionally added;

and step i, hardening the seedlings in a culture room, and removing the bottle stoppers of the triangular bottles or the wide-mouth bottles 7 days before planting to harden the seedlings to obtain the tissue culture seedlings.

6. The method for producing potato seeds of claim 5, wherein: in the step f, in the cutting and cuttage, when the nontoxic seedlings in the bottles are cut and taken out, a leaf is reserved at the base of each seedling, and 1/3 culture medium is added; and g, in the step g of encapsulation culture, setting the temperature of the culture chamber to be 25-28 ℃, setting the illumination condition to be continuously irradiated for 7 days under 1000-1500lx, and repeating the step 6 to expand and propagate until the required number is reached after the plantlets with 3-5 leaves grow.

7. The method of producing a detoxified seed potato as set forth in claim 1, characterized in that: in the step 7, in the transplanting, the hardening time is 4-6 days, the temperature is set to be 15-20 ℃, and the distance between vessels is set to be 5 cm;

meanwhile, the seedbed is treated, namely phoxim and carbendazim are sprayed, 80-mesh insect-proof nets and 8cm vermiculite are paved, two bags of organic fertilizer and 40kg of compound fertilizer are added into the vermiculite in each shed, and the mixture is uniformly stirred; the watering amount reaches 95 percent, and the water is leveled for use;

then, cutting, namely soaking the tweezers in potassium permanganate, disinfecting the arms of operators by potassium permanganate, soaking the tissue culture seedlings in rooting powder for 1 minute for cutting, wherein the cutting depth is 2-2.5 cm; after cuttage, agricultural streptomycin and Kelu spraying are used, and a small arched shed is built for heat preservation and moisture preservation; meanwhile, setting up a sunshade net for 5-7 days, setting the temperature in the shed to be 22-25 ℃, the relative humidity to be 95 percent and the temperature in the small arch shed to be 25-28 ℃; removing the arched shed after rooting for 7 to 12 days;

watering and nutrient solution every 7-10 days after the arched shed is removed, so as to keep the water content of the matrix at 60-70%; when the plant is raised to 7-8 leaves, vermiculite with the thickness of 5-6cm is cultured.

8. The method of producing a detoxified seed potato as set forth in claim 1, characterized in that: in the step 9, in the original seed storage, the harvested potato blocks are pre-stored in a ventilated and dried storehouse for 15 to 20 days; simultaneously, before storage, the storage is fumigated by formaldehyde, spread with quicklime and sprayed with a bactericide for disinfection treatment; the temperature of the storage is set to be 2-4 ℃, the humidity is 75-85%, and the interior of the storage is ventilated regularly and kept clean.

9. The method of producing a detoxified seed potato as set forth in claim 6, characterized in that: in the step f, in the cutting and cuttage, firstly, the basic seedling bottle and the culture medium bottle are opened, the bottle caps cannot be used together, and the cover openings are upward when the basic seedling bottle and the culture medium bottle are placed on the table top;

then, shearing basic seedlings in the basic seedling bottles by using scissors after disinfection and cooling, and placing the basic seedlings in the culture medium bottles, wherein the scissors need to disinfect once when each basic seedling is sheared, and the tweezers need to disinfect once when each tissue culture seedling is cut, and the disinfection time is 30 seconds;

during cuttage, each section is 1CM long with at least one leaf, 25 leaves are placed in each bottle of the stem tip, and 25 leaves are placed in each bottle of the stem section; after the cuttage is finished, the bottle body is marked, and the written date, variety and shearer number are sent into a culture room in a unified way.