CN115136769B - Sweet potato latent virus removal method - Google Patents
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- CN115136769B CN115136769B CN202210885110.5A CN202210885110A CN115136769B CN 115136769 B CN115136769 B CN 115136769B CN 202210885110 A CN202210885110 A CN 202210885110A CN 115136769 B CN115136769 B CN 115136769B
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- 241001429316 Sweet potato latent virus Species 0.000 title claims abstract description 24
- 238000000034 method Methods 0.000 title claims abstract description 19
- 244000017020 Ipomoea batatas Species 0.000 claims abstract description 40
- 235000002678 Ipomoea batatas Nutrition 0.000 claims abstract description 40
- 238000002791 soaking Methods 0.000 claims abstract description 30
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000011259 mixed solution Substances 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- IWUCXVSUMQZMFG-AFCXAGJDSA-N Ribavirin Chemical compound N1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 IWUCXVSUMQZMFG-AFCXAGJDSA-N 0.000 claims abstract description 13
- 239000005708 Sodium hypochlorite Substances 0.000 claims abstract description 13
- 238000004140 cleaning Methods 0.000 claims abstract description 13
- 229960000329 ribavirin Drugs 0.000 claims abstract description 13
- HZCAHMRRMINHDJ-DBRKOABJSA-N ribavirin Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1N=CN=C1 HZCAHMRRMINHDJ-DBRKOABJSA-N 0.000 claims abstract description 13
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims abstract description 13
- 230000035784 germination Effects 0.000 claims abstract description 8
- 238000012258 culturing Methods 0.000 claims abstract description 7
- 238000011010 flushing procedure Methods 0.000 claims abstract description 7
- 239000002689 soil Substances 0.000 claims abstract description 7
- 239000008223 sterile water Substances 0.000 claims abstract description 7
- 210000002615 epidermis Anatomy 0.000 claims abstract description 5
- 238000005286 illumination Methods 0.000 claims description 24
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 claims description 3
- 230000000052 comparative effect Effects 0.000 description 17
- 238000001784 detoxification Methods 0.000 description 11
- 238000001514 detection method Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000007850 degeneration Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 2
- USSIUIGPBLPCDF-KEBDBYFISA-N pyriminobac-methyl Chemical group CO\N=C(/C)C1=CC=CC(OC=2N=C(OC)C=C(OC)N=2)=C1C(=O)OC USSIUIGPBLPCDF-KEBDBYFISA-N 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 241000700605 Viruses Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/08—Immunising seed
-
- 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
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/48—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with two nitrogen atoms as the only ring hetero atoms
- A01N43/54—1,3-Diazines; Hydrogenated 1,3-diazines
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/64—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with three nitrogen atoms as the only ring hetero atoms
- A01N43/647—Triazoles; Hydrogenated triazoles
- A01N43/653—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Agronomy & Crop Science (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Soil Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a sweet potato latent virus removal method, which comprises the following steps: A. selecting sweet potato blocks without worm damage and mildew, and insolating in the sun; B. cleaning sweet potato blocks with clear water, and soaking the sweet potato blocks in a mixed solution containing ribavirin and sodium hypochlorite; C. planting sweet potato blocks in an incubator filled with sandy soil for germination accelerating culture; cutting a part 1cm from the top of the bud seedling, flushing with sterile water, and then cutting stem tip meristems; E. cutting a triangular notch at one end of the surface of the shoot apex meristem away from the leaf primordia by using an dissecting needle, and sliding a linear opening on the epidermis between the triangular notch and the leaf primordia; F. e, immersing the stem tip meristem treated in the step E into a mixed solution of hydrogen peroxide and pyrimidomycin for treatment; G. and culturing stem tip meristem, and cutting after the bud grows to 3cm to obtain the detoxified seedling. The invention effectively improves the removal rate of the sweet potato latent virus.
Description
Technical Field
The invention belongs to the technical field of sweet potato breeding, and particularly relates to a sweet potato latent virus removal method.
Background
After the sweet potato fine variety is planted for many years, obvious seed degeneration problems can occur, the yield of the sweet potato is seriously affected, and the seed degeneration reasons of the sweet potato are mainly caused by virus infection. Sweet Potato Latent Virus (SPLV) is a major virus that causes the sexual degeneration of sweet potato, and the removal rate of the traditional detoxification means for sweet potato latent virus is not high.
Disclosure of Invention
The invention aims to provide a sweet potato latent virus removal method, which can solve the defects in the prior art and effectively improve the removal rate of sweet potato latent viruses.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows.
A sweet potato latent virus removal method, comprising the following steps:
A. selecting sweet potato blocks without worm damage and mildew, and insolating for 1-2 days in the sun;
B. cleaning sweet potato blocks with clear water, and soaking the sweet potato blocks in a mixed solution containing ribavirin and sodium hypochlorite;
C. cleaning sweet potato blocks with clear water again, and then planting the sweet potato blocks in an incubator filled with sandy soil for germination accelerating culture;
D. when the bud grows to 3-5 cm, cutting a part of 1cm at the top of the bud, flushing for 2-3 min by using sterile water, and then cutting stem tip meristems with 1-2 leaf primordia;
E. cutting a triangular notch at one end of the surface of the shoot apex meristem away from the leaf primordia by using an dissecting needle, and sliding a linear opening on the epidermis between the triangular notch and the leaf primordia;
F. e, immersing the stem tip meristem treated in the step E into a mixed solution of hydrogen peroxide and pyrimidomycin for treatment;
G. and (3) placing the stem tip meristem subjected to soaking treatment in an MS minimal medium for culturing, and cutting after the bud grows to 3cm to obtain the detoxified seedling.
Preferably, in the step B, the mixed solution comprises 0.005wt% of ribavirin and 1.5wt% of sodium hypochlorite, and the soaking time is 30-40 min.
Preferably, in the step C, the culture temperature is 28-30 ℃, the illumination time is 12-13 h/D, and the illumination intensity is 2000Lx.
Preferably, in the step F, the mixed solution comprises 1.5wt% of hydrogen peroxide and 2wt% of pyrimidomycin, and the soaking time is 5-8 h.
Preferably, in the step G, the culture temperature is 32-35 ℃, the illumination time is 12-13 h/D, and the illumination intensity is 3000Lx.
The beneficial effects brought by adopting the technical scheme are as follows: according to the invention, the stem tip meristem is cut and streaked, and then is soaked in low-concentration hydrogen peroxide, so that the removal rate of the sweet potato latent virus is effectively improved.
Detailed Description
Example 1
A sweet potato latent virus removal method, comprising the following steps:
A. selecting sweet potato blocks without worm damage and mildew, and insolating for 1 day in the sun;
B. cleaning sweet potato blocks with clear water, and soaking the sweet potato blocks in a mixed solution containing ribavirin and sodium hypochlorite; the mixed solution comprises 0.005wt% of ribavirin and 1.5wt% of sodium hypochlorite, and the soaking time is 40min;
C. cleaning sweet potato blocks with clear water again, and then planting the sweet potato blocks in an incubator filled with sandy soil for germination accelerating culture; the culture temperature is 28-30 ℃, the illumination time is 12h/D, and the illumination intensity is 2000Lx;
D. when the bud grows to 3-5 cm, cutting a part of 1cm at the top of the bud, flushing for 2-3 min by using sterile water, and then cutting stem tip meristems with 1-2 leaf primordia;
E. cutting a triangular notch at one end of the surface of the shoot apex meristem away from the leaf primordia by using an dissecting needle, and sliding a linear opening on the epidermis between the triangular notch and the leaf primordia;
F. e, immersing the stem tip meristem treated in the step E into a mixed solution of hydrogen peroxide and pyrimidomycin for treatment; the mixed solution comprises 1.5 weight percent of hydrogen peroxide and 2 weight percent of pyriminobac-methyl, and the soaking time is 7 hours;
G. placing the stem tip meristem subjected to soaking treatment in an MS basic culture medium for culturing, and cutting after the bud grows to 3cm to obtain detoxified seedlings; the culture temperature is 32-35 ℃, the illumination time is 12h/D, and the illumination intensity is 3000Lx.
Comparative example 1
A sweet potato latent virus removal method, comprising the following steps:
A. selecting sweet potato blocks without worm damage and mildew, and insolating for 1 day in the sun;
B. cleaning sweet potato blocks with clear water, and soaking the sweet potato blocks in a mixed solution containing ribavirin and sodium hypochlorite; the mixed solution comprises 0.005wt% of ribavirin and 1.5wt% of sodium hypochlorite, and the soaking time is 40min;
C. cleaning sweet potato blocks with clear water again, and then planting the sweet potato blocks in an incubator filled with sandy soil for germination accelerating culture; the culture temperature is 28-30 ℃, the illumination time is 12h/D, and the illumination intensity is 2000Lx;
D. when the bud grows to 3-5 cm, cutting a part of 1cm at the top of the bud, flushing for 2-3 min by using sterile water, and then cutting stem tip meristems with 1-2 leaf primordia;
E. immersing the shoot apex meristem into a mixed solution of hydrogen peroxide and pyrimidomycin for treatment; the mixed solution comprises 7wt% of hydrogen peroxide and 2wt% of pyrimidomycin, and the soaking time is 2h;
F. placing the stem tip meristem subjected to soaking treatment in an MS basic culture medium for culturing, and cutting after the bud grows to 3cm to obtain detoxified seedlings; the culture temperature is 32-35 ℃, the illumination time is 12h/D, and the illumination intensity is 3000Lx.
Comparative example 2
A sweet potato latent virus removal method, comprising the following steps:
A. selecting sweet potato blocks without worm damage and mildew, and insolating for 1 day in the sun;
B. cleaning sweet potato blocks with clear water, and soaking the sweet potato blocks in a mixed solution containing ribavirin and sodium hypochlorite; the mixed solution comprises 0.005wt% of ribavirin and 1.5wt% of sodium hypochlorite, and the soaking time is 40min;
C. cleaning sweet potato blocks with clear water again, and then planting the sweet potato blocks in an incubator filled with sandy soil for germination accelerating culture; the culture temperature is 28-30 ℃, the illumination time is 12h/D, and the illumination intensity is 2000Lx;
D. when the bud grows to 3-5 cm, cutting a part of 1cm at the top of the bud, flushing for 2-3 min by using sterile water, and then cutting stem tip meristems with 1-2 leaf primordia;
E. immersing the shoot apex meristem into a mixed solution of hydrogen peroxide and pyrimidomycin for treatment; the mixed solution comprises 1.5 weight percent of hydrogen peroxide and 2 weight percent of pyriminobac-methyl, and the soaking time is 7 hours;
F. placing the stem tip meristem subjected to soaking treatment in an MS basic culture medium for culturing, and cutting after the bud grows to 3cm to obtain detoxified seedlings; the culture temperature is 32-35 ℃, the illumination time is 12h/D, and the illumination intensity is 3000Lx.
Comparative example 3
A sweet potato latent virus removal method, comprising the following steps:
A. selecting sweet potato blocks without worm damage and mildew, and insolating for 1 day in the sun;
B. cleaning sweet potato blocks with clear water, and soaking the sweet potato blocks in a mixed solution containing ribavirin and sodium hypochlorite; the mixed solution comprises 0.005wt% of ribavirin and 1.5wt% of sodium hypochlorite, and the soaking time is 40min;
C. cleaning sweet potato blocks with clear water again, and then planting the sweet potato blocks in an incubator filled with sandy soil for germination accelerating culture; the culture temperature is 28-30 ℃, the illumination time is 12h/D, and the illumination intensity is 2000Lx;
D. when the bud grows to 3-5 cm, cutting a part of 1cm at the top of the bud, flushing for 2-3 min by using sterile water, and then cutting stem tip meristems with 1-2 leaf primordia;
E. cutting a triangular notch at one end of the surface of the shoot apex meristem away from the leaf primordia by using an dissecting needle, and sliding a linear opening on the epidermis between the triangular notch and the leaf primordia;
F. e, immersing the stem tip meristem treated in the step E into a mixed solution of hydrogen peroxide and pyrimidomycin for treatment; the mixed solution comprises 7wt% of hydrogen peroxide and 2wt% of pyrimidomycin, and the soaking time is 2h;
G. placing the stem tip meristem subjected to soaking treatment in an MS basic culture medium for culturing, and cutting after the bud grows to 3cm to obtain detoxified seedlings; the culture temperature is 32-35 ℃, the illumination time is 12h/D, and the illumination intensity is 3000Lx.
In order to verify the effectiveness of the present invention (example 1), three comparative examples (comparative example 1 is a conventionally known detoxification method, comparative example 2 is a detoxification method of immersing in a low concentration solution for a long time based on comparative example 1, comparative example 3 is a detoxification method of immersing in a detoxification solution in a conventional detoxification method for a long time based on example 1), and comparative example 1 was subjected to comparative experiments (experimental raw materials are a Ipomoea batatas 18 for detecting sweet potato latent virus), 1000 strain of each of the experimental raw materials were selected, and after tissue fluid extraction of 1000 strain of each of the experimental raw materials, sweet potato latent virus was detected using fluorescent PCR detection, as follows:
| group of | Detection result |
| Example 1 | Negative of |
| Comparative example 1 | Positive and negative |
| Comparative example 2 | Positive and negative |
| Comparative example 3 | Negative of |
From the table, the sweet potato latent virus can be effectively removed by cutting and scribing the stem tip meristem and then carrying out detoxification soaking.
Comparative experiments on yield of detoxified seedlings were carried out using the above example 1 and comparative example 3, shoot apical meristems were prepared according to the procedures of example 1 and comparative example 3, respectively, 500 shoot apical meristems were selected for shoot culture, and the number of finally obtained detoxified seedlings was as shown in the following table:
| group of | Number of detoxified seedlings |
| Example 1 | 496 |
| Comparative example 3 | 453 |
From the table, after the stem tip meristem treatment mode is used, the germination rate of the detoxified seedlings can be effectively improved by adopting low-concentration and long-time detoxified soaking.
In order to further explore the relation of low-concentration detoxification soaking time to detoxification stability, on the basis of the embodiment 1, the soaking time in the step F is changed to 1h, 2h, 3h, 4h, 5h and 6h respectively, then fluorescence PCR detection is carried out on the obtained detoxification seedlings, and the detection result shows that the detoxification seedlings can be ensured to contain no sweet potato latent virus as long as the soaking time is greater than or equal to 5 h.
The foregoing has shown and described the basic principles and main features of the present invention and the advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that the above embodiments and descriptions are merely illustrative of the principles of the present invention, and various changes and modifications may be made without departing from the spirit and scope of the invention, which is defined in the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (4)
1. The sweet potato latent virus removing method is characterized by comprising the following steps:
A. selecting sweet potato blocks without worm damage and mildew, and insolating for 1-2 days in the sun;
B. cleaning sweet potato blocks with clear water, and soaking the sweet potato blocks in a mixed solution containing ribavirin and sodium hypochlorite;
C. cleaning sweet potato blocks with clear water again, and then planting the sweet potato blocks in an incubator filled with sandy soil for germination accelerating culture;
D. when the bud grows to 3-5 cm, cutting a part of 1cm at the top of the bud, flushing for 2-3 min by using sterile water, and then cutting stem tip meristems with 1-2 leaf primordia;
E. cutting a triangular notch at one end of the surface of the shoot apex meristem away from the leaf primordia by using an dissecting needle, and sliding a linear opening on the epidermis between the triangular notch and the leaf primordia;
F. e, immersing the stem tip meristem treated in the step E into a mixed solution of hydrogen peroxide and pyrimidomycin for treatment; the mixed solution comprises 1.5 weight percent of hydrogen peroxide and 2 weight percent of pyrimidomycin, and the soaking time is 5-8 hours;
G. and (3) placing the stem tip meristem subjected to soaking treatment in an MS minimal medium for culturing, and cutting after the bud grows to 3cm to obtain the detoxified seedling.
2. The sweet potato latent virus removal method according to claim 1, characterized in that: in the step B, the mixed solution comprises 0.005wt% of ribavirin and 1.5wt% of sodium hypochlorite, and the soaking time is 30-40 min.
3. The sweet potato latent virus removal method according to claim 2, characterized in that: in the step C, the culture temperature is 28-30 ℃, the illumination time is 12-13 h/D, and the illumination intensity is 2000Lx.
4. The method for removing sweet potato latent virus according to claim 3, wherein: in the step G, the culture temperature is 32-35 ℃, the illumination time is 12-13 h/D, and the illumination intensity is 3000Lx.
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| JPH05260869A (en) * | 1992-02-10 | 1993-10-12 | Iseki & Co Ltd | Adventitious root of sweet potato, its inducing method and its culture method |
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