CN110250113B - Method for culturing Meloidogyne incognita by using Eupatorium adenophorum - Google Patents
Method for culturing Meloidogyne incognita by using Eupatorium adenophorum Download PDFInfo
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- 244000062748 Eupatorium adenophorum Species 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 27
- 238000012258 culturing Methods 0.000 title claims abstract description 11
- 241000243786 Meloidogyne incognita Species 0.000 title claims abstract description 6
- 241000196324 Embryophyta Species 0.000 claims abstract description 46
- 241000243787 Meloidogyne hapla Species 0.000 claims abstract description 39
- 239000000463 material Substances 0.000 claims abstract description 35
- 241000244206 Nematoda Species 0.000 claims abstract description 17
- 238000011081 inoculation Methods 0.000 claims abstract description 12
- 210000003250 oocyst Anatomy 0.000 claims abstract description 8
- 239000002689 soil Substances 0.000 claims description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 26
- 239000010455 vermiculite Substances 0.000 claims description 16
- 229910052902 vermiculite Inorganic materials 0.000 claims description 16
- 235000019354 vermiculite Nutrition 0.000 claims description 16
- 239000012153 distilled water Substances 0.000 claims description 12
- 210000004209 hair Anatomy 0.000 claims description 10
- 229920000742 Cotton Polymers 0.000 claims description 9
- 241001143352 Meloidogyne Species 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 6
- 230000007935 neutral effect Effects 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- 238000002791 soaking Methods 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 241000208292 Solanaceae Species 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 238000005286 illumination Methods 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 5
- 230000035699 permeability Effects 0.000 claims description 5
- 230000021749 root development Effects 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 239000002352 surface water Substances 0.000 claims description 5
- 238000009736 wetting Methods 0.000 claims description 5
- 235000007688 Lycopersicon esculentum Nutrition 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- 239000002244 precipitate Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 239000000758 substrate Substances 0.000 claims description 4
- 229960004793 sucrose Drugs 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 3
- 239000008055 phosphate buffer solution Substances 0.000 claims description 3
- 238000005507 spraying Methods 0.000 claims description 3
- 238000003306 harvesting Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 238000005057 refrigeration Methods 0.000 claims description 2
- 238000007873 sieving Methods 0.000 claims description 2
- 240000003768 Solanum lycopersicum Species 0.000 claims 1
- 238000009395 breeding Methods 0.000 abstract description 7
- 230000001488 breeding effect Effects 0.000 abstract description 7
- 241000243785 Meloidogyne javanica Species 0.000 abstract description 4
- 239000011159 matrix material Substances 0.000 abstract 1
- 241000227653 Lycopersicon Species 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 235000017060 Arachis glabrata Nutrition 0.000 description 2
- 244000105624 Arachis hypogaea Species 0.000 description 2
- 235000010777 Arachis hypogaea Nutrition 0.000 description 2
- 235000018262 Arachis monticola Nutrition 0.000 description 2
- 235000003143 Panax notoginseng Nutrition 0.000 description 2
- 241000180649 Panax notoginseng Species 0.000 description 2
- 235000020232 peanut Nutrition 0.000 description 2
- 235000003181 Panax pseudoginseng Nutrition 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000009328 dry farming Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 230000003032 phytopathogenic effect Effects 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
Classifications
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- 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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Botany (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Cultivation Of Plants (AREA)
Abstract
A method for culturing Meloidogyne incognita by using Eupatorium adenophorum belongs to the field of biotechnology. The method comprises the following steps: collecting or purchasing Meloidogyne incognita or oocysts, refrigerating for later use, preparing culture material of Eupatorium Adenophorum, preparing planting matrix, and inoculating. The invention has the advantages that: the eupatorium adenophorum has strong vitality and adaptability, is easy to survive and plant, is not sensitive to most varieties of nematodes, has higher specific correspondence to the northern root-knot nematodes and is easy to obtain single northern root-knot nematodes; the cost for breeding the root-knot nematode by the eupatorium adenophorum is low; the breeding period is longer than that of other crops, and the living body of the northern root-knot nematode can be continuously obtained; can be used repeatedly after one-time inoculation.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for culturing northern meloidogyne.
Background
Root-knot nematodes (Meloidogyne spp.) are highly obligate parasitic phytopathogenic nematodes, whereas northern root-knot nematodes (Meloidogyne hapla) are among the nematodes mainly affecting crops. Northern root-knot nematodes mainly parasitize crops at roots, resulting in tumors. In the field with large population density, a plurality of tumors are formed at the root, and when the tumors are serious, the root is rotted and falls off, so that the leaves are yellowed, the growth is stopped, and the plants wither and die, especially the false planting bed and the seedbed of the long runners are seriously damaged. The harm range of the meloidogyne hapla in north is very wide, and almost all dry crops are harmed except a few crops, and the harm to annual solanaceae crops and annual cruciferous crops is the most serious. In recent years, the northern root knot nematode disease also becomes one of the important diseases of the traditional Chinese medicine pseudo-ginseng. In the research process of preventing and controlling northern root-knot nematode diseases, people need a large amount of northern root-knot nematodes. Although the panax notoginseng can be directly used for breeding the meloidogyne hapla, the cost is high, and the panax notoginseng has high requirements on environmental conditions and is often influenced by seasons, geographical positions and the like. If the tomatoes are used for breeding, as the tomatoes are sensitive to most root-knot nematodes, multiple nematodes are easy to infect simultaneously in the breeding process, so that a single variety of nematodes is difficult to separate. Peanut roots are also used for culturing northern root-knot nematodes, but the peanut roots can only be used once. No method for culturing Meloidogyne haplocalyx by using Eupatorium adenophorum is available so far.
Disclosure of Invention
The invention aims to provide a method for culturing Meloidogyne haplocalyx by using Eupatorium adenophorum, based on the problems in the prior art.
The method of the invention can comprise the following steps:
1. collecting or purchasing the meloidogyne hapla for refrigeration and standby. The northern meloidogyne can be purchased and collected by the prior art.
If the northern root-knot nematode is collected by self, the optimal method is as follows:
digging complete plant roots of the solanaceae crops at the last stage of harvesting, taking root knots on the root roots, washing with water, soaking for 10-18 hours at room temperature by using distilled water, collecting soaking liquid with precipitates, preserving for later use at 2-6 ℃, repeating the process for 2-4 times on the soaked root knots, discarding the root knots, combining the soaking liquids with the precipitates collected for several times, sieving by using a 15-20-mesh separation sieve, standing the sieved substances for 3-5 hours, slightly removing liquid with the upper part accounting for 75-85% of the total volume of the standing substances, centrifuging the residues at the lower part for 1-3 minutes at 1500-2500 r/min, discarding supernatant, adding cane sugar water with the concentration of 480-500 g/L into the residues, wherein the volume of the added cane sugar water is 3-5 times of the volume of the residues, mixing uniformly, centrifuging for 1-3 minutes at 1500-2500 r/min, and centrifuging the supernatant for 700-900-mesh separation sieve, collecting the polypide on the separation sieve, and refrigerating the polypide at 3-5 ℃ for later use after the polypide is determined as the meloidogyne hapla through microscopic examination, wherein the refrigerating time is not more than 72 hours. The solanaceae crops in the step are preferably tomatoes, the distilled water is preferably single distilled water, and the root knots are preferably soaked for 3 times.
2. Preparing culture material of Eupatorium adenophorum
Selecting healthy Eupatorium adenophorum plants with the growth time of not more than 2 months, the plant height of not more than 1.0 meter and the root length of more than 10cm, washing with water, placing in a ventilated place, removing the upper parts and main buds of the plants after the surface water is dried, wherein the removed upper parts account for 25-35% of the total length of the plants, and obtaining the culture material of the Eupatorium adenophorum for later use.
3. Preparing planting substrate
Taking soil suitable for planting Eupatorium adenophorum Spreng, drying at 80-120 deg.C for 1.5-2.5 hr to kill other nematodes and ova possibly contained in the soil, and uniformly spreading the soil recovered to room temperature on the bottom of a container with good air permeability, wherein the thickness of the soil is 1.5-2.0 cm. Taking vermiculite particles with the diameter of 2-8mm for later use after sterilization.
4. Inoculation of
Spreading the eupatorium adenophorum culture material prepared in the step 2 on sterile neutral filter paper, wetting the filter paper with distilled water, uniformly spraying a phosphate buffer solution with the pH value of 6.5-8.0 on the culture material, recovering the northern root-knot nematode prepared in the step 1 to room temperature, performing microscopic examination on the activity of the nematode, inoculating the nematode to a root hair with the diameter of more than 1.0mm, inoculating 10-20 head worms per 2.0cm of the root hair, covering the surface of the culture material with wet absorbent cotton after inoculation, transferring the degreased cotton to an artificial climate box with the temperature of 25-30 ℃ and the relative humidity of 50-70 percent, standing for 4-6 h under the condition of no light, taking out, paving the culture material on the surface of the soil prepared in the step 3, filling the vermiculite prepared in the step 3, completely burying the culture material into the vermiculite, planting under the normal day at the temperature of 25-32 ℃, and watering once every 3-4 days to keep the eupatorium adenophorum leaves from wilting, controlling the height of the plant to be 1.2-1.5m, carrying out bud picking treatment once every 8-12 days to promote root development, culturing for 28-45 days to generate root knots, obtaining a living body of the northern root-knot nematode after the root knots are separated, reserving a small number of root knots of the plant, and re-planting to continuously obtain the northern root-knot nematode.
The method of the invention can also comprise the following steps:
1. preparing culture material of Eupatorium adenophorum
Selecting healthy Eupatorium adenophorum plants with the growth time of not more than 2 months, the plant height of not more than 1.0 meter and the root length of more than 10cm, washing with water, placing in a ventilated place, removing the upper parts and main buds of the plants after the surface water is dried, wherein the removed upper parts account for 25-35% of the total length of the plants, and obtaining the culture material of the Eupatorium adenophorum for later use.
2. Preparing planting substrate
Taking soil suitable for planting Eupatorium adenophorum Spreng, drying at 80-120 deg.C for 1.5-2.5 hr to kill other nematodes and ova possibly contained in the soil, and uniformly spreading the soil recovered to room temperature on the bottom of a container with good air permeability, wherein the thickness of the soil is 1.5-2.0 cm. Taking vermiculite particles with the diameter of 2-8mm for later use after sterilization.
3. Inoculation of
Spreading the eupatorium adenophorum culture material prepared in the step 1 on sterile neutral filter paper, wetting the filter paper with distilled water, inoculating 10-20 oocysts of northern root-knot nematodes to root hairs with the diameter larger than 1.0mm of the culture material, wherein the root hairs are inoculated per 1.0cm, covering the surface of the culture material with wet absorbent cotton after inoculation, transferring the inoculated culture material to an artificial climate box with the temperature of 25-30 ℃ and the relative humidity of 50-70% for standing for 12-16 h under the condition of no illumination, taking out the culture material, paving the culture material on the surface of the soil prepared in the step 2, filling the vermiculite prepared in the step 3, completely burying the culture material into the vermiculite, planting the culture material at the temperature of 25-32 ℃ under normal sunlight, watering once every 3-4 days during the period, controlling the plant height to be 1.2-1.5m during the planting process, and C, bud picking treatment is carried out every 8-12 days to promote root development, root knots appear in 35-45 days after culture, the living body of the northern root-knot nematode can be obtained after the root knots are taken and separated, a small number of root knots of the plants are reserved, and the northern root-knot nematode can be continuously obtained after re-planting.
The soil suitable for planting the eupatorium adenophorum is known by the professional, for example, dry farming soil meeting the national standard GB15618-2018 can be used as test soil.
The northern root-knot nematode oocysts can be purchased or collected by self, and the collection method is the prior art.
The invention has the advantages that: the eupatorium adenophorum has strong vitality and adaptability, is easy to survive and plant, is not sensitive to most varieties of nematodes, has higher specific correspondence to the northern root-knot nematodes and is easy to obtain single northern root-knot nematodes; the cost for breeding the root-knot nematode by the eupatorium adenophorum is low; the breeding period is longer than that of other crops, and the living body of the northern root-knot nematode can be continuously obtained; can be used repeatedly after one-time inoculation.
Detailed Description
Example 1.
Selecting healthy plants with the eupatorium adenophorum which are grown in the current year, the growth time of less than or equal to 2 months, the plant height of less than or equal to 1.0m and the root length of more than 10cm, cleaning the root system and the plants by using tap water, placing the plants to a ventilated place, cutting off the upper part of the plants to about 1/3 degrees after the surface water is dried, removing main buds, and reserving the leaves of the rest part 2/3 for later use.
Taking soil suitable for planting, drying at 100 ℃ for 2 hours, killing other nematodes and ova possibly contained in the soil, recovering to normal temperature, uniformly paving in a container with good air permeability, and keeping the soil thickness of about 1.8cm for later use. Taking vermiculite with 5mm diameter particles for standby after sterilization.
Spreading the root system of the prepared eupatorium adenophorum plant on sterile neutral filter paper, wetting the filter paper with distilled water, preferably, no water can be accumulated under the filter paper by naked eyes, uniformly spraying a phosphate buffer solution with the pH value of 7.5 on the root system, recovering the prepared northern root knot nematode body to the room temperature, inoculating the seedling body to the root hair with the diameter of the eupatorium adenophorum root of more than 1.0mm after microscopic examination of the activity of the larva body, inoculating 15 larvae per 2.0cm, covering the wet (no water is extruded by hands) absorbent cotton on the surface of the root system after inoculation, transferring the seedling to an artificial climate box with the temperature of 27 ℃ and the relative humidity of 65 percent, standing for 5 hours under the condition of no illumination, taking out, laying the tail end root system of the plant on the surface of soil, burying the inoculated part as little as possible in the soil, filling the prepared vermiculite, burying the rest roots of the plant into the soil, planting at the temperature of 25-32 ℃ and under normal sunshine, watering once per 3-4 days, the leaves of the eupatorium adenophorum are not wilted, the height of the plant is controlled to be about 1.2-1.5m in the planting process, and bud picking treatment is carried out every 10 days to promote root development. Root knots appear after about 35 days of culture, and 2-year-old living bodies of the northern root-knot nematodes can be obtained after the root knots are taken and separated.
Example 2.
The root system of the eupatorium adenophorum plant is prepared according to the method of example 1, the root system is laid on sterile neutral filter paper, and the filter paper is moistened by distilled water, preferably, no water is accumulated under the filter paper by naked eyes. Inoculating purchased northern root-knot nematode oocysts to root hairs of the eupatorium adenophorum with the root diameter of more than 1.0mm, and inoculating 8 oocysts to every 1.0cm of root hairs. After inoculation, covering wet (until no water is extruded by hands), absorbent cotton on the surface of a root system, transferring the cotton to an artificial climate box with the temperature of 36 ℃ and the relative humidity of 60% for standing for 24 hours under the condition of illumination, taking out the cotton, paving the root system at the tail end of a plant on the surface of the soil prepared according to the method in the embodiment 1, filling vermiculite prepared according to the method in the embodiment 1 into the inoculated part as far as possible, completely burying the rest roots of the plant into the vermiculite, planting the plant at 30 ℃ in normal sunshine, watering the plant once every 4 days during the period, and controlling the height of the plant to be about 1.3m during the planting process, and performing bud picking treatment every 10 days to promote the development of the root. Root knots can appear after 40 days of culture. And (4) obtaining the 2-year-old living body of the northern root-knot nematode after taking and separating root knots. A small amount of root knots of the plants are reserved and planted again to continuously obtain the northern root-knot nematodes.
Claims (4)
1. A method for culturing Meloidogyne incognita by using Eupatorium adenophorum is characterized by comprising the following steps:
(1) collecting or purchasing northern root-knot nematodes for refrigeration for later use;
(2) preparing a culture material of eupatorium adenophorum: selecting healthy Eupatorium adenophorum plants with the growth time of not more than 2 months, the plant height of not more than 1.0 meter and the root length of more than 10cm, washing with water, placing in a ventilated place, removing the upper parts and main buds of the plants after the surface water is dried, wherein the removed upper parts account for 25-35% of the total length of the plants, and obtaining a culture material of the Eupatorium adenophorum for later use;
(3) preparing a planting substrate: taking soil suitable for planting Eupatorium adenophorum Spreng, drying at 80-120 deg.C for 1.5-2.5 hr to kill other nematodes and ova possibly contained in the soil, and uniformly spreading the soil recovered to room temperature on the bottom of a container with good air permeability, wherein the thickness of the soil is 1.5-2.0 cm; taking vermiculite particles with the diameter of 2-8mm, and sterilizing for later use;
(4) inoculation: spreading the eupatorium adenophorum culture material prepared in the step (2) on sterile neutral filter paper, wetting the filter paper with distilled water, uniformly spraying a phosphate buffer solution with the pH value of 6.5-8.0 on the culture material, recovering the northern root-knot nematodes prepared in the step (1) to room temperature, inoculating 10-20 heads of nematodes to root hairs with the diameter of more than 1.0mm after microscopic examination of the activity of the nematodes, inoculating the nematodes to the root hairs with the diameter of more than 1.0mm, inoculating 10-20 heads of nematodes to the root hairs every 2.0cm, covering the surface of the culture material with wet absorbent cotton after inoculation, transferring the culture material to an artificial climate box with the temperature of 25-30 ℃ and the relative humidity of 50-70% for standing for 4-6 h under the condition of no illumination, laying the culture material on the surface of the soil prepared in the step (3) after taking out, filling the vermiculite prepared in the step (3), completely burying the culture material into vermiculite, planting the culture material at, watering every 3-4 days to keep the leaves of the eupatorium adenophorum from wilting, controlling the height of the plant to be 1.2-1.5m, carrying out bud picking treatment every 8-12 days to promote root development, culturing for 28-45 days to generate root knots, taking the root knots and separating to obtain the living body of the northern root-knot nematode, reserving a small number of root knots of the plant and replanting the plant to continuously obtain the northern root-knot nematode.
2. The method for cultivating Meloidogyne haplocalyx using Eupatorium adenophorum of claim 1, wherein the Meloidogyne haplocalyx is collected by itself according to the following method: digging complete plant roots of the solanaceae crops at the last stage of harvesting, taking root knots on the root roots, washing with water, soaking for 10-18 hours at room temperature by using distilled water, collecting soaking liquid with precipitates, preserving for later use at 2-6 ℃, repeating the process for 2-4 times on the soaked root knots, discarding the root knots, combining the soaking liquids with the precipitates collected for several times, sieving by using a 15-20-mesh separation sieve, standing the sieved substances for 3-5 hours, slightly removing liquid with the upper part accounting for 75-85% of the total volume of the standing substances, centrifuging the residues at the lower part for 1-3 minutes at 1500-2500 r/min, discarding supernatant, adding cane sugar water with the concentration of 480-500 g/L into the residues, wherein the volume of the added cane sugar water is 3-5 times of the volume of the residues, mixing uniformly, centrifuging for 1-3 minutes at 1500-2500 r/min, and centrifuging the supernatant for 700-900-mesh separation sieve, collecting the polypide on the separation sieve, and refrigerating the polypide at 3-5 ℃ for later use after the polypide is determined as the meloidogyne hapla through microscopic examination, wherein the refrigerating time is not more than 72 hours.
3. The method for cultivating Meloidogyne haplocalyx by using Eupatorium adenophorum of claim 2, wherein when collecting Meloidogyne haplocalyx by itself, the Solanaceae crop is tomato, distilled water is single distilled water, and the root knot is soaked for 3 times.
4. A method for culturing Meloidogyne incognita by using Eupatorium adenophorum is characterized by comprising the following steps:
(1) preparing a culture material of eupatorium adenophorum: selecting healthy Eupatorium adenophorum plants with the growth time of not more than 2 months, the plant height of not more than 1.0 meter and the root length of more than 10cm, washing with water, placing in a ventilated place, removing the upper parts and main buds of the plants after the surface water is dried, wherein the removed upper parts account for 25-35% of the total length of the plants, and obtaining a culture material of the Eupatorium adenophorum for later use;
(2) preparing a planting substrate: taking soil suitable for planting Eupatorium adenophorum Spreng, drying at 80-120 deg.C for 1.5-2.5 hr to kill other nematodes and ova possibly contained in the soil, and uniformly spreading the soil recovered to room temperature on the bottom of a container with good air permeability, wherein the thickness of the soil is 1.5-2.0 cm; taking vermiculite particles with the diameter of 2-8mm, and sterilizing for later use;
(3) inoculation: spreading the eupatorium adenophorum culture material prepared in the step (1) on sterile neutral filter paper, wetting the filter paper with distilled water, inoculating 10-20 oocysts per 1.0cm of the diameter of the culture material to purchased or automatically collected northern root-knot nematode oocysts, covering the surface of the culture material with wet absorbent cotton after inoculation, transferring the inoculated or automatically collected northern root-knot nematode oocysts to an artificial climate box with the temperature of 25-30 ℃ and the relative humidity of 50-70 percent, standing for 12-16 h under the condition of no illumination, taking out, paving the culture material on the surface of the soil prepared in the step (2), filling the vermiculite prepared in the step (2), completely burying the culture material into the vermiculite, planting the culture material at the temperature of 25-32 ℃ under normal sunlight, watering once every 3-4 days during the period, preventing the leaves of the eupatorium adenophorum from wilting, and controlling the plant height to be 1.2-1.5m during the planting process, and C, bud picking treatment is carried out every 8-12 days to promote root development, root knots appear in 35-45 days after culture, the living body of the northern root-knot nematode can be obtained after the root knots are taken and separated, a small number of root knots of the plants are reserved, and the northern root-knot nematode can be continuously obtained after re-planting.
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| CN116171864B (en) * | 2023-03-27 | 2024-01-19 | 湖南大学 | Method for synchronously infecting arabidopsis thaliana on root-knot nematode dish and application |
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