Method for rapidly identifying fusarium wilt resistance of large-batch cowpea resources

Abstract

The invention relates to a method for rapidly identifying fusarium wilt resistance of a large batch of cowpea resources, which comprises the following steps: (1) preparing bacterium-carrying seedling culture soil, obtaining pathogenic bacteria strains of cowpea fusarium wilt, activating and culturing the pathogenic bacteria strains to prepare a pathogenic bacteria inoculation bacterium solution, and preparing the bacterium-carrying seedling culture soil by using the pathogenic bacteria inoculation bacterium solution; (2) sowing cowpeas, selecting strong cowpea seeds, and culturing the seeds until the seeds are white by taking a disease-susceptible variety with known resistance expression as a contrast; the seeds with white buds are sown in bacterium-carrying seedling culture soil and irrigated; (3) result investigation and evaluation: the disease investigation is started when the disease of the control variety to be infected is obvious and the disease development tends to be stable, and the whole-period investigation is not less than one time; and recording disease indexes and carrying out resistance evaluation. The cowpea resource fusarium wilt resistance identification method provided by the invention is simple to operate, high in accuracy and short in identification period, is suitable for identifying the fusarium wilt resistance of large-batch cowpea resources, and is beneficial to the development of cowpea resource fusarium wilt resistance breeding work.

Description

Technical Field

[0001] The present invention relates to the technical field of plant protection, and particularly to a method for rapidly identifying the fusarium wilt resistance of a large number of cowpea resources. Background Art

[0002] Cowpea fusarium wilt caused by infection with Fusarium oxysporum is one of the important diseases in cowpea production. The pathogen mainly infects the vascular bundles of plants, ultimately leading to the wilting and death of the whole plant, causing serious losses to cowpea producers. This disease is a soil-borne disease, and the high-temperature and high-humidity weather in South China throughout the year is likely to cause the occurrence of the disease. Once the plants are infected, there is no cure. The main control measures for this disease are water-flood rotation, planting disease-resistant (tolerant) varieties, high-ridge and deep-gutter cultivation, and medicament irrigation of the roots around the diseased plants. Among them, planting disease-resistant (tolerant) varieties is the most economical and effective measure. An accurate and reliable disease resistance identification method is the basis for cowpea disease-resistant breeding. Currently, there is an urgent need for a method that can identify the fusarium wilt resistance of cowpea resources in large quantities and quickly in cowpea disease-resistant breeding and related molecular biology research on resistance.

[0003] In the identification of cowpea resources resistant to fusarium wilt, the method of artificial inoculation at the seedling stage indoors is mainly used. The commonly used methods are the method of wounding the roots of seedlings and soaking them in bacteria and the method of soaking the radicles in bacteria after seed germination. The method of wounding the roots of seedlings and soaking them in bacteria is widely used by scientific researchers. After procedures such as pulling out the seedlings to wound the roots (cutting the roots), washing the roots, transplanting after soaking in bacteria, and cultivating at high temperature and high humidity until the disease occurs when the cowpea grows to two leaves and one heart, the investigation is carried out. This method is simple and feasible, and the identification result is reliable. However, a large amount of labor and time are required in the links such as pulling out the seedlings, washing the roots, and transplanting after soaking in bacteria. It is very time-consuming and laborious when dealing with the identification of the fusarium wilt resistance of hundreds or even hundreds of cowpea resources. Although the method of soaking the radicles in bacteria is simple to operate and suitable for the disease resistance identification of a large number of cowpea breeding materials, due to the differences in germination rate and seedling growth, the results often have large errors and cannot accurately reflect the resistance of the plants at the seedling stage. Therefore, exploring a rapid identification method for cowpea fusarium wilt resistance that is simple to operate, convenient to manage, has a short identification period, and is suitable for a large number of breeding materials or varieties has important application value in cowpea disease-resistant breeding and variety layout. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for rapidly identifying the fusarium wilt resistance of a large number of cowpea resources, so as to at least partially solve the technical problems mentioned in the above background art.

[0005] To achieve the above purpose, the present invention provides the following technical solution: A method for rapidly identifying the fusarium wilt resistance of a large number of cowpea resources, comprising the following steps:

[0006] (1) Prepare soil for raising seedlings with bacteria

[0007] Obtain the pathogenic strain of cowpea wilt, activate and culture the pathogenic strain to prepare a pathogenic bacteria inoculation solution, and use the pathogenic bacteria inoculation solution to prepare a pathogen-infected seedling-raising soil;

[0008] (2) Cowpea sowing

[0009] Select strong cowpea seeds, use a susceptible variety with known resistance performance as a control, and culture until the seeds show white tips;

[0010] Sow the seeds with white tips in the pathogen-infected seedling-raising soil and irrigate;

[0011] (3) Result investigation and evaluation

[0012] Start disease investigation when the disease of the susceptible control variety is obvious and the disease development tends to be stable, and conduct at least 1 full-period investigation;

[0013] Record the disease index and conduct resistance evaluation.

[0014] In a preferred embodiment, in step (1), the culture method of the pathogenic bacteria inoculation solution is as follows:

[0015] After activating and culturing the pathogenic strain, place it in a PD liquid medium for shaking culture, filter and then let it stand, take the precipitate solution and centrifuge to obtain the pathogenic bacteria spore precipitate, add clear water to the precipitate to obtain the pathogenic bacteria spore suspension, and continue to add water to obtain the pathogenic bacteria inoculation solution.

[0016] In a preferred embodiment, the spore concentration range of the pathogenic bacteria inoculation solution is: 1×10 6 spores / mL to 4×10 6 spores / mL.

[0017] In a preferred embodiment, pour the pathogenic bacteria inoculation solution into a preset volume of rice, soak it, add the seedling-raising substrate soil, and mix evenly to obtain the pathogen-infected seedling-raising soil.

[0018] In a preferred embodiment, the mixing ratio of the rice to the seedling-raising substrate soil is 1:3 by volume.

[0019] In a preferred embodiment, the seedling-raising substrate soil does not contain microbial agents and fungicides.

[0020] In a preferred embodiment, the seedling-raising substrate soil contains an agricultural and forestry water-retaining agent.

[0021] In a preferred embodiment, the volume of the agricultural and forestry water-retaining agent is 0.1% - 0.5% of the volume of the seedling-raising substrate soil.

[0022] In a preferred embodiment, when conducting experiments in a plant culture room or a plant incubator, place the seedling plug trays in the plug tray holders, add clear water to the plug tray holders, and cover them with a transparent plastic cover to maintain moisture for inoculation.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0024] (1) The method for rapidly identifying the fusarium wilt resistance of a large number of cowpea resources provided by the present invention is simple to operate, and the identification results are accurate and reliable. After adding the agroforestry water-retaining agent that can absorb water and retain fertilizer, there is no need to apply fertilizer, and almost no watering is required during the experimental process of maintaining moisture with a transparent plastic cover, saving labor and effort.

[0025] (2) The method for identifying the fusarium wilt resistance of cowpea provided by the present invention can test one material in one plug tray, and the maximum number of test plants can reach 144 plants / plug tray. The inoculation process does not require pulling out seedlings and soaking roots and re-transplanting operations, and is suitable for large-scale inoculation.

[0026] (3) Using the method for identifying the fusarium wilt resistance of cowpea provided by the present invention can ensure the consistency of the inoculated bacterial amount, light intensity, soil temperature and humidity, etc., and can be carried out in a plant incubator, with little influence of environmental factors and high repeatability, which is beneficial to reducing the error between experimental batches.

[0027] (4) The method for identifying the fusarium wilt resistance of cowpea of the present invention can provide food for the activity of pathogenic bacteria by adding rice grains, can better simulate the situation of the roots of field plants contacting the pathogenic bacteria and getting sick, and can better reflect the resistance level of the plants themselves. Description of the Drawings

[0028] Figure 1 It is a flow chart of the method for rapidly identifying the fusarium wilt resistance of a large number of cowpea resources provided by the embodiment of the present invention. Detailed Embodiments

[0029] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0030] As Figure 1 shown, the present invention discloses a method for rapidly identifying the fusarium wilt resistance of a large number of cowpea resources, including the following steps:

[0031] Step 1, prepare the disease-carrying seedling soil:

[0032] (1) Material preparation: Prepare cowpea seeds, ordinary rice, agricultural and forestry water retainer, seedling trays, tray for seedling trays and transparent seedling covers, seedling substrate soil, balanced compound fertilizer, PD solution;

[0033] (2) Bacterial liquid preparation: Obtain the pathogenic strain of cowpea wilt disease through collecting typical diseased samples of cowpea wilt disease in the field, isolating and culturing diseased tissues, purifying the pathogen, and verifying Koch's postulates. After activating and culturing the strain, place it in a PD solution medium for shaking culture. After filtering with gauze and standing, take the precipitate solution for centrifugation to obtain the pathogen spore precipitate. Add clear water to the precipitate to obtain the pathogen spore suspension. Then, calculate the concentration of the spore suspension mother liquor by dilution and counting under a microscope, and calculate the water addition amount according to the target of the pathogen spore suspension. Finally, obtain the standby pathogen inoculation bacterial liquid;

[0034] (3) Preparation of pathogen-infected seedling soil: Measure a certain volume of rice, pour in the pathogen inoculation bacterial liquid, soak it, and then add the seedling substrate soil containing a small amount of agricultural and forestry water retainer. Mix well to obtain the standby pathogen-infected seedling soil.

[0035] Step two, sowing cowpeas:

[0036] (4) Seed germination acceleration: Select strong cowpea seeds, use a susceptible variety with known resistance performance as a control. After soaking the seeds in warm water, wrap them with a wet gauze and place them in a constant temperature incubator for germination acceleration. After the seeds show white tips, use them for sowing, with 1 - 2 seeds per hole;

[0037] (5) Sowing and management: Place a small amount of pathogen-infected seedling soil at the bottom of the seedling tray, then sow the seeds with white tips in the tray, cover them with the seedling substrate soil containing a small amount of agricultural and forestry water retainer, and then water with a balanced compound fertilizer aqueous solution using a flower sprinkler or sprayer to ensure that the seedling substrate soil is moist but no water droplets overflow from the bottom. When conducting experiments in a plant culture room or plant incubator, place the seedling tray in the tray for seedling trays, add an appropriate amount of clear water to the tray for seedling trays, cover it with a transparent plastic cover for moisturizing and inoculation, and maintain a high-temperature environment and the moistness of the substrate soil after inoculation.

[0038] Step three, result investigation and evaluation:

[0039] (6) Result investigation: Start the investigation when the susceptible control variety shows obvious disease symptoms and the disease development tends to be stable. The whole-period investigation is not less than 1 time.

[0040] The method for classifying diseased plants is as follows: Grade 0, no disease symptoms; Grade 1, the two cotyledons turn yellow, but the growth is normal; Grade 3, the cotyledons turn yellow and wither, affecting growth; Grade 5, the cotyledons wither, and a small part of the plant wilts; Grade 7, part of the plant wilts or stops growing; Grade 9, the whole plant wilts, topples or withers.

[0041] The formula for calculating the disease index is: ∑(number of diseased plants at each level × relative level) / (total number of plants investigated × highest level value) × 100.

[0042] (7) Resistance evaluation: Determine the resistance level of cowpea resources based on the size of the disease index.

[0043] Method for resistance classification: Highly resistant (HR), 0 < disease index ≤ 15; Resistant (R), 15 < disease index ≤ 35; Moderately resistant (MR), 35 < disease index ≤ 55; Susceptible (S), 55 < disease index ≤ 75; Highly susceptible (HS), 75 < disease index.

[0044] Furthermore:

[0045] The seedling-raising plug tray described in step (1) is preferably 52-hole or 72-hole; the seedling-raising substrate soil does not contain microbial agents and fungicides; the transparent moisture-holding cover is preferably 20 cm in height; the balanced compound fertilizer is preferably Norwegian ternary compound fertilizer (N-P-K: 15-15-15).

[0046] The oscillation culture conditions described in step (2) are preferably 28 °C, 150 rpm, dark culture for 7 d; the filter gauze is preferably 3 layers, the precipitation time of the filtrate is preferably 3 h; the centrifugation speed is preferably 6000 rpm; the method for measuring the concentration of the pathogen spore suspension is to dilute the mother liquor of the pathogen spore suspension by 10-fold gradient by 1000 times or 10000 times, then suck 1 μL and count the number of spores under a 10-fold eyepiece microscope (the counting of 1 μL of the diluted bacterial solution is preferably 5 - 30 spores), count at least 10 times, and take the average as the concentration of the mother liquor of the spore suspension; the target inoculation concentration of the pathogen is: 1×10 6 spores / mL to 4×10 6 spores / mL. For inoculation in summer at high temperature, the lower limit of the acceptable concentration can be taken, and for inoculation with heating in winter, the upper limit of the concentration can be taken.

[0047] When the pathogen inoculation liquid soaks the rice in step (3), it is preferably about 1 cm higher than the rice, and the soaking time is 1 h; the volume of the agricultural and forestry water-retaining agent added to the seedling-raising substrate soil is 0.1% - 0.5% of the volume of the seedling-raising substrate soil; the mixing ratio of the bacteria-containing rice and the seedling-raising substrate soil containing the agricultural and forestry water-retaining agent is preferably 1:3 by volume; the height of filling into the plug tray holes is preferably half of the hole height.

[0048] The warm water soaking method described in step (4) is to soak the seeds in warm water at 55 - 60 °C for 30 s, wrap them with wet gauze and place them in a covered plastic box with a hollow bottom; the germination temperature is preferably 30 °C, and the bud length after the seeds show white is preferably no more than 0.5 cm. The seeds that germinate first can be taken out and placed in an incubator at 15 °C to wait for other seeds to germinate; when sowing, the bud heads are facing down or placed flat, and 1 or 2 seeds are sown in each hole.

[0049] In step (5), the preferred amount of clear water added to the seedling tray is 1 cm in depth; the preferred concentration of the balanced compound fertilizer aqueous solution is 1:1000 in mass-volume ratio; after inoculation, the indoor temperature is set at 25-35°C, preferably 28-33°C. For experiments inoculated in a plant culture room or a plant incubator, in summer with high temperature, it can be set to the air-conditioning cooling mode at 30°C, and in winter with low temperature, it can be set to the air-conditioning heating mode at 28°C. The relative humidity of the seedling substrate soil is preferably maintained at about 60% (dry and wet alternately); for experiments inoculated in a greenhouse, natural light can be used, and for a plant culture room or a plant incubator, a full-spectrum plant supplementary light can be used. The preferred light intensity is 15000 Lux, and the plants are cultured with 24-hour lighting.

[0050] The following further elaborates on the present invention in combination with embodiments. It should also be understood that the following embodiments are only used to further illustrate the present invention and cannot be construed as limiting the protection scope of the present invention. Some non-essential improvements and adjustments made by those skilled in the art based on the above content of the present invention all fall within the protection scope of the present invention.

[0051] Example 1

[0052] I. Test and identification method (identification of the resistance of cowpea resources to Fusarium wilt at the seedling stage in a greenhouse)

[0053] This example discloses a method for rapidly identifying the resistance of a large number of cowpea resources to Fusarium wilt, including the following steps:

[0054] Step 1, preparation of pathogen-infected seedling soil:

[0055] (1) Material preparation: Prepare 7 cowpea germplasm resources (provided by the Vegetable Research Institute of Guangdong Academy of Agricultural Sciences), silk rice (purchased in the market), agricultural and forestry water-retaining agent (Zhongshan Saipu Technology Co., Ltd.), 52-hole seedling plug trays (purchased in the market), seedling substrate soil (Guangzhou Shengsheng Agriculture Co., Ltd.), balanced compound fertilizer (Yara International ASA, Norway), PD solution (self-made);

[0056] (2) Bacterial liquid preparation: Culture the cowpea Fusarium wilt pathogen on a PDA plate in the dark at 28°C for 3 days, then pick the edge mycelial blocks and place them in a PD solution medium. After culturing in the dark at 28°C, 150 rpm for 7 days, filter with three layers of gauze. After the filtrate stands at room temperature for 3 hours, pour off the supernatant, then centrifuge the precipitated filtrate (room temperature, centrifuge speed 6000 rpm), pour off the supernatant, and mix the precipitate with clear water to obtain the mother liquid of the inoculated pathogen spore suspension. The mother liquid of the inoculated pathogen spore suspension is diluted 10-fold to 10000-fold in gradient. Then, suck 1 μL of the diluted bacterial liquid and read the spore count under a 10-fold eyepiece microscope. Count 10 times and take the average to obtain the concentration of the mother liquid of the pathogen spore suspension (i.e., the concentration is the average number of spores × 10 7spores / mL), and then inoculated with the target concentration of pathogen spore suspension (2×10 6 spores / mL) to calculate the amount of clean water to be added, and after mixing, obtain an inoculated pathogen spore suspension for later use;

[0057] (3) Preparation of contaminated seedling soil: Use a 15L plastic bucket to measure 90L of seedling matrix soil (not compacted), add 90mL of agricultural and forestry water retaining agent with a volume ratio of 0.1%, and mix thoroughly to obtain the spare seedling matrix soil. Use a plastic beaker to measure 1L of silk rice, pour the target concentration of pathogen spore suspension into the rice, and the bacterial liquid should be about 1cm above the rice. After soaking for 1h, add 3 times the volume of seedling matrix soil containing agricultural and forestry water retaining agent, and mix thoroughly to obtain the contaminated seedling soil. Measure 10mL of contaminated seedling soil for each hole in the hole tray (a total of 520mL), and then spread it evenly in each hole in the hole tray, waiting for sowing.

[0058] Step 2: Sowing cowpea:

[0059] (4) Seed germination: Select strong cowpea seeds, soak them in warm water at 55-60℃ for 30s, take them out and rinse them with cold water, then wrap them with wet gauze and place them in a plastic box with a lid and a hollow bottom. Place the plastic box in a 30℃ constant temperature incubator for germination. Sow them immediately after 1-2 days of whitening, with one seed per hole. After sowing the seeds, cover them with seedling substrate soil containing a small amount of agricultural and forestry water retaining agent. Then wet the substrate soil with a balanced compound fertilizer solution with a mass volume ratio of 0.1%, in small amounts and multiple times, to prevent water from overflowing from the bottom of the substrate soil. Use cowpea materials known to be susceptible to the disease as the disease control;

[0060] (5) Sowing and management: After sowing, place the plug tray in a temperature-controlled room at 28-33°C (the experiment was conducted in July, with shade nets used for shade and temperature control). Water the soil from time to time to keep it moist (relative humidity of about 60%, with a dry and wet state).

[0061] Step 3: Result investigation and evaluation:

[0062] (6) Result investigation: When the disease of the susceptible control varieties becomes obvious (more than 50% of the plants show symptoms), we will start to strengthen the observation. When the disease of the susceptible control varieties becomes obvious and the condition tends to be stable, we will conduct a diseased plant investigation.

[0063] The diseased plant grading method is as follows: Grade 0, no symptoms; Grade 1, two cotyledons turn yellow, but the growth is normal; Grade 3, the cotyledons turn yellow and die, affecting the growth; Grade 5, the cotyledons die, and a small part of the plant wilts; Grade 7, part of the plant wilts or stops growing; Grade 9, the whole plant wilts, falls over or dies.

[0064] (7) Resistance evaluation: After obtaining the survey data, calculate the disease index of the tested cowpea resources according to the disease index calculation formula, and judge the resistance level of the cowpea resources based on the size of the disease index.

[0065] Resistance classification method: Highly resistant (HR), 0 < disease index ≤ 15; Resistant (R), 15 < disease index ≤ 35; Moderately resistant (MR), 35 < disease index ≤ 55; Susceptible (S), 55 < disease index ≤ 75; Highly susceptible (HS), 75 < disease index.

[0066] II. Test results and analysis

[0067] The test results of this example are shown in Table 1. After 15 days of sowing, the disease situation of cowpea wilt tended to be stable. The survey results showed that among the 7 cowpea resources identified, 1 cowpea resource was highly susceptible, 4 cowpea resources were susceptible, 1 cowpea resource was moderately resistant, and 1 cowpea resource was resistant. In this example, the known cowpea control variety that was susceptible in the field was susceptible in the test, and the identification result of the control variety was consistent with its resistance performance in the field. The results indicate that the method for rapidly identifying the wilt resistance of a large number of cowpea resources provided in this example is applicable to the resistance evaluation and screening of cowpea germplasm resources to wilt.

[0068] Table 1 Identification results of the resistance of 7 cowpea materials to wilt

[0069]

[0070] Note: CK is the susceptible control (S)

[0071] Example 2

[0072] I. Test identification method (Seedling stage resistance identification of cowpea resources to wilt in a plant culture room)

[0073] This example discloses a method for rapidly identifying the wilt resistance of a large number of cowpea resources, including the following steps:

[0074] Step 1, Prepare inoculated nursery soil:

[0075] (1) Material preparation: Prepare 5 cowpea germplasm resources (provided by the Vegetable Research Institute of Guangdong Academy of Agricultural Sciences), plug trays and transparent plastic covers for seedling raising, and the rest is the same as in Example 1;

[0076] (2) Bacterial solution preparation: The same as in Example 1;

[0077] (3) Prepare inoculated nursery soil: The same as in Example 1.

[0078] Step 2, Sow cowpeas:

[0079] (4) Seed germination acceleration: The same as in Example 1;

[0080] (5) Sowing and management: After sowing, place the seedling-raising plug trays in the plug tray trays, wet the substrate soil (the specific steps are the same as in Example 1), add clear water to the plug tray trays to a height of about 2 cm, then cover with a transparent plastic cover for seedling raising to keep moisture, and finally place the sown plug trays in a plant culture room in an air-conditioned room with a temperature of 28 °C (test in November, the air conditioner is in heating mode) and illuminated by plant supplementary lighting (24-hour lighting). Add clear water to the plug tray trays once after inoculation and cultivation for 7 days.

[0081] Step 3, Result investigation and evaluation:

[0082] (6) Result investigation: The same as in Example 1.

[0083] (7) Resistance evaluation: The same as in Example 1.

[0084] II. Test results and analysis

[0085] The test results of this example are shown in Table 2. After 15 days of sowing, the disease situation of cowpea fusarium wilt tended to be stable. The investigation results showed that among the 5 cowpea resources identified, 4 cowpea resources showed susceptibility, and 1 cowpea resource showed moderate resistance. In this example, the known cowpea control variety that showed susceptibility in the field showed susceptibility in the test, and the identification result of the control variety was consistent with its resistance performance in the field. The results indicate that the method provided in this example for rapidly identifying the fusarium wilt resistance of a large number of cowpea resources is applicable to the resistance evaluation and screening of cowpea germplasm resources against fusarium wilt.

[0086] Table 2 Identification results of the resistance of 5 cowpea materials to fusarium wilt

[0087]

[0088] Note: CK is the susceptible control (S)

[0089] The method provided by the present invention for rapidly identifying the fusarium wilt resistance of a large number of cowpea resources is simple to operate. The inoculation link does not require operations such as pulling out seedlings and soaking roots and re-transplanting, which is suitable for large-scale inoculation. The identification results are accurate and reliable; at the same time, it can ensure the consistency of the inoculated bacterial amount, light intensity, soil temperature and humidity, etc., and can be carried out in a plant incubator with little influence from environmental factors and high repeatability, which is beneficial to reducing the error between test batches.

[0090] The foregoing has shown and described the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments, and the above embodiments and the descriptions in the specification are only preferred examples of the present invention and are not used to limit the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed. The scope of the present invention claimed is defined by the appended claims and their equivalents.

Claims

1. A method for rapidly identifying resistance to Fusarium wilt in large quantities of cowpea resources, characterized in that: The following steps are involved: (1) Preparation of bacterial seedling soil Obtaining a cowpea wilt pathogen strain, activating and culturing the pathogen strain to prepare a pathogen inoculation solution, and using the pathogen inoculation solution to prepare a fungus-carrying seedling soil; (2) Cowpea sowing Select healthy cowpea seeds, use susceptible varieties with known resistance as controls, and culture until the seeds turn white; Sow the seeds after they turn white in the soil with bacteria and water them; (3) Result investigation and evaluation When the disease of the susceptible control varieties becomes obvious and the disease progresses steadily, the disease investigation shall be started, and the investigation shall be conducted at least once during the whole period; The disease index was recorded and the resistance was evaluated.

2. The method for rapidly identifying resistance to Fusarium wilt of large quantities of cowpea resources according to claim 1, characterized in that: In step (1), the method for culturing the pathogen inoculated bacterial solution is: The pathogenic strain is activated and cultured in a PD liquid culture medium for shaking culture, filtered and then allowed to stand, the precipitate is centrifuged to obtain a pathogenic bacteria spore precipitate, clear water is added to the precipitate to obtain a pathogenic bacteria spore suspension, and water is continued to be added to obtain a pathogenic bacteria inoculation solution.

3. The method for rapidly identifying resistance to Fusarium wilt of large quantities of cowpea resources according to claim 2, characterized in that: The spore concentration range of the pathogen inoculation solution is: 1×10 6 spores / mL~4×10 6 spores / mL.

4. The method for rapidly identifying resistance to Fusarium wilt of large quantities of cowpea resources according to claim 2, characterized in that: The pathogen inoculated bacterial solution is poured into a preset volume of rice, and after soaking, seedling medium soil is added, and the seedling soil with bacteria is obtained after mixing.

5. The method for rapidly identifying resistance to Fusarium wilt of large quantities of cowpea resources according to claim 4, characterized in that: The mixing ratio of the rice to the seedling medium soil is 1:3 by volume.

6. The method for rapidly identifying resistance to Fusarium wilt of large quantities of cowpea resources according to claim 4, characterized in that: The seedling raising matrix soil does not contain microbial agents and fungicides.

7. The method for rapidly identifying resistance to Fusarium wilt of large quantities of cowpea resources according to claim 4, characterized in that: The seedling raising matrix soil contains an agricultural and forestry water retaining agent.

8. The method for rapidly identifying resistance to Fusarium wilt of large quantities of cowpea resources according to claim 7, characterized in that: The volume of the agricultural and forestry water-retaining agent is 0.1% to 0.5% of the volume of the seedling-raising matrix soil.

9. The method for rapidly identifying resistance to Fusarium wilt of large quantities of cowpea resources according to claim 1, characterized in that: When the test is conducted in a plant culture room or a plant culture box, the seedling hole tray is placed in a hole tray, clean water is added to the hole tray, and a transparent plastic cover is covered to keep moisture and inoculation.

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