CN115777462A - Artificial inoculation method of indoor corn seedling-stage northern leaf blight - Google Patents
Artificial inoculation method of indoor corn seedling-stage northern leaf blight Download PDFInfo
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Abstract
The invention provides an artificial inoculation method for indoor corn seedling-stage northern leaf blight, which comprises the following steps: 1) Planting and managing indoor corns; 2) Artificial inoculation treatment of northern leaf blight in corn seedling stage; 3) The three parts of moisture preservation management after the corn seedlings are artificially inoculated, namely selecting the corn seedlings sown in a plastic bowl (the diameter is 11.5 cm) as an inoculation object under indoor conditions, soaking sanitary paper extraction strips (3.0 multiplied by 1.0 cm, the length is multiplied by the width) with hypha and conidium suspension of the corn northern leaf blight obtained by culturing 10 d on a PDA culture medium to inoculate the corn seedlings in 3-4 leaf stages, and optimizing the moisture preservation conditions of the corn seedlings after the artificial inoculation, thereby obtaining ideal disease attack effect. The invention has simple operation and low cost, can obtain ideal disease attack effect and has good practicability.
Description
Technical Field
The invention relates to an artificial inoculation method for plant diseases, in particular to an artificial inoculation method for indoor corn seedling-stage northern leaf blight.
Background
Cordycephem (A. Maydis)Exserohilum turcicum) Is an important leaf pathogenic fungus in the production of corn and sorghum in China. The withered disease spot area caused by the infection of the leaves by the northern leaf blight of corn is large, the influence on the yield and the quality of corn and sorghum is very obvious, the yield loss caused generally is 20 to 30 percent, and the yield loss can reach more than 50 percent in serious cases. With the development of fresh-eating sweet (waxy) corn industry, transformation of planting system and high multiple cropping index, favorable conditions are provided for the prevalence and harm of corn northern leaf blight, so that the field morbidity and the disease severity become increasingly serious. The outbreak and disaster-forming of the alternaria turcicola severely restrict the safe production of corn and sorghum.
The occurrence condition of the corn northern leaf blight has high requirements on the humidity and the temperature of the environment, and the high-humidity and warm environmental condition is favorable for the occurrence of diseases. The development of maculopathy is significantly inhibited when the ambient temperature is below 15 ℃ or above 30 ℃ and the relative humidity is less than 60%. Spraying of high-efficiency and low-toxicity bactericide is one of important strategies for preventing and controlling field corn northern leaf blight. However, excessive dependence on chemical fungicides can easily cause resistance to northern leaf blight. Planting disease-resistant varieties is another important measure for controlling the northern leaf blight of corn. Researches show that the resistance of corn varieties carrying different disease-resistant genes to the northern leaf blight has obvious difference. Due to the continuous emergence of new physical races of the northern leaf blight, the genetic variation of pathogenic bacteria is fast, and disease resistance of disease-resistant varieties is often lost, so that the northern leaf blight is widely outbreaked and epidemic. Therefore, developing the resistance identification of new corn varieties to the alternaria turciosa and monitoring the development and the change of the new corn varieties to the disease resistance in the field have important significance for guiding the reasonable layout of the different resistant new corn varieties.
Because the fresh corn in south China has high multiple cropping index, the corn northern leaf blight is infected and damaged in the field in a circulating way, and therefore, the determination of the disease resistance of the new corn variety in the seedling stage to the northern leaf blight is very important. The invention provides an artificial inoculation method of alternaria maculosa in the seedling stage of indoor corn by improving the artificial inoculation technology in the seedling stage of indoor corn, and the method is simple to operate and low in cost and can obtain an ideal disease attack effect.
Disclosure of Invention
The invention aims to provide an artificial inoculation method for indoor corn seedling-stage northern leaf blight, which is simple to operate, low in cost, capable of obtaining an ideal disease effect and good in practicability.
In order to achieve the purpose, the invention adopts the following technical scheme:
an artificial inoculation method of alternaria alternata in an indoor corn seedling stage comprises three parts of indoor corn planting management, an artificial inoculation method of alternaria alternata in a corn seedling stage and moisture preservation management after artificial inoculation of corn seedlings; the operation of each part is as follows:
1) Planting management of indoor corns: sowing healthy and plump corn seeds in seedling raising trays filled with Topu peat soil indoors, sowing 2 seeds in each hole, placing the seedling raising trays in plastic trays filled with half-tray water, and raising the seedlings on an illumination rack (60 cm in each layer and 4 incandescent lamps with 48W cold light sources arranged at the top) to 2-leaf stage; then, transplanting the corn seedlings and the peat soil at the roots in each hole into a white plastic bowl (the diameter is 11.5 cm), transplanting 2 holes (4 seedlings in total) in each bowl, placing the plastic bowl into a plastic tray filled with half-tray water, and growing the seedlings on an illumination frame until the seedling stage reaches 3-4 leaves, wherein the indoor temperature is controlled to be 25-30 ℃ during the seedling growing period;
2) Artificial inoculation treatment of northern leaf blight in corn seedling stage: inoculating rejuvenated northern corn northern leaf blight in a PDA culture medium (fresh peeled potato 200 g + glucose 20 g + agar powder 18 g + deionized water 1000 mL), culturing 10 d in dark at 25 ℃, adding 10mL of sterile water into each dish, slightly brushing northern corn northern leaf blight hypha and conidia on the surface of the culture medium by using a sterile brush, preparing hypha + conidia suspension by using the sterile water, and adjusting the concentration of spores in the hypha + conidia suspension to 20-30 spores per visual field (10 multiplied by 20 times of lens); when indoor maize seedlings are cultivated to 3-4 leaf stages, cutting sanitary paper (Vida brand paper, three layers) into paper strips of 3.0 cm x 1.0 cm (length x width), immersing the paper strips into the hypha + conidium suspension liquid 30 s, clamping 1 paper strip from the suspension liquid by using sterile tweezers, attaching the paper strips to the middle of maize seedling leaves, and inoculating 2 leaves to each maize seedling;
3) Moisture preservation management after artificial inoculation of corn seedlings: uniformly spraying water on the inner wall of a plastic barrel (the inner diameter is 31 cm, the height is 32 cm) by using a manual pressure spraying pot, transferring the inoculated corn seedlings into the barrel, wherein 4 bowls are arranged in each barrel, the opening of each barrel is covered by a soaked plastic film, performing moisture preservation culture until the inoculated corn leaves show symptoms, and spraying water on the inner wall of the barrel and the plastic film by using the manual pressure spraying pot for moisture preservation, and simultaneously keeping the indoor temperature at 20-25 ℃;
wherein the corn variety is the 'top grade' of susceptible super-sweet corn.
The method is suitable for indoor seedling stage artificial inoculation of the northern leaf blight of corn at any time, and is also suitable for indoor seedling stage resistance identification of northern leaf blight of different corn varieties.
The invention has the advantages that:
(1) The invention provides an artificial inoculation method of indoor corn seedling-stage northern leaf blight by utilizing tissue strip bacteria carrying (having a moisturizing effect), indoor temperature control and moisturizing in a moisturizing barrel, and under the condition of ensuring normal growth of corn seedlings, through scientific artificial inoculation method selection, optimization of a post-inoculation moisturizing method and the like, the artificial inoculation of the indoor corn seedling-stage northern leaf blight can achieve an ideal and stable disease attack effect.
(2) The artificial inoculation method of the indoor corn seedling stage northern leaf blight, provided by the invention, can provide important technical support for indoor pathogenicity determination of the corn northern leaf blight, identification of seedling stage resistance of corn varieties to northern leaf blight and prevention and control research of the corn northern leaf blight, and has a wide application prospect.
Drawings
FIG. 1: the cultivation of the maize seedlings on the indoor illumination rack is realized in the embodiment of the invention. (A): culturing the corn seedlings in the seedling raising tray to the 2-leaf stage; (B): transplanting the seedlings into a plastic bowl to cultivate the corn seedlings in 3-4 leaf stage.
FIG. 2: the bacterial filament and spore suspension of the northern leaf blight strain for artificial inoculation in the embodiment of the invention. The spore concentration is 20-30 spores/visual field (A) and 2-3 spores/visual field (B) under a 10X 20 times microscope respectively.
FIG. 3: different vaccination methods in the examples of the present invention. (A): carrying bacteria on a tissue strip and inoculating; (B): filter paper strip bacteria-carrying inoculation method; (C): spray inoculation method.
FIG. 4: the invention provides a method for moisturizing a plastic barrel covered film after inoculation.
FIG. 5: the disease onset effect of different inoculation methods in the examples of the invention. (A): carrying bacteria on a tissue strip and inoculating; (B): filter paper strip bacteria-carrying inoculation method; (C): spray inoculation method.
Detailed Description
In order to make the content of the present invention easier to understand, the technical solutions of the present invention are further described below with reference to specific embodiments, but the present invention is not limited thereto.
The first embodiment is as follows: effect of different inoculation methods on the onset of disease
1. Test strains: the northern leaf blight NJ08 with better spore yield is used for the inoculation experiment of the invention after indoor living inoculation rejuvenation;
2. test PDA medium: fresh peeled potatoes 200 g, glucose 20 g, agar powder 18 g, deionized water 1000 mL;
3. the corn variety to be tested: the disease-susceptible variety super-sweet corn 'Shangping' (agriculture development Co., ltd. Of Fujian province).
4. The test method comprises the following steps:
(1) Planting corn: in the room, healthy corn seeds are sown in seedling raising trays filled with topiraw peat soil, 2 seeds are sown in each hole, the seedling raising trays are placed in plastic trays filled with half-tray water, and the seedlings are raised to the 2-leaf stage on illumination racks (4 layers of stainless steel frames, 60 cm per layer of racks, 4 incandescent lamps with 48W cold light sources are installed at the top of each layer to provide light sources) (fig. 1A). Then, transplanting the corn seedlings and the peat soil at the roots in each hole into a white plastic bowl (diameter is 11.5 cm), transplanting 2 holes (3-4 seedlings in total) in each bowl, placing the plastic bowl into a plastic tray filled with half tray water, and culturing the seedlings on a lighting frame to 3-4 leaf stages (figure 1B). The indoor temperature is controlled to be 25-30 ℃ during the seedling raising period.
(2) Preparation of hypha + spore suspension: inoculating rejuvenated northern leaf blight of maize to a PDA culture medium, culturing in the dark at 25 ℃ for 10 d, adding 10mL sterile water into each dish, gently brushing hyphae and conidia on the surface of the culture medium by using a sterile brush, preparing a suspension by using the sterile water, and adjusting the concentration of the spores in the suspension to 20-30 spores per visual field (10X 20 times of a mirror) (figure 2A). The hypha + spore suspension was aliquoted into 3 portions for subsequent inoculation experiments.
(3) The inoculation method comprises the following steps: the invention adopts 3 inoculation methods, namely (1) a tissue strip bacteria-carrying inoculation method (figure 3A), (2) a neutral filter strip bacteria-carrying inoculation method (figure 3B), and (3) a spray inoculation method (figure 3C). When indoor maize seedlings are cultivated to 3-4 leaf stages, respectively cutting sanitary paper (Vida brand paper, three layers) and neutral filter paper into strips of 3.0 cm × 1.0 cm (length × width), respectively immersing the paper extraction strips and the filter paper strips into hypha + spore suspension liquid of 20-30 spores/visual field (10 × 20 times of lens) with 30 s, clamping 1 paper strip from the suspension liquid by using sterile tweezers, adhering the paper strip to the middle of leaves of the maize seedlings, inoculating 2 leaves to each maize seedling, and inoculating 4 bowls (12-16 seedlings in total) for each treatment; the remaining 1 part of the hypha and spore suspension is uniformly sprayed on the leaves of the corn seedlings by a manual pressure spray can until the leaves drip water, and the treatment is also carried out for 4 bowls (12-16 seedlings in total).
(4) And (3) moisturizing management after maize seedling inoculation: uniformly spraying the inner wall of a plastic barrel (with the inner diameter of 31 cm and the height of 32 cm) with sterile water by using a manual pressure spraying pot, preferably dripping water on the inner wall of the plastic barrel, transferring the inoculated corn seedlings into the barrel, wherein 4 bowls are arranged in each barrel, the opening of each barrel is covered with a plastic film which is soaked in the sterile water for 5 min, and performing moisture preservation culture until the inoculated corn leaves show symptoms (figure 4). During the period, the inner wall of the barrel and the plastic film can be sprayed with sterile water by a manual pressure spray can to keep moisture, the inner wall of the plastic barrel/the plastic film is preferably dripped, and the indoor temperature is kept at 20 to 25 ℃.
(5) And (4) disease investigation: after inoculation of 2 d, observing the disease occurrence condition of the plant every day, investigating the disease level of each inoculated leaf after the disease is stable, and counting the disease occurrence rate and disease index. The severity of the disease was calculated according to the disease classification standard of corn northern leaf blight in Wang Xiaoming handbook of corn field Manual, disease and pest identification and resistance identification (2010, chinese agricultural science and technology Press). The disease grading standard of the corn northern leaf blight is as follows:
level 0: no obvious scab is generated on the leaves;
level 1: the lesion area on the leaves accounts for less than 5 percent of the leaf area;
and 3, level: the lesion area on the leaves accounts for 6 to 10 percent of the leaf area;
and 5, stage: the lesion area on the leaves accounts for 11 to 30 percent of the leaf area;
and 7, stage: the lesion area on the leaves accounts for 30 to 70 percent of the leaf area;
and 9, stage: the lesion spots on the leaves account for more than 70 percent of the area of the leaves.
5. Results and analysis:
as can be seen from Table 1 and FIG. 5, 3 methods of bacteria-carrying inoculation and spray inoculation of paper strips and filter paper strips can cause diseases, but the degree of the diseases (disease index) is significantly different (P< 0.05). The tissue strip bacteria-carrying inoculation method has the best effect, the disease index is more than 60, the error is minimum, and the disease level is 1-9; the spray inoculation method has secondary effect, the disease index is between 45.5 and 53.7, and the disease level is 0 to 9; the filter paper strip has the worst bacteria-carrying inoculation effect, the disease index is less than 43, the error is large, and the disease level is 1-9.
TABLE 1 Effect of different methods for inoculating northern leaf blight of corn
Note: different lower case letter representationsPThe difference is obvious below the level of 0.05, and the same is true below.
From the onset time, the tissue extraction and filter paper strip bacteria-carrying inoculation method is obviously faster than the spray inoculation method, water stain-shaped disease spots can be seen after 3 d is inoculated by the first 2 methods, 5-7 d disease spots can expand rapidly, the blades covered by the paper strips are died after 7 d, and the whole inoculated blades are died after 15 d. After 7 d is inoculated by the spray inoculation method, a strip-shaped typical lesion appears, and the disease condition tends to be stable after 15 d. According to Wang Xiaoming, a corn plant disease and insect pest field manual, namely, plant disease identification and resistance identification (2010, chinese agricultural science and technology publishing Co.), when the disease level of a susceptible variety reaches 7 grades or above, the resistance identification of a new corn variety to the corn northern leaf blight is regarded as effective, and the inoculation method for carrying bacteria by using the tissue strips meets the disease condition required by the resistance identification of the corn northern leaf blight.
In addition, the paper extraction strip has excellent water absorption and moisture retention, and the texture is soft, and the adhesion of the paper extraction strip to the corn leaves is extremely strong. Although the filter paper strips have strong water absorption and moisture retention performances, the filter paper strips have strong hardness and poor adhesion with corn leaves, and are easy to fall off in the processes of inoculation and subsequent moisture retention culture. Therefore, the invention takes the tissue paper strip bacteria-carrying inoculation as the best embodiment.
Example two: influence of different bacterial receiving amount on disease onset effect
1. Corn variety: 'Top grade'
2. Planting corn: the same as the first embodiment.
3. Hypha + spore suspension: a hypha + spore suspension was prepared as in example one. The spore concentration in the hyphal + spore suspension was adjusted to 20-30 spores/field (10 × 20 fold mirror) with sterile water (fig. 2A). The hypha + spore suspension was divided into 3 portions, and each portion was diluted 10-fold to make the spore concentration in the hypha + spore suspension 2 to 3 spores per visual field (10 × 20-fold mirror) (fig. 2B). The diluted hyphal + spore suspension was used for subsequent inoculation experiments.
4. Inoculation method and moisture preservation management: the same as the first embodiment.
5. Results and analysis
TABLE 2 Effect of the onset of disease Using different inoculation methods and inoculum sizes
As can be seen from Table 2, 20-30 spores/visual field (10X 20 times of lens) of hypha + spore suspension are diluted by 10 times, and the corn seedling can be attacked by 3 inoculation methods, and the disease index of the diluted solution is obviously lower than that of the mother solution. Wherein, the disease grades of the hypha + spore suspension with 2-3 spores/visual field by adopting filter paper strip bacteria carrying and spray inoculation methods are 3 grades and 5 grades respectively, and the disease onset requirement of disease resistance identification can not be met. Hypha + spore suspension with 2-3 spores/visual field adopts a tissue strip bacteria-carrying inoculation method, although the disease condition of 9 grades can be caused, the overall disease index is lower (less than 34). Therefore, the present invention uses hypha + spore suspension of 20-30 spores/visual field (10 × 20 times of mirror) as the best example for the subsequent experiments.
Example three: influence of moisturizing and non-moisturizing treatment on disease onset effect
1. Corn variety: 'Top grade'
2. Planting corn: the same as the first embodiment.
3. The test method comprises the following steps: the other methods are the same as the first embodiment. The moisturizing treatment method of the inoculated corn seedlings is designed with 2 treatments, namely, wetted plastic films are adopted to cover the opening of the barrel for moisturizing (moisturizing group) and unwetted plastic films are adopted to cover the opening of the barrel for moisturizing (non-moisturizing group), and other treatments are unchanged.
4. And (4) analyzing results:
TABLE 3 onset of post-inoculation moisturization and non-moisturization treatments
The results are shown in table 3, and the moisturizing treatment of maize seedlings has a significant effect on the onset of northern leaf blight. After the corns are inoculated in the 3-4 leaf stage, the disease attack effect of the moisturizing treatment group is the best by covering the bucket opening with a soaked plastic film, the disease index is between 58.9 and 64.4, and the highest disease grade reaches 9. The group treated without moisturizing can also have diseases, but the disease onset effect is very poor, the disease index is only 18.5 at the highest, and the highest disease grade is only 3. Therefore, the wetting treatment method for covering the opening of the bucket by the wetted plastic film ensures the humidity requirements required by germination and infection of the corn northern leaf blight spores, and effectively improves the infection rate of the corn northern leaf blight and the severity of diseases.
Example four: effect of inoculating different corn varieties on disease onset
1. The corn variety to be tested: yuexingnuo No. 6 (institute of agricultural sciences, guangdong province), shangpin (agricultural development Co., ltd. Of Fujian province), caiyounuo No. 168 (institute of Dongyang corn, zhejiang province), taixianuo No. 1 (Shanlong agricultural science and technology Co., ltd. Of Jianou City), minsunuo No. 816 (institute of agricultural sciences, fujian province) and Yuexingnuo No. 32 (institute of agricultural sciences, guangdong province).
2. The test method comprises the following steps: the same as in example 1.
3. Results and analysis
TABLE 4 Effect of the invention on the inoculation of different maize varieties using the best mode of practice of the invention
The results are shown in table 4, the disease indexes of 6 fresh-eating corn varieties to be tested are between 55.6 to 77.8, wherein the disease index is remarkably higher than the disease indexes of 'Yueyangnuo No. 6', 'Shangping', 'Caiguan 168', 'Taxianuan No. 1' and 'Minweinuo 816' by taking the highest degree of disease incidence of 'Yueynuo 32', which indicates that the method provided by the embodiment 1 can achieve ideal disease incidence effect and distinguish the resistance of different corn varieties to the northern leaf blight after being inoculated with the northern leaf blight.
The above description is only for the preferred embodiment of the present invention, and all the equivalent changes and modifications made according to the claims of the present invention are included in the scope of the present invention.
Claims (5)
1. An artificial inoculation method of indoor corn seedling-stage northern leaf blight, which is characterized by comprising the following steps: comprises three parts of indoor corn seedling planting management, artificial inoculation treatment of northern leaf blight in a corn seedling stage and moisture preservation management after artificial inoculation of corn seedlings; the specific operation is as follows:
(1) Planting management of indoor maize seedlings: sowing corn seeds in a seedling raising tray indoors, sowing 2 seeds in each hole, placing the seedling raising tray in a plastic tray filled with half tray water, raising the seedlings to a 2-leaf stage on an illumination frame, then transplanting the seedlings into a white plastic bowl, transplanting 3-4 seedlings in 2 holes in each bowl, placing the plastic bowl in the plastic tray filled with half tray water, raising the seedlings to a 3-4-leaf stage on the illumination frame, and controlling the indoor temperature to be 25-30 ℃ during seedling raising;
(2) Artificial inoculation treatment of northern leaf blight in corn seedling stage: inoculating rejuvenated maize northern leaf blight which is inoculated in a living body to a PDA culture medium, culturing 10 d in dark at 25 ℃, adding 10mL sterile water into each dish, slightly brushing maize northern leaf blight hypha and conidia on the surface of the culture medium by using a sterile brush, and preparing hypha + conidia suspension by using the sterile water for inoculation; when indoor maize seedlings are cultivated to 3-4 leaf stages, cutting sanitary paper extracts into strips, immersing the strips into the hypha and conidium suspension for 30 s, clamping 1 paper extract strip from the hypha and conidium suspension by using sterile forceps, adhering the paper extract strip to the middle of leaves of the maize seedlings for inoculation, and inoculating 2 leaves to each maize seedling;
(3) Moisture preservation management after artificial inoculation of corn seedlings: uniformly spraying water on the inner wall of a plastic barrel by using a manual pressure spraying pot, transferring the inoculated corn seedlings into the barrel, covering 4 bowls of each barrel by using a soaked plastic film at the barrel opening, performing moisture preservation culture until inoculated corn leaves show symptoms, spraying water on the inner wall of the barrel and the plastic film by using the manual pressure spraying pot for moisture preservation in the period, and simultaneously keeping the indoor temperature at 20 to 25 ℃.
2. The artificial inoculation method of indoor northern leaf blight in a seedling stage of corn according to claim 1, characterized in that: the size of the white plastic bowl is 11.5 cm; the illumination frame is a 4-layer stainless steel frame, each layer is 60 cm in height, and 4 incandescent lamps with 48W cold light sources are arranged at the top to provide light sources.
3. The artificial inoculation method of indoor northern leaf blight in a seedling stage of corn according to claim 1, characterized in that: the specification of the paper extraction strip is 3.0 cm × 1.0 cm.
4. The artificial inoculation method of indoor northern leaf blight in a seedling stage of corn according to claim 1, characterized in that: the spore concentration in the hypha + spore suspension is 20-30 spores per visual field under a microscope with the magnification of 10 multiplied by 20.
5. The artificial inoculation method of indoor northern leaf blight in a seedling stage of corn according to claim 1, characterized in that: the plastic bucket is 31 cm in inside diameter and 32 cm in height.
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| CN104164472A (en) * | 2014-07-15 | 2014-11-26 | 武汉市蔬菜科学研究所 | Indoor identification method for anti-phomopsis blight |
| CN105925522A (en) * | 2016-07-19 | 2016-09-07 | 福建省农业科学院植物保护研究所 | Setosphaeria turcica spore production medium and application thereof |
| CN109042156A (en) * | 2018-07-18 | 2018-12-21 | 福建省农业科学院植物保护研究所 | A kind of Inoculation Method of the temperature canopy facility leaf blight of corn |
| US20200362425A1 (en) * | 2019-05-13 | 2020-11-19 | Shenyang Agricultural University | Bacillus having effect of inducing corn to resist northern corn leaf blight (exserohilum turcicum) of corn and application |
| CN113597917A (en) * | 2021-08-05 | 2021-11-05 | 嘉兴市农业科学研究院 | Artificial inoculation method for ustilaginoidea virens |
| CN115058486A (en) * | 2022-06-23 | 2022-09-16 | 贵州省植物保护研究所 | Disease resistance identification method for corn leukoderma |
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104164472A (en) * | 2014-07-15 | 2014-11-26 | 武汉市蔬菜科学研究所 | Indoor identification method for anti-phomopsis blight |
| CN105925522A (en) * | 2016-07-19 | 2016-09-07 | 福建省农业科学院植物保护研究所 | Setosphaeria turcica spore production medium and application thereof |
| CN109042156A (en) * | 2018-07-18 | 2018-12-21 | 福建省农业科学院植物保护研究所 | A kind of Inoculation Method of the temperature canopy facility leaf blight of corn |
| US20200362425A1 (en) * | 2019-05-13 | 2020-11-19 | Shenyang Agricultural University | Bacillus having effect of inducing corn to resist northern corn leaf blight (exserohilum turcicum) of corn and application |
| CN113597917A (en) * | 2021-08-05 | 2021-11-05 | 嘉兴市农业科学研究院 | Artificial inoculation method for ustilaginoidea virens |
| CN115058486A (en) * | 2022-06-23 | 2022-09-16 | 贵州省植物保护研究所 | Disease resistance identification method for corn leukoderma |
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