CN110607345A - Method for identifying resistance of wheat in seedling stage of sheath blight - Google Patents
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Abstract
The invention provides a method for identifying resistance of wheat in a seedling stage of sheath blight, which relates to the technical field of plant protection and breeding, and comprises the following steps: s1: preparing a suspension of wheat sheath blight hyphae; s2: taking wheat seeds as a material, and carrying out seed germination to obtain germinated seeds; s3: culturing the germinated seeds in the dark at the temperature of 22 ℃ to obtain wheat sprouts; s4: soaking the wheat sprouts in the wheat sheath blight hypha suspension liquid for inoculation to obtain inoculated wheat sprouts; s5: culturing the inoculated wheat plumule to obtain a wheat seedling; s6: investigating the outbreak situation of the sheath blight of the wheat seedlings, and predicting the resistance of different wheat varieties in the sheath blight seedling stage. The method for identifying the resistance of the wheat sheath blight in the seedling stage is simple to operate, convenient, rapid and efficient, can accurately identify the resistance level of the wheat sheath blight in the seedling stage in a short time, is low in cost, labor-saving and suitable for popularization and application.
Description
Technical Field
The invention relates to the technical field of plant protection and breeding, in particular to a method for identifying resistance of wheat in a seedling stage of sheath blight.
Background
Wheat sharp eyespot is a fungal disease mainly caused by Rhizoctonia cerealis (Rhizoctonia cerealis) and Rhizoctonia solani (Rhizoctonia solani) and is mainly transmitted by soil-borne bacteria. Under natural conditions, the sheath blight of wheat in seedling stage mainly occurs on the leaf sheath of wheat, and after the wheat is pulled out of the node, the symptoms gradually get worse along with the rise of temperature and sufficient rainwater. At the initial stage of disease attack, tawny elliptic or fusiform disease spots are generated on leaf sheaths on the surface of the earth or near the surface of the earth, then pathogenic bacteria are gradually infected into the inner stalks, the stalks form grayish brown in the middle and brown near-circular or elliptic eye spots on the periphery, when the disease attack is serious, the stalk walls are dehydrated and necrotized, finally, the diseased wheat plants die due to insufficient supply of nutrients and water, and dead wheat plants are formed, so that thousand seed weight is reduced, and the yield is sharply reduced.
In recent years, due to the lack of disease-resistant varieties in production, excessive use of nitrogen fertilizers, change of farming systems such as straw returning and the like, the incidence rate of wheat sharp eyespot is increased year by year. According to the data of Ministry of agriculture, the annual incidence area of wheat sharp eyespot in China reaches ten million mu, and the number of retransmitted year can reach hundreds of millions of mu.
Although the harm of the wheat sharp eyespot can be reduced to a certain degree by the control of the medicament, the production cost of the wheat is increased and the environment is polluted, so that the cultivation and the utilization of disease-resistant varieties are the most economic, effective and environment-friendly means for reducing the harm of the wheat sharp eyespot.
The establishment of an efficient and reliable identification method for the resistance of the wheat sharp eyespot is an important basis for cultivating and utilizing disease-resistant varieties. At present, the resistance identification methods of wheat sheath blight in the adult stage are more, and are divided into natural identification and artificial inoculation identification. However, the identification in the adult plant stage is greatly influenced by the field environment, the identification time is long, the phenotypic stability is poor, or the method is complex, the workload is large, the identification time is long, and the method is not suitable for large-scale resistance identification (Yunberg, any Yan, Houweixiu, and the like, improvement of the identification method for the banded sclerotial blight resistance in the seedling stage of wheat and screening of disease-resistant varieties [ J/OL ]. Phytopathology report: 1-8).
Relatively speaking, the resistance identification of the wheat sheath blight in the seedling stage in the greenhouse has the advantages of easy environment control, short period and strong repeatability. At present, some researches are carried out on the identification method of the seedling stage of the sheath blight of the wheat in a greenhouse, the Zijuan and the like are lightly embedded into stems and sheaths of the wheat seedlings by utilizing toothpicks with bacteria in a 3-4 leaf stage (the greenhouse) of the wheat and are inoculated by winding wetted medical absorbent cotton balls at the inoculation positions, and results show that the identification and evaluation method of the method has good disease consistency and stability (the Zijuan, the Yajinbao, the Chenazol and the Mahongxiang). The method is characterized in that Ergberg and the like utilize plastic small box greenhouse soil to plant wheat to be identified, diseased wheat grains are inoculated in the 2-leaf stage of wheat seedlings, and the results of identifying the resistance to banded sclerotial blight of 96 wheat varieties show that the method is stable in banded sclerotial blight morbidity and good in repeatability (Ergberg, Nioyan, Renwei, Renweixiu, and the like, the improvement of the identification method for the banded sclerotial blight in the seedling stage of the wheat and screening of the disease-resistant varieties [ J/OL ]. the plant pathologist: 1-8), but the method needs the processes of diseased wheat grain preparation, high-temperature soil sterilization and the like, and has the problems of relatively large workload and low efficiency.
At present, the disease area of wheat sharp eyespot in China is continuously enlarged, the harm degree is increasingly serious, and a variety for resisting sharp eyespot is urgently needed in production. Therefore, the establishment of a rapid, accurate and efficient method for identifying the sheath blight of wheat is necessary, and the establishment of the method can provide a basis for the research of screening of sheath blight resistant germplasm resources, breeding of new disease resistant wheat varieties, disease resistance mechanisms and the like.
Disclosure of Invention
The invention solves the problem of how to quickly, accurately and efficiently identify the resistance of wheat sheath blight at the seedling stage.
In order to solve the problems, the invention provides a method for identifying the resistance of wheat sheath blight in the seedling stage, which comprises the following steps:
s1: preparing a suspension of wheat sheath blight hyphae;
s2: taking wheat seeds as a material, and carrying out seed germination to obtain germinated seeds;
s3: carrying out dark culture on the germinated seeds at 22 ℃ to obtain wheat sprouts;
s4: soaking the wheat sprouts in the wheat sheath blight hypha suspension liquid for inoculation to obtain inoculated wheat sprouts;
s5: culturing the inoculated wheat plumule to obtain a wheat seedling;
s6: investigating the outbreak situation of the sheath blight of the wheat seedlings, and predicting the sheath blight seedling stage resistance of the wheat seedlings of different wheat varieties.
The identification method for the resistance of the wheat sheath blight in the seedling stage is not limited by the growing season and the external environment by carrying out seed germination and culture in a greenhouse, and meanwhile, because the conditions such as indoor temperature, humidity and the like in the greenhouse environment are easy to control, compared with the traditional method for identifying the resistance of the wheat sheath blight in the seedling stage in the field, the method is more convenient, time-saving and labor-saving.
In order to improve the consistency of the identification results, the invention further selects the seeds with consistent germination degree for culture in step S3, specifically, the step is that the seeds with consistent germination degree are selected and transferred into a germination box for continuous dark culture at 22 ℃ to obtain the wheat sprouts.
In addition, after the seeds germinate, dark culture is carried out, so that compared with wheat sprouts cultured by illumination, the wheat sprouts cultured by the germinated seeds have yellow plants and relatively weak growth, and are easier to inoculate in the subsequent process, thereby being beneficial to shortening the time for identifying the resistance of the wheat in the seedling stage of sheath blight, reducing the difficulty for identifying the resistance of the wheat in the seedling stage of sheath blight and improving the reliability and stability of identification results.
Optionally, step S1 includes:
s1-1: inoculating Rhizoctonia cerealis strain stored at-20 deg.C on PDA plate, culturing at 22 deg.C for 3d, and activating Rhizoctonia cerealis to obtain activated mycelium;
s1-2: inoculating the activated hyphae on a PDA liquid culture medium, performing shake culture at 22 ℃ for 5d, and performing hyphae culture to obtain a bacterial liquid;
s1-3: and transferring the bacterial liquid into a centrifugal tube, adding steel balls, and performing vortex oscillation to obtain the wheat sheath blight hypha suspension.
Wherein the rhizoctonia solani strain in the process of preparing the wheat sharp eyespot hypha suspension is 29-1 rhizoctonia solani strain stored at the temperature of-20 ℃, and is stored in key laboratories of Hubei province in the prevention and control of major diseases and pests of crops.
Culturing Rhizoctonia solani strain stored at-20 deg.C at 22 deg.C for 3d, activating Rhizoctonia solani, and shake culturing activated mycelium at 22 deg.C for 5d for mycelium culture; specifically, the shake culture process is that shake culture is carried out in a shaking table at 22 ℃ at the frequency of 150r/min for 5d to obtain a bacterial liquid; in order to facilitate the inoculation of the wheat seedlings, the hyphae in the bacterial liquid are further crushed by vortex oscillation; in the vortex oscillation treatment process, the specification of a centrifugal tube is 50ml, the number of steel balls added is 15-20, and the diameter of the steel balls is 3 mm; after vortex oscillation, beating the rhizoctonia solani hyphae into small segments with the length of 1-2 mm, and uniformly mixing to form hypha suspension.
In addition, during the culture in step S1-1, in order to prevent distilled water present on the lid of the culture dish from flowing onto the culture dish during the culture, resulting in a sheet of colonies that cannot be counted or separated, and to reduce the possibility of contamination, the present invention performs an inverted culture of the strain of Rhizoctonia solani in the incubator in S1-1.
Optionally, step S2 includes:
s2-1: sterilizing the wheat seeds to obtain sterilized wheat seeds;
s2-2: and (3) placing the sterilized wheat seeds in sterile water, and germinating for 2d in the dark at the temperature of 22 ℃ to obtain the germinated seeds.
In order to improve the germination rate of the seeds, the seeds with better plumpness are selected in the process of germinating the seeds, and the selected seeds are sterilized and then germinated so as to avoid influencing the accuracy of the identification result due to other problems such as seed vitality and the like.
The seeds are germinated under the dark condition, so that the wheat sprouts cultured by the germinated seeds are weaker in growth and easier to inoculate, and the time for identifying the resistance of the wheat in the seedling stage of the sheath blight is shortened.
Alternatively, step S2-1 includes: and (3) sequentially washing the wheat seeds with an ethanol solution and sterile water, and soaking the washed wheat seeds in hydrogen peroxide to obtain the sterilized wheat seeds.
Optionally, the concentration of the hydrogen peroxide is 0.6%.
Optionally, the ethanol solution has a concentration of 70%.
Washing the selected wheat seeds with 70% ethanol for 1min, and washing the washed seeds with sterile water at a high speed to remove ethanol on the surface layers of the wheat seeds; and (3) placing the wheat seeds washed by the sterile water into hydrogen peroxide with the concentration of 0.6% in a culture dish, and soaking overnight to obtain the sterilized wheat seeds.
When the sterilized wheat seeds germinate, pouring out hydrogen peroxide in a culture dish for soaking the wheat seeds, and washing the wheat seeds for 1 time by using 5ml of sterile water to remove the hydrogen peroxide on the surfaces of the wheat seeds; adding a proper amount of sterile water into a culture dish containing the wheat seeds, and germinating for 2d at the temperature of 22 ℃ to obtain germinated seeds.
Optionally, the dark culture time in step S3 is 2 d.
Specifically, the germination seeds are cultured in the dark in a sterilized germination box, and the length, width and height of the germination box are 11.5cm, 11.5cm and 9.8cm respectively; firstly, filling filter paper on the bottom of a sterilized germination box, preferably filling three layers of filter paper on the bottom of the germination box; and secondly, adding 10ml of sterile water into a germination box filled with filter paper, selecting germinated seeds with better growth vigor, selecting 20 seeds with better growth vigor and consistent germination degree, transferring the seeds into the germination box filled with sterile water, and carrying out dark culture for 2d at 22 ℃.
After 2d of dark culture, the sheath of the wheat germ in the germination box grows to about 2cm to obtain wheat sprouts; and soaking the wheat sprouts in the wheat sheath blight hypha suspension for inoculation to obtain the inoculated wheat sprouts.
The inoculation process in the invention is specifically that 5ml of the wheat sharp eyespot hypha suspension obtained in the step S1 is evenly inoculated in each germination box with wheat sprouts through a liquid transfer gun to obtain the inoculated wheat sprouts.
Optionally, step S5 includes: and (3) carrying out dark culture on the inoculated wheat plumule for 5d, and then carrying out culture for 10d under the conditions of the illumination time length of 16h and the dark time length of 8 h.
Optionally, the temperature for culturing the inoculated wheat sprouts is 22 ℃.
The inoculated wheat sprouts are continuously cultured in the germination box, and the wheat sprouts are continuously kept soaked in the wheat sharp eyespot hypha suspension liquid in the culture process, so that the success rate of inoculation of the wheat sprouts is convenient to improve, the wheat sprouts are prevented from being moved to other positions from the germination box, the operation difficulty is favorably reduced, the damage to the wheat sprouts in the transplanting process of the wheat sprouts can be avoided, and the accuracy of an identification result is improved.
In addition, the wheat sprouts are cultured in the germination box, and the control of the culture environment is facilitated; specifically, in the process of culturing the wheat sprouts, the wheat sprouts in the germination box are covered by the cover of the germination box, so that the wheat sprouts are cultured for 5 days in a dark environment; and then opening the cover of the germination box, placing the germination box in a greenhouse with the temperature of 22 ℃, the illumination for 16h and the darkness for 8h for continuous culture for 10d, and adding a proper amount of sterile water into the germination box every day during the whole culture period to finish the culture of the wheat plumule and obtain the wheat seedling.
Because the wheat sprouts are firstly cultured for 5 days in the dark environment, the wheat sprouts are yellow, the growth vigor is weaker, the inoculation is easier, and the dark culture of 5 days after the inoculation is also favorable for the occurrence of sheath blight, so the time for identifying the resistance of the wheat at the seedling stage of the sheath blight is favorably shortened, for example, the process of culturing the wheat sprouts into the wheat seedlings can be completed only by 20 days.
Optionally, step S6 includes:
s6-1: according to the disease severity of the wheat seedlings, dividing sheath blight into the following grades:
level 0: no sheath blight disease; level 1: sheath has sheath blight disease spots, and the length of the disease spots on the stem is less than or equal to 1 cm; and 2, stage: the length of the disease spot on the stem is more than 1 cm; and 3, level: the first leaf shows wilting symptoms; 4, level: death of the seedling;
s6-2: calculating the disease index of the wheat seedling according to the following formula:
the disease index [ (Σnumber of diseased plants at each stage × representative values at each stage)/(total number of plants × highest representative value) ] × 100;
s6-3: according to the disease index, identifying the resistance of the wheat seedlings in the sheath blight seedling stage according to the following standards:
disease index less than or equal to 40 is 'anti'; the disease index is more than 40 and less than or equal to 60, and the disease resistance is obtained; the disease index is more than 60 and less than or equal to 100, which is the feeling.
In order to identify the resistance of the wheat variety in the seedling stage of the sheath blight, the wheat seedling is taken out from a germination box, the sheath blight is classified according to the disease severity of the wheat seedling, a calculation formula of disease indexes is provided, the disease indexes of the wheat seedlings of different wheat varieties are calculated, the resistance of the wheat seedlings of different varieties in the seedling stage of the sheath blight is identified according to the calculated disease indexes and corresponding standards, so that the identification standards are unified, and the aim of objectively evaluating the resistance of the wheat seeds of different varieties in the seedling stage of the sheath blight is fulfilled.
Compared with the prior art, the method for identifying the resistance of the wheat sheath blight in the seedling stage has the following advantages:
the method for identifying the resistance of the wheat sharp eyespot in the seedling stage is simple to operate, convenient, rapid and efficient, can accurately identify the resistance level of the wheat sharp eyespot in the seedling stage in a short time, does not need expensive instruments and equipment, is low in cost, saves labor and time, and is suitable for popularization and application.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, specific embodiments thereof are described in detail below.
Example 1
The embodiment provides a method for identifying resistance of wheat in a seedling stage, which comprises the following steps:
s1-1: inoculating Rhizoctonia solani strain 29-1 stored at-20 deg.C on Potato Dextrose Agar (PDA) plate, culturing in an incubator at 22 deg.C for 3 days, and activating Rhizoctonia solani to obtain activated mycelium;
s1-2: inoculating the activated hyphae on a PDA liquid culture medium, performing shake culture in a shaking table at 22 ℃ at a rotating speed of 150r/min for 5d, and performing hyphae culture to obtain a bacterial liquid;
s1-3: transferring the obtained bacterial liquid into a centrifugal tube with the specification of 50ml, adding 20 steel balls with the diameter of 3mm, carrying out vortex oscillation until the rhizoctonia solani hyphae are beaten into small segments with the diameter of 1-2 mm, and then uniformly mixing to obtain a wheat sharp eyespot hypha suspension;
s2-1: selecting 50 seeds with good plumpness from wheat seeds of Mianmai 37 as material, and washing with 70% ethanol for 1 min; after the wheat seeds are quickly washed by sterile water, the wheat seeds are placed in a culture dish, and 5ml of 0.6% hydrogen peroxide is added into the culture dish to be soaked overnight, so that sterilized wheat seeds are obtained;
s2-2: and (3) pouring out hydrogen peroxide in the culture dish, washing the wheat seeds once by using 5ml of sterile water, adding a proper amount of sterile water, placing the sterilized wheat seeds in the sterile water, and germinating for 2d at the temperature of 22 ℃ to obtain germinated seeds.
S3: sterilizing the germination box with an autoclave, filling three layers of filter paper on the bottom of the sterilized germination box (length, width and height of 11.5cm, 11.5cm and 9.8cm respectively), and adding 10ml of sterile water; selecting 20 germinated seeds with consistent growth vigor, transferring to a germination box, enabling the back of each seed to face upwards, carrying out dark culture at 22 ℃ in a greenhouse for 2 days, and enabling the sheath of the wheat germ to grow to be about 2cm to obtain the wheat germ;
s4: uniformly inoculating 5ml of wheat sheath blight hypha suspension into each germination box through a liquid transfer gun to obtain inoculated wheat sprouts;
s5: shielding the germination box through a cover of the germination box, carrying out dark culture on the inoculated wheat sprouts in a greenhouse at 22 ℃ for 5 days, opening the cover of the germination box, placing the germination box in the greenhouse at 22 ℃ for 16h under light and 8h in darkness, continuously culturing for 10 days, and adding a proper amount of sterile water every day during the period to carry out culture to obtain wheat seedlings;
s6: taking out the wheat seedlings from the germination box, investigating the banded sclerotial blight morbidity of the wheat seedlings, and predicting the banded sclerotial blight seedling stage resistance of the wheat seedlings according to the following method:
dividing the sheath blight disease into 5 grades, 0 grades and no sheath blight disease according to the severity of the disease; grade 1, sheath of the leaf has sheath blight disease spots, and the length of the disease spots on the stem is less than or equal to 1 cm; grade 2, the length of the disease spot on the stem is more than 1 cm; grade 3, the first leaf shows wilting symptoms; grade 4, seedling death.
Calculating disease index according to the total number of the investigated seedlings; the disease index [ (Σnumber of diseased plants at each stage × representative values at each stage)/(total number of plants × highest representative value) ] × 100.
Judgment standard of banded sclerotial blight resistance: the disease index is less than or equal to 30 and is High Resistance (HR), the disease index is less than or equal to 30 and is Medium Resistance (MR), the disease index is less than or equal to 50 and is medium feeling (MS), the disease index is less than or equal to 70 and is high feeling (HS), and the disease index is less than or equal to 70 and is 100.
In order to ensure the accuracy of the identification result, the present example continuously performed 3 independent seedling sheath blight resistance identifications for all wheat varieties, and 20 plants were investigated each time. The results of the study are shown in table 1; the results show that the disease resistance identification result has good repeatability (the correlation coefficient among the repeats is all above 0.9).
The method for identifying the resistance of the wheat sheath blight in the seedling stage is simple to operate, convenient, rapid and efficient, can accurately identify the resistance level of the wheat sheath blight in the seedling stage in a short time, does not need expensive instruments and equipment, is low in cost, saves labor and time, and is suitable for popularization and application.
Examples 2 to 10
In examples 2 to 10, the materials were all selected from the Erlenmeyer 1, Erlenmeyer 5, Erlenmeyer 426, Zheng wheat 9023, Erlenmeyer 170, Yu wheat 70, Yu wheat 52, Ning wheat 9 and Erlenmeyer 577 in this order, and the resistance to sheath blight of wheat was identified by the method provided in example 1, and the results are shown in Table 1.
TABLE 1
| Name of wheat Material | Repeat 1 (finger of disease) | Repeat 2 (finger of disease) | Repeat 3 (finger of disease) | Mean (finger disease) | Evaluation of resistance |
| Sponge wheat 37 | 24.3 | 27.5 | 29.6 | 27.1 | HR |
| Een No. 1 | 33.5 | 38.4 | 39.3 | 37.1 | MR |
| Een No. 5 | 41.4 | 48.8 | 31.8 | 40.7 | MR |
| Jaw 426 | 59.4 | 57.5 | 62.5 | 59.8 | MS |
| Zheng Mai 9023 | 68.4 | 53.3 | 51.3 | 57.7 | MS |
| Hubei 170 | 61.6 | 54.8 | 56.9 | 57.8 | MS |
| Yumai 70 | 68.6 | 54.9 | 58.4 | 60.6 | MS |
| Yumai 52 | 82.7 | 78.9 | 77.4 | 79.7 | S |
| Ning Mai No. 9 | 88.5 | 92.4 | 70.1 | 83.7 | S |
| Ebei 577 | 88.2 | 92.7 | 86.4 | 89.1 | S |
As can be seen from the data in Table 1, the resistance to sheath blight in seedling stage was the strongest in Mianmai No. 37 and the disease resistance was the worst in Eanmai 577 among the reference varieties.
The embodiment proves that the method for identifying the resistance of the wheat in the seedling stage of the sheath blight can identify the resistance of the wheat seedlings of different varieties in the seedling stage of the sheath blight according to corresponding standards, so that the identification standards are unified, and the aim of objectively evaluating the resistance of the wheat seeds of different varieties in the seedling stage of the sheath blight is fulfilled.
Although the present disclosure has been described above, the scope of the present disclosure is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present disclosure, and such changes and modifications will fall within the scope of the present invention.
Claims (10)
1. A method for identifying resistance of wheat in a seedling stage of sheath blight, which is characterized by comprising the following steps:
s1: preparing a suspension of wheat sheath blight hyphae;
s2: taking wheat seeds as a material, and carrying out seed germination to obtain germinated seeds;
s3: carrying out dark culture on the germinated seeds at 22 ℃ to obtain wheat sprouts;
s4: soaking the wheat sprouts in the wheat sheath blight hypha suspension liquid for inoculation to obtain inoculated wheat sprouts;
s5: culturing the inoculated wheat plumule to obtain a wheat seedling;
s6: investigating the outbreak situation of the sheath blight of the wheat seedlings, and predicting the sheath blight seedling stage resistance of the wheat seedlings of different wheat varieties.
2. The method for identifying resistance to sheath blight of wheat as claimed in claim 1, wherein step S1 comprises:
s1-1: inoculating Rhizoctonia cerealis strain stored at-20 deg.C on PDA plate, culturing at 22 deg.C for 3d, and activating Rhizoctonia cerealis to obtain activated mycelium;
s1-2: inoculating the activated hyphae on a PDA liquid culture medium, performing shake culture at 22 ℃ for 5d, and performing hyphae culture to obtain a bacterial liquid;
s1-3: and transferring the bacterial liquid into a centrifugal tube, adding steel balls, and performing vortex oscillation to obtain the wheat sheath blight hypha suspension.
3. The method for identifying resistance to sheath blight of wheat as claimed in claim 1, wherein step S2 comprises:
s2-1: sterilizing the wheat seeds to obtain sterilized wheat seeds;
s2-2: and (3) placing the sterilized wheat seeds in sterile water, and germinating for 2d in the dark at the temperature of 22 ℃ to obtain the germinated seeds.
4. The method for identifying resistance to sheath blight of wheat as claimed in claim 3, wherein the step S2-1 comprises: and (3) sequentially washing the wheat seeds with an ethanol solution and sterile water, and soaking the washed wheat seeds in hydrogen peroxide to obtain the sterilized wheat seeds.
5. The method for identifying the resistance of wheat sheath blight at the seedling stage as claimed in claim 4, wherein the concentration of the hydrogen peroxide is 0.6%.
6. The method for identifying resistance to wheat sheath blight at the seedling stage of claim 4, wherein the concentration of the ethanol solution is 70%.
7. The method for identifying resistance to sheath blight of wheat in the seedling stage of claim 1, wherein the dark culture time in the step S3 is 2 d.
8. The method for identifying resistance to sheath blight of wheat as claimed in claim 1, wherein step S5 comprises: and (3) carrying out dark culture on the inoculated wheat plumule for 5d, and then carrying out culture for 10d under the conditions of the illumination time length of 16h and the dark time length of 8 h.
9. The method for identifying resistance to wheat sharp eyespot in seedling stage of claim 8, characterized in that the temperature for culturing said inoculated wheat sprouts is 22 ℃.
10. The method for identifying resistance to sheath blight of wheat as claimed in claim 1, wherein step S6 comprises:
s6-1: according to the disease severity of the wheat seedlings, dividing sheath blight into the following grades:
level 0: no sheath blight disease; level 1: sheath has sheath blight disease spots, and the length of the disease spots on the stem is less than or equal to 1 cm; and 2, stage: the length of the disease spot on the stem is more than 1 cm; and 3, level: the first leaf shows wilting symptoms; 4, level: death of the seedling;
s6-2: calculating the disease index of the wheat seedling according to the following formula:
the disease index [ (Σnumber of diseased plants at each stage × representative values at each stage)/(total number of plants × highest representative value) ] × 100;
s6-3: according to the disease index, identifying the resistance of the wheat seedlings in the sheath blight seedling stage according to the following standards:
disease index less than or equal to 40 is 'anti'; the disease index is more than 40 and less than or equal to 60, and the disease resistance is obtained; the disease index is more than 60 and less than or equal to 100, which is the feeling.
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