CN109526443B - Method for rapidly identifying disease resistance of radish - Google Patents
Method for rapidly identifying disease resistance of radish Download PDFInfo
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- CN109526443B CN109526443B CN201910031999.9A CN201910031999A CN109526443B CN 109526443 B CN109526443 B CN 109526443B CN 201910031999 A CN201910031999 A CN 201910031999A CN 109526443 B CN109526443 B CN 109526443B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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Abstract
The invention belongs to the technical field of plant resistance identification, and particularly relates to a method for quickly identifying disease resistance of radish. Collecting radishes with diseases to obtain disease pollutants; planting the variety to be detected in a field; inoculating disease pollutants when the radish grows to 3-8 true leaves, respectively inoculating the disease pollutants once in 3-4 leaf stages and 7-8 leaf stages, and investigating disease indexes 10 days after inoculating the disease pollutants in 7-8 leaf stages; and judging the plant resistance according to the disease index. The method of the invention increases the operation of artificial inoculation of disease pollutants on the basis of natural disease garden identification, thereby not only accelerating the infection probability of the natural disease garden, but also improving the screening accuracy of radish resistant varieties.
Description
Technical Field
The invention belongs to the technical field of plant resistance identification, and particularly relates to a method for quickly identifying disease resistance of radish.
Background
Diseases such as black rot and black spot are common diseases for radish planting, and the diseases not only reduce the yield and variety of radish, but also influence the economic income of growers. The method solves the problems that on one hand, the disease resistance and elimination work is required to be done in the radish planting process, and on the other hand, new radish disease-resistant varieties are required to be actively cultivated.
In the prior art, the new species is mostly planted in a disease nursery and is naturally induced to carry out resistance identification, but the current or local climatic conditions are suitable for inducing diseases, so that the identification work can not be smoothly carried out. In order to solve the problem, research on an identification method for disease resistance of radish black rot in seedling stage, Gancaoxia, Hubei agricultural science, volume 49, stage 12 discloses an identification method for indoor artificial inoculation of radish black rot in seedling stage, and by controlling factors such as inoculation concentration, temperature and the like, the disease resistance of radish varieties can be identified smoothly. However, the test strains adopted in the paper are obtained by sampling, separating and identifying disease-resistant strains from a disease area, and only three strains of HB1, WHHB1 and JZHB1 are adopted, so the identified disease-resistant strains are only resistant to the three strains, and radish strains resistant to other microorganisms cannot be screened out. Furthermore, the paper adopts artificial inoculation of specific strains to induce diseases, which is greatly different from a naturally induced nursery identification mode, and the number of the strains causing radish black rot is not only three, but also can be the result of the combined action of a plurality of strains, so that the research method of cabbages and the like has the situation of omitting screening of radish disease-resistant varieties.
Disclosure of Invention
According to the rapid identification method for the disease resistance of the radish, provided by the invention, the operation of artificially inoculating disease pollutants is added on the basis of identification of a natural disease garden, so that the disease infection probability of the natural disease garden is accelerated, and the screening accuracy of radish resistant varieties is improved.
The invention provides a method for rapidly identifying the disease resistance of radish, which comprises the following steps:
s1, collecting diseased radishes, cutting the radishes into radish dices, adding the radish dices into an acetic acid solution, soaking, filtering, and collecting filtrate to obtain disease pollutants;
s2, planting the variety to be detected in the field in the following way: planting by adopting a hole digging and seed dibbling mode, wherein 1-2 seeds are planted in each hole, the plant spacing is 10-20 cm, and the row spacing is 10-30 cm;
s3, performing field management, inoculating disease pollutants when the radish grows to 3-8 true leaves, wherein the disease pollutants are inoculated once in the 3-4 leaf stage and the 7-8 leaf stage respectively, and the disease index is investigated on the 10 th day after the disease pollutants are inoculated in the 7-8 leaf stage;
the disease survey was graded as follows: grade 0, no disease; 0.5 grade, several disease groups are on the leaf surface, and the area of the disease area is less than 1/4 of the leaf area; the disease area on the 1-level leaf is more than or equal to 1/4 leaf area and less than 3/8 leaf area; the area of the disease area on the 2-level leaf is more than or equal to 3/8 of the area of the leaf and less than 1/2 of the area of the leaf; the disease area on the 3-level leaves is more than or equal to 1/2 of the area of the leaves and less than or equal to 3/4 of the area of the leaves, and the disease area on the 4-level leaves is more than 3/4 of the area of the leaves;
disease index of 100X Sigma (number of disease plants at each stage x disease level value)/(total number of investigated plants x 4)
When the disease index is more than 40, the variety to be detected is a susceptible variety; when the disease index is more than or equal to 40 and less than 20, the variety to be detected is a disease-resistant variety; when the disease index is more than or equal to 20 and less than 10, the variety to be detected is a disease-resistant variety; and when the disease index is less than or equal to 10, determining that the variety to be detected is a high disease-resistant variety.
Preferably, in the method for rapidly identifying the disease resistance of radishes, the acetic acid solution accounts for 1-3% by mass, and the soaking time is 5-20 min.
Preferably, in the method for rapidly identifying the disease resistance of the radish, the mass ratio of the diced radish to the acetic acid solution is 1: 1-2.
Preferably, the method for rapidly identifying the disease resistance of the radish comprises the following steps of: and (3) spraying the diluted solution obtained by diluting the disease pollutants by 1000 times to the leaf surfaces, wherein 20-30L of the diluted solution is used per mu.
Preferably, in the method for rapidly identifying the disease resistance of the radish, the side length of the diced radish after dicing of the radish is 0.5-1 cm.
Preferably, in the method for rapidly identifying the disease resistance of radish, the tool used for filtering in the step 1 is a mesh screen, and the diameter of the mesh screen is 0.5-2 mm.
Preferably, in the method for rapidly identifying disease resistance of radish, the step S1 includes the steps of: taking a disease-resistant radish variety as a standard strain A; taking a susceptible radish variety as a standard strain B;
the step of S2 further includes the steps of: planting the standard plant A, the standard plant B and the variety to be detected in a field, and sequentially and repeatedly arranging and planting the radishes according to the sequence of a line of the standard plant A, a line of the standard plant B and a line of the variety to be detected;
s3 further includes the steps of: disease incidence is investigated on the 10 th day after disease pollutants are inoculated in the 7-8 leaf stage;
the incidence rate is the total number of plants with the disease/the total number of investigated plants;
the morbidity of the standard strain A is recorded as KA, the morbidity of the standard strain B is recorded as KB, and the morbidity of the variety to be detected is recorded as KD; when KD is less than or equal to KA, the variety to be detected is a disease-resistant variety; when KD is larger than or equal to KB, the variety to be detected is a susceptible variety.
Preferably, in the method for rapidly identifying disease resistance of radish, the disease index of the standard strain A identified by the method of claim 1 is 35-39, and the disease index of the standard strain B identified by the method of claim 1 is 40-45; when KB is more than KD and less than KA, the variety to be detected is a susceptible variety.
Compared with the prior art, the method for rapidly identifying the disease resistance of radish has the following beneficial effects:
the invention adds the operation of artificial inoculation of disease pollutants on the basis of natural disease nursery identification, the inoculated strain is from radish with diseases, when the disease needs to be identified, the radish with the disease is selected to prepare the disease pollutants, if the selected radish has more than two diseases, the resistance of the variety to various diseases can be simultaneously determined, and the screening accuracy of radish resistant varieties is improved. The method of the invention can also accelerate the infection probability of natural disease gardens.
The acetic acid solution is adopted to improve the survival rate and the propagation rate of microorganisms, and research shows that when the acetic acid solution with the mass fraction of 1-3% is adopted to preserve the Xanthomonas campestris causing the radish black rot, the order of magnitude of the Xanthomonas campestris is 10 days later2Increased to 106(ii) a When the xanthomonas campestris causing radish black rot is preserved by using sterile water, the order of magnitude of the xanthomonas campestris is 10 after 3 days2Increased to 105。
Detailed Description
The present invention is described in detail below with reference to specific examples, but the present invention should not be construed as being limited thereto. The test methods in the following examples, which are not specified in specific conditions, are generally conducted under conventional conditions, and the steps thereof will not be described in detail since they do not relate to the invention.
Example 1
A method for rapidly identifying disease resistance of radish comprises the following steps:
s1, collecting the radishes with the black rot disease, cutting the radishes into radish dices, adding the radish dices into an acetic acid solution with the mass fraction of 1%, wherein the side length (maximum width meter) of the radish dices after dicing is 0.5-0.7 cm, the mass ratio of the radish dices to the acetic acid solution is 1:2, soaking for 5min, filtering, and collecting filtrate, wherein a mesh screen is used as a tool for filtering, and the diameter of a hole of the mesh screen is 1mm, so that the disease pollutant is obtained;
s2, planting the variety outcrop green radish (known to resist black rot and strong comprehensive disease resistance) to be detected in a field in the following manner: planting in a hole digging and seed dibbling mode, wherein 2 seeds are planted in each hole, the plant spacing is 10cm, and the row spacing is 10 cm;
s3, field management, wherein when the radish grows to 3-8 true leaves, disease pollutants are inoculated, and the disease pollutants are inoculated once in the 3-4 leaf stage and the 7-8 leaf stage respectively, the method for inoculating the disease pollutants comprises the following steps: spraying the diluted solution of disease pollutants 1000 times to the leaf surface, wherein 20L of the diluted solution is used per mu;
disease indexes are investigated on the 10 th day after disease pollutants are inoculated in the 7-8 leaf stage;
the disease survey was graded as follows: grade 0, no disease; 0.5 grade, several disease groups are on the leaf surface, and the area of the disease area is less than 1/4 of the leaf area; the disease area on the 1-level leaf is more than or equal to 1/4 leaf area and less than 3/8 leaf area; the area of the disease area on the 2-level leaf is more than or equal to 3/8 of the area of the leaf and less than 1/2 of the area of the leaf; the disease area on the 3-level leaves is more than or equal to 1/2 of the area of the leaves and less than or equal to 3/4 of the area of the leaves, and the disease area on the 4-level leaves is more than 3/4 of the area of the leaves;
in this example, a total of 100 strains were examined, including 24 strains at grade 0, 40 strains at grade 0.5, 17 strains at grade 1, 13 strains at grade 2, 5 strains at grade 3, and 1 strain at grade 4.
Disease index of 100 ×. sigma (number of disease-treated plants at each stage × disease-grade value)/(total number of investigated plants × 4) × 100 × (24 × 0+40 × 0.5+17 × 1+13 × 2+5 × 3+1 × 4)/(100 × 4) × 20.5
The identification of the method shows that the outcrop green radish is a disease-resistant variety and is consistent with the currently recognized result.
Example 2
A method for rapidly identifying disease resistance of radish comprises the following steps:
s1, collecting radishes with black rot diseases, cutting the radishes into radish dices, adding the radish dices into an acetic acid solution with the mass fraction of 3%, soaking for 20min, filtering, wherein the side length (maximum width meter) of each radish dice is 0.8-1 cm, the mass ratio of each radish dice to the acetic acid solution is 1:1, and a tool used for filtering is a mesh screen with the diameter of each mesh screen being 2mm, and collecting filtrate to obtain disease pollutants;
s2, planting the varieties to be detected, namely Mantang red, Xinlimei and 91-1 radish (known to resist turnip mosaic virus and downy mildew but not black rot), in a field in the following way: planting in a hole digging and seed dibbling mode, wherein 1 seed is planted in each hole, the plant spacing is 15cm, and the row spacing is 30 cm;
s3, field management, wherein when the radish grows to 3-8 true leaves, disease pollutants are inoculated, and the disease pollutants are inoculated once in the 3-4 leaf stage and the 7-8 leaf stage respectively, the method for inoculating the disease pollutants comprises the following steps: spraying the diluted solution of disease pollutants 1000 times to the leaf surface, wherein 30L of the diluted solution is used per mu;
disease indexes are investigated on the 10 th day after disease pollutants are inoculated in the 7-8 leaf stage;
the disease survey was graded as follows: grade 0, no disease; 0.5 grade, several disease groups are on the leaf surface, and the area of the disease area is less than 1/4 of the leaf area; the disease area on the 1-level leaf is more than or equal to 1/4 leaf area and less than 3/8 leaf area; the area of the disease area on the 2-level leaf is more than or equal to 3/8 of the area of the leaf and less than 1/2 of the area of the leaf; the disease area on the 3-level leaves is more than or equal to 1/2 of the area of the leaves and less than or equal to 3/4 of the area of the leaves, and the disease area on the 4-level leaves is more than 3/4 of the area of the leaves;
in this example, 100 strains were investigated in total, including grade 0 15, grade 0.5 14, grade 1 16, grade 2 19, grade 3 21 and grade 4 15.
Disease index of 100 ×. sigma (number of disease-treated plants at each stage × disease-grade value)/(total number of investigated plants × 4) × 100 × (15 × 0+14 × 0.5+16 × 1+19 × 2+21 × 3+15 × 4)/(100 × 4) × 46.0
The full heart red beautiful 91-1 radish is identified by the method as a susceptible variety, and is consistent with the currently recognized result.
Example 3
A method for rapidly identifying disease resistance of radish comprises the following steps:
s1, collecting radishes with black spot diseases, cutting the radishes into radish dices, adding the radish dices into an acetic acid solution with the mass fraction of 4%, soaking for 15min, filtering, wherein the side length (maximum width meter) of each radish dice after dicing is 1.1-1.3 cm, the mass ratio of each radish dice to the acetic acid solution is 1:3, and a tool used for filtering is a mesh screen with the diameter of 0.5mm, and collecting filtrate to obtain disease pollutants;
s2, planting the varieties to be detected, namely Mantang red, Xinlimei and 91-1 radish (known to resist against turnip mosaic virus and downy mildew but not resist against black spot) in a field in the following way: planting in a hole digging and seed dibbling mode, wherein 2 seeds are planted in each hole, the plant spacing is 20cm, and the row spacing is 20 cm;
s3, field management, wherein when the radish grows to 3-8 true leaves, disease pollutants are inoculated, and the disease pollutants are inoculated once in the 3-4 leaf stage and the 7-8 leaf stage respectively, the method for inoculating the disease pollutants comprises the following steps: spraying the diluted solution of disease pollutants 1000 times to the leaf surface, wherein 25L of the diluted solution is used per mu;
disease indexes are investigated on the 10 th day after disease pollutants are inoculated in the 7-8 leaf stage;
the disease survey was graded as follows: grade 0, no disease; 0.5 grade, several disease groups are on the leaf surface, and the area of the disease area is less than 1/4 of the leaf area; the disease area on the 1-level leaf is more than or equal to 1/4 leaf area and less than 3/8 leaf area; the area of the disease area on the 2-level leaf is more than or equal to 3/8 of the area of the leaf and less than 1/2 of the area of the leaf; the disease area on the 3-level leaves is more than or equal to 1/2 of the area of the leaves and less than or equal to 3/4 of the area of the leaves, and the disease area on the 4-level leaves is more than 3/4 of the area of the leaves;
in this example, 100 strains were investigated in total, including 10 strains at grade 0, 16 strains at grade 0.5, 21 strains at grade 1, 28 strains at grade 2, 16 strains at grade 3, and 9 strains at grade 6.
Disease index of 100 ×. sigma (number of disease-treated plants at each stage × disease-grade value)/(total number of investigated plants × 4) × 100 × (10 × 0+16 × 0.5+21 × 1+28 × 2+16 × 3+9 × 4)/(100 × 4) × 42.2
The full heart red beautiful 91-1 radish is identified by the method as a susceptible variety, and is consistent with the currently recognized result.
Example 4
A method for rapidly identifying disease resistance of radish comprises the following steps:
s1, collecting radishes with black rot diseases, cutting the radishes into radish dices, adding the radish dices into an acetic acid solution with the mass fraction of 1%, wherein the side length (maximum width meter) of the radish dices after dicing is 0.5-0.7 cm, the mass ratio of the radish dices to the acetic acid solution is 1:1, soaking for 30min, filtering, and collecting filtrate, wherein a mesh screen is used as a tool for filtering, and the diameter of a hole of the mesh screen is 0.4mm, so that disease pollutants are obtained;
taking disease-resistant radish variety outcrop green radish as a standard plant A; using susceptible radish varieties of Mantang hong, Xinlimei and 91-1 radish as a standard strain B;
s2, planting the standard plant A, the standard plant B and the variety Xinuo spring green jade fruit green radish (known to resist black rot and strong in comprehensive disease resistance) to be detected in a field, and sequentially and repeatedly arranging and planting the radish according to the sequence of one line of the standard plant A, one line of the standard plant B and one line of the variety to be detected; the planting mode is as follows: planting in a hole digging and seed dibbling mode, wherein 2 seeds are planted in each hole, the plant spacing is 25cm, and the row spacing is 35 cm;
s3, field management, wherein when the radish grows to 3-8 true leaves, disease pollutants are inoculated, and the disease pollutants are inoculated once in the 3-4 leaf stage and the 7-8 leaf stage respectively, the method for inoculating the disease pollutants comprises the following steps: spraying the diluted solution of disease pollutants 1000 times to the leaf surface, wherein 20L of the diluted solution is used per mu;
disease indexes are investigated on the 10 th day after disease pollutants are inoculated in the 7-8 leaf stage;
the disease survey was graded as follows: grade 0, no disease; 0.5 grade, several disease groups are on the leaf surface, and the area of the disease area is less than 1/4 of the leaf area; the disease area on the 1-level leaf is more than or equal to 1/4 leaf area and less than 3/8 leaf area; the area of the disease area on the 2-level leaf is more than or equal to 3/8 of the area of the leaf and less than 1/2 of the area of the leaf; the disease area on the 3-level leaves is more than or equal to 1/2 of the area of the leaves and less than or equal to 3/4 of the area of the leaves, and the disease area on the 4-level leaves is more than 3/4 of the area of the leaves;
in this example, a total of 100 strains were investigated.
The incidence of the standard strain A is 75 percent of total strains with incidence/total strains with investigation
The incidence of the standard strain B is 85 percent of total strains with incidence/total strains with investigation
The disease incidence of the variety to be tested is 71 percent of total disease incidence plants/total survey plants
The evaluation by the method shows that the Xinuo spring green jade fruit green radish is a disease-resistant variety and is consistent with the currently recognized result.
It should be noted that the "field management" described in embodiments 1 to 4 of the present invention is as follows: the conventional operations of fertilizing, weeding, turning over the land, irrigating and the like are adopted.
It should be noted that when numerical ranges are given herein, it is understood that both endpoints of each of the numerical ranges and any number between the endpoints are optional unless the invention otherwise specifically states. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (8)
1. A method for rapidly identifying disease resistance of radish is characterized by comprising the following steps:
s1, collecting diseased radishes, cutting the radishes into radish dices, adding the radish dices into an acetic acid solution, soaking, filtering, and collecting filtrate to obtain disease pollutants;
s2, planting the variety to be detected in the field in the following way: planting by adopting a hole digging and seed dibbling mode, wherein 1-2 seeds are planted in each hole, the plant spacing is 10-20 cm, and the row spacing is 10-30 cm;
s3, performing field management, inoculating disease pollutants when the radish grows to 3-8 true leaves, wherein the disease pollutants are inoculated once in the 3-4 leaf stage and the 7-8 leaf stage respectively, and the disease index is investigated on the 10 th day after the disease pollutants are inoculated in the 7-8 leaf stage;
the disease survey was graded as follows: grade 0, no disease; 0.5 grade, several disease groups are on the leaf surface, and the area of the disease area is less than 1/4 of the leaf area; the disease area on the 1-level leaf is more than or equal to 1/4 leaf area and less than 3/8 leaf area; the area of the disease area on the 2-level leaf is more than or equal to 3/8 of the area of the leaf and less than 1/2 of the area of the leaf; the disease area on the 3-level leaves is more than or equal to 1/2 of the area of the leaves and less than or equal to 3/4 of the area of the leaves, and the disease area on the 4-level leaves is more than 3/4 of the area of the leaves;
disease index = 100 × ∑ (number of diseased plant at each stage × disease value)/(total number of investigated plants × 4)
When the disease index is more than 40, the variety to be detected is a susceptible variety; when the disease index is more than or equal to 40 and less than 20, the variety to be detected is a disease-resistant variety; when the disease index is more than or equal to 20 and less than 10, the variety to be detected is a disease-resistant variety; and when the disease index is less than or equal to 10, determining that the variety to be detected is a high disease-resistant variety.
2. The method for rapidly identifying the disease resistance of radish according to claim 1, wherein the acetic acid solution is used in an amount of 1-3% by weight and the soaking time is 5-20 min.
3. The method for rapidly identifying the disease resistance of radish according to claim 2, wherein the mass ratio of the diced radish to the acetic acid solution is 1: 1-2.
4. The method for rapidly identifying the disease resistance of radish according to claim 3, wherein the method for inoculating disease contaminants comprises the following steps: and (3) spraying the diluted solution obtained by diluting the disease pollutants by 1000 times to the leaf surfaces, wherein 20-30L of the diluted solution is used per mu.
5. The method for rapidly identifying the disease resistance of radish according to claim 1, wherein the side length of the diced radish after dicing is 0.5-1 cm.
6. The method as claimed in claim 5, wherein the means used in the filtering in step 1 is a mesh screen, and the diameter of the mesh screen is 0.5-2 mm.
7. The method for rapidly identifying disease resistance of radish according to claim 1, wherein the step of S1 comprises the steps of: taking a disease-resistant radish variety as a standard strain A; taking a susceptible radish variety as a standard strain B;
the step of S2 further includes the steps of: planting the standard plant A, the standard plant B and the variety to be detected in a field, and sequentially and repeatedly arranging and planting the radishes according to the sequence of a line of the standard plant A, a line of the standard plant B and a line of the variety to be detected;
s3 further includes the steps of: disease incidence is investigated on the 10 th day after disease pollutants are inoculated in the 7-8 leaf stage;
incidence = total number of plants in the disease/total number of plants investigated;
the morbidity of the standard strain A is recorded as KA, the morbidity of the standard strain B is recorded as KB, and the morbidity of the variety to be detected is recorded as KD; when KD is less than or equal to KA, the variety to be detected is a disease-resistant variety; when KD is larger than or equal to KB, the variety to be detected is a susceptible variety.
8. The method for rapidly identifying the disease resistance of radish according to claim 7, wherein the disease index of standard strain A identified by the method of claim 1 is 35 to 39, and the disease index of standard strain B identified by the method of claim 1 is 40 to 45; when KB is more than KD and less than KA, the variety to be detected is a susceptible variety.
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