CN112293182A - Method and system for identifying disease resistance of new rice variety - Google Patents
Method and system for identifying disease resistance of new rice variety Download PDFInfo
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Abstract
The invention belongs to the technical field of new rice variety identification, and discloses a method and a system for identifying disease resistance of a new rice variety, wherein natural induction is combined with inoculation identification in a field adult stage; collecting, separating and storing a rice blast disease specimen; performing field identification by using a regional test; grading disease record, evaluating resistance and calculating standard; the disease resistance identification system for the new rice variety comprises: the rice blast standard sample collection and updating system comprises a rice blast natural induction module, a rice blast standard sample collection module, a rice blast standard sample separation module, a rice blast standard sample storage module, a physiological race identification module, a central control module, a data fusion processing module, an index calculation module, a data storage module and an updating display module. The method collects the rice blast standard samples of different rice planting areas and different rice varieties, completes the separation, purification and preservation of strains; the composition and distribution of physiological races of rice blast germs are proved by infecting and differentiating hosts, and effective resistance genes of the rice blast germs are clarified.
Description
Technical Field
The invention belongs to the technical field of new rice variety identification, and particularly relates to a method and a system for identifying disease resistance of a new rice variety.
Background
At present, rice is one of the most important grain crops in China, and plays a significant role in guaranteeing national grain safety. The source of grain production lies in the breeding of good varieties, while the traditional breeding of varieties depends on the phenotype of plants, and the cultivation of a good variety usually takes a time period of up to ten years.
The safety of the grain is related to social stability and economic development, is related to civil problems, and plays a unique role in agricultural production development, wherein the rice yield is close to 40 percent of the total grain yield in China and approximately accounts for 30 percent of the total rice yield in the world. The safe production of grains is closely related to the occurrence of crop diseases, and the breeding and the utilization of disease-resistant varieties are the most economic, most effective and most safe technologies and measures in the comprehensive prevention and control of the crop diseases, so that the diseases can be effectively controlled, the method is a sustainable ecological protection measure, and the identification of the disease resistance of crops is a necessary way for obtaining disease-resistant crop varieties. At present, the breeding, resistance identification evaluation and utilization of crop disease-resistant varieties are highly regarded by government departments at all levels. The Ministry of agriculture in China also develops a high-susceptibility variety 'one-vote rejection control' to prevent serious loss caused by production and application of a variety with poor resistance.
When the disease resistance of the rice is identified, different rice varieties need to be sown in the resistance identification nursery, the confusion of the seeds of the different varieties cannot occur in the process, and the identification result is inaccurate once the confusion is caused, so that the reasonable utilization of the sowing space of the identification nursery and the sowing of the different rice varieties in mutually isolated spaces are particularly important.
The traditional sowing mode generally adopts a common seedling tray to sow or sow after square is drawn on slurry, adopts the common seedling tray to sow, before sowing, slurry is filled into a sowing cup, then the sowing tray is put into the slurry to enable the tray surface to be level with the slurry surface, and seeds are sown on the slurry of the sowing cup during sowing. The sowing method can reasonably utilize the space of the resistance identification nursery and better distinguish different rice varieties, but the defects are also obvious: 1. because the slurry is softer, the surface is difficult to be absolutely smooth, more water accumulation space exists, and the sown rice seeds are easy to float and mix; 2. when the disease resistance of rice is identified, the number of varieties to be tested is more than hundreds, and more than thousands of varieties are provided, during sowing, rice seeds are manually grabbed and then placed on slurry in a sowing cup, about 20 seeds need to be planted in each hole, and because the seeds are small and smooth, the seeds in hands are easy to fall into other sowed seeds in the grabbing process, the variety confusion occurs, and the accuracy and the scientificity of the identification result are influenced; 3. each variety needs to be sown repeatedly (more than 20), each repetition needs to be sown in 3 holes, the workload is large, and the efficiency is low; 4. after the rice variety is sown, water is required to be supplemented regularly along with the evaporation of water in the slurry, the water is required to be supplemented for many times before the rice seed is germinated and rooted, and the seed drift is easy to occur after the water is sprayed, so that different varieties are mixed up, and the accuracy of an identification result is also influenced. 5. The bottom of the common seedling raising plate is only provided with a water permeable hole which is in contact with soil, so that the growth of later-stage rice plants is not facilitated, rice leaves are narrow, the attachment of pathogens during disease resistance identification is not facilitated, and the accuracy of identification results is influenced. The sowing mode of drawing squares on the slurry is beneficial to the growth of rice plants in the later period, but is also limited by the factors, and the utilization of the space of the appraisal nursery is poor.
The prior art discloses a seeding device for identifying disease resistance of rice varieties, which comprises a seeding disc and a split charging assembly; the seeding tray comprises a tray and a plurality of seeding cups distributed on the tray in an array manner, the seeding cups are through up and down, and the peripheral wall of the seeding cups is provided with water permeable holes; the tray is connected to the middle part of the outer wall of the seeding cup; the sub-packaging assembly comprises a plurality of sub-packaging pieces which are used for sub-packaging seeds and matched with the seeding cup, and the sub-packaging pieces are made of water-soluble materials.
Through the above analysis, the problems and defects of the prior art are as follows:
(1) in the prior art, the separation, purification and preservation effects on rice blast standard samples of different rice varieties are poor.
(2) The prior art can not find out the composition and distribution of physiological races of rice blast germs by infecting and differentiating hosts, and can not define effective resistance genes of the rice blast germs.
(3) The prior art has the advantages of small quantity of disease-resistant varieties for identifying new rice strains and high cost.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a method and a system for identifying the disease resistance of a new rice variety.
The invention is realized in such a way, and the method for identifying the disease resistance of the new rice variety comprises the following steps:
step one, identifying the field plant growth period by combining natural induction with inoculation: using field disease straws as an inducing material, applying N fertilizer to create a disease condition, combining artificial inoculation to spray mixed spore liquid on rice leaves by a sprayer at a tillering peak stage in the middle and last ten days of June, assisting in sun-shading treatment, creating a condition beneficial to rice blast by a rice blast natural inducing module, investigating a blast disease condition index in the last ten days of June, and investigating a neck blast disease condition in 1 day of June from 8 Yue 15 to 9 Yue;
step two, collecting the rice blast standard samples of the rice planting area in the season of rice blast occurrence from the first 7 months to the first 10 months through a rice blast standard sample collecting module, and collecting rice blast specimens: the collection places are widely distributed, the collected specimens comprise ear blast and node blast specimens, and the specimens are uniformly numbered and classified for storage;
performing tissue separation and purification on a rice blast specimen in a laboratory through a rice blast standard sample separation module to obtain a single spore strain, performing moisture preservation culture on the rice blast specimen in a constant temperature box at 25 ℃ for 24-48h, observing whether rice blast exists or not in an ultraclean environment through microscopic examination, shaking the spores on a water agar plate culture medium when spores are generated on the scab, and performing moisture preservation culture in the constant temperature box at 25 ℃ for 16 h;
step four, selecting single spores by using a scalpel under an ultraclean environment microscope, inoculating the single spores to a PDA culture medium, carrying out moisture-preserving culture at 25 ℃ for 3-5 days until the single spores grow to 2cm in diameter, transferring the single spores to a rice flour culture medium again, carrying out moisture-preserving culture at 25 ℃ for 3-5 days, similarly removing the polluted strains, and separating 3-5 single spore strains from each sample; co-separating and purifying 1136 parts of strains, eliminating the strains with pollution and low spore production capacity, and co-preserving 771 parts of effective strains;
inoculating the separated monospore strain to a spore production culture medium, storing the monospore strain in a constant-temperature incubator until the monospore strain is dried, putting the monospore strain into a sterilized cryopreservation tube in an aseptic environment, putting the cryopreservation tube into a sealed container according to the number, and temporarily storing the monospore strain in a refrigerator at the temperature of-20 ℃ for later use through a rice blast standard sample storage module;
sixthly, identifying physiological races of rice blast germs; respectively carrying out regional test and quality ratio test of an experiment base through a physiological race identification module to determine the dominant physiological races in the region; after purification and rejuvenation, different types of collected differential hosts are utilized by single spores to respectively carry out species and pathogenicity determination on different varieties of rice blast germs in different areas, and physiological race compositions and distribution of the rice blast germs are proved by infecting the differential hosts, so that dominant physiological races in the areas are determined;
controlling the normal operation of each module of the new rice variety disease resistance identification system by a central control module through a central processing unit; fusing the acquired natural rice blast inducing data, the rice blast standard sample data and the physiological race identification result by using a data fusion processing module through a data fusion program to obtain rice blast data;
step eight, disease condition recording grading, resistance evaluation and standard calculation: respectively calculating the disease grade of the seedling leaf blast, the incidence rate of the panicle blast, the loss rate of the panicle blast and the comprehensive rice blast index according to the obtained rice blast data by using an index calculation module through a calculation program;
storing the obtained natural rice blast inducing data, rice blast standard sample data, physiological race identification results, fused rice blast data and calculation results by using a memory through a data storage module;
and step ten, updating the acquired natural rice blast inducing data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result by using an updating program through an updating display module, and displaying the data in real time through a display.
Further, in the first step, in the 3-leaf stage of the rice seedlings, the test variety and the differential host are inoculated in a seed inoculation box by spraying, the inoculated seed is placed in the seed inoculation box and is moved into a shading shed for 24 hours after inoculation, water is sprayed for 5min every 1 hour in the daytime for moisture preservation, a film is covered at night for heat preservation, and the disease occurrence condition is analyzed 8 days after inoculation.
Further, in the second step, the collection method of the rice blast disease specimen comprises the following steps: the collection sites are distributed in 43 sites of the irrigation area, 531 parts of the collected specimens are collected, and the collected specimens of the panicle blast and the nodular cast blast are marked with numbers and stored in a classified mode.
Further, in the sixth step, the field identification method for physiological race of rice blast fungus in the area test further comprises the following steps:
utilizing 24 monogenes to identify rice blast germs, utilizing different types of identifying hosts to infect and identify in a three-leaf one-heart environment under the sterile high-temperature high-humidity environment, and determining physiological races and anti-infection reaction on the monogenes identifying hosts; the identification process needs to pay attention to the grasp of the propagation time of the strain spores and the growth time of the differential hosts, multiple times of seedling culture are carried out in multiple stages, seeds are soaked by 402 during seedling culture, bed soil is disinfected, and the standard establishment of infection environment is carried out.
Further, in the eighth step, the grade of the seedling leaf blast is calculated according to GLB ═ Σ (NDL × GDL)/TNL;
in the formula: GLB-grade of seedling blight;
NDL-number of diseased leaves at each level;
GDL-representative value of each disease grade;
TNL-total leaf number investigated.
Further, in step eight, the incidence rate of the panicle blast is calculated as IDP ═ TNDP/TNP × 100;
in the formula: IDP-panicle blast incidence rate, and the unit is percentage (%);
TNDP-number of panicles affected;
TNP-total number of ears investigated;
resistance index corresponding to GIDP-IDP.
Further, in the eighth step, the panicle blast loss rate (grade) is calculated as GLRP ═ Σ (NDP × GDP)/TNP;
in the formula: GLRP-panicle blast loss rate (grade);
NDP-the number of diseased ears at each level;
GDP-loss rate grade of each grade;
TNP-total number of ears investigated;
the rice blast integrated index is calculated as IB ═ GLB × 25% + GIDP × 25% + GLRP × 50%.
Another object of the present invention is to provide a new rice disease resistance identification system for carrying out the method for identifying a new rice disease resistance, the system comprising:
the rice blast standard sample collection and updating system comprises a rice blast natural induction module, a rice blast standard sample collection module, a rice blast standard sample separation module, a rice blast standard sample storage module, a physiological race identification module, a central control module, a data fusion processing module, an index calculation module, a data storage module and an updating display module.
The rice blast natural induction module is connected with the central control module and is used for spraying the mixed spore liquid on rice leaves through a sprayer, applying N fertilizer at high level and assisting in sun-shading treatment to create disease conditions of rice blast;
the rice blast standard sample collection module is connected with the central control module and is used for collecting neck blast and node blast samples of the rice planting area, marking the samples with uniform numbers and storing the samples in a classified manner;
the rice blast standard sample separation module is connected with the central control module and is used for carrying out tissue separation and purification on a rice blast specimen in a laboratory to obtain a monospore strain;
the rice blast standard sample storage module is connected with the central control module and is used for temporarily storing the monospore strain in a refrigerator for later use;
the physiological race identification module is connected with the central control module and is used for respectively carrying out field identification on physiological races of rice blast germs in a regional test through a regional test and a quality test of an experimental base;
the central control module is connected with the rice blast natural induction module, the rice blast standard sample collection module, the rice blast standard sample separation module, the rice blast standard sample storage module, the physiological race identification module, the data fusion processing module, the index calculation module, the data storage module and the updating display module and is used for controlling the normal operation of each module of the new rice variety disease resistance identification system through a central processing unit;
the data fusion processing module is connected with the central control module and is used for carrying out fusion processing on the acquired natural rice blast inducing data, the rice blast standard sample data and the physiological race identification result through a data fusion program to obtain rice blast data;
the index calculation module is connected with the central control module and is used for calculating the disease level of the seedling-leaf blast, the incidence rate of the panicle blast, the loss rate of the panicle blast and the comprehensive index of the rice blast through a calculation program according to the obtained rice blast data;
the data storage module is connected with the central control module and used for storing the acquired rice blast natural induction data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result through the memory;
and the updating display module is connected with the central control module and used for updating the acquired rice blast natural induction data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result through an updating program and displaying the data in real time through a display.
Another objective of the present invention is to provide a computer program product stored on a computer readable medium, comprising a computer readable program, which when executed on an electronic device, provides a user input interface to implement the method for identifying disease resistance of a new variety of rice.
Another object of the present invention is to provide a computer-readable storage medium storing instructions which, when executed on a computer, cause the computer to perform the method for identifying disease resistance of a new variety of rice.
By combining all the technical schemes, the invention has the advantages and positive effects that:
the invention collects the standard rice blast samples of different rice planting areas and different rice varieties, and the separation, purification and storage of the strains in the last year are completed in a laboratory.
The invention discloses a method for detecting physiological race composition and distribution of rice blast germs by infecting differential hosts, and a rice blast effective resistance gene is defined.
The invention establishes a rice blast disease-resistant identification nursery and an anti-source screening nursery, and identifies the disease resistance of 450 new rice lines per year on average.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the embodiments of the present application will be briefly described below, and it is obvious that the drawings described below are only some embodiments of the present application, and it is obvious for those skilled in the art that other drawings can be obtained from the drawings without creative efforts.
FIG. 1 is a flowchart of a method for identifying disease resistance of a new variety of rice according to the present invention.
FIG. 2 is a block diagram showing the structure of a system for identifying disease resistance of a new variety of rice according to an embodiment of the present invention;
in the figure: 1. a rice blast natural induction module; 2. a rice blast standard sample collection module; 3. a rice blast standard sample separation module; 4. a rice blast standard sample storage module; 5. a physiological race identification module; 6. a central control module; 7. a data fusion processing module; 8. an index calculation module; 9. a data storage module; 10. and updating the display module.
FIG. 3 is a graph showing the trend of the rice resistance gene over time according to the present invention.
FIG. 4 is a graph showing the frequency of distribution of physiological races of rice blast disease in some province 2018 provided by the embodiment of the present invention.
Fig. 5 is a transition diagram of dominant physiological races in the whole region 2015-2018 provided by the embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Aiming at the problems in the prior art, the invention provides a method and a system for identifying the disease resistance of a new rice variety, and the invention is described in detail with reference to the accompanying drawings.
As shown in fig. 1, the method for identifying disease resistance of a new rice variety provided by the embodiment of the present invention includes the following steps:
s101, spraying mixed spore liquid on rice leaves by a sprayer through a rice blast natural induction module, and creating disease conditions of rice blast through high application of N fertilizer and auxiliary sun-shading treatment;
s102, collecting neck blast and node blast specimens of a rice planting area through a rice blast standard sample collecting module, marking the specimens with uniform numbers, and storing the specimens in a classified manner;
s103, performing tissue separation and purification on a rice blast specimen in a laboratory through a rice blast standard sample separation module to obtain a monospore strain; the monospore strain is put into a refrigerator for storage for later use through a rice blast standard sample storage module;
s104, respectively carrying out field identification on physiological races of rice blast germs in a regional test by using a regional test and a quality ratio test of an experimental base through a physiological race identification module;
s105, controlling the normal operation of each module of the new rice variety disease resistance identification system by using a central processing unit through a central control module;
s106, fusing the acquired natural rice blast inducing data, the rice blast standard sample data and the physiological race identification result by using a data fusion program through a data fusion processing module to obtain rice blast data;
s107, calculating the disease grade of the seedling-leaf blast, the incidence rate of the panicle blast, the loss rate of the panicle blast and the comprehensive rice blast index according to the obtained rice blast data by using an index calculation module and a calculation program;
s108, storing the acquired natural rice blast inducing data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result by using a memory through a data storage module;
and S109, updating the acquired natural rice blast inducing data, rice blast standard sample data, physiological race identification results, fused rice blast data and calculation results by using an updating program through an updating display module, and displaying the data in real time through a display.
As shown in fig. 2, the identification system of the method for identifying disease resistance of a new rice variety provided by the embodiment of the present invention includes: the rice blast standard sample collection system comprises a rice blast natural induction module 1, a rice blast standard sample collection module 2, a rice blast standard sample separation module 3, a rice blast standard sample storage module 4, a physiological race identification module 5, a central control module 6, a data fusion processing module 7, an index calculation module 8, a data storage module 9 and an update display module 10.
The rice blast natural induction module 1 is connected with the central control module 6 and is used for spraying the mixed spore liquid on rice leaves through a sprayer, applying N fertilizer at high level and assisting in sun-shading treatment to create disease conditions of rice blast;
the rice blast standard sample collection module 2 is connected with the central control module 6 and is used for collecting neck blast and node blast samples of a rice planting area, marking the samples with uniform numbers and storing the samples in a classified manner;
the rice blast standard sample separation module 3 is connected with the central control module 6 and is used for carrying out tissue separation and purification on a rice blast specimen in a laboratory to obtain a monospore strain;
the rice blast standard sample storage module 4 is connected with the central control module 6 and is used for temporarily storing the monospore strain in a refrigerator for later use;
the physiological race identification module 5 is connected with the central control module 6 and is used for respectively carrying out field identification on physiological races of rice blast germs in a regional test through a regional test and a quality test of an experiment base;
the central control module 6 is connected with the rice blast natural induction module 1, the rice blast standard sample collection module 2, the rice blast standard sample separation module 3, the rice blast standard sample storage module 4, the physiological race identification module 5, the data fusion processing module 7, the index calculation module 8, the data storage module 9 and the update display module 10, and is used for controlling the normal operation of each module of the new rice variety disease resistance identification system through a central processing unit;
the data fusion processing module 7 is connected with the central control module 6 and is used for carrying out fusion processing on the obtained rice blast natural induction data, the rice blast standard sample data and the physiological race identification result through a data fusion program to obtain rice blast data;
the index calculation module 8 is connected with the central control module 6 and used for calculating the seedling blast disease grade, the panicle blast incidence, the panicle blast loss rate and the rice blast comprehensive index according to the obtained rice blast data through a calculation program;
the data storage module 9 is connected with the central control module 6 and used for storing the acquired rice blast natural induction data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result through a memory;
and the updating display module 10 is connected with the central control module 6 and used for updating the acquired rice blast natural induction data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result through an updating program and displaying the data in real time through a display.
The present invention will be further described with reference to specific examples and experiments.
Test contents and results
Collecting and identifying physiological race of rice blast bacteria: in the season of rice blast occurrence (from the beginning of 7 months to the beginning of 10 months), collecting rice blast standard samples in certain provincial rice planting areas for separation.
1. Collecting rice blast disease specimens: the collecting places are distributed in 43 places counted in each city and county and each large farm in the irrigation district, 531 parts of specimens are collected, wherein 189 parts of Yongning, 48 parts of Wuzhong, 26 parts of bronze gorges, 48 parts of Lingwu, 9 parts of Zhongwei, 15 parts of Nanliang farm, 18 parts of warm spring farm, 25 parts of advancing farm, 26 parts of Zhongning, 86 parts of plain and 41 parts of Helan collected neck blast and festoon blast specimens are numbered uniformly and stored in a classified manner.
2. Isolation and preservation of Rice blast Strain: performing tissue separation and purification on a rice blast specimen in a laboratory to obtain a monospore strain, performing moisture preservation culture on the rice blast specimen in a constant temperature box at 25 ℃ for 24-48 hours, observing whether rice blast exists or not in an ultraclean environment through microscopic examination, shaking the spores onto a water agar plate culture medium when the disease spots generate the spores, performing moisture preservation culture in the constant temperature box at 25 ℃ for 16 hours, picking out the single spores under the ultraclean environment microscope by using a scalpel, inoculating the single spores onto a PDA culture medium, performing moisture preservation culture at 25 ℃ for about 3-5 days, paying attention to the polluted strain in the period, removing the polluted strain in time, culturing the polluted strain until the diameter is 2cm, transferring the polluted strain into a rice flour culture medium again, performing moisture preservation culture at 25 ℃ for about 3-5 days, removing the polluted strain in the same way, and placing the monospore strain into a refrigerator at 4 ℃ for temporary storage for later use. 3-5 single spore strains were isolated from each specimen. Inoculating the separated monospore strain to a spore-producing culture medium, storing in a constant-temperature incubator until the monospore strain is dried, placing in a sterilized freezing storage tube in an aseptic environment, placing the freezing storage tube in a sealed container according to the number, and storing at-20 ℃. And (3) co-separating and purifying 1136 parts of strains, eliminating the strains with pollution and low spore production capacity in the strains, and co-preserving 771 parts of effective strains, wherein the preservation rate reaches 67.9%. The monospore of rice blast collected in 2019 is being separated in the pathological laboratory of crops.
3. The physiological race identification research of rice blast germs comprises the following steps: the dominant physiological races in this area are clarified. The physiological race species and pathogenicity of different varieties of rice blast germs in different areas of a certain province are respectively measured by using different types of differential hosts collected in the years after purification and rejuvenation of single spores, and the composition and distribution of the physiological race species of the rice blast germs in the areas are proved by infecting the differential hosts, so that the dominant physiological race species in the areas are determined.
3.1 identification of Magnaporthe grisea by use of 24 single genes
The determination method comprises the steps of utilizing different types of differential hosts to infect and discriminate in a three-leaf environment under a sterile high-temperature high-humidity environment, (when the rice seedling is in a 3-leaf period, carrying out spray inoculation on a test variety and the differential hosts in an inoculation box, placing the inoculated host in the inoculation box after inoculation for 24 hours, transferring the inoculated host into a shading shed, spraying water for 5min for moisturizing every 1 hour in the day, covering a film for heat preservation at night, and investigating the disease condition after inoculation for 8 days) to determine physiological races and the anti-infection reaction to the single-gene differential hosts. The identification process needs to pay attention to the grasp of the propagation expanding time of the spores of the strains and the growth time of the differential hosts, seedlings are raised for multiple times in multiple stages, seeds need to be soaked by 402 during seedling raising, bed soil is disinfected, and infection environment specifications are established, errors of experimental links are eliminated as much as possible, 7 batches of physiological microspecies of rice blast germs are detected from 5 months to 10 months at present, 145 parts of single spore strains are detected, through analysis of resistance frequency of detection results, the resistance frequency of the strains Pia, Pizt, Pit, Pi9 and Pita2 separated from rice blast specimens collected in 2018 is more than 25 percent and shows high resistance, while the resistance frequency of other resistance genes is less than 20 percent, and the effect on certain provinces of rice blast is relatively small; the resistance frequency of the resistance gene Pia is on the trend of rising year by year, and attention should be paid to the utilization of the resistance gene; the Pikh resistance frequency of the resistance gene is reduced from 41.67% to 17.69% in 2016, the resistance frequency is reduced year by year, the effect of the resistance gene is weakened, and the effect of the resistance gene on rice blast of a certain province is continuously identified and observed in 2019. The change frequency of other genes is not obvious, and the change frequency is shown in a change trend graph among years of main resistance genes of rice in tables 1, 2 and 3.
TABLE 1 frequency of resistance of the major resistance genes to rice blast to 147 Rice blast strains of a certain province
TABLE 2 Table of annual resistance frequency of major resistance genes against rice blast
3.2 naming and grouping 147 rice blast germs identified by using selected single gene differential hosts of a province
The 147 identified strains were named according to the names of physiological races of rice blast, and 44 races were divided, as shown in Table 3 and FIG. 4. The proportion of dominant race is shown in Table 4, wherein the dominant race is NA01 39.5%, NC01 10.2%, NA33 5.44%, NB01 4.08%, and NA09 2.74%. When the distribution frequency of dominant physiological races appearing in 2014-2016 is analyzed, some dominant physiological races vary greatly within the year, particularly the NA01 accounts for 39.5%, so that the dominant physiological races are strong, and the dominant physiological races are also the main reasons for serious rice blast of varieties which cannot be prevented and controlled timely in production in this year, and the detailed results are shown in tables 4, 5 and 5.
TABLE 3 host response to differential host infection and nomenclature of strains in certain provinces
TABLE 4 list of dominant physiological races of rice blast in some province 2018
Table 52013-2018 dominant physiological race proportion summary table
(II) the present invention is further described below with reference to various breeding identifications, quality ratios, and identification of disease resistance of the test material in the region.
1. Test materials:
461 parts of materials participating in disease resistance identification.
The number of the early maturing group, the late maturing group, the high quality rice group and the production test in the regional test is 40 parts in total.
The test article in this area is 34 parts of the early-maturing group and the late-maturing group.
Breeding new varieties of high-yield and stable-yield rice: 41 parts of high-generation breeding materials.
Breeding new varieties of high-quality special rice: 26 parts of high-generation breeding materials.
Green and efficient breeding of new rice varieties: 51 parts of high-generation breeding materials.
And (3) identification of excellent germplasm resources of rice: and 9 parts of a quality ratio test, and 109 parts of introduced seed resources.
The new technical application and material innovation of rice breeding are as follows: and identifying and screening 30 parts of parent materials.
21 parts of united body area test material and reference material.
Other stable lines and varieties: 54 parts of the components.
2. Test design and method:
2.1 experimental design:
and (3) rice blast disease resistance identification nursery: the rice blast inducing material is arranged in a perennial rice field of a Taiwang breeding base of crop institute, the area of the test field is 2.0 mu, 18 rows are formed, 40 materials are planted in each row, 2 rows (10-15 plants in each row) are planted in each material, 4-5 plants are planted in each hole, the walking path between the rows is 50 cm, and rice blast inducing materials (white rice) are planted in each material, walking path between the rows and the periphery. The periphery of the test area is planted with 1 meter wide inducing belt (white-bark rice).
2.2 test strains:
the Pyricularia oryzae selects the monospora oryzae strain of the dominant population collected and identified from different rice varieties in the whole rice planting area stored in 2018, is cultured and activated in a water agar culture medium, hypha in the monospora culture medium is moved into a single spore and a culture dish in a PDA culture medium for culture, and is moved onto a spore production culture medium after one week to produce spores, and finally mixed bacteria of 36 monospora strains are obtained. The concentration of the mixed spore liquid is based on that more than 30 spores are seen in each visual field under a 100-fold microscope.
2.3 identification method:
(1) and (3) identifying the field plant growth period by combining natural induction and inoculation: the field disease rice straw is used as an inducing material, the disease condition is created by applying N fertilizer to the field disease rice straw, the mixed spore liquid is sprayed on the rice leaves by a sprayer in the tillering peak period in the middle and last days of June in combination with artificial inoculation, sun shading is assisted, the disease condition beneficial to rice blast is created, the leaf blast condition index is investigated in the last ten days of June, and the neck blast condition is investigated in 1 day of 8-15-9.
(2) Field identification in a regional test: regional test and quality ratio test of a crop research institute of academy of agriculture and forestry of a certain province.
(3) Disease record grading and resistance evaluation criteria: the identification and evaluation standard adopts agricultural industry standard NY/T2646-2014 newly issued in 2015 of the national republic of China, which specifies the relevant definitions of the rice blast resistance identification of the rice variety test, an identification method investigation method, data calculation, resistance evaluation and summary report format. The standard is suitable for the national and provincial rice variety tests; the comparison test of the disease resistance of the variety and the monitoring of the disease resistance of the dominant variety can be executed by reference.
Calculating the disease level of the seedling leaf blast according to GLB ═ Sigma (NDL multiplied by GDL)/TNL;
in the formula: GLB-grade of seedling blight;
NDL-number of diseased leaves at each level;
GDL-representative value of each disease grade;
TNL-total leaf number investigated.
Calculating the incidence rate of the panicle blast according to IDP (trinitrotoluene) TNDP/TNP multiplied by 100;
in the formula: IDP-panicle blast incidence rate, and the unit is percentage (%);
TNDP-number of panicles affected;
TNP-total number of ears investigated;
GIDP is the resistance index (grade) corresponding to IDP; the classification standard of the resistance of the rice blast morbidity population is shown in an attached table A.
Calculating the panicle blast loss rate (grade) according to GLRP ═ sigma (NDP multiplied by GDP)/TNP;
in the formula: GLRP-panicle blast loss rate (grade);
NDP-the number of diseased ears at each level;
GDP-loss rate grade of each grade;
TNP-total ear number investigated.
The rice blast comprehensive index is calculated as IB-GLB × 25% + GIDP × 25% + GLRP × 50%; the comprehensive evaluation grading standard of the rice blast resistance is shown in an attached table B.
TABLE A grading Standard of population resistance of incidence of Rinderpest in Rice
| Reactance stage | Type of anti-influenza | Ear rate (%) |
| 0 | High Resistance (HR) | 0 |
| 1 | Anti (R) | ≤5.0 |
| 3 | Moderate (MR) | 5.1～10.0 |
| 5 | Middle Sensing (MS) | 10.1～25.0 |
| 7 | Feel (S) | 25.1～50.0 |
| 9 | High Sensitivity (HS) | ≥50.1 |
The method comprises the steps of investigating the ear neck blast when the ear neck blast exists, investigating the branch and stalk blast when the ear neck blast does not exist, converting the branch and stalk blast into a grading standard of the ear neck blast, wherein the grade 1 is that the incidence rate of the branch and stalk blast is less than or equal to 10%, the grade 3 is that the incidence rate is 11% -30%, and the grade 5 is that the incidence rate is greater than 31%. Stalk blast refers to the onset of the first stalk of the cob (including the cob part of 2/3 at the upper end) and the grain is full.
When there is no ear blast and there is node blast, the node blast is counted as ear blast.
TABLE B comprehensive evaluation grading Standard for Rice blast resistance
| Reactance stage | Type of anti-influenza | |
| 0 | High Resistance (HR) | ＜0.1 |
| 1 | Anti (R) | 0.1～2.0 |
| 3 | Moderate (MR) | 2.1～5.0 |
| 5 | Middle Sensing (MS) | 5.1～6.0 |
| 7 | Feel (S) | 6.1～7.5 |
3. Field management:
applying 4 kg/mu of base fertilizer NEB and 15 kg/mu of organic fertilizer in 19 days in 5 months, simultaneously matching with butachlor and oxyfluorfen for sealing, transplanting seedlings in 23 days in 5 months, applying 10 kg/mu of urea in 24 days in 5 months, soaking the rice in 23 days in 5 months, sowing the rice in 25 days in 5 months, applying 10 kg/mu of urea in 5 days in 6 months, applying 10 kg/mu of herbicide in the 2-3 leaf stage of barnyard grass in 9 days in 6 months, and spraying 20kg of water by using 80 ml of 2.5 percent butachlor oil suspending agent in each mu. The rice is planted in soil for 22 days in 6 months, 10 kg/mu of urea is applied for 12 days in 6 months, 7.5 kg/mu of urea is applied in the whole growth period, 2.3 parts of organic fertilizer is applied for one mu of base fertilizer, and the rice straw with the rice core diseases is planted for 6 days in 7 months. Investigating the leaf blast in the middle 7 th month, and investigating the neck blast in 8 months, 15 days to 9 months and 3 days.
4. And (3) test results:
the artificial inoculation induction under natural conditions is adopted to carry out rice disease resistance identification, 461 parts of rice varieties are subjected to leaf blast resistance identification in the tillering stage, and ear blast resistance identification in the heading stage. According to the identification result IB (comprehensive disease index), the following conditions are shown: the disease resistance difference of different varieties is obvious, wherein 49 parts of the variety with the comprehensive disease resistance index of less than or equal to 2.0 accounts for 10.6%, 276 parts of the material with the comprehensive disease resistance index of between 2.01 and 4.0 accounts for 59.9%, 66 parts of the material with the comprehensive disease resistance index of between 4.01 and 5.0 accounts for 14.3%, 33 parts of the material with the comprehensive disease resistance index of between 5.01 and 6.0 accounts for 7.2%, 23 parts of the material with the comprehensive disease resistance index of between 6.01 and 7.5 accounts for 5.0%, and 14 parts of the material with the comprehensive disease resistance index of more than or equal to 7.51 accounts for 3.0%.
4.1 identification of disease resistance of regional test products compared with precocity group:
the method comprises the following steps of (1) testing 17 parts of materials, and comprehensively resisting 12 parts of a medium-resistant (MR) variety in disease-resistant type expression; 4 shares of variety expressed as susceptible (MS) and 1 share of variety expressed as susceptible (S); the contrast rich source No. 4 comprehensive disease-resistant index is 3.23, the highest single panicle loss rate is 7 grade, wherein the comprehensive disease-resistant index of the Yidao 1, 2017QX-561, flower 147, flower 150, flower 152, HR-18, HR-19, Ningshi 14, Ningshi 47, good care 205, SD-22 and SD-24 is less than 5, the panicle neck blast loss rate is less than 5, and the highest panicle neck blast loss rate is lower than the contrast index. MY-2, 2018KF-42, super-excellent No. 8, a comprehensive disease resistance index of less than 5, a resistance loss rate of less than 5 and a single spike loss rate of the highest level of the resistance loss rate of less than 5. Ningyuan HX6 has comprehensive disease resistance index higher than 5. The specific identification results are shown in Table 6.
4.2 disease resistance identification of regional test products compared with late-maturing groups:
the method comprises the following steps of (1) testing 17 parts of materials, and comprehensively resisting 12 parts of a medium-resistant (MR) variety in disease-resistant type expression; 4 shares of variety expressed as susceptible (MS) and 1 share of variety expressed as susceptible (S); the control Ningjing No. 41 comprehensive disease resistance index is 4.99, the highest single panicle loss rate is 5 grade, wherein the comprehensive disease resistance index of flowers 148, HR-20, excellent breeding 113, No. 11 grain and No. 638 Zhonghe is less than 5, the panicle neck blast loss rate is less than 5, and the highest grade of the panicle neck blast loss rate is lower than the control index. Ningzi 438, HR-21, Ningzi 20, Ningzi 46, sound care 206, SD-23, Ningda No. 9, Jk-436 comprehensive disease resistance index less than 5, resistance loss rate less than 5 and single spike loss rate higher than the highest. Control index. The comprehensive disease resistance index of 2017QX-3032016NF-657 Ningyuan HX7 is more than 5, and the specific identification result is shown in Table 7.
4.3 regional test early-maturing group disease resistance identification:
the material is tested by 10 parts, and the comprehensive disease-resistant type is represented by 6 parts of an anti-disease (MR) variety, 3 parts of an anti-disease (MS) variety and 1 part of a disease-resistant (R) variety; the control rich source No. 4 comprehensive disease-resistant index is 3.04, the highest single panicle loss rate is grade 5, wherein the combined disease-resistant indexes of Jiyan 46, 2014QX-295, Ningyuan HX-5, HR-15, good rearing 203, SD-18, good rearing 107 and Ningda No. 6 are less than 5, the panicle neck blast loss rate is less than 5, and the highest panicle neck blast loss rate is lower than the control index. HR-14 comprehensive disease resistance index is less than 5, resistance loss rate is less than 5, but single spike loss rate is higher than the control index at the highest level. The specific identification results are shown in Table 8.
4.4 regional test late-maturing group disease resistance identification:
referring to 11 parts of materials, the comprehensive disease-resistant type shows 1 part of a disease-resistant (R) variety, shows 8 parts of a medium-resistant (MR) variety, shows 2 parts of a medium-sensitive (MS) variety, shows a contrast Ningjing No. 41 comprehensive disease-resistant index of 4.08 and has a highest grade of 3 of the single head loss rate of the neck blast, wherein the comprehensive disease-resistant index of Chinese medicine No. 6, HR-16, HR-17, excellent growth 108, Ningyuan No. 8, Ningzi 235 comprehensive disease-resistant index is less than 5, the neck blast loss rate is less than 5, and the highest grade of the neck blast loss rate is lower than the contrast index. Ninghua No. 5, 2015QX-165, 2014QX-296 and Ninghua No. 2 have comprehensive disease resistance indexes of less than 5, resistance loss rate of less than 5 and highest single spike loss rate higher than control indexes. The results of the specific identification of the disease nursery are shown in Table 9.
4.5 regional test for disease resistance identification of high-quality rice group:
the materials are tested by 11 parts, and 3 parts of disease-resistant (R) varieties are comprehensively expressed by disease-resistant types; 6 portions of an anti-virus (MR) variety in the expression, 2 portions of an intermediate (MS) variety in the expression, 3.33 of the control Ningjing No. 57 comprehensive disease resistance index and 3-grade highest grade of the single head loss rate of the neck blast, wherein 2015QX-169, 1, RS-16, 108 and 207 comprehensive disease resistance index are less than 5, the head blast loss rate is less than 5, and the highest grade of the head blast loss rate is lower than the control index. The comprehensive disease resistance index of the flower 145, the flower 146, the 2016NF-618, the Ningda No. 10 and the 2014NF-744 is less than 5, the resistance loss rate is less than 5, and the highest single spike loss rate is higher than the control index. The results of the specific identification of the disease nursery are shown in Table 10.
4.6 regional test early-maturing group production test disease resistance identification:
referring to 4 parts of materials, the comprehensive disease resistance type expression shows 2 parts of a Medium Resistant (MR) variety, the expression shows 2 parts of a Medium Sensitive (MS) variety, the contrast rich source 4 comprehensive disease resistance index is 5.03, the highest grade of the single head loss rate of the neck blast is 5 grade, wherein the HR-10 and Ningyuan flower 4 comprehensive disease resistance index is less than 5, the neck blast loss rate is less than 5, and the highest grade of the neck blast loss rate is lower than the contrast index. The combined disease resistance index of the Jiyan 108 is less than 5, the resistance loss rate is less than 5, but the single spike loss rate is highest higher than the control index. The specific identification results of disease nursery are shown in Table 11.
4.7 regional test late-maturing group production test disease resistance identification:
referring to 4 parts of materials, the comprehensive disease resistance type expression shows 3 parts of a Medium Resistant (MR) variety, the expression shows 1 part of a medium resistant (MS) variety, the contrast Ningjing No. 41 comprehensive disease resistance index is 3.51, the highest grade of the single panicle loss rate of the neck blast is 3 grade, wherein the section 15 and HR-9 comprehensive disease resistance index is less than 5, the neck blast loss rate is less than 5, and the highest grade of the neck blast loss rate is lower than the contrast index. 2012XW-239 comprehensive disease resistance index is less than 5, resistance loss rate is less than 5, but single spike loss rate is highest higher than control index. The specific identification results of disease nursery are shown in Table 12.
TABLE 6 resistance identification of test specimen in this region compared with early-maturing group
Note: HR: high resistance; r: disease resistance; MR: resisting; MS: sensing in the middle; s: infection; HS is high feeling. (ii) Miao Ye Wen disease grade GLB; incidence rate IDP of panicle blast; panicle blast incidence Grade (GIDP); the loss rate of panicle blast GLRP;
the integrated index of rice blast IB ═ GLB 25% + GIDP 25% + GLRP 50%.
TABLE 7 resistance identification of pre-test late-maturing group reference varieties in local area test
Note: as shown in Table 6
Table 8 resistance identification of early-maturing group reference varieties in the regional test:
as shown in Table 6
Table 9 resistance identification of late-maturing group reference varieties in the regional test of this region:
as shown in Table 6
Table 10 resistance identification of high-quality rice group reference varieties in the regional test:
note: as shown in Table 6
TABLE 11 resistance identification of variety to be tested in regional test early-maturing group production test
Note: as shown in Table 6
TABLE 12 resistance identification of test varieties from late-maturing group production test in local area
Note: as shown in Table 6
(III) disease resistance research of introduced resources:
the test design, field management, strain inoculation and identification recording methods are the same as above.
The subject of identifying variety resources was introduced into 108 parents, 8 parts of material with resistance reaching R grade accounting for 7.4%, 49 parts of material reaching MR grade level accounting for 45.4%, 34 parts of material reaching MS grade level accounting for 31.5%, 9 parts of material reaching S grade level, and 8 parts of material reaching HS grade. Provides the basis of disease resistance for the selection and the utilization of rice breeding parents, and the resistance results are shown in a table 13;
TABLE 13 evaluation of blast resistance by introduction of germplasm resources
In the description of the present invention, "a plurality" means two or more unless otherwise specified; the terms "upper", "lower", "left", "right", "inner", "outer", "front", "rear", "head", "tail", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are only for convenience in describing and simplifying the description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, should not be construed as limiting the invention. Furthermore, the terms "first," "second," "third," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the above embodiments, the implementation may be wholly or partially realized by software, hardware, firmware, or any combination thereof. When used in whole or in part, can be implemented in a computer program product that includes one or more computer instructions. When loaded or executed on a computer, cause the flow or functions according to embodiments of the invention to occur, in whole or in part. The computer may be a general purpose computer, a special purpose computer, a network of computers, or other programmable device. The computer instructions may be stored in a computer readable storage medium or transmitted from one computer readable storage medium to another, for example, the computer instructions may be transmitted from one website site, computer, server, or data center to another website site, computer, server, or data center via wire (e.g., coaxial cable, fiber optic, Digital Subscriber Line (DSL), or wireless (e.g., infrared, wireless, microwave, etc.)). The computer-readable storage medium can be any available medium that can be accessed by a computer or a data storage device, such as a server, a data center, etc., that includes one or more of the available media. The usable medium may be a magnetic medium (e.g., floppy Disk, hard Disk, magnetic tape), an optical medium (e.g., DVD), or a semiconductor medium (e.g., Solid State Disk (SSD)), among others.
The above description is only for the purpose of illustrating the present invention and the appended claims are not to be construed as limiting the scope of the invention, which is intended to cover all modifications, equivalents and improvements that are within the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A method for identifying disease resistance of a new rice variety is characterized by comprising the following steps:
step one, identifying the field plant growth period by combining natural induction with inoculation: using field disease straws as an inducing material, applying N fertilizer to create a disease condition, combining artificial inoculation to spray mixed spore liquid on rice leaves by a sprayer at a tillering peak stage in the middle and last ten days of June, assisting in sun-shading treatment, creating a condition beneficial to rice blast by a rice blast natural inducing module, investigating a blast disease condition index in the last ten days of June, and investigating a neck blast disease condition in 1 day of June from 8 Yue 15 to 9 Yue;
step two, collecting the rice blast standard samples of the rice planting area through a rice blast standard sample collecting module in the season of rice blast occurrence from the first 7 months to the first 10 months, and collecting and identifying physiological microspecies of rice blast germs;
performing tissue separation and purification on a rice blast specimen in a laboratory through a rice blast standard sample separation module to obtain a single spore strain, performing moisture preservation culture on the rice blast specimen for 24-48h at a constant temperature, observing whether rice blast exists or not under a microscopic examination in an ultraclean environment, shaking spores on a water agar plate culture medium when spores are generated on scabs, and performing moisture preservation culture for 16h in a constant temperature box at 25 ℃;
step four, selecting single spores by using a scalpel under an ultraclean environment microscope, inoculating the single spores to a PDA culture medium, carrying out moisture-preserving culture at 25 ℃ for 3-5 days until the single spores grow to 2cm in diameter, transferring the single spores to a rice flour culture medium again, carrying out moisture-preserving culture at 25 ℃ for 3-5 days, similarly removing the polluted strains, and separating 3-5 single spore strains from each sample; separating and purifying a plurality of strains, eliminating the strains with pollution and low spore production capacity in the strains, and storing effective strains;
inoculating the separated monospore strain to a spore production culture medium, storing the monospore strain in a constant-temperature incubator until the monospore strain is dried, putting the monospore strain into a sterilized cryopreservation tube in an aseptic environment, putting the cryopreservation tube into a sealed container according to the number, and temporarily storing the monospore strain in a refrigerator at the temperature of-20 ℃ for later use through a rice blast standard sample storage module;
sixthly, field identification of physiological races of rice blast germs by regional tests: respectively carrying out regional test and quality ratio test of an experiment base through a physiological race identification module to determine the dominant physiological races in the region; respectively carrying out species and pathogenicity determination on different varieties of rice blast germs in different areas by utilizing different types of collected differential hosts in the years after purification and rejuvenation of single spores, and detecting the composition and distribution of the physiological races of the rice blast germs by infecting the differential hosts to determine the dominant physiological races in the areas;
controlling the normal operation of each module of the new rice variety disease resistance identification system by a central control module through a central processing unit; fusing the acquired natural rice blast inducing data, the rice blast standard sample data and the physiological race identification result by using a data fusion processing module through a data fusion program to obtain rice blast data;
step eight, disease condition recording grading, resistance evaluation and standard calculation: respectively calculating the disease grade of the seedling leaf blast, the incidence rate of the panicle blast, the loss rate of the panicle blast and the comprehensive rice blast index according to the obtained rice blast data by using an index calculation module through a calculation program;
storing the obtained natural rice blast inducing data, rice blast standard sample data, physiological race identification results, fused rice blast data and calculation results by using a memory through a data storage module;
and step ten, updating the acquired natural rice blast inducing data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result by using an updating program through an updating display module, and displaying the data in real time through a display.
2. The method for identifying disease resistance of a new rice variety as claimed in claim 1, wherein in the first step, at the 3-leaf stage of a rice seedling, the test variety and the differential host are inoculated in a spray manner in an inoculation box, the inoculated variety and the differential host are placed in the inoculation box after inoculation and are moved into a shading shed for 24h, water is sprayed for 5min every 1h in the daytime for moisture preservation, a film is covered at night for heat preservation, and the disease occurrence is analyzed 8d after inoculation.
3. The method for identifying disease resistance of a new rice variety according to claim 1, wherein in the second step, the method for collecting the rice blast disease specimen comprises: the collection sites are distributed in 43 sites of the irrigation area, 531 parts of the collected specimens are collected, and the collected specimens of the panicle blast and the nodular cast blast are marked with numbers and stored in a classified mode.
4. The method for identifying disease resistance of a new rice variety according to claim 1, wherein in the sixth step, the field identification method for physiological races of the field test Pyricularia oryzae further comprises:
utilizing 24 monogenes to identify rice blast germs, utilizing different types of identifying hosts to infect and identify in a three-leaf one-heart environment under the sterile high-temperature high-humidity environment, and determining physiological races and anti-infection reaction on the monogenes identifying hosts; the identification process needs to pay attention to the grasp of the propagation time of the strain spores and the growth time of the differential hosts, multiple times of seedling culture are carried out in multiple stages, seeds are soaked by 402 during seedling culture, bed soil is disinfected, and the standard establishment of infection environment is carried out.
5. The method for identifying disease resistance of a new rice variety according to claim 1, wherein in the eighth step, the disease grade of the seedling leaf blast is calculated as GLB ═ Σ (NDL × GDL)/TNL;
in the formula: GLB-grade of seedling blight;
NDL-number of diseased leaves at each level;
GDL-representative value of each disease grade;
TNL-total leaf number investigated.
6. The method for identifying disease resistance of a new rice variety according to claim 1, wherein in the eighth step, the incidence rate of panicle blast is calculated as IDP ═ TNDP/TNP x 100;
in the formula: IDP-panicle blast incidence rate, and the unit is percentage (%);
TNDP-number of panicles affected;
TNP-total number of ears investigated;
resistance index corresponding to GIDP-IDP.
7. The method for identifying disease resistance of a new rice variety according to claim 1, wherein in the eighth step, the panicle blast loss rate (grade) is calculated as GLRP ═ Σ (NDP × GDP)/TNP;
in the formula: GLRP-panicle blast loss rate (grade);
NDP-the number of diseased ears at each level;
GDP-loss rate grade of each grade;
TNP-total number of ears investigated;
the rice blast integrated index is calculated as IB ═ GLB × 25% + GIDP × 25% + GLRP × 50%.
8. A new rice disease resistance identification system for performing the method for identifying new rice disease resistance according to any one of claims 1 to 7, comprising:
the rice blast standard sample collection module, the rice blast standard sample separation module, the rice blast standard sample storage module, the physiological race identification module, the central control module, the data fusion processing module, the index calculation module, the data storage module and the update display module;
the rice blast natural induction module is connected with the central control module and is used for spraying the mixed spore liquid on rice leaves through a sprayer, applying N fertilizer at high level and assisting in sun-shading treatment to create disease conditions of rice blast;
the rice blast standard sample collection module is connected with the central control module and is used for collecting neck blast and node blast samples of the rice planting area, marking the samples with uniform numbers and storing the samples in a classified manner;
the rice blast standard sample separation module is connected with the central control module and is used for carrying out tissue separation and purification on a rice blast specimen in a laboratory to obtain a monospore strain;
the rice blast standard sample storage module is connected with the central control module and is used for temporarily storing the monospore strain in a refrigerator for later use;
the physiological race identification module is connected with the central control module and is used for respectively carrying out field identification on physiological races of rice blast germs in a regional test through a regional test and a quality test of an experimental base;
the central control module is connected with the rice blast natural induction module, the rice blast standard sample collection module, the rice blast standard sample separation module, the rice blast standard sample storage module, the physiological race identification module, the data fusion processing module, the index calculation module, the data storage module and the updating display module and is used for controlling the normal operation of each module of the new rice variety disease resistance identification system through a central processing unit;
the data fusion processing module is connected with the central control module and is used for carrying out fusion processing on the acquired natural rice blast inducing data, the rice blast standard sample data and the physiological race identification result through a data fusion program to obtain rice blast data;
the index calculation module is connected with the central control module and is used for calculating the disease level of the seedling-leaf blast, the incidence rate of the panicle blast, the loss rate of the panicle blast and the comprehensive index of the rice blast through a calculation program according to the obtained rice blast data;
the data storage module is connected with the central control module and used for storing the acquired rice blast natural induction data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result through the memory;
and the updating display module is connected with the central control module and used for updating the acquired rice blast natural induction data, the rice blast standard sample data, the physiological race identification result, the fused rice blast data and the calculation result through an updating program and displaying the data in real time through a display.
9. A computer program product stored on a computer readable medium, comprising a computer readable program, which when executed on an electronic device, provides a user input interface for implementing the method of identifying disease resistance of a new variety of rice as claimed in any one of claims 1 to 7.
10. A computer-readable storage medium storing instructions which, when executed on a computer, cause the computer to execute the method for identifying disease resistance of a new rice variety according to any one of claims 1 to 7.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107190049A (en) * | 2017-06-07 | 2017-09-22 | 袁隆平农业高科技股份有限公司 | A kind of natural infection induces method of the rice blast seedling pest to identify or screen disease resisting rice material |
| CN109596788A (en) * | 2019-01-30 | 2019-04-09 | 湖南农业大学 | A kind of double season super hybridization rice water and fertilizer coupling measuring methods |
| CN109618896A (en) * | 2019-01-22 | 2019-04-16 | 温州科技职业学院 | A kind of rapid screening method of tomato Evaluation of Cold Tolerance and application |
| CN110607346A (en) * | 2019-11-01 | 2019-12-24 | 宁夏农林科学院农作物研究所（宁夏回族自治区农作物育种中心） | Identification method and application of pathogenic bacteria of weedy rice |
-
2020
- 2020-10-29 CN CN202011185424.1A patent/CN112293182A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107190049A (en) * | 2017-06-07 | 2017-09-22 | 袁隆平农业高科技股份有限公司 | A kind of natural infection induces method of the rice blast seedling pest to identify or screen disease resisting rice material |
| CN109618896A (en) * | 2019-01-22 | 2019-04-16 | 温州科技职业学院 | A kind of rapid screening method of tomato Evaluation of Cold Tolerance and application |
| CN109596788A (en) * | 2019-01-30 | 2019-04-09 | 湖南农业大学 | A kind of double season super hybridization rice water and fertilizer coupling measuring methods |
| CN110607346A (en) * | 2019-11-01 | 2019-12-24 | 宁夏农林科学院农作物研究所（宁夏回族自治区农作物育种中心） | Identification method and application of pathogenic bacteria of weedy rice |
Non-Patent Citations (4)
| Title |
|---|
| 史延丽等: "64个水稻品种(系)对稻瘟病混合菌的抗性鉴定", 《甘肃农业科技》 * |
| 史延丽等: "宁夏稻瘟病菌致病型单基因鉴别寄主筛选研究", 《宁夏农林科技》 * |
| 无: "2017年水稻品种稻瘟病抗病鉴定总结", 《豆丁网》 * |
| 陈锦文等: "人工注射接种及田间病圃鉴定水稻对稻瘟病的抗病性", 《广西植保》 * |
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