CN114410731B - Grate screen for screening rice blast resistance material and application thereof - Google Patents
Grate screen for screening rice blast resistance material and application thereof Download PDFInfo
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Abstract
The invention discloses a grate screen for screening rice blast (gene) resistant materials and application thereof, belonging to the technical field of resistant breeding. The application screens out the standard dominant non-toxic gene strain grate corresponding to the rice blast disease resistance genes through a large number of experiments, can directionally give the rice blast resistance pressure, screens out specific rice blast resistant rice varieties (materials) and lays a foundation for the broad-spectrum resistance breeding of the rice blast.
Description
Technical Field
The invention belongs to the technical field of resistance breeding, and particularly relates to a grate screen for screening rice blast resistance resources (materials) and application thereof.
Background
Rice is a staple food for 70% of the population on earth. Rice blast is a worldwide epidemic destructive disease which seriously affects the yield and quality of rice, and directly threatens the safe production of the rice. The planting of disease-resistant varieties is an effective measure for effectively controlling the occurrence of diseases, reducing the investment of chemical pesticides, protecting the ecological environment and improving the quality of life. However, the breeding of disease-resistant varieties requires broad-spectrum disease-resistant resources, and the virulence level of local rice blast bacteria needs to be mastered, so that the disease-resistant resources are selected in a targeted manner and used as donors to improve the resistance level of the varieties at home. The excavation of broad-spectrum resources for resisting rice blast is a bottleneck problem in breeding work, and the premise is that pathogenicity of pathogenic bacteria of contradictory main bodies needs to be mastered. The physiological race is used for identifying hosts to develop pathogenic differentiation research of rice blast bacteria, a plurality of systems are established at home and abroad, and positive effects are exerted for pathogenic genetic differentiation of pathogenic bacteria and guiding disease-resistant breeding research. 23 rice blast resistant broad-spectrum single-gene varieties cultivated by using the International Rice institute (IRRI) and the New Congress black cereal (LTH) of japonica rice Lijiang as receptors provide an effective tool for systematically developing pathogenic differentiation research of rice blast pathogenic bacteria, and are convenient for developing cooperation and communication of disease resistant germplasm resources internationally.
The rice variety interacts with the rice blast bacteria, and follows the theory of gene pair, namely, if one disease-resistant gene in the variety corresponds to one nontoxic gene in pathogenic bacteria or exists in parallel, the variety and the pathogenic bacteria do not show symptoms and show incompatibility even under the most proper disease condition. If there are a plurality of such avirulence genes in the pathogen, there is exactly the corresponding disease resistance gene in the variety, and such variety exhibits broad spectrum resistance. This is the ultimate goal and effective way to cultivate disease resistant varieties. In the breeding work of rice blast resistance, the identification and the utilization of a single broad-spectrum disease resistance gene are focused, and the rice blast bacteria have higher spore yield and mutation characteristics, so that the varieties relying on the single disease resistance gene are extremely easy to lose resistance.
Therefore, how to provide a tool and method capable of screening multiple disease resistant/broad spectrum varieties (materials) is a problem to be solved in the art.
Disclosure of Invention
The invention discloses a grate screen for screening rice blast resistant materials and application thereof.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
use of AvrPi12 (t), avrPi19, avrPi9, avrPiz or AvrPi20 in or in the preparation of a Jilin province rice blast resistant screening, evaluating and excavating rice blast resistant rice varieties (resources);
a grate screen for screening rice blast resistance grate screen, comprising:
rice blast screening evaluation vectors containing any one or more genes of AvrPi12 (t), avrPi19, avrPi9, avrPiz or AvrPi 20; and
rice blast screening evaluation vector containing any one gene of AvrPi12 (t), avrPi19, avrPi9, avrPiz or AvrPi20z, and any combination of two or more thereof.
As a preferable technical scheme, the rice blast screening and evaluating carrier is rice blast pathogenic bacteria;
a grate screen for screening rice blast resistance gene material comprising one or more of strain 201932, strain 201729, strain 201948, strain 20020817, strain 201782 and strain 201640;
the 201932 strain is preserved in China general microbiological culture collection center (CGMCC) with the address of China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.23255 in the North-ward, yangyang area of Beijing, at the 9 th month 13 of 2021, and is classified and named as Pyricularia oryzae (Pyricularia oryzae);
the 201729 strain is preserved in China general microbiological culture collection center (CGMCC) with the address of China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.23253 in the North-ward, yangyang area of Beijing, at the 9 th month 13 of 2021, and is classified and named as Pyricularia oryzae (Pyricularia oryzae);
the 201948 strain is preserved in China general microbiological culture collection center (CGMCC) with the address of China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.23256 in the North-ward, yangyang area of Beijing, at the 9 th month 13 of 2021, and is classified and named as Pyricularia oryzae (Pyricularia oryzae);
the 20020817 strain is preserved in China general microbiological culture Collection center (CGMCC) with the address of China general microbiological culture Collection center (CGMCC) with the collection number of CGMCC No.23251 in the North-ward-sun area of Beijing city at 9 and 13 days of 2021, and is classified and named as Pyricularia oryzae (Pyricularia oryzae);
the 201782 strain is preserved in China general microbiological culture collection center (CGMCC) with the address of China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.23254 in the North-ward, yangyang area of Beijing, at the 9 th month 13 of 2021, and is classified and named as Pyricularia oryzae (Pyricularia oryzae);
the 201640 strain is preserved in China general microbiological culture collection center (CGMCC) with the address of China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.23252 in the North-ward, yankee area of Beijing, at 9 and 13 days of 2021;
according to the gene-to-gene hypothesis, dominant non-toxic gene strains corresponding to broad-spectrum disease-resistant genes are obtained through screening: 201932, 201729, 201948, 20020817, 201782 and 201640 strains
The 201932 strain contains AvrPi12 (t), the 201729 strain contains AvrPi19, the 201948 strain contains AvrPi9, the 20020817 strain contains AvrPiz, the 201782 strain contains AvrPi20, and the 201640 strain contains AvrPi120, avrPi12 (t) and AvrPi19 simultaneously;
as an optimal technical scheme, the grate screen is used for screening broad-spectrum disease resistance genotypes of paddy rice in Jilin province;
the application of the grate screen in screening, evaluating and excavating rice blast resistance or in preparing a product for screening, evaluating and excavating rice blast resistance;
a method for using a grate screen for screening rice blast resistance gene materials comprises the following steps:
regulating the rice blast fungus spores of the Gracilaria verrucosa as claimed in claim 3 or 4 to 10 5 Inoculating 5-leaf-age rice, and determining the resistance of the rice according to the disease condition after 7 days;
as a preferable technical scheme, the method is characterized in that the culture humidity is 95-100%, and the culture temperature is 26-30 ℃;
as a preferable technical scheme, the method is also matched with gene level detection.
In summary, the invention discloses a grate for screening rice blast resistance and application thereof, wherein a large number of experiments are adopted to screen a standard dominant non-toxic gene strain grate corresponding to rice blast disease resistance genes, so that the rice blast resistance pressure can be given to rice in a directed manner, rice varieties with directed rice blast resistance can be screened out, and a foundation is laid for broad-spectrum resistance breeding of rice blast.
Drawings
FIG. 1 is a diagram of a standard strain pattern template of a grate screen;
FIG. 2 shows an example of a grate screen made of dominant virulent gene strains of Pyricularia oryzae in Jilin province;
FIG. 3 shows the interaction affinity of rice varieties Jie Japonica 88, jiu Dao 44 and Qing Lin 511 with Pyricularia oryzae 20020817 (avrPiz); the screening conditions of the rice blast bacterial strain are formed by sequencing the broad-spectrum disease-resistant gene resistance spectrum from high to low, namely A > B > C > D > E > F > G; the target gene in the grate screen shows unaffinity (disease resistance R, resistance) and the resistance spectrum is reduced, and the pathogenicity of other strains is as affinity as possible;
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1 grate Screen calibration "Standard Strain" screening procedure
From 1996, the Jilin agricultural academy of sciences rice disease team collected 100 or more representative strain standards (leaves or stems) of the rice blast frequent region in the northern japonica rice region each year, to 2020, approximately 2500. Separating monospore, and storing. In 2001, 31 parts of the team cultivated by International Rice institute (IRRI) contained 23 rice blast resistant single-gene line varieties (respectively containing disease-resistant genes Pia, pib, pii, pik, pik-h, P ik-m, pik-P, pik-s, pish, pit, pita, pita-2, piz-t, pi1, piz-5, pi3, pi5 (t), pi7 (t), pi9, pin 2 (t), pi19 and Pi 20), which were artificially inoculated and interacted with rice blast single-spore cultures collected in the past, pathotype of pathogenic bacteria was confirmed by affinity phenotype, specifically as follows:
(1) Collecting rice blast standard samples, and carrying out pathogen monospore separation, preservation and propagation in a laboratory;
single spore isolation of rice blast fungus: typical single rice blast fusiform plaque standard samples were selected, and 1 effective single spore was isolated for each plaque. The method comprises the following specific steps: the disease sample is rinsed in sodium hypochlorite solution for 2 minutes, rinsed for three times by sterile water, and is spread on a 2% water agar plate containing tetracycline after being dried by sterile water absorbing paper, and is cultivated for 2-5 days by illumination at 28 ℃, and a large number of conidia can grow around the edge of the cultivated leaf segment or stalk and the disease spot. Shaking monospore onto 5% agar culture medium, culturing at 25deg.C for 24 hr, picking monospore with picking needle under microscope, and transferring into PDA culture medium for culturing.
Preparation of oat agar medium: 45g oat flour and 18g agar powder were weighed and tap water was used to determine 1L volume. Sterilizing at 1 deg.C for 60min, and pouring into flat plate. The monospore isolate was transferred to oat plate medium with a transplanting needle and induced to produce spores at 28℃under black light and fluorescent lamps (12 h/12 h).
Preservation of Magnaporthe grisea monospore isolate: 17g of straw powder and 18g of agar powder are weighed, and distilled water is 1L.12 Sterilizing at 1deg.C for 60min, and pouring into flat plate. Sterile filter paper sheets are spread on the flat plate, and the monospore isolate is transferred onto a straw agar medium by using a transplanting needle. After the filter paper is full of hypha, the paper is completely dried, and then the filter paper is put into a collecting pipe and a preservation box for preservation at a low temperature of-20 ℃ (or normal temperature drying).
Artificial inoculation and resistance evaluation: compounding 10 with 0.1% Gelatin in sterile solution 5 The rice blast fungus spores with the concentration of/mL are sprayed in a greenhouse at 26-30 ℃ by a manual sprayer with the dosage of 30mL/m 2 The 5-leaf-age rice blast-resistant monogenic variety is inoculated manually, and after 7 days of full development, the disease spots (or disease spot areas) are investigated. The disease fraction is 0-9, wherein 0-3 is disease-resistant (R), and 4-9 is disease-sensitive (S). The evaluation of the resistance may be performed by evaluating the degree of interaction with the disease spots, or by evaluating the disease area. Selection is made according to specific conditions, such as screening of resistant resources or pathogenic analysis of pathogenic bacteria, and can be evaluated according to the type of the lesion (table 1); if resistance genetic analysis is performed, the disease area is evaluated (Table 2).
Table 1. The criteria were investigated according to the lesion type and the extent of damage.
Table 2. According to the area of the lesions.
The broad-spectrum disease-resistant genotype and the dominant nontoxic genotype of pathogenic bacteria are determined by the proportion of the strains with affinity (establishing the disease relationship). Each inoculation test is independently carried out at least 2 times, and each time, 2 times of repeated tests are carried out, investigation data of the occurrence of the disease is taken as interaction results, and the repeatability of evaluation data is ensured.
(2) Selection of "standard" strains of grate screen: selecting rice blast strains which have the interaction expression non-affinity ratio (pathogen dominant non-toxic genotype) from large to small, are non-pathogenic to a variety containing a specific disease-resistant single gene, are pathogenic to other disease-resistant single gene varieties at the same time, and have the most spore yield compared with the rice blast strains, and selecting the rice blast strains as 'standard' strains for the grate screening model.
More than 2000 rice blast bacteria with definite non-toxic genotype characteristics are analyzed, and avrPi12 (t), avrPi19, avr Pi9, avrPiz and avrPi20 are sequentially provided with dominant non-toxic genotypes of rice blast bacteria in Jilin province, in other words, pi12 (t), pi19, pi9, piz, pi20 and the like are sequentially provided with broad-spectrum disease resistant genotypes of local varieties. The strain meeting the characteristic requirements in a large number of strain groups and in the local region is selected as a candidate strain of the grate screen, and the strain obtained by screening only shows no toxicity for rice blast bacteria containing a specific disease resistance gene variety and shows toxicity for other disease resistance genotypes. Based on the above conditions, the "standard" strains were constituted to be in the grate screen mode (as shown in FIG. 1).
Example 2 screening of Magnaporthe grisea Graded screening Standard Strain for Pyricularia oryzae in Jilin province
Determining dominant non-toxic genotypes (or variety broad-spectrum disease-resistant genotypes) of the rice blast bacteria according to the interaction evaluation result of the rice blast monospore isolate and the monogenic varieties; and then the non-toxic genotype 'standard' strain of the non-toxic genotype is screened by sequencing the grate from large to small according to the dominant non-toxic genotype of pathogenic bacteria: the grid screening patterns were ordered in this order by the interaction data, avrPi12 (t), avrPi19, avrPi9, avrPiz, avrPi20 Jilin province rice broad-spectrum disease resistant genotypes.
Screening a rice blast fungus nontoxic genotype strain which contains specific disease resistance genes and is toxic to other disease resistance genotypes to form a grate screen. According to the sequence from high to low of the resistance spectrum of the broad-spectrum disease-resistant genotype, corresponding non-toxic gene standard strains are arranged, and strain number 201932 strain containing AvrPi12 (t), strain number 201729 containing AvrPi19, strain number 201948 strain containing AvrPi9, strain number 20020817 containing A vrPiz and strain number 201782 strain containing AvrPi20 are obtained through screening; strain numbers 201640 containing both AvrPi120, avrPi12 (t) and AvrPi19, the results are shown in fig. 2.
And selecting the strain meeting the conditions as a candidate standard strain (avoiding the introduction of exogenous pathogenic bacteria, using a combined strainer strain and avoiding the generation of a strain with super-toxic rice blast by diffusion). Finally, according to the characteristics of strong spore-producing capability and the like of the strain, the standard strain of the grate screen is selected and used for screening disease-resistant resource materials.
Aiming at different regions, such as different provinces or different countries, a grate is constructed according to the mode to screen and select a standard strain for screening rice blast resistant variety resources (materials).
The preservation condition of the standard strain is screened by the screening grate:
the standard strain of the rice blast fungus nontoxic gene grate screen corresponding to the broad-spectrum disease-resistant genotype of Jilin japonica rice area is delivered to the China general microbiological culture Collection center for preservation in 2021, 9 and 13 days and registered in a book. Specific strain codes and deposit numbers are as follows:
201932 strain: the preservation number is CGMCC No.23255;
201729 strain: the preservation number is CGMCC No.23253;
201948 strain: the preservation number is CGMCC No.23256;
20020817 strain: the preservation number is CGMCC No.23251;
201782 strain: the preservation number is CGMCC No.23254;
strains 201640: the preservation number is CGMCC No.23252.
The center responsible person issues acceptance notices and survival reports (see annex specifically) at day 10 and 18 of 2021. The center will hold the grate screen "standard" strain for 30 years.
Example 3 use of grate Screen
The Jijing No. 88 is a hybrid combination prepared by taking Japanese high-yield high-quality variety Aojiu No. 346 as a female parent and taking long white No. 9 with strong stress resistance, large ears and few tillers as a male parent. A record of the birth date was created. During 2005-2008, the planting area of Jijing 88 variety in Jilin province exceeds 50% of the total planting area, and the resistance is gradually lost due to the increase of the planting area and the extension of the popularization time. The resistance gene background is analyzed to find that the strain lacks certain broad-spectrum disease-resistant genes, such as Piz, pi9 and the like, based on the gene, jijing 88 is taken as a female parent, jiudao 44 is taken as a male parent, the resistance of the strain to rice blast is improved through hybridization combination by assisting genotype and phenotype identification, the resistance of Jijing 88 strain is accurately improved, and the Qinglin 511 disease-resistant high-quality strain is cultivated. In addition to Qinglin 511, qinglin 611, qinglin 711 and Qinglin 811 are also characterized in that Jijing 88 is used as a female parent, and resistance of Jijing 88 to rice blast is improved through hybrid family selection, genotype and phenotype identification auxiliary screening, and no genetic background and breeding process are reported.
Qinglin 511:
the Qinglin 511 variety is obtained by taking a non-toxic rice blast-containing strain JLAvr20020817 (containing AvrPiz) as a screening strain and assisting Piz molecular linkage markers.
(1) According to the dominant non-toxic gene group of Pyricularia oryzae in Jilin province, selecting a rice blast strain 20020817 (containing AvrPiz) containing non-toxic genes as a grate screen;
(2) Preserving 20020817 strain filter paper sheets, and propagating through oat in a laboratory;
(3) F1 generation seedling raising in a greenhouse by taking Jijing 88 as a female parent and Jiudao 44 as a male parent;
(4) At a temperature of between 26 and 30 ℃ and a humidity of between 95 and 100 percent, at a temperature of between 10 percent 5 Inoculating a rice blast resistant single-gene line variety of the rice with 5 leaf ages at the concentration of the rice blast fungus spores;
(5) After 7 days of disease development, the number of affinities (disease incidence relation) is determined through the disease spot type, the anti-susceptibility type of variety resources is determined, plants with strong resistance are selected, and molecular markers of corresponding disease resistance genes are used for auxiliary breeding. The disease degree was evaluated by using the disease spots (table 1), the disease resistance (R, R-resistance) was rated from 0 to 3, and the disease susceptibility (S, upper) was rated from 4 to 9.
The method comprises the following steps:
and (3) selecting and combining: work researches show that the Jijing 88 rice variety is a Jilin province super rice variety, the planting area reaches 50% of the area of a planting area in the whole province, the Jilin province technological progress first prizes are obtained, the Jilin province super rice variety is high-yield and high-quality, the initial disease resistance evaluation is MS (sense of middle-sized), the population of the infectable strain is increased rapidly along with the expansion of the planting area, and the variety resistance is gradually lost. Therefore, many breeders seek to improve their resistance. The nine rice 44 varieties have strong resistance, and the identification analysis shows that the Piz-containing gene plays a positive role. The parent is Jijing 88 (female parent) and Jiudao 44 (male parent) for hybridization combination, and the selfing mode is adopted for continuous propagation after hybridization.
Identification of disease resistance phenotype: the F3 generation material is planted in a seed selection garden from 2008-2013, an ideal single plant type is screened by combining a disease resistance phenotype and a genotype, and the disease resistance phenotype is identified by using a standard strain JLAV RPIZ20020817 with stronger pathogenicity (a strain which is non-pathogenic to Piz-containing varieties). Manual seed identification at seedling stage was performed by reference to method Wang Jichun (2015,crop prote ction): after the strain is subjected to spore production by an oat culture medium, spores are washed by using 0.1% of a solution of Gelatin, and the spore concentration is adjusted to be 2 multiplied by 10 5 And inoculating 4-5 leaf age generation material leaves per mL. After artificial inoculation, the temperature is controlled at 25 ℃ and the temperature is kept in the dark for 16 hours, and the temperature is kept at 25 ℃ and the relative humidity is 90% for developing symptoms in the greenhouse. The disease resistance phenotype evaluation was performed using the susceptibility evaluation method of Wang Jichun et al (2013,crop protection) (grade 0-9 plaque type evaluation, see Table 1 herein). After inoculation with standard avirulence gene strains, the resistance and the susceptibility ratios of the F2-3 material were counted (128:39). The crossing generation material is subjected to resistance phenotype and genotype screening from F2, and positive plants are reserved.
Screening target genes: leaf DNA is extracted from the plant with anti-influenza and the band of target gene is amplified and detected by PCR. The specific linkage marker primer of the disease resistance gene is JCPIz-F: 5'-CGTTGTATAGGACA GTTTCATT-3', JCPPiz-R5'-AATCTAGGCACTCAAGTGTTC-3'. The amplified positive products were sent to the company for sequencing and product information was analyzed by comparison with the NCBI database.
Pre-test, area test and production test: screening test of middle-early maturing of rice in the Jilin province of 2016, test of middle-early maturing area of rice in the Jilin province of 2017 and 2018, and test of suitable planting area; and 2018, the rice medium early ripening production test of Jilin province is participated. Through disease resistance, high yield and comprehensive agronomic character assessment and evaluation, the material is approved by the rice variety approval committee of Jilin province in the early 2020 and is named as Qinglin 511. The resistance tracking detection resistance spectrum in the east three provinces in the current year is proved to be more than 90%, and the stable resistance phenotype is shown. The strain JLAvr20020817 is utilized to perform greenhouse artificial inoculation identification on parent varieties Jijing 88 (female parent), jiudao 44 (male parent) and Qinglin 511 again in the rice 5-leaf stage, jijing 88 (female parent) presents disease resistance, and Jiudao 44 (male parent) and Qinglin 511 presents disease resistance, and the resistance situation is shown in figure 3.
The specific process and method for the research are as follows: assisted genotypic and phenotypic identification improves resistance of varieties to rice blast, wang Jichun et al, molecular plant breeding, 2020, 18 (20): 6762-6768.
Qinglin 611 (target gene is Pib, and the grate screening strain contains avirulence gene AvrPib), qinglin 711 (Pikh, avrPikh) and Qinglin 811 (Pizt, avrPizt,2014-45 strains are satisfied), which are materials obtained by taking Jijing 88 variety as a receptor, respectively taking different resistant background materials as donors, carrying out standard strain phenotype identification and target gene molecule linkage marker double screening, and being examined as a new variety by the Jilin province rice variety examination committee in the year 2020.
Example 3
The standard strain of Pyricularia oryzae in Jilin province is used for forming a grate screen, and in 2019, different disease-resistant materials are screened from 640 parts of materials in a rice computing resource library of Jilin province academy of agricultural sciences, wherein 60 parts of materials show broad-spectrum resistance and can be used for local rice blast resistance breeding.
The standard strain grate sieves are formed by using Jilin, liaoning and Heilongjiang representative strains, the resistance evaluation is carried out on 430 more generation materials entrusted by the crop research institute of China academy of agricultural science by using the grate sieves in 2021, 56 parts of disease-resistant materials are screened out by using Jilin strain grate sieves, 51 parts of disease-resistant materials are screened out by using Heilongjiang province strain grate sieves, 146 parts of disease-resistant materials are screened out by using Liaoning strain composition grate sieves, and the screening materials respectively account for 12.9 percent, 11.8 percent and 33.7 percent of the total number of the materials. Meanwhile, 20 parts of materials which simultaneously show disease resistance are screened out by three different grate screen strains, and the total weight of the materials is 4.6 percent. The disclosure of specific methods and results is inconvenient due to the involvement of business collaboration contracts (privacy guidelines).
In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to the embodiments described above will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (7)
1. A grate screen for screening rice blast resistant variety resources, comprising:
rice blast screening evaluation vector containing AvrPiz gene; the evaluation vector comprises a rice blast pathogen 20020817 strain with a preservation number of CGMCC No.23251.
2. A grate screen for screening rice blast resistance target genetic material, characterized by further comprising one or more of 201932 strain, 201729 strain, 201948 strain, 201782 strain and 201640 strain;
the 201932 strain has a preservation number of CGMCC No.23255, the 201729 strain has a preservation number of CGMCC No.23253, the 201948 strain has a preservation number of CGMCC No.23256, the 201782 strain has a preservation number of CGMCC No.23254, and the 201640 strain has a preservation number of CGMCC No.23252.
3. A grate screen for screening rice blast resistance according to claim 1 or 2, wherein said grate screen is used for screening broad spectrum disease resistance genotypes of rice in Jilin province.
4. The use of the grate screen of any one of claims 1-3 in or in the preparation of a rice blast resistant screening, evaluating and excavating product.
5. A method of using a grate screen for screening rice blast resistant material, comprising the steps of:
regulating the rice blast fungus spores of the Gracilaria verrucosa of claim 1 or 2 to 10 5 And/ml, inoculating 5-leaf-age rice, and determining the resistance of the rice according to the disease condition after 7 days.
6. The method of using a grate for screening rice blast resistance according to claim 5, wherein said cultivation is carried out at a temperature of 26-30℃and a cultivation humidity of 95-100%.
7. The method of claim 5, further comprising detecting the level of a gene.
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