NL2031181A - Method for cultivating scab - resistant and high - yield wheat in Huanghuai wheat area based on multi - gene polymerization - Google Patents

Method for cultivating scab - resistant and high - yield wheat in Huanghuai wheat area based on multi - gene polymerization Download PDF

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NL2031181A
NL2031181A NL2031181A NL2031181A NL2031181A NL 2031181 A NL2031181 A NL 2031181A NL 2031181 A NL2031181 A NL 2031181A NL 2031181 A NL2031181 A NL 2031181A NL 2031181 A NL2031181 A NL 2031181A
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resistance
wheat
scab
ljj
yield
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NL2031181A
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Hu Wenjing
Zi Yan
Chu Xiusheng
Song Jianmin
Cheng Dungong
Wang Xiaolu
Liu Cheng
Li Faji
Liu Jianjun
Guo Jun
Zhai Shengnan
Liu Aifeng
han Ran
Chen Xueyan
Li Haosheng
Cao Xinyou
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Crop Res Inst Shandong Acad Agricultural Sciences
Jiangsu Lixiahe Regional Agricultural Res Institute
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G22/00—Cultivation of specific crops or plants not otherwise provided for
    • A01G22/20—Cereals
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
    • A01H1/045—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection using molecular markers
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/12—Processes for modifying agronomic input traits, e.g. crop yield
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/12—Processes for modifying agronomic input traits, e.g. crop yield
    • A01H1/122—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • A01H1/1245—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, e.g. pathogen, pest or disease resistance
    • A01H1/1255—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, e.g. pathogen, pest or disease resistance for fungal resistance
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/13—Plant traits
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Abstract

Disclosed is a method for cultivating scab-resistant and high-yield wheat in wheat regions, in particular in the Huanghuai wheat region, based on polygene polymerization. In the method, the scab-resistant intermediate material NMASOZO with excellent comprehensive agronomic characters carrying a plurality of scab-resistant genes is used as the resistance source, and the large-area varieties and excellent strains in wheat region are used as the background parents. Using molecular markers linked markers LJJ-1 of th1, linked markers LJJ-2 and LJJ-3 of th2, and linked markers LJJ-4 and LJJ-5 of th5 assisted selection combined with identification techniques for resistance to expansion and infection of scab, the semi-winter breeding materials or lines with good adaptability and comprehensive characters were created, which provided a material source for cultivating high-yield wheat varieties with scab resistance suitable for planting in the wheat area.

Description

Method for cultivating scab - resistant and high - yield wheat in Huanghuai wheat area based on multi - gene polymerization
TECHNICAL FIELD The invention belongs to the technical field of wheat breeding methods, and relates to a method for cultivating wheat with scab - resistance and high vield in Huanghuai wheat area based on multi polymerization.
BACKGROUND Fusarium head blight (FHB, also called Fusarium ear blight (FEB) or scab) has always been the most serious disease affecting wheat yield and quality in the middle and lower reaches of the Yangtze River, northeast spring wheat region and south China wheat region. In recent years, due to the influence of climate warming, the popularization of dwarf varieties, the improvement of multiple cropping index and straw returning, the epidemic area of wheat scab is constantly expanding, rapidly expanding to Huanghuai winter wheat area in the main wheat producing areas and even the northern winter wheat area, and changing from occasional diseases to recurrent diseases and major diseases. Due to the lack of inducing environment of scab in the history of Huanghuai wheat area, breeders paid late attention to disease - resistant varieties, which caused the climate change and the change of farming system, and the scab hazard in this wheat area spread rapidly and aggravated. Therefore, it has become one of the main breeding objectives in Huanghuai wheat area to cultivate wheat varieties with scab resistance of moderate or above level. The cultivation of wheat varieties with scab resistance cannot be separated from the utilization of scab resistance genes. In recent years, the research on wheat scab resistance genes at home and abroad has made rapid progress, which has laid a theoretical foundation for wheat scab resistance genetic improvement. At present, there are more than 250 QTLs related to scab resistance, which are distributed on 21 chromosomes of wheat, but there are only 7 clear scab resistance genes, namely Fhb1 - Fhb7, among them, Fhb1 (References: Cuthbert P A, Somers D J, Thomas J, et al. Fine mapping Fhb1, a major gene controlling Fusarium head blight resistance in bread wheat (Triticum aestivum L.) [J]. Theoretical & Applied Genetics, 20086, 112(8):1465-1472), Fhb2 (References: Cuthbert PA, Somers DJ, Brulé-Babel A. Mapping of Fhb2 on chromosome 6BS: a gene controlling Fusarium head b light field resistance in bread wheat (Triticum aestivum L.)[J]. Theoretical & Applied Genetics, 2007, 114: 429 - 437) is a scab resistant expansion type gene derived from common wheat, Fhb4 (References: Xue SL, Li G Q,JiaHY, etal. Fine mapping Fhb4, a major QTL conditioning resistance to Fusarium infection in bread wheat (Triticum aestivum L.) [J]. Theoretical & Applied Genetics, 2010, 121(1):147-156) and Fhb5 (References: Xue S L, Xu F, Tang M Z, et al. Predise mapping Fhb5, a major QTL conditioning resistance to Fusarium infection in bread wheat (Triticum aestivum L.)
[J]. Theoretical & App lied Genetics, 2011, 123(8): 1055 - 1063) is a scab - resistant infection type derived from common wheat. Studies have shown that the resistance to scab of main wheat varieties in Huanghuai wheat area is generally poor. With the development of molecular biology, the mapping of disease resistance genes and molecular marker technology are becoming more and more perfect, and molecular marker - assisted breeding technology has been widely used. Developed countries such as the United States, Canada, Australia and Japan have selected a number of new wheat varieties assisted by Fhb1 molecular markers, and their resistance to scab has been significantly improved. Germplasm resources are the basis of disease - resistant gene mining and disease - resistant breeding. Varieties and germplasm materials with good resistance to scab in the middle and lower reaches of the Yangtze River are of poor spring and cold tolerance, which are difficult to be directly applied in the breeding of scab - resistant varieties in Huanghuaibei area. Germplasm materials with resistance to scab in winter and semi - winter wheat are scarce. Therefore, based on the large - scale varieties in this wheat area, it is an important way to breed scab resistance in Huanghuai wheat region to introduce disease resistance genes by using existing molecular markers for scab resistance and to select comprehensive agronomic characters by conventional breeding. At present, there is no precedent for using molecular marker - assisted selection to breed scab resistant varieties in this wheat area.
In order to improve wheat scab resistance, the existing technology is to select varieties (lines) with scab resistance in the middle and lower reaches of the Yangtze River or cross existing resistance sources with varieties in Huanghuai wheat area, naturally select materials with poor agronomic characters in the low generation field, and select phenotypic choices such as large - scale inoculation and identification of scab in the high generation field, and measure the yield after harvest to select varieties with good scab resistance and high yield. However, 1. The varieties and germplasm materials with good resistance to scab in the middle and lower reaches of the Yangtze River are poor in spring and cold tolerance, so it is difficult to directly apply them in the breeding of scab - resistant varieties in Huanghuaibei area. Because of the differences in genetic background of breeding materials, various traits of offspring are separated greatly, and the stability of some traits takes a long time. When there are more excellent traits to be aggregated, the above - mentioned problems become more prominent, and often lead to too long breeding cycle or even failure. 2. Treating hybrid progeny by traditional hybrid method or pedigree method: hybrid method usually starts to select plants in F4 generation, and only the inferior plants are eliminated in the early stage, without effective selection of target characters.
However, the resistance to scab infection of wheat scab is often linked with some unfavourable agronomic characters, such as high plant height, late heading date, low ear density and low grain weight, so many materials carrying disease resistance genes will be eliminated in the lower generation. However, pedigree method is very purposeful in F2 generation, but the workload is too large, and the identification environment of wheat scab is very demanding. However, the environment in Huanghuai wheat area is not conducive to large - scale disease, and the selection pressure is not strong, which leads to low breeding efficiency and even breeding failure. Therefore, there is an urgent need for a breeding method that can cultivate wheat with scab resistance and high yield in Huanghuai wheat area.
SUMMARY The objective of the present invention is to overcome the defects existing in the prior art and provide a method for cultivating high - yield wheat resistant to scab, in particular in the Huanghuai wheat region based on polygene polymerization. The method uses scab resistant intermediate materials with excellent comprehensive agronomic characters carrying multiple scab resistant genes as the resistance source, and large - scale varieties and excellent lines in wheat regions, in particular in the Huanghuai wheat region, as the background parents, and uses molecular marker - assisted selection to combine identification technology of scab resistance to spread and infection to create semi - winter scab resistant breeding materials or lines with good adaptability and excellent comprehensive characters for cultivation. It provides a material source for cultivating wheat varieties with high yield and scab resistance suitable for planting in Huanghuai wheat area.
The invention provides a method for cultivating wheat with scab resistance and high yield in wheat areas, in particular in the Huanghuai wheat area, based on polygene polymerization, which comprises the following steps: step 1, crossing intermediate materials carrying multiple scab resistance genes as female parent and widely popularized varieties with an average yield of over 500 kg per mu as male parent to obtain hybrid seeds of the first generation; step 2, using the first hybrid seed in step 1 as the female parent, using the wheat strain with short plant, compact plant type, strong cold resistance and high powdery mildew resistance as the male parent, and harvesting the hybrid F: seed; step 3, planting F+, mixing and threshing to generate F; step 4, F is planted in the field, single plants with excellent characteristics such as cold resistance, short height, compact plant type, powdery mildew resistance, square spike and the like are selected, and the selected single plants are harvested and threshed to generate the seeds of the compound F3 single plant, and the red peel seeds are eliminated, step 5, in the multiple crossing Fs generation, planting in the field according to the plant row, selecting the plants with consistent plant row traits and excellent comprehensive disease resistance and comprehensive agronomic traits, the linked markers LJJ-1 of Fhb1, the linked markers LJJ-2 and LJJ-3 of Fhb2, the linked markers LJJ-4 and LJJ-5 of Fhb5 of the selected single plant in detection and screening, select the lines with pasitive molecular markers; at the later growth stage of wheat, according to the detection results of molecular markers, plants with good comprehensive agronomic traits, powdery mildew resistance and consistent phenotypes were selected from plants containing three target genes of scab resistance genes Fhb1, Fhb2 and Fhb5. The selected plants in the field are threshed according to the single - plant harvest (whole - row single - plant full - pull) when harvested, and the single plants with small grains, poor plumpness and red skin grains are eliminated, step 6, in the backcross F4 generation, molecular marker detection is carried out, and the linkage markers LJJ-1, LJJ-2 and LJJ-3 of Fhb2 and LJJ-4 and LJJ-5 of Fhb5 of the scab resistant gene Fhb1 in the selected plant rows are detected and screened, and the plant rows with positive molecular marker detection are selected; plant rows with positive molecular markers at heading and flowering stage of wheat were inoculated with single flower drip method to identify scab resistance.
At the later growth stage of wheat, select the plants with consistent phenotypes of plant row characters, better resistance to scab than the middle resistance control, comprehensive disease resistance and good agronomic characters to harvest and thresh, measure the yield of plant row, and select the plants with the highest yield rank to become Fs strain; step 7, the selected Fs strain is planted into an identification garden according to the plot, with two replicates, and the first replicate uses the single flower drip method to identify the strain's anti - expansibility.
The second repeated application of diseased wheat grain method identified the scab resistance, and the harvest resistance to infection and expansion were better than or equal to the middle resistance strain.
The yield of the first repeated application was identified, and the strain with an average yield of more than 500 kg per mu was selected, which was recommended to enter the production test, and a new wheat variety (line) with scab resistance and high yield with polygenic polymerization in Huanghuai wheat area was bred.
Furthermore, the intermediate materials carrying multiple scab resistance genes are wheat NMAS020 and NMAS018. Furthermore, the varieties with an average yield of over 500 kg per mu that are widely popularized are Jimai 22, Jimai 44, Jimai 23 and Luyuan 502. Furthermore, the wheat strain with short plant, compact plant type, strong cold resistance and high powdery mildew resistance is Shi HO83-366. Furthermore, the primer sequences of the linkage marker LJJ-1 of the scab resistance gene Fhb1 are shown in SEQ ID NO:1 and SEQ ID NO:2. Furthermore, the primer sequences of the Fhb2 linkage marker LJJ-2 of the scab resistance gene are shown in SEQ ID No:3 and SEQ ID No:4, and the primer sequences of LJJ- 3 are shown in SEQ ID No:5 and SEQ ID No:6. Furthermore, the primer sequences of the linkage marker LJJ-4 of the scab resistance gene Fhb5 are shown in SEQ ID No:7 and SEQ ID No:8, and the primer sequences of LJJ-5 are shown in Seq ID No.9 and SEQ ID NO:10.
Furthermore, the corresponding linkage marker LJJ1 of the scab resistant major gene Fhb1 is detected by PCR amplification, which comprises the following steps: the PCR amplification system is 10 pl, including 30 ng/ul wheat genomic DNA 1.0 pl, 10 x PCR buffer 1.0 yl, 10 Mm dNTP 0.2 pl, 10 Mm MgCl: 1.0 ul, 5U Taq polymerase 0.2 pl, 5 uM upstream primer 0.4 pl, 5 5 pM downstream primer 0.4 pl and sterile deionized water 5.8 pl; the PCR amplification procedure is: pre - denaturation at 94°C for 5min; denaturation at 94°C for 30 s, annealing at 64°C for 30 s, extension at 72°C for 45 s, 35 cycles; 72°C for 10 minutes; store at 4°C.
LJJ1 primer was used to detect the materials involved in this study together with their parents in 1% agarose electrophoresis solution.
The target genotype was the same as the intermediate material carrying multiple scab resistance genes, and it was the selected material.
Furthermore, the corresponding linkage markers LJJ-2 and LJJ-3 of FHB major gene Fhb2 and LJJ-4 and LJJ-5 of Fhb5 are detected by PCR amplification.
The PCR amplification method is as follows: the PCR amplification system is 10 pl, including 30 ng/ul wheat genomic DNA 1.0 pl, 10 x PCR buffer 1.0 pl, 10 Mm dNTP 0.2 pl, 10 Mm MgCl: 1.0 pl, 5U Tag polymerase 0.2 pl, 5 uM upstream primer 0.4 ul, 5 uM downstream primer 0.4 pl and sterile deionized water 5.8 pl; the PCR amplification procedure is: (1) pre - denaturation at 94°C for 8min, (2) denaturation at 94°C for 30 s, (3) annealing of SDAAS2 primer at 60°C for 40 s; LJJ-3 primer annealed at 61°C for 40 s; LJJ-4 primer annealed at 52°C for 40 s; LJJ-5 primer annealed at 61°C for 40 s (4) extended at 72°C for 30 s, 36 cycles, (5) extended at 72°C for 10 Min; (8) store at 4°C.
LJJ-2 and LJJ-3 primers, LJJ-4 and LJJ-5 primers were used in 8% non - denatured polyacrylamide gel electrophoresis solution, arc: bis = 19 : 1, 200 volts for 1 hour and 40 minutes - 2 hours and 30 minutes.
The materials involved in this study, together with their parents, were detected.
The target genotype was the same as the intermediate material with multiple scab resistance genes, which was regarded as positive and was the selected material.
Furthermore, in the identification of scab resistance, Jimai 22 was used as the susceptible control, Su Mai No.3 as the resistant control and Zheng 9023 as the middle resistant control.
Further, the strain used in the single - flower drip method is Fusarium graminearum (F.
G) 15 - ADON type, which was donated by the Institute of Food Quality, Safety and Detection of Jiangsu Academy of Agricultural Sciences, propagated and provided by Chu Xiusheng of Crop Research Institute of Shandong Academy of Agricultural Sciences, and artificially inoculated and identified by single - flower drip method; take 20 microliters of conidia suspension with micropipette and inject it into the floret just blossoming in the middle spikelet of wheat ear; Each variety was inoculated with 5 ears, and wheat ears were inoculated with fresh - keeping bags to keep moisture for 3 days.
The number of diseased spikelets and total spikelets per inoculated ear was investigated 21 days after inoculation, and the average number of diseased spikelets and the average rate of diseased spikelets (%) were calculated as the evaluation index of scab resistance. (Xue SL, Xu F ‚Tang M Z ‚et al.
Predise mapping Fhb5 ‚a major QTL conditioning resistance to Fusarium infection in bread wheat (Triticum aestivum L .) [J] . Theoretical & Applied Genetics, 2011,123(6):1055 -1063). Compared with the prior art, the present invention has the following technical effects: (1) The present invention analyses and verifies that it contains different disease resistance genes and different disease resistance gene combinations, and its scab resistance is different. Among the four types of strains containing only one disease resistance gene, the average number of diseased spikelets of strains containing Fhb1 and Fhb4 was significantly lower than that of strains containing Fhb2 and Fhb5 and the control Jimai 22, showing good resistance to scab. In the combination containing two disease - resistant genes, the strain containing (Fhb1+Fhb2) showed the lowest number of susceptible spikelets and the highest scab resistance, which was significantly lower than that of the strain containing (Fhb1+Fhb4). It can be seen that the disease resistance of any combination of two disease resistance genes is not predictable, and there is a significant difference between the performance of a single disease resistance gene and that of two disease resistance genes of the same type. At the same time, the lines with three disease - resistant gene combinations (Fhb1+Fhb2+Fhb5) showed the lowest average number of susceptible spikelets and the best scab resistance. According to the invention, molecular marker - assisted selection and conventional breeding technology are utilized, wherein the molecular markers adopted are the linkage markers LJJ-1 of Fhb1, LJJ-2 and LJJ-3 of Fhb2, LJJ-4 and LJJ- 5 of Fhb5, and the specific combination can improve the breeding efficiency of FHB1.
(2) Through the method of the invention, a semi - winter scab resistant breeding material Jimai 8803 with prominent target characters and excellent comprehensive characters was created, which laid a material foundation for cultivating wheat varieties with high yield and scab resistance suitable for planting in Huanghuai wheat area.
BRIEF DESCRIPTION OF THE FIGURES In order to more clearly explain the specific embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the description of the specific embodiments will be briefly introduced below.
Fig. 1 Disease performance of new wheat strain Jimai 8803 and control Jimai 22 after 21 days of single flower drip inoculation.
Fig. 2 Performance of comprehensive agronomic characters of Jimai 8803.
DESCRIPTION OF THE INVENTION Embodiments of the technical scheme of the present invention will be described in detail below with reference to the accompanying drawings. The following examples are only used to more clearly illustrate the technical scheme of the present invention, so they are only examples, and cannot be used to limit the scope of protection of the present invention. It should be noted that unless otherwise stated, the technical terms or scientific terms used in this application should have the ordinary meaning understood by the technical personnel in the field to which this invention belongs.
Example 1 Construction of breeding population for resistance to scab and breeding method of hybrid progeny Hybrid F1 was obtained by crossing with NMAS020 as the female parent and Jimai 22 as the male parent.
Taking the hybrid F as the female parent, taking Shi HO83 - 366 containing the powdery mildew resistance gene Pm21 as the male parent recrossing, the recrossing purpose is mainly to improve the powdery mildew resistance of the hybrid offspring; the recrossed progeny were bred for cold tolerance, plant height, plant type, panicle traits, grain color and powdery mildew resistance by pedigree method, and molecular markers were used to track and detect scab resistance genes; 53 strains with basically stable characters and excellent comprehensive characters were inoculated by single flower drip method to identify scab resistance, and Jimai 22 was the susceptible control.
Marker - specific primer sequences closely linked with scab resistance genes Fhb1, Fhb2 and Fhb5 are shown in Table 1. Table 1 Fhb1 and Fhb2 linked marker primer sequence information to mer BEE GK AUTOR AATAS UAT: SEG Wy Ne oO LIRA CGUAGAT THAUCGROTEGAY REQ MF HERR a sans WARE CUTGAGAGUIRITIIYTTG BEST JJ Nes or “as LJ SERSDGOTNMNTLACTE SED UI Mins ws IJE TUCTSEERSSTDOUITTURD BEI Us NES or SEE yaa FOTTICICCCOTTOUAATLT SEO IE MDS LE CRATTFTAGATOAR AAR BELG ID Nw ay re BAR JNM EVER CURA BEE HES NDS
The amplification method is as follows: the PCR amplification system is 10 pl, including 30 ng/pl wheat genomic DNA 1.0 yl, 10 x PCR buffer 1.0 pl, 10 Mm dNTP 0.2 pl, 10 Mm MgCl, 1.0 ul, SU Taq polymerase 0.2 pl, 5 HM upstream primer 0.4 ul and 5 pM downstream primer 0.4 pl and sterile deionized water 5.8 pl.
The PCR amplification procedure is: pre - denaturation at 84°C for Smin; denaturation at 94°C for 30 s, annealing at 64°C for 30 s, extension at 72°C for 45 s, 35 cycles; 72°C for 10 minutes; store at 4°C.
LJJ-1 primers were used to detect the materials involved in this study together with their parents in 1% agarose electrophoresis solution.
The target genotype was the same as NMAS020, which was the selected material.
Adopt the method for PCR amplification to detect the corresponding linkage markers LJJ-2 and LJJ-3 of the scab resistance major gene Fhb2, and the method for PCR amplification described in the corresponding linkage markers LJJ-4 and LJJ-5 of Fhb5 is: the PCR amplification system is 10 pl, containing 30 ng/ul wheat genomic DNA 1.0 pl, 10 x PCR buffer
1.0 pl, 10 Mm dNTP 0.2 pl, 10 Mm MgCI2 1.0 pl, 5UTaq polymerase 0.2 pl, 5 uM upstream primer 0.4 pl, 5 HM downstream primer 0.4 pl and sterile deionized water 5.8 ul. The PCR amplification procedure is: (1) pre - denaturation at 94°C for 8 min, (2) denaturation at 94°C for 30 s, (3) SDAAS2 primer annealed at 60°C for 40 s; LJJ-3 primer annealed at 61°C for 40 s; LJJ-4 primers were annealed at 52°C for 40 s; LJJ-5 primers were annealed at 61°C for 40 s (4) 72°C extension for 30 s, 36 cycles, (5) 72°C extension for 10 minutes; (6) store at 4°C. LJJ - 2 and LJJ-3 primers, LJJ-4 and LJJ-5 primers were used for electrophoresis in 8% non - denatured polyacrylamide gel electrophoresis solution, arc: bis = 19 : 1, 200 volts for 1 hour and 40 minutes - 2 hours and 30 minutes. The materials involved in this study and their parents were detected, and the target genotype was the same as that of NMAS020, which was regarded as positive, and it was the selected material.
Tightly linked marker - specific primers and amplification of Fhb4 refer to the article (Xue SL Li GQ, Jia HY, et al. Fine mapping Fhb4, a major QTL conditioning resistance to Fusarium infection in bread wheat (Triticum aestivum L.) [J]. Theoretical & Applied Genetics, 2010, 121(1):147 - 156 .). Detection and judgment method 53 strains and control Jimai 22 were planted in 15 fields of Crop Research Institute of Shandong Academy of Agricultural Sciences, arranged in sequence, with one repetition, 6 rows of plots, plot length of 4m and width of 1.5m, mechanical drilling, and basic seedlings of 150,000/mu. For the investigation of agronomic traits such as seedling habit, plant type, grain type and plant height, refer to the technical specification of regional test of crop varieties (NY/T 1301 - 2007).
Fusarium graminearum 15 - ADON was used for identification of scab, which was donated by Institute of Food Quality, Safety and Detection of Jiangsu Academy of Agricultural Sciences, propagated and provided by Chu Xiusheng of Crop Research Institute of Shandong Academy of Agricultural Sciences, and was artificially inoculated and identified by single flower drip method. Take 20 microliters of conidia suspension with micropipette, and inject it into the newly blossoming florets of the middle spikelets of wheat ears; each variety was inoculated with 5 ears, inoculated with wheat ears in fresh - keeping bags for 3 days, and the number of diseased spikelets and total spikelets per inoculated ear was investigated 21 days after inoculation, and the average number of diseased spikelets and the average rate of diseased spikelets (%) were calculated as the evaluation index of scab resistance.
The breeding practice shows that although Su Mai No.3 and Wangshuibai and their derivative lines have good resistance to scab, their agronomic traits, resistance to other diseases and high yield are poor. Because of gene linkage, the target genes of scab resistance are often lost when comprehensive traits are selected. According to the scab - resistant multiple cross breeding population of the present invention, 53 strains are finally retained after successive generations of selection of excellent single plant and agronomic characters of strains in the field, and through detection of scab - resistant molecular markers, 33 strains out of 56 strains contain 1 - 3 disease - resistant genes, accounting for 62.3%; 20 (37.7%) did not contain any loci (Table 2). Although the hybrids were detected by molecular markers in each generation, and there were 15 types of genome combinations (including 4 types of gene combinations, Fhb1+Fhb2+Fhb4+Fhb5) in the early generation, there were only 7 types of gene combinations due to the selection of comprehensive agronomic characters in the field.
Table 2 Number of strains of different genes and their combinations ed Gene Sivan quantity {pieces} of diseased, | spikelet vate {25} diseased u spikelets {pieces} & spikelet rate ERIN 3 4 BES 31083 On the whole, there are significant differences in the average number of diseased spikelets and the average rate of diseased spikelets with different numbers of scab resistance genes.
The average number and average rate of diseased spikelets with three scab resistance genes are significantly lower than those with two scab resistance genes, those with two scab resistance genes are significantly lower than those with one scab resistance gene, and those with one scab resistance gene are significantly lower than the strain and control variety Jimai 22 without any scab resistance genes (Table 2). According to the grading standard and evaluation standard of scab severity (agricultural industry standard), the varieties (lines) containing one major resistance gene can reach the middle level, two genes can reach the middle level or above, and three resistance genes can reach the high level, and the varieties (lines) without any major genes (including the control Jimai 22) show high sensitivity to scab.
Table 3 Average expression of scab of strains containing different numbers of disease resistance genes
{ isd 32 Example 2 Step 1: in April of the first year, in the greenhouse, NMA S020, an intermediate material carrying multiple scab resistance genes, was used as the female parent, and Jimai 22, a variety with an average yield of more than 500 kg per mu and a cumulative popularization of more than 100 million mu, was used as the male parent to obtain hybrid F1 seeds. NMASO20(NIL/PH691}, the donor material of scab resistance gene, is an intermediate material for scab resistance created by Nanjing Agricultural University by crossing the near - isogenic line of the major QTL of Shuishuibai with PH691. It contains four scab resistance genes such as Fhb1 (located on chromosome 3B), Fhb2(8B), Fhb4(4B) and Fhb5(5A).
Jimai 22, a wheat variety bred by Shandong Academy of Agricultural Sciences, took part in Shandong provincial test from 2004 to 2006, ranking first in both years, with an average yield of
536.81 kg/mu, an extremely significant increase of 10.79% compared with the control, and an average yield of 519.1 kg/mu in production test, an increase of 4.05% compared with the control. From 2004 to 2006, the average yield per mu was 518.08 kg, which was significantly increased by 4.67% compared with the control, and the average yield of production test was increased by 2.05% compared with the control.
Step 2: in April of the next year, the hybrid F1 was used as the female parent in the greenhouse, and the wheat strain HO83 -366 with short plant, compact plant type, strong cold resistance and high powdery mildew resistance was used as the male parent, and the hybrid F+ seeds were harvested.
ShiH083 -366 comes from Shijiazhuang Academy of Agricultural Sciences, (References: Fu Xiaoyi, Li Caihua, Zhao Yankun, Shi Zhanliang, Guo Jinkao, He Minggi. Analysis of high yield, stable yield and adaptability of new wheat variety’ Shimai 22', chinese agricultural science bulletin, 2016 (32):21:38 -43).
Step 3: In October of the third year, multiple cross F: is planted in the field, mixed harvest and threshing are carried out to generate multiple cross F2 seeds.
Step 4: in October of the fourth year, F2 was planted in the field, and a single plant with cold tolerance of Grade 2 or above, plant height of 70 - 85 cm, compact plant type, 400 - 450,000 ears per mu, 36 - 42 grains per ear, thousand - grain weight of 40 - 45 g and white grains was selected.
Step 5: in October of the fifth year, Fs was planted in the field according to plant rows (double row area, 3m in length, 30cm in row spacing and 5.2 cm in plant spacing), and the control variety Jimai 22 in Huanghuai wheat area was set every 20 rows as the reference of agronomic traits. At the rising stage of wheat, leaves are taken by plant row (5 leaves of each plant row are mixed), and molecular marker detection is carried out according to the method described in Example 1. Linkage markers LJJ1, LJJ-2 and LJJ-3 of Fhb1, Fhb2 and Fhb5 genes inthe selected plant row are detected and screened, and the plant rows with positive molecular marker detection (including heterozygosity) are selected. At the later growth stage of wheat, according to the detection results of molecular markers, select the plant rows with good comprehensive agronomic characters, powdery mildew resistance and consistent phenotypes among the plant rows containing the three target genes of scab resistance genes Fhb1, Fhb2 and Fhb5, and thresh the selected plant rows in the field according to the single - plant harvest (whole - row single - plant full - pull) when harvesting, and eliminate the single plants with small grains, poor plumpness and red skin grains into Fa.
Step 6: in October of the sixth year, the compound cross Fa is planted in the field by row (double row area, 3 m in length, 30 cm in row spacing and 5.2 cm in plant spacing), and a row of Huanghuai wheat area control variety Jimai 22 is set every 20 rows as reference for agronomic traits. At the rising stage of wheat, leaves were taken by plant row (5 leaves from each plant row were mixed). Molecular marker detection was carried out according to the method described in Example 1. Linkage markers LJJ-1 of Fhb1 gene, LJJ-2 and LJJ-3 of Fhb2, LJJ-4 and LJJ-5 of Fhb5 in the selected plant row were detected and screened. Select the lines with positive molecular markers; the plants with positive molecular markers in wheat heading and flowering stage were inoculated into the selected plants by single flower drip method. After inoculation, fresh - keeping bags were used to keep moisture for 3 days. After 21 days, the incidence rate of scab spikelets was investigated to identify scab resistance. Jimai 22 was the susceptible control, Su Mai 3 was the disease - resistant control and Zheng 9023 was the medium - resistant control. At the later growth stage of wheat, select the plants with consistent phenotypes of plant row characters, the average diseased spikelet rate less than Zheng 2023 after 21 days of inoculation, and good comprehensive disease resistance and agronomic characters to harvest and thresh, measure the yield of plant row, and select the plants with the highest yield to become Fs strain.
The steps of scab identification are the same as in Example 1. In the present invention, the average diseased spikelet rate of the strain after 21 days of inoculation is less than that of the middle resistance control, which is regarded as the selected material.
Step 7: in October of the seventh year, the selected Fs strain was planted in the field as an identification garden according to the plot, and the control variety Jimai 22 in Huanghuai wheat area was taken as the reference of agronomic traits, and the yield identification and disease resistance identification were carried out comprehensively, with one repetition, six rows of plots, plot length of 4m and width of 1.5m, mechanical drilling, and basic seedlings of 150,000/mu. For the investigation of agronomic traits such as seedling habit, plant type, grain type and plant height, refer to the technical specification of regional test of crop varieties (NY/T 1301 -2007). The single flower drip method was used to identify the anti - spreading ability of scab. After inoculation, fresh - keeping bags were used to keep moisture for 3 days. After 21 days, the average disease spikelet rate of the strain was less than that of the middle resistance control, which was regarded as the selected strain. Identification of scab resistance by spreading diseased wheat grains, inoculation by spreading diseased wheat grains, identification of specific methods, randomly listing 100 ears of selected strains in the field, spreading diseased wheat grains at 4 - 6 kg/mu at booting stage (one week before flowering), and spraying water to keep moisture for 0.8 - 1.2 hours, stop spraying water on wheat 15 days after flowering, and immediately investigate the diseased panicle rate of listed ears. Jimai 22 is the susceptible control, Su Mai 3 is the disease - resistant control, Zheng 9023 is the middle resistance control, and the diseased panicle rate less than or equal to the middle resistance control is regarded as the selected scab - resistant strain. Harvest strains with better infection resistance and expansibility than Zheng 9023, identify the yield, and select strains Jimai 8803 with an average yield of more than 500 kg per mu (Table 4).
Characteristics of Jimai 8803: Table 4 Test results of resistance to scab of Jimai 8803 Name Padt FWY jj Average ear disease rate (99) GD A Jimar 32 Gy YE Zheng 9023 (Middle antibody RN _ . 394 38 controll {THeease resistanne + ik sd IRS zó poniral Jimai 8803 has good comprehensive agronomic characters, resistance to scab and high yield (Table 4, 5 and Figure 2). Its seedlings are semi - creeping, dark in color and good in cold tolerance (Grade 2). The plant type is compact, with a plant height of 82cm and lodging resistance. The heading date is about 2 days earlier than that of the control Jimai 22, and the maturity is similar. Square ear, long awn, white shell, white grain, oval grain, 1000 - grain weight 43g; high powdery mildew resistance, containing powdery mildew resistance gene Pm21. The yield of experimental base plot in Jiyang District, Jinan is 561 .48 kg per mu, which is 9.38% higher than that of control Jimai 22 (the yield of control Jimai 22 is 513.33 kg/mu). Jinan experimental base, the plot yield is 624.4 kg/mu, compared with Jimai 22, the lodging is serious, soit is impossible to calculate the yield.
Table 5 Comparison of agronomic characters and yield between Jimai 8803 and Jimai 22 Toma pa u Perfonma Type Period of Height 2? = cold Kilogram FHB 40e 7 and n Yance watoht duration om weight ken The heading date is about 3 days earlier Conmact plant, Jimai than the conwal wy good cold tolerance eis Modembr 21 46 3803 Jimai 22 andthe a: {Grade 7} 3D geoistacee mature period is similar. Jima 3 40.4 S133 Mid -late maturity, . nasa the maturity is one Compant plant, Diseased ay day later than that poor cold tolerance Hk of control stone {Grade 23
4185.
To sum up, through the molecular marker polymerization breeding method of wheat scab resistance of the present invention, in the breeding generation, the linkage markers LJJ-1 of molecular marker scab resistance gene Fhb1, LJJ-2 and LJJ-3 of Fhb2, LJJ-4 and LJJ-5 of Fhb5 are used to assist selection and conventional breeding techniques, Jimai 8803, a semi - winter wheat breeding material with outstanding target traits and excellent comprehensive traits, was created, which laid a material foundation for cultivating wheat varieties with high yield and scab resistance suitable for planting in Huanghuai wheat area.
Unless otherwise specified, the numerical values set forth in these examples do not limit the scope of the present invention. In all examples shown and described here, unless otherwise specified, any specific value should be interpreted as exemplary only, not as a limitation, so other examples of exemplary embodiments may have different values.
SEQUENCE LISTING <110> Crop Research Institute, Shandong Academy of Agricultural Sciences <110> Jiangsu Lixiahe Regional Agricultural Research Institute <120> Method for cultivating scab-resistant and high-yield wheat in Huanghuai wheat area based on multi-gene polymerization <130> SHX-ScapRes Wheat NL <150> CN202110471350.6 <151> 2021-04-29 <160> 10 <170> PatentIn version 3.3 <210> 1 <211> 20 <212> DNA <213> Artificial sequence <400> 1 attcctacta gccgcctggt 20 <210> 2 <211> 20 <212> DNA <213> Artificial sequence <400> 2 actggggcaa gcaaacattg 20 <210> 3 <211> 19 <212> DNA <213> Artificial sequence <400> 3 ggagattgac cgagtggat 19 <210> 4 <211> 18 <212> DNA <213> Artificial sequence <400> 4 cgtgagagcg gttctttg 18 <210> 5 <211> 18 <212> DNA <213> Artificial sequence <400> 5 atgcctgctt gctcactg 18
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<212> DNA
<213> Artificial sequence
<400> 10 tctagagagt ctttttcccg agc 23

Claims (8)

CONCLUSIESCONCLUSIONS 1. Een werkwijze voor het in tarwegebieden kweken van tarwe met resistentie tegen tarweschurft met een hoge opbrengst op basis van polygene polymerisatie, waarbij de werkwijze de volgende stappen omvat: stap 1: het kruisen van tarwerassen die meerdere schurftresistentiegenen dragen als vrouwelijke ouder tarwerassen met een grote areaalspreiding met een gemiddelde opbrengst van meer dan 500 kg per mu als mannelijke ouder om hybride zaden van de eerste generatie te verkrijgen; stap 2: het toepassen van het hybride zaad van de eerste generatie van stap 1 als vrouwelijke ouder, en het toepassen van een kort tarweras van het compacte planttype, met sterke koude bestendigheid en hoge resistentie tegen echte meeldauw als mannelijke ouder, en het oogsten van het hybride F+-zaad; stap 3: het planten van de F: planten, mengen en dorsen om de F: te genereren stap 4: het planten van de F: op het veld, het selecteren van afzonderlijke planten met uitstekende eigenschappen zoals koude bestendigheid, korte hoogte, compact planttype, weerstand tegen echte meeldauw, vierkantige aar en dergelijke, het oogsten en dorsen van de geselecteerde afzonderlijke planten om F3 zaden van de samengestelde afzonderlijke planten te genereren, waarbij zaden met rood kaf worden geëlimineerd; stap 5: het bij de meervoudige kruising van de F3 generatie in het veld planten van de plantlijnen, het selecteren van de planten met consistente plantlijn - kenmerken en uitstekende uitgebreide resistentie tegen ziekten en uitgebreide agronomische kenmerken, en het in de geselecteerde individuele planten detecteren en screenen van de koppelingsmerker LJJ1 van het schurft resistentie gen Fhb1, koppelingsmerkers LJJ-2 en LJJ-3 van Fhb2 en LJJ-4 en LJJ-5 van Fhb5, en het selecteren van de planten met positieve moleculaire merker detectie, het in het latere groeistadium van de tarwe op basis van de detectieresultaten van moleculaire merkers, plantenlijnen met goede uitgebreide agronomische kenmerken, resistentie tegen echte meeldauw en consistente fenotypes selecteren op plantenlijnen die drie doelgenen van schurftresistentiegenen Fhb1, Fhb2 en Fhb5 bevatten, en het afzonderlijk oogsten en dorsen van planten met kleine zaden, weinig volronde zaden en zaden met rood kaf.A method for cultivating wheat scab resistance in wheat fields with a high yield based on polygenic polymerization, the method comprising the following steps: step 1: crossing wheat varieties carrying multiple scab resistance genes as female parent wheat varieties having a large area spread with an average yield of more than 500 kg per mu as a male parent to obtain first generation hybrid seeds; step 2: applying the first generation hybrid seed of step 1 as the female parent, and applying a short wheat variety of the compact plant type, with strong cold resistance and high resistance to powdery mildew as the male parent, and harvesting the hybrid F+ seed; step 3: planting the F: planting, mixing and threshing to generate the F: step 4: planting the F: in the field, selecting individual plants with excellent characteristics such as cold resistance, short height, compact plant type , resistance to powdery mildew, square spike and the like, harvesting and threshing the selected single plants to generate F3 seeds of the composite single plants, eliminating seeds with red chaff; step 5: planting the plant lines in the field at the multiple cross of the F3 generation, selecting the plants with consistent plant lineage traits and excellent extensive disease resistance and extensive agronomic traits, and detecting and screening the linkage marker LJJ1 of the scab resistance gene Fhb1, linkage markers LJJ-2 and LJJ-3 of Fhb2 and LJJ-4 and LJJ-5 of Fhb5, and selecting the plants with positive molecular marker detection, the later growth stage of the wheat based on the detection results of molecular markers, select plant lines with good comprehensive agronomic traits, resistance to powdery mildew and consistent phenotypes on plant lines containing three target genes of scab resistance genes Fhb1, Fhb2 and Fhb5, and separate harvesting and threshing of plants with small seeds, little rounded seeds and seeds with red chaff. stap 8: het uitvoeren van een detectie van moleculaire merkers in de terugkruisingsgeneratie F4, en detectie en screening van de koppelingsmerker LJJ1, koppelingsmerkers LJJ-2 en LJJ-3 van Fhb2, LJJ-4 en LJJ-5 van Fhb5 van het schurft resistentie gen Fhb1 van de geselecteerde plantenlijn, en het selecteren van plantenlijnen met positieve detectie van moleculaire merkers; het inoculeren van plantenlijnen met positieve moleculaire merkers in het kop- en bloeistadium van tarwe met behulp van de enkele bloemdruppelmethode om schurftresistentie te identificeren; het in een later groeistadium van tarwe selecteren van planten met consistente fenotypen van de plantenljnkarakteristieken, een betere schurftresistentie dan de gemiddelde resistente controle, een betere algemene ziekteresistentie en agronomische karakteristieken, het oogsten en dorsen, het meten van de opbrengst van de plantenlijn en het selecteren van de planten met de hoogste opbrengst als Fs-stam selecteren; stap 7: het tweemaal herhaald planten van de geselecteerde Fs-stam in een perceel in een identificatietuin, waarbij in de eerste herhaling de enkele bloemdruppelmethode wordt toegepast om de weerstand van de stam tegen schurftinfectie te identificeren, en in de tweede herhaling zieke tarwekorrels worden toegepast om de weerstand tegen schurftinfectie te identificeren, waarbij de weerstand tegen infectie en de weerstand tegen uitbreiding van de oogst beter waren dan of gelijk waren aan de gemiddelde resistente stam, het identificeren van de eerste herhaalde opbrengst, het selecteren van de stam met een gemiddelde opbrengst van meer dan 500 kg per mu, en het aanbevelen om een productie test te starten, om zodoende met behulp van polygene polymerisatie in het tarwegebied te kunnen telen met hoge opbrengst en hoge weerstand tegen tarweschurft.step 8: performing a detection of molecular markers in the backcross generation F4, and detection and screening of the linkage marker LJJ1, linkage markers LJJ-2 and LJJ-3 of Fhb2, LJJ-4 and LJJ-5 of Fhb5 of the scab resistance gene Fhb1 from the selected plant line, and selecting plant lines with positive detection of molecular markers; inoculating plant lines with positive molecular markers at the head and flowering stage of wheat using the single flower droplet method to identify scab resistance; later stage wheat selection of plants with consistent plant line trait phenotypes, better scab resistance than the average resistant control, better overall disease resistance and agronomic characteristics, harvesting and threshing, measuring plant line yield, and selecting select from the plants with the highest yield as Fs strain; step 7: replanting the selected Fs strain twice in a plot in an identification garden, applying the single flower drop method in the first repetition to identify resistance of the strain to scab infection, and diseased wheat grains in the second repetition to identify resistance to scab infection, where resistance to infection and resistance to crop extension were better than or equal to the average resistant strain, identify the first repeat yield, select the strain with an average yield of more than 500 kg per mu, and recommending to start a production trial, so as to cultivate in the wheat area with high yield and high resistance to wheat scab using polygenic polymerization. 2. De werkwijze volgens conclusie 1, waarbij de tarwerassen met meerdere schurftresistentiegenen tarwe NMAS020 en NMAS018 zijn.The method of claim 1, wherein the wheat varieties having multiple scab resistance genes are wheat NMAS020 and NMAS018. 3. De werkwijze volgens conclusie 1, waarbij de wijd verbreide variëteiten met een gemiddelde opbrengst van meer dan 500 kg per mu Jimai 22, Jimai 44, Jimai 23 en Luyuan 502 zijn.The method of claim 1, wherein the widespread varieties having an average yield of more than 500 kg per mu are Jimai 22, Jimai 44, Jimai 23 and Luyuan 502. 4. De werkwijze volgens conclusie 1, waarbij het korte tarweras van het compacte planttype, met sterke koude bestendigheid en hoge resistentie tegen echte meeldauw Shi HO83 -366 is.The method according to claim 1, wherein the short wheat variety of compact plant type, with strong cold resistance and high powdery mildew resistance is Shi HO83-366. 5. De werkwijze volgens conclusie 1, waarbij de primersequenties van de koppelingsmerker LJJ-1 van het schurft resistentie gen Fhb1 zijn weergegeven in SEQ ID NO:1 en SEQ ID NO:2.The method of claim 1, wherein the primer sequences of the linkage marker LJJ-1 of the scab resistance gene Fhb1 are set forth in SEQ ID NO:1 and SEQ ID NO:2. 6. De werkwijze volgens conclusie 1, waarbij de primersequenties van de Fhb2- koppelingsmerker LJJ-2 van het schurf tresistentie gen zijn weergegeven in SEQ ID NO:3 en SEQ ID NO:4, en de primersequenties van LJJ-3 zijn weergegeven in SEQ ID NO:5 en SEQ ID NO:6.The method of claim 1, wherein the primer sequences of the Fhb2 linkage marker LJJ-2 of the scabies resistance gene are shown in SEQ ID NO:3 and SEQ ID NO:4, and the primer sequences of LJJ-3 are shown in SEQ ID NO:5 and SEQ ID NO:6. 7. De werkwijze volgens conclusie 1, waarbij de primersequenties van de Fhb5- koppelingsmerker LJJ-4 van het schurft resistentie gen zijn weergegeven in SEQ ID NO:7 en SEQ ID NO:8, en de primersequenties van LJJ-5 zijn weergegeven in SEQ ID NO:9 en SEQ ID NO:10.The method of claim 1, wherein the primer sequences of the Fhb5 splice marker LJJ-4 of the scab resistance gene are shown in SEQ ID NO:7 and SEQ ID NO:8, and the primer sequences of LJJ-5 are shown in SEQ ID NO:9 and SEQ ID NO:10. 8. De werkwijze volgens conclusie 1, waarbij de voor het identificeren van schurftresistentie als vatbare controle Jimai 22, als resistente controle Su Mai No.3 en als gemiddelde resistente controle Zheng 9023 wordt genomen.The method according to claim 1, wherein the susceptible control is Jimai 22, Su Mai No.3 resistant control, and Zheng 9023 intermediate resistant control is taken to identify scab resistance.
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