Breeding method and application of wheat capable of resisting gibberellic disease and resisting herbicide for starter propagation

Abstract

The invention belongs to the technical field of crop variety innovation, and particularly relates to a breeding method and application of wheat capable of resisting gibberellic disease and herbicide-tolerant starter propagation. According to the method disclosed by the invention, through combination of molecular marker-assisted selection and backcross breeding, excellent characters from different donors are rapidly introduced into a genetic background of an excellent koji-making variety, so that accurate and efficient polymerization of target characters is realized, and the screening efficiency and accuracy can be improved; meanwhile, the screening index of starter propagation quality runs through the breeding process, so that the bred material is not only excellent in resistance and agronomic character, but also can meet the specific requirements of the terminal industry. The variety bred by the method provided by the invention can reduce the use of pesticides for preventing and treating gibberellic disease and reduce the pollution risk of fungaltoxin; the herbicide-resistant characteristic facilitates field weed management, and the yield and quality stability are improved; the special quality guarantees the high additional value in the starter propagation industry, and creates the value of the whole industrial chain from the field to the workshop.

Description

Technical Field

[0001] This invention belongs to the field of crop variety innovation technology, specifically relating to a breeding method and application of wheat resistant to Fusarium head blight and herbicide-tolerant for koji making. Background Technology

[0002] Wheat is an important food crop in my country, and its yield and quality are crucial to national food security. However, wheat production faces two major challenges: First, there is Fusarium head blight. Fusarium head blight is mainly caused by Fusarium graminearum, which not only drastically reduces yields but also produces mycotoxins (such as deoxynivalenol, or DON toxin) that severely contaminate grains, threatening human and animal health. Developing disease-resistant varieties is the most economical, environmentally friendly, and effective strategy for controlling Fusarium head blight.

[0003] Secondly, there is the issue of weed infestation. Weeds compete with wheat for water, fertilizer, and sunlight, leading to reduced yields. Chemical weed control is the primary method, but traditional herbicides can cause some phytotoxicity to wheat itself, and the range of herbicides to choose from is narrow. Developing wheat varieties tolerant to specific, highly effective herbicides (such as glyphosate and glufosinate) can broaden the range of herbicides to choose from, improve weed control efficiency, and reduce management costs.

[0004] Furthermore, some wheat varieties are specifically used for making koji (fermentation starter), which is the starting point of the brewing industry. Koji-making wheat requires moderate protein content, soft texture, high starch content, and ease of microbial colonization. While wheat varieties with resistance to Fusarium head blight or herbicide tolerance have been reported in existing technologies, a breeding method that combines these three key traits—resistance to Fusarium head blight, herbicide tolerance, and koji-making-specific qualities—is still lacking. Therefore, there is an urgent need for an efficient and precise breeding method to rapidly cultivate new multi-trait hybrid wheat varieties that meet the needs of modern agricultural production and specific processing. Summary of the Invention

[0005] The purpose of this invention is to provide a method and application for breeding wheat resistant to Fusarium head blight and tolerant to herbicides for koji making. The wheat bred by the method has Fusarium head blight resistance, herbicide tolerance, and excellent koji making quality.

[0006] This invention provides a method for breeding wheat resistant to Fusarium head blight and tolerant to herbicides for koji making, comprising the following steps: using a wheat variety resistant to Fusarium head blight as the female parent and a wheat variety tolerant to herbicides as the male parent, hybridizing them to obtain an F1 generation population; and identifying the herbicide tolerance of the F1 generation population to obtain a herbicide-tolerant F1 generation population. Using the herbicide-tolerant F1 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the F1-P312 generation population; the F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant F1-P312 generation population. Using the herbicide-tolerant and Fusarium head blight-resistant F1-P312 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the BC1F1-P312 generation population; the BC1F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population. Using the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the BC2F1-P312 generation population; the BC2F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population. The herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population was subjected to self-pollination and low-generation screening to obtain BC2F2-P312~BC2F6-P312 generation populations with the target traits in sequence. The low-generation screening includes herbicide resistance identification, Fusarium head blight resistance screening, Fusarium head blight resistance identification, and preliminary screening of koji quality; the target traits include herbicide resistance, Fusarium head blight resistance of moderate to high, cuticle ratio ≤10%, and protein content of 13%~16%; the traits for preliminary screening of koji quality include cuticle ratio and protein content. The BC2F6-P312 generation population with the target traits was subjected to high-generation screening to obtain stable wheat lines resistant to Fusarium head blight and tolerant to herbicides for koji making; the high-generation screening included secondary screening of koji making quality; the traits of the secondary screening of koji making quality included test weight, hardness index, protein content and crude starch content.

[0007] As a preferred embodiment, the wheat variety with Fusarium head blight resistance contains a major Fusarium head blight resistance gene. Fhb1 wheat varieties.

[0008] As a preferred option, the wheat variety with resistance to Fusarium head blight includes at least one of Xumai DH9, Jiayuanmai 1, Nannong 999 and Xumai DH2.

[0009] As a preferred embodiment, the herbicide-tolerant wheat varieties include Fumai 2000 and / or Huamai 299.

[0010] As a preferred option, the wheat varieties with excellent koji-making and processing characteristics include Fumai 666 and / or Fanmai 8.

[0011] As a preferred embodiment, the herbicide tolerance identification includes: spraying herbicides on wheat seedlings, eliminating sensitive individual plants, and obtaining herbicide-tolerant wheat.

[0012] As a preferred embodiment, the herbicide includes imidazolinone herbicides.

[0013] As a preferred option, the resistance identification for Fusarium head blight is conducted in accordance with the technical specification “NY / T 1443.4-2007”.

[0014] As a preferred embodiment, the stable wheat variety for koji making resistant to Fusarium head blight and tolerant to herbicides has a test weight ≥790g / L; a hardness index ≤45; a protein content of 13%~16%; and a crude starch content ≥60%.

[0015] This invention also provides the application of stable wheat lines for fusarium head blight resistance and herbicide tolerance obtained by the breeding method described above in brewing and food production.

[0016] Beneficial effects: This invention provides a method for breeding wheat resistant to Fusarium head blight and tolerant to herbicides for koji making, which has the following advantages: High-efficiency aggregation of multiple traits: By combining molecular marker-assisted selection with backcross breeding, superior traits from different donors are rapidly introduced into the genetic background of superior koji making varieties, achieving precise and efficient aggregation of target traits.

[0017] Early targeted screening shortens the breeding cycle: Using herbicides for live screening in the early stages of breeding significantly reduces the size of later populations, lowering workload and costs. Molecular marker selection is unaffected by the environment and can be performed at any growth stage, greatly improving selection efficiency and accuracy.

[0018] With a clear quality orientation, the selection indicators for koji production quality (grain hardness, protein content, and crude starch content) are integrated into the breeding process, ensuring that the selected materials are not only excellent in terms of resistance and agronomic traits, but also meet the specific needs of the end-user industry, thus realizing the targeted breeding of "specialized" varieties.

[0019] Significant environmental and economic benefits: The developed varieties can reduce the use of pesticides for Fusarium head blight control and reduce the risk of fungal toxin contamination; the herbicide-resistant characteristics facilitate field weed management and improve yield and quality stability; the special quality guarantees its high added value in the koji-making industry, creating full-chain value from field to workshop. Attached Figure Description

[0020] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0021] Figure 1 This is a flowchart of the breeding process. Detailed Implementation

[0022] This invention provides a method for breeding wheat resistant to Fusarium head blight and tolerant to herbicides for koji making, comprising the following steps: using a wheat variety resistant to Fusarium head blight as the female parent and a wheat variety tolerant to herbicides as the male parent, hybridizing them to obtain an F1 generation population; and identifying the herbicide tolerance of the F1 generation population to obtain a herbicide-tolerant F1 generation population. Using the herbicide-tolerant F1 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the F1-P312 generation population; the F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant F1-P312 generation population. Using the herbicide-tolerant and Fusarium head blight-resistant F1-P312 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the BC1F1-P312 generation population; the BC1F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population. Using the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the BC2F1-P312 generation population; the BC2F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population. The herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population was subjected to self-pollination and low-generation screening to obtain BC2F2-P312~BC2F6-P312 generation populations with the target traits in sequence. The low-generation screening includes herbicide resistance identification, Fusarium head blight resistance screening, Fusarium head blight resistance identification, and preliminary screening of koji quality; the target traits include herbicide resistance, Fusarium head blight resistance of moderate to high, cuticle ratio ≤10%, and protein content of 13%~16%; the traits for preliminary screening of koji quality include cuticle ratio and protein content. The BC2F6-P312 generation population with the target traits was subjected to high-generation screening to obtain stable wheat lines resistant to Fusarium head blight and tolerant to herbicides for koji making; the high-generation screening included secondary screening of koji making quality; the traits of the secondary screening of koji making quality included test weight, hardness index, protein content and crude starch content.

[0023] This invention uses a wheat variety resistant to Fusarium head blight as the female parent and a wheat variety resistant to herbicides as the male parent for hybridization to obtain the F1 generation. As one embodiment, the wheat variety resistant to Fusarium head blight contains a major gene for resistance to the disease. Fhb1 Wheat varieties. As another embodiment, the wheat varieties resistant to Fusarium head blight include at least one of Xumai DH9, Jiayuanmai 1, Nannong 999, and Xumai DH2. As another embodiment, the wheat varieties resistant to herbicides include Fumai 2000 and / or Huamai 299.

[0024] After obtaining the F1 generation population, this invention conducts herbicide tolerance identification on the F1 generation population to obtain a herbicide-tolerant F1 generation population. As one implementation method, the herbicide tolerance identification includes: spraying herbicides on wheat seedlings, eliminating susceptible individual plants, and obtaining herbicide-tolerant wheat. As one implementation method, the seedling stage includes the four-leaf stage. At the four-leaf stage, wheat plants are of moderate size, facilitating even spraying of the herbicide solution onto the plants. Furthermore, the plants at the four-leaf stage exhibit enhanced resistance, and spraying herbicides at this time avoids false death caused by the wheat being too young and fragile. Simultaneously, weeds have emerged and are at a good time for control (weeds are at the 2-4 leaf stage), allowing for simultaneous evaluation of wheat resistance and weed control effectiveness. As one implementation method, the spraying site includes stems and leaves; the spraying amount is 25 L / acre. As one implementation method, the symptoms of susceptible individual plants include: inhibited growth, yellowing and browning of stems and leaves, and death. As one implementation method, the herbicide includes imidazolinone herbicides. As one implementation, the imidazolinone herbicide includes imidazoline and / or methoxyfenozide. In a specific embodiment of the present invention, the herbicide is methoxyfenozide.

[0025] This invention uses the herbicide-tolerant F1 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents for hybridization to obtain the F1-P312 generation population. As one embodiment, the excellent koji-making and processing characteristics include: grain hardness index ≤ 45; protein content 13%~16%; crude starch content ≥ 60%. As one embodiment, the grain hardness index can be 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, or 35; as one embodiment, the protein content can be any value within the range of 13%~16%, for example 13%, 13.5%, 14%, 14.5%, 15%, or 16%; as one embodiment, the crude starch content can be 60%, 61%, 62%, 63%, 65%, 68%, 70%, 72%, or 75%. As one embodiment, the wheat varieties with excellent koji-making and processing characteristics include Fumai 666 and / or Fanmai 8.

[0026] After obtaining the F1-P312 generation population, herbicide tolerance was assessed on the F1-P312 generation population to obtain herbicide-tolerant F1-P312 generation population; the herbicide tolerance assessment method has been described above and will not be repeated here.

[0027] After obtaining the herbicide-tolerant F1-P312 generation population, the herbicide-tolerant F1-P312 generation population was screened for Fusarium head blight resistance to obtain a herbicide-tolerant and Fusarium head blight-resistant F1-P312 generation population. As one implementation method, the Fusarium head blight resistance screening includes utilizing a major Fusarium head blight resistance gene. Fhb1 Molecular markers were used to screen for single plants resistant to Fusarium head blight. In one specific embodiment of the invention, a major Fusarium head blight resistance gene was used. Fhb1 The diagnostic molecular marker His-InDel was used to detect herbicide-resistant F1-P312 generation populations, and those without the marker were eliminated. Fhb1 Individual plants containing the disease-resistant gene were used to obtain the F1-P312 generation population, which is herbicide-tolerant and resistant to Fusarium head blight. The Fusarium head blight major resistance gene described in this invention... Fhb1 For the detection method of the diagnostic molecular marker His-InDel, please refer to [Zhu Zhanwang et al., Fusarium head blight resistance gene in Chinese wheat varieties]. Fhb1 Identification and Origin Tracing, Acta Agronomica Sinica, 2018, 44(4): 473-482.

[0028] After obtaining the herbicide-tolerant and Fusarium head blight-resistant F1-P312 generation population, crosses were performed using this population as the male parent and a wheat variety with excellent koji-making and processing characteristics as the recurrent parent to obtain the BC1F1-P312 generation population. The characteristics of the wheat variety with excellent koji-making and processing characteristics described in this invention have been explained above and will not be repeated here. After obtaining the BC1F1-P312 generation population, herbicide tolerance identification and Fusarium head blight resistance screening were performed sequentially on the BC1F1-P312 generation population to obtain the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population. Using the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population as the male parent, and a wheat variety with excellent koji-making and processing characteristics as the recurrent parent, a hybridization process was conducted to obtain the BC2F1-P312 generation population. After obtaining the BC2F1-P312 generation population, herbicide tolerance identification and Fusarium head blight resistance screening were performed sequentially on the BC2F1-P312 generation population to obtain the herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population. The screening method for the herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population of this invention is the same as that for the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population, and will not be repeated here.

[0029] The herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population was subjected to self-pollination and low-generation screening to obtain BC2F2-P312~BC2F6-P312 generation populations with the target trait. As one implementation method, the herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population was self-pollinated to obtain the BC2F2-P312 segregating population; after obtaining the BC2F2-P312 segregating population, low-generation screening was performed on the BC2F2-P312 segregating population to obtain the BC2F2-P312 generation population; the low-generation screening included herbicide tolerance identification, Fusarium head blight resistance screening, Fusarium head blight resistance identification, and initial screening for koji-making quality.

[0030] In one implementation method, after obtaining the BC2F2-P312 segregating population, herbicide tolerance is assessed to obtain a herbicide-tolerant BC2F2-P312 segregating population. Then, the herbicide-tolerant BC2F2-P312 segregating population is screened for Fusarium head blight resistance to obtain a herbicide-tolerant and Fusarium head blight-resistant BC2F2-P312 segregating population. This invention, through herbicide tolerance assessment and Fusarium head blight resistance screening, allows for large-scale, low-cost screening of individual plants with the target trait in the early stages of wheat growth. The herbicide tolerance assessment and Fusarium head blight resistance screening have been described above and will not be repeated here.

[0031] After obtaining the herbicide-resistant and Fusarium head blight-resistant BC2F2-P312 segregating population, its Fusarium head blight resistance was identified, yielding a BC2F2-P312 segregating population with herbicide resistance and Fusarium head blight resistance at the moderate resistance level or above. As one implementation method, the Fusarium head blight resistance identification was conducted according to the technical specification “NY / T 1443.4-2007”. In a specific embodiment of the present invention, Fusarium head blight strain conidial solution was inoculated using the single-flower drip inoculation method during the heading and flowering stage. The number of diseased spikelets and the severity of the disease were investigated during the grain-filling stage, and individual plants with Fusarium head blight resistance reaching the moderate resistance level or above were screened.

[0032] After obtaining the BC2F2-P312 segregating population with herbicide resistance and moderate or higher resistance to Fusarium head blight, the BC2F2-P312 segregating population with herbicide resistance and moderate or higher resistance to Fusarium head blight is subjected to initial screening for koji-making quality, resulting in a BC2F2-P312 generation population with the target traits. The traits screened for koji-making quality include corneum percentage and protein content. The target traits include herbicide resistance and moderate or higher resistance to Fusarium head blight, corneum percentage ≤10%, and protein content of 13%~16%. As one implementation method, the corneum percentage is determined by randomly selecting 100 grains from each plant, cutting them cross-sections, observing the percentage of transparent portion in the cross-section, and taking the average value as the corneum percentage of that plant. As another implementation method, the protein content is determined using a near-infrared grain quality analyzer. The corneum percentage of wheat grains is an important appearance indicator for judging the quality of wheat. Wheat with a high keratin content is generally considered hard wheat, while wheat with a low keratin content is considered soft wheat. A keratin content of less than 10% indicates that there are relatively few hard grains per plant, increasing the chance of selecting plants with soft grains during further generations of breeding. Wheat with a low keratin content generally has lower grain hardness, making it easier to break into a plum blossom shape during koji making, achieving a state where the core rots but the husk remains intact. Wheat varieties with a protein content of 13%–16% are more conducive to microbial growth and enzymatic degradation into small soluble molecules during koji making. These molecules participate in the Maillard reaction, generating various aroma and flavor compounds, resulting in a mellow, smooth, and delicate liquor.

[0033] After obtaining the BC2F2-P312 generation population with the target trait, self-crossing and low-generation screening were performed to obtain the BC2F3-P312~BC2F6-P312 generation populations with the target trait. The methods of self-crossing and low-generation screening have been described above and will not be repeated here.

[0034] After obtaining the BC2F6-P312 generation population with the target trait, the BC2F6-P312 generation population with the target trait is subjected to high-generation screening to obtain stable wheat lines for koji making that are resistant to Fusarium head blight and tolerant to herbicides. The high-generation screening includes a second screening of koji making quality to obtain stable wheat lines for koji making that are resistant to Fusarium head blight and tolerant to herbicides.

[0035] In one implementation method, after obtaining the BC2F6-P312 generation population, the BC2F6-P312 generation population is subjected to herbicide resistance identification, Fusarium head blight resistance screening, and Fusarium head blight resistance identification to obtain a BC2F6-P312 generation population with herbicide resistance and Fusarium head blight resistance of moderate resistance or above. The BC2F6-P312 generation population with herbicide resistance and Fusarium head blight resistance of moderate resistance or above is then subjected to a second screening for koji-making quality to obtain a stable wheat line resistant to Fusarium head blight and herbicide-resistant for koji-making. The methods for herbicide resistance identification, Fusarium head blight resistance screening, and Fusarium head blight resistance identification in the screening of the BC2F6-P312 generation population of this invention have been discussed above and will not be repeated here. In one implementation method, the traits for the second screening of koji-making quality include test weight, hardness index, protein content, and crude starch content. In one implementation method, the test weight is ≥790 g / L; the hardness index is ≤45; the protein content is 13%~16%; and the crude starch content is ≥60%. As one implementation, the bulk density can be any value ≥790 g / L, such as 790 g / L, 792 g / L, 793 g / L, 795 g / L, 796 g / L, 798 g / L, 800 g / L, 803 g / L, 805 g / L, 807 g / L, 809 g / L, 810 g / L, 815 g / L, 816 g / L, 819 g / L, 820 g / L, 825 g / L, 828 g / L, or 830 g / L. L; the hardness index can be any value ≤45, such as 45, 44, 43, 42, 41, 40, 39, 38, 37, 36 or 35; the protein content can be any value between 13% and 16%, such as 13%, 13.5%, 14%, 14.5%, 15% or 16%; as one embodiment, the crude starch content can be 60%, 61%, 62%, 63%, 65%, 68%, 70%, 72% or 75%. This invention integrates the screening indicators for koji quality (grain hardness, protein content, crude starch content) into the breeding process, ensuring that the selected materials not only excel in resistance and agronomic traits, but also meet the specific needs of the end-user industry, thus achieving targeted breeding of "specialized" varieties.

[0036] As one embodiment, the Fusarium head blight-resistant herbicide-tolerant stable wheat strain for koji production has a test weight ≥790 g / L; a hardness index ≤45; a protein content of 13%~16%; and a crude starch content ≥60%. As another embodiment, the test weight can be any value ≥790 g / L, such as 790 g / L, 792 g / L, 793 g / L, 795 g / L, 796 g / L, 798 g / L, 800 g / L, 803 g / L, 805 g / L, 807 g / L, 809 g / L, 810 g / L, 815 g / L, 816 g / L, 819 g / L, 820 g / L, 825 g / L, 828 g / L, or 830 g / L. L; the hardness index can be any value ≤45, such as 45, 44, 43, 42, 41, 40, 39, 38, 37, 36 or 35; the protein content can be any value between 13% and 16%, such as 13%, 13.5%, 14%, 14.5%, 15% or 16%; as one embodiment, the crude starch content can be 60%, 61%, 62%, 63%, 65%, 68%, 70%, 72% or 75%. The wheat described in this invention has a bulk density ≥790g / L, meeting the bulk density requirements for first-grade wheat in the GB1351-2008 standard. The hardness index ≤45 meets the hardness index requirements for soft wheat in the GB1351-2008 standard, ensuring that the wheat, when crushed, achieves the "heart rots but the husk remains intact" requirement for koji-making raw materials. The wheat protein content is 13%~16%, which, during koji-making, facilitates microbial growth and enzymatic degradation into small soluble molecules, participating in the Maillard reaction to generate various aroma and flavor compounds, resulting in a mellow, smooth, and delicate liquor. The crude starch content ≥60% provides sufficient sugar for fermentation, promoting microbial growth and metabolism, and producing more alcohol and aroma compounds.

[0037] As one implementation method, after obtaining the stable line of Fusarium head blight-resistant and herbicide-tolerant wheat for koji making, the method further includes verification of target traits and yield. As one implementation method, the verification of target traits and yield includes: conducting small-plot comparison trials on the stable line to comprehensively evaluate its agronomic traits, yield, disease resistance, herbicide tolerance stability, and koji-making quality, thereby obtaining a new Fusarium head blight-resistant and herbicide-tolerant wheat variety for koji making. The flowchart of the breeding method described in this invention is as follows: Figure 1 As shown, the method of this invention combines molecular marker-assisted selection with backcross breeding to rapidly introduce superior traits from different donors into the genetic background of superior koji-making varieties, achieving precise and efficient aggregation of target traits. Furthermore, the use of herbicides for live screening in the early stages of breeding significantly reduces the size of the later population, lowering workload and costs. Molecular marker selection is unaffected by the environment and can be performed at any growth stage, greatly improving selection efficiency and accuracy.

[0038] This invention also provides the application of stable wheat varieties resistant to Fusarium head blight and tolerant to herbicides, obtained using the breeding method described above, in brewing and food production. The wheat varieties obtained by the breeding method described in this invention can reduce the use of pesticides for Fusarium head blight control and lower the risk of fungal toxin contamination; their herbicide tolerance facilitates field weed management, improving yield and quality stability; and their specialized qualities ensure high added value in the koji-making industry, creating value across the entire industrial chain from field to workshop.

[0039] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0040] Example 1 Selected genes carrying the major resistance gene against Fusarium head blight Fhb1 The parent lines are Jiayuanmai 1 (P1), a wheat variety resistant to Fusarium head blight; Fumai 2000 (P2), a herbicide-tolerant wheat variety; and Fanmai 8 (P3), a dual-purpose wheat variety for brewing and eating.

[0041] (1) Using Jiayuanmai No. 1 as the female parent and Fumai 2000 as the male parent, hybridization was carried out, and F1-P12 seeds were harvested at maturity.

[0042] (2) Plant F1-P12 and Panmai 8.

[0043] Herbicide resistance identification: At the four-leaf stage, apply 4% (w / w) methoxyfenozide at a concentration of 4 ml / L to the foliar spray of the F1-P12 population, with a spraying rate of 25 L per acre. Plants in the F1-P12 population exhibiting growth inhibition, yellowing, browning, or death are considered susceptible false hybrids. These susceptible false hybrids are eliminated, and the insensitive F1-P12 population is retained.

[0044] Using the stable true hybrid F1-P12 population as the male parent and Fanmai 8 as the female parent, hybridization was carried out during the flowering period, and F1-P312 seeds were harvested at maturity.

[0045] (3) Plant Fanmai No. 8 and F1-P312.

[0046] Germplasm screening: A. Herbicide resistance identification: Herbicide resistance was identified in the F1-P312 population using the same method as step (2). Sensitive false hybrids were eliminated to obtain an insensitive F1-P312 population. B. Fusarium head blight resistance screening: Fusarium head blight major resistance genes were used. Fhb1 His-InDel, a diagnostic molecular marker (Zhu Zhanwang et al., a gene for resistance to Fusarium head blight in Chinese wheat varieties). Fhb1 Identification and traceability of the virus, Acta Agronomica Sinica, 2018, 44(4): 473-482), and detection was performed on insensitive F1-P312 individual plants to eliminate those that did not carry the virus. Fhb1Individual plants with disease-resistant genes were used to obtain the F1-P312 population, which possesses both herbicide resistance and Fusarium head blight resistance.

[0047] Hybridization: During the flowering period, Fanmai 8 was used as the recurrent parent, and F1-P312, which has herbicide resistance and Fusarium head blight resistance, was used as the male parent to obtain BC1F1-P312.

[0048] (4) Plant Panmai No. 8 and BC1F1-P312.

[0049] Germplasm screening: A. Herbicide resistance identification; B. Fusarium head blight resistance screening process is the same as step (3), to obtain the BC1F1-P312 population with herbicide resistance and Fusarium head blight resistance.

[0050] Hybridization: During the flowering period, Fanmai 8 was used as the recurrent parent and BC1-P312, which has herbicide resistance and Fusarium head blight resistance, was used as the male parent for hybridization to obtain BC2F1-P312.

[0051] (5) Planting BC2F1-P312 Germplasm screening: A. Herbicide resistance identification; B. Fusarium head blight resistance screening process is the same as step (3), to obtain the BC2F1-P312 population with herbicide resistance and Fusarium head blight resistance.

[0052] Self-pollination: The herbicide-resistant and Fusarium head blight-resistant BC2F1-P312 population was self-pollinated to obtain BC2F2-P312.

[0053] (6) Plant 1200 individual BC2F2-P312 plants. Germplasm screening: A. Herbicide resistance identification; B. Fusarium head blight resistance screening process is the same as step (3), resulting in 123 plants of the BC2F2-P312 population with herbicide resistance and Fusarium head blight resistance.

[0054] Fusarium head blight resistance identification: During the heading and flowering stage, single-flower drip inoculation with conidial solution of Fusarium head blight strains was performed. Disease resistance was identified according to the "Technical Specification for Evaluation of Wheat Resistance to Fusarium Head Blight" (NY / T 1443.4-2007), yielding 16 plants with moderate resistance. The Fusarium head blight strain was published in ["Screening of Wheat Germplasm Resistant to Fusarium Head Blight and Study on the Distribution of Fhb1 Gene", Zhang Shanlei et al., Jiangxi Journal of Agricultural Sciences, 2021].

[0055] Quality screening for koji production: After the wheat plants with intermediate resistance levels matured, 100 grains were randomly selected from each plant, cross-sectioned, and the percentage of transparent portion in the cross-section was observed. Eleven plants with a corneum rate ≤10% were selected. FOSS Infratec, a analyte manufactured by FOSS Analytical Instruments Ltd. of Denmark, was used. TMA small sample was analyzed using a 1241 Grain An alyzer analyzer, and nine individual plants with crude protein content of 13% to 16% were selected, namely BC2F2-P312, which has the composite target traits (herbicide resistance, Fusarium head blight resistance, and good koji quality).

[0056] (7) Self-pollination and generation: Nine BC2F2-P312 plants with herbicide resistance, Fusarium head blight resistance and koji-making quality selected in step (6) were self-pollinated and generation was carried out. Herbicide resistance screening, Fusarium head blight resistance identification and koji-making quality selection were carried out using the method in step (6). Plants with composite target traits were screened out and generation was continued to obtain BC2F3-P312.

[0057] The same selection process was carried out for generations BC2F3-P312 to BC2F5-P312 as for BC2F2-P312. Single plants with the composite target trait were screened out and continued to be multiplied to obtain BC2F6-P312.

[0058] (8) Eight plots of BC2F6-P312 single plants were planted. Herbicide resistance was identified by foliar spraying with 4% methoxyfenozide solution at the four-leaf stage. Fusarium head blight inoculation was conducted at the heading and flowering stage. Three plots with uniform plant growth, herbicide tolerance, and good Fusarium head blight resistance were obtained.

[0059] After maturity, grains were harvested in separate plots. The bulk density of grains in each plot was measured using a GHCS-1000 electronic grain bulk density meter manufactured by Zhejiang Top Cloud Agricultural Technology Co., Ltd., and the hardness index of grains in each plot was measured using a BLH-1100 wheat hardness index meter manufactured by Zhejiang Bethlehem Instrument Equipment Co., Ltd. Finally, the hardness index of grains in each plot was measured using a FOSSInfratec analytical instrument manufactured by FOSS Analytical Instruments Ltd. of Denmark. TM The crude protein and crude starch content of the grains in each plot were determined using a 1241 Grain Analyzer. Based on the combined results, one stable plot was selected and named QM01, with a bulk density ≥790g / L, hardness index ≤45, crude protein content ≤16% (13%≤), and crude starch content ≥60%.

[0060] (9) Verify the target traits and yield performance of QM01 QM01, Jiayuanmai 1, Fumai 2000, and Fumai 666 were planted in separate plots, with 8 rows per plot, each row 8.4m long and 0.2m apart, and 453 plants per row. The experiment was repeated three times. Herbicide tolerance and Fusarium head blight resistance were assessed at the four-leaf stage and the heading and flowering stage, respectively, to obtain herbicide-tolerant and Fusarium head blight-resistant populations. After heading, yield was measured and koji-making quality was analyzed.

[0061] The results showed that QM01 had a bulk density of 800 g / L, a hardness index of 43, a crude protein content of 15.2%, a crude starch content of 64.7%, and a yield that was 10.3% higher than Jiayuanmai No. 1, 8.8% higher than Fumai 2000, and 5.9% higher than Fumai 666.

[0062] Example 2 Selected genes carrying the major resistance gene against Fusarium head blight Fhb1 The parent lines were Xumai DH9 (P1), a wheat variety resistant to Fusarium head blight; Huamai 299 (P2), a wheat variety tolerant to herbicides; and Fumai 666 (P3), a wheat variety suitable for both brewing and eating.

[0063] (1) Using Xumai DH9 as the female parent and Huamai 299 as the male parent, hybridization was carried out, and F1-P12 seeds were obtained by harvesting at maturity.

[0064] (2) Plant F1-P12 and Fumai 666.

[0065] Herbicide resistance identification: At the four-leaf stage, apply 4% (w / w) methoxyfenozide at a concentration of 4 ml / L to the foliar spray of the F1-P12 population, with a spraying rate of 25 L per acre. Plants in the F1-P12 population exhibiting growth inhibition, yellowing, browning, or death are considered susceptible false hybrids. These susceptible false hybrids are eliminated, and the insensitive F1-P12 population is retained.

[0066] During the flowering period, Fumai 666 was used as the female parent and the stable true hybrid F1-P12 population was used as the male parent for hybridization. F1-P312 seeds were harvested at maturity.

[0067] (3) Planting Fumai 666 and F1-P312 Germplasm screening: A. Herbicide resistance identification: The operation method is the same as step (2), sensitive false hybrids are eliminated, and the insensitive F1-P312 population is obtained. B. Fusarium head blight resistance screening: Fusarium head blight major resistance gene is used. Fhb1 His-InDel, a diagnostic molecular marker (Zhu Zhanwang et al., a gene for resistance to Fusarium head blight in Chinese wheat varieties). Fhb1 Identification and traceability of the virus, Acta Agronomica Sinica, 2018, 44(4): 473-482), and detection was performed on insensitive F1-P312 individual plants to eliminate those that did not carry the virus. Fhb1 Individual plants with disease-resistant genes were used to develop the F1-P312 population, which possesses herbicide resistance and Fusarium head blight resistance.

[0068] Hybridization: During the flowering period, Fumai 666 was used as the recurrent parent and F1-P312 was used as the male parent to obtain BC1F1-P312.

[0069] (4) Planting Fumai 666 and BC1F1-P312.

[0070] Germplasm screening: A. Herbicide resistance identification; B. Fusarium head blight resistance screening process is the same as step (3), to obtain the BC1F1-P312 population with herbicide resistance and Fusarium head blight resistance.

[0071] Hybridization: During the flowering period, Fumai 666 was used as the recurrent parent and BC1-P312, which has herbicide resistance and Fusarium head blight resistance, was used as the male parent to hybridize and obtain BC2F1-P312.

[0072] (5) Plant BC2F1-P312.

[0073] Germplasm screening: A. Herbicide resistance identification; B. Fusarium head blight resistance screening process is the same as step (3), to obtain the BC2F1-P312 population with herbicide resistance and Fusarium head blight resistance.

[0074] Self-pollination: The BC2F1-P312 population, which has herbicide resistance and Fusarium head blight resistance, was self-pollinated to obtain BC2F2-P312.

[0075] (6) Plant 1200 individual BC2F2-P312 plants.

[0076] Germplasm screening: A. Herbicide resistance identification; B. Fusarium head blight resistance screening process is the same as step (3), resulting in 201 plants of the BC2F2-P312 population with herbicide resistance and Fusarium head blight resistance.

[0077] Fusarium head blight resistance identification: Fusarium head blight resistance was identified using the single-flower drip method during the heading and flowering stage, yielding 30 plants with moderate resistance. The identification method was the same as step (6) in Example 1.

[0078] Screening for koji-making quality: After the wheat plants with moderate resistance levels matured, 19 individual plants with a vitreous content ≤10% were screened out, using the same screening method as step (6) in Example 1. A small number of samples were analyzed using a near-infrared quality analyzer, and 12 individual plants with a crude protein content of 13%-16% were screened out, namely BC2F2-P312, which has the composite target traits (herbicide resistance, Fusarium head blight resistance, and good koji-making quality).

[0079] (7) Self-pollination and generation: The 12 BC2F2-P312 single plants with herbicide resistance, Fusarium head blight resistance and koji-making quality selected in step (6) were self-pollinated and generation was carried out. Herbicide resistance screening, Fusarium head blight resistance identification and koji-making quality selection were carried out using the method in step (6). The single plants with the composite target traits were screened out and generation was continued to obtain BC2F3-P312.

[0080] The same selection process was carried out for generations BC2F3-P312 to BC2F5-P312 as for BC2F2-P312. Single plants with the composite target trait were screened out and continued to be multiplied to obtain BC2F6-P312.

[0081] (8) Five plots of BC2F6-P312 single plants were planted. At the four-leaf stage, 4 ml / L of 4% methoxyfenozide was sprayed on the stems and leaves. At the heading and flowering stage, Fusarium head blight was inoculated and identified. Three plots with uniform plant growth, herbicide tolerance and good resistance to Fusarium head blight were obtained.

[0082] After maturity, the grains were harvested in separate plots, and the bulk density, hardness index, crude protein content, and crude starch content were measured using the same methods as in Example 1. Finally, one stable plot was selected with a bulk density ≥790 g / L, hardness index ≤45, crude protein content ≤16% (13% ≤ crude protein), and crude starch content ≥60%, and named QM02.

[0083] (9) Verify the target traits and yield performance of QM02: QM02, Xumai DH9, Huamai 299, and Fanmai 8 were planted in separate plots, with 8 rows per plot, each row 8.4m long and 0.2m apart, and 453 plants per row. The experiment was repeated three times. Herbicide tolerance and Fusarium head blight resistance were assessed at the four-leaf stage and the heading and flowering stage, respectively. After heading, the yield was measured and the quality of the koji (fermented rice) was analyzed.

[0084] The results showed that QM02 had a bulk density of 823 g / L, a hardness index of ≤45, a crude protein content of 13.9%, a crude starch content of 66.5%, and a yield that was 4.7% higher than Xumai DH9, 9.1% higher than Huamai 299, and 5.6% higher than Fanmai 8.

[0085] In summary, the wheat bred using the method described in this invention is resistant to herbicides, has resistance to Fusarium head blight, and produces high-quality koji (fermented wheat).

[0086] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for breeding wheat resistant to Fusarium head blight and tolerant to herbicides for koji production, characterized in that, The process includes the following steps: using a wheat variety resistant to Fusarium head blight as the female parent and a wheat variety tolerant to herbicides as the male parent, hybridizing them to obtain the F1 generation population; and identifying the herbicide tolerance of the F1 generation population to obtain the herbicide-tolerant F1 generation population. Using the herbicide-tolerant F1 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the F1-P312 generation population; the F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant F1-P312 generation population. Using the herbicide-tolerant and Fusarium head blight-resistant F1-P312 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the BC1F1-P312 generation population; the BC1F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population. Using the herbicide-tolerant and Fusarium head blight-resistant BC1F1-P312 generation population as the male parent and wheat varieties with excellent koji-making and processing characteristics as recurrent parents, a hybridization was carried out to obtain the BC2F1-P312 generation population; the BC2F1-P312 generation population was subjected to herbicide tolerance identification and Fusarium head blight resistance screening to obtain the herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population. The herbicide-tolerant and Fusarium head blight-resistant BC2F1-P312 generation population was subjected to self-pollination and low-generation screening to obtain BC2F2-P312~BC2F6-P312 generation populations with the target traits in sequence. The low-generation screening includes herbicide resistance identification, Fusarium head blight resistance screening, Fusarium head blight resistance identification, and preliminary screening of koji quality; the target traits include herbicide resistance, Fusarium head blight resistance of moderate to high, cuticle ratio ≤10%, and protein content of 13%~16%; the traits for preliminary screening of koji quality include cuticle ratio and protein content. The BC2F6-P312 generation population with the target traits was subjected to high-generation screening to obtain stable wheat lines resistant to Fusarium head blight and tolerant to herbicides for koji making; the high-generation screening included secondary screening of koji making quality; the traits of the secondary screening of koji making quality included test weight, hardness index, protein content and crude starch content.

2. The breeding method according to claim 1, characterized in that, The wheat variety with resistance to Fusarium head blight contains a gene that has a major effect in resisting Fusarium head blight. Fhb1 wheat varieties.

3. The breeding method according to claim 1 or 2, characterized in that, The wheat varieties with resistance to Fusarium head blight include at least one of Xumai DH9, Jiayuanmai 1, Nannong 999, and Xumai DH2.

4. The breeding method according to claim 1, characterized in that, The wheat varieties with herbicide tolerance characteristics include Fumai 2000 and / or Huamai 299.

5. The breeding method according to claim 1, characterized in that, The wheat varieties with excellent koji-making and processing characteristics include Fumai 666 and / or Fanmai 8.

6. The breeding method according to claim 1, characterized in that, The herbicide tolerance identification process includes: spraying herbicides on wheat seedlings, eliminating sensitive individual plants, and obtaining herbicide-tolerant wheat.

7. The breeding method according to claim 1 or 6, characterized in that, The herbicides include imidazolinone herbicides.

8. The breeding method according to claim 1, characterized in that, The resistance to Fusarium head blight was determined in accordance with the technical specification "NY / T1443.4-2007".

9. The breeding method according to claim 1, characterized in that, The stable wheat lines for koji making resistant to Fusarium head blight and tolerant to herbicides have a test weight ≥790g / L; hardness index ≤45; protein content of 13%~16%; and crude starch content ≥60%.

10. The application of stable wheat lines for koji making that are resistant to Fusarium head blight and tolerant to herbicides, obtained by the breeding method according to any one of claims 1 to 9, in brewing and food use.

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