WO2021003879A1 - 一种小麦产量与品质协同提高的劣汰选优选育方法 - Google Patents

一种小麦产量与品质协同提高的劣汰选优选育方法 Download PDF

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WO2021003879A1
WO2021003879A1 PCT/CN2019/112083 CN2019112083W WO2021003879A1 WO 2021003879 A1 WO2021003879 A1 WO 2021003879A1 CN 2019112083 W CN2019112083 W CN 2019112083W WO 2021003879 A1 WO2021003879 A1 WO 2021003879A1
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yield
plant
generation
quality
less
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PCT/CN2019/112083
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English (en)
French (fr)
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曹新有
刘建军
李豪圣
程敦公
王利彬
王灿国
宋健民
刘爱峰
赵振东
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山东省农业科学院作物研究所
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Priority to AU2019438435A priority Critical patent/AU2019438435B2/en
Priority to US17/066,568 priority patent/US11716942B2/en
Priority to ZA2020/06385A priority patent/ZA202006385B/en
Publication of WO2021003879A1 publication Critical patent/WO2021003879A1/zh

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    • 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
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • 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
    • A01H1/12—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
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/46—Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
    • A01H6/4678—Triticum sp. [wheat]
    • 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
    • C12Q2600/158—Expression markers
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture

Definitions

  • the present invention relates to the field of biotechnology, in particular to a method for synergistically improving wheat yield and quality by selection and breeding of the fittest.
  • the purpose of the present invention is to provide a method for breeding target wheat varieties with improved yield and quality.
  • the method provided by the present invention includes the following steps:
  • 4)-1 a single plant containing high-quality related protein
  • 4)-2 a single plant has no less than 5 ears, sticks or square ears, and flag leaves (the angle between flag leaves is 10-30
  • 4)-3 The protein content of a single plant's grain is greater than or equal to 13%;
  • the winter freezing damage grade is less than or equal to 3; 5)-2, powdery mildew disease and stripe rust disease are both less than 4; 5)-3, plant height is 75-80 cm; 5)-4 thousand grain weight Plant rows greater than or equal to 38g; 5)
  • the peak time of the kneading instrument in the identification of the kneading characteristics of the individual plants in the 5 rows is not less than 2 minutes, and the 8-minute bandwidth is not less than 10%;
  • the cell screening criteria are as follows 6)-1 to 6)-3:
  • winter freezing damage is less than or equal to grade 3
  • the number of ears per mu is greater than the quality control or greater than or equal to the high-yielding control, and the lodging is less than or equal to level 2;
  • the stability time of dough characteristics in the community quality identification is not less than 8 minutes, and the bread score is not less than 80;
  • the plot yield is higher than the high-quality control or greater than or equal to the high-yield control variety, or the yield is less than 5% lower than the high-yield control variety;
  • the selection criteria for the strain population are as follows 6)-4 to 6)-5:
  • the target wheat variety whose yield and quality are improved is a high-yield control or a high-quality control whose yield and quality are higher.
  • the above-mentioned high-yield control is a high-yield regional test control variety in the local wheat area, which is specifically Jimai 22 in the embodiment of the present invention; the above-mentioned high-quality control is a high-quality regional test control variety in the local wheat area, which is specifically in the embodiment of the present invention Jinan 17.
  • the cell seeding is all drill seeding
  • the on-demand sowing is all plant-by-line seeding.
  • step 1) the method of selecting a parent combination from multiple parent combinations that meets the following criteria is: performing protein level identification and molecular level identification on multiple parent combinations, and selecting any parent combination in the combination contains At least one high-quality related protein, and the cDNA of any parent in the combination contains a DNA molecule encoding the high-quality related protein;
  • step 4) and step 6) that contain high-quality related proteins have passed the molecular level identification;
  • the protein level is identified as the molecular weight of the high-quality related protein detected by SDS electrophoresis; the specific method is as follows: extract the total protein of the sample, and detect the molecular weight by electrophoresis to obtain a band with the size of the high-quality related protein, then the sample contains high-quality Related protein
  • the identification at the molecular level is the amplification of the high-quality related protein with a molecular marker corresponding to the high-quality related protein.
  • the specific method is as follows: amplify the sample to be tested with the molecular marker corresponding to the high-quality related protein to obtain the size of the PCR product corresponding to the molecular marker, then the sample to be tested contains the DNA molecule encoding the high-quality related protein, thereby determining that the sample to be tested contains High-quality related proteins.
  • the high-yield control is a high-yield regional test control variety in the local wheat area
  • the high-quality control is a high-quality regional test control variety in a local wheat area.
  • step 2) the on-demand F 0 hybrid grains is to sow 40 F 0 hybrid grains in a row length of 2-4 meters;
  • step 3 the cell used for seeding the cell is 4 meters long, 1.5 meters wide, and 6 rows are seeded;
  • step 4 the on-demand is 60 rows per ear, 80 grains per row;
  • step 5 the on-demand is 60 rows per ear with 80 grains per row.
  • the high-quality related proteins are glutenin subunit 11 (Genbank No: LT626208.1.2016.10.13), 7+8 (Genbank No: JF736013.1.2011.9.19), JF736014.1.2011.9.19) and 5+ 10 (Genbank No: AY804129.2.2007.3.8, AY863056.1.2005.1.19) any of them.
  • the molecular marker corresponding to the 7+8 high molecular weight glutenin subunit is the By8 molecular marker
  • the molecular marker corresponding to the 5+10 high molecular weight glutenin subunit is Dx5 molecular marker
  • the By8 molecular marker is composed of the single-stranded DNA molecule or its derivative shown in sequence 1 and the single-stranded DNA molecule or its derivative shown in sequence 2;
  • the Dx5 molecular marker is composed of the single-stranded DNA molecule shown in sequence 3 or its derivative and the single-stranded DNA molecule shown in sequence 4 or its derivative.
  • the single-stranded DNA molecule or its derivative is a single-stranded DNA molecule that undergoes one or several nucleotide substitutions and/or deletions and/or additions and has the same function as the single-stranded DNA molecule Single-stranded DNA molecule.
  • the parent combination is a combination of Ji 954072 and Jimai 17 or a combination of Ji 954072 and Jimai 19.
  • the yield of the plot is calculated according to the following method: first select a single plant from a plot with a stripe rust disease level of less than or equal to 1, a single ear length of not less than 8 cm, stick or square ear, plant height 70-85 cm The individual plant is recorded as a single-ear selected single plant, the grain weight of all single-ear selected single plants in the district is counted, and the grains of other single-plants except the single-ear selected single plant in the district are harvested, and the grain weights of other single plants are counted; Then add the grain weight of the selected single plant from a single ear to the grain weight of other single plants to obtain the yield of the plot;
  • the production of F 1 plant population is one-line single-line groups of F 1 Yield / F 1 generation of single row sowing grains;
  • the grain protein content is measured by DA7200 multifunctional near-infrared analyzer.
  • the present invention combines the advantages of pedigree method and hybrid method, and provides a method that uses molecular markers combined with phenotypic identification to select all generations, and each generation is selected according to different standards. It is different from the derivative system method, that is, it retains the pedigree method to examine the strain. The traceability of line history and genetic relationship retains the characteristics of simple operation and rich genetic diversity of the hybrid method.
  • the parent background is clear, the combination combination combination has a strong purpose, the selection process is simple, the target is clear, and it is easy to operate, especially for those with no breeding experience, it is easier to grasp, and provides technical support for the rapid selection of high-quality and high-yield wheat varieties.
  • Figure 1 shows the SDS-PAGE electrophoresis map of HMW-GS, from left to right: Chuanmai 56 (standard control), China Spring (standard control), Shumai 482 (standard control), Jinan 17, Jimai 954072.
  • Figure 2 shows the molecular marker detection of Dx5 subunit; from left to right: 1: Marker; 2: Ji 954072; 3: Jinan 17; 4: Jimai 19; 5-13: other varieties.
  • Figure 3 shows the molecular marker detection of By8 subunits; from left to right: 1: Marker; 2: Ji 954072; 3: Jinan 17; 4-7: other varieties; 8: Jimai 19; 9-15: other varieties.
  • Figure 4 shows an 8 mesh iron sieve.
  • Figure 5 is a schematic diagram of ear selection with 3M tape.
  • Figure 6 shows that the counting plate counts 80 tablets.
  • Figure 7 shows the HMW-GS SDS-PAGE electrophoresis chart of Jimai 44.
  • Test materials Ji 954072, Jinan 17, Jimai 19 and other parent materials.
  • Jinan 17 was sold by Shandong Luyan Seed Co., Ltd.;
  • Jimai 19 was sold by Shandong Luyan Seed Co., Ltd.;
  • Example 1 A method for breeding wheat with improved yield and quality
  • Combination 2 The combination of Ji 954072 and Ji Mai 19.
  • the glutenin subunits related to high quality are 1 (Genbank No: LT626208.1.2016.10.13), 7+8 (Genbank No: JF736013.1.2011.9.19), JF736014.1.2011.9.19) and 5+10 (Genbank No: AY804129.2.2007.3.8, AY863056.1.2005.1.19).
  • Genomic DNA from leaf tissues of Ji 954072, Jinan 17, and Jimai 19 were extracted respectively, and the 5+10 high molecular weight glutenin subunit molecules in Table 1 were used to label Dx5 and the By8 molecular label of 7+8 high molecular weight glutenin subunits (Table 1). 1) Perform PCR amplification.
  • the combination of Ji 954072 and Jinan 17, and the combination of Ji 954072 and Jimai 19 are all expected to breed individual plant combinations with 1, 7+8, and 5+10 high molecular weight glutenin subunits, that is, the combination of Ji 954072
  • the 5+10 high molecular weight gluten subunits were transferred to Jinan 17 and Jimai 19 to replace the 4+12 and 2+12 subunits, respectively.
  • Plant yield were calculated to obtain the above-described two kinds of groups of F 1 in combination a single row, select high control is greater than the yield per plant species (Jinan, 17) is greater than or equal to yield control variety (Jimai 22), or control variety yield than the yield in production within 5% of the F 1 plant, referred to as the F1 plant selected.
  • the above-mentioned seeds of the selected F1 generation individual plants from the two combinations are sieved with an 8 mesh sieve (as shown in Figure 4) to obtain F1 generation individual seeds larger than 8 mesh (the purpose is to remove the seeds with light weight of 1,000 grains);
  • Stripe rust is generally divided into the following levels (NY/T 1443.1-2007): "0" near immunity: completely asymptomatic, or occasionally with very small light spots, various types of dead groups, no spore piles; "1" height Resistance: There are few urinifera piles and there are obvious dead spots around; “2" Moderate resistance: There are few and scattered uremic spore piles, but the appearance is normal, and there are chlorosis or dead spots around; “3” Moderate infection: urinifera piles There are many, normal appearance, and there is chlorosis around; "4" high infection: there are many urinifera piles, the appearance is normal, and there is no chlorosis around.
  • the stripe rust grade of the F 2 generation community population derived from the combination of Ji 954072 and Jimai 17 is 3;
  • the stripe rust grade of the F 2 generation community population derived from the combination of Ji 954072 and Jimai 19 is 3.
  • the winter freezing damage level of the F 2 generation community group derived from the combination of Ji 954072 and Jimai 17 is 3;
  • the F 2 generation community population derived from the combination of Ji 954072 and Jimai 19 has a freezing injury level of 3.
  • the F 2 generation cell population derived from the 2 combinations obtained in the above 1) is selected for single ear, and the individual plant stripe rust disease grade is less than or equal to 1, the single ear length is not less than 8 cm, the stick or square ear, the plant height is 70-85 A single plant in centimeters is recorded as a single-ear selected single plant (using 3M blue label to select the ear, Figure 5), and the grain weight of all single-ear selected single plants in the F2 generation plot is counted.
  • the F2 generation plots were mechanically harvested to remove the grains of other single plants from the single ear selected single plants.
  • the yield based on 2-generation cell F F2 generation cell in the grain weight per panicle selected all the individuals of the F2 + removes other plant cell selected panicle grain per plant.
  • the output of the control Jimai 22 plot and the control Jinan 17 plot were 5.46 kg and 5.02 kg respectively.
  • the results of the F2 generation community derived from the combination of Ji 954072 and Jimai 17 are consistent and will be retained.
  • the result of the F2 generation community derived from the combination of Ji 954072 and Jimai 19 is not consistent and will be eliminated.
  • the result is selected from Ji 954072 and F2 Jimai cell 17 for selecting a set of the F 2 population of cells cell production.
  • the plots that meet the above (1), (2) and (3) that is, the plots that meet the stripe rust disease grade less than 4 and the winter freeze damage is less than 4, the plots whose output is greater than or equal to the high-yield control varieties or high-quality control varieties,
  • a cell with a 1,000-grain weight equal to or greater than 35 grams is the F2 generation cell derived from the combination of Ji 954072 and Jimai 17; it is recorded as the selected F 2 generation cell;
  • the F3 generation population is listed on a single plant. Take the leaves of a single plant, and extract DNA as a template by conventional methods. Use Dx5 molecular marker (target fragment is 450bp), By8 molecular marker (target fragment is 527bp) for detection, containing Dx5 molecular marker target fragment
  • the individual plants that mark the target fragment with the By8 molecular marker are the individual plants that contain 1, 7+8, and 5+10 subunits, which are recorded as individual plants selected in the F3 generation population by molecular identification, and a total of 382 plants were selected.
  • the individual plants of the F3 generation population selected by the above molecular identification are selected according to the following criteria: select a single plant with no less than 5 ears, stick or square ears, and flag leaves raised (the angle between the flag leaves is between 10-30 degrees) Individual plants with powdery mildew less than grade 4, stripe rust less than grade 4, and plant height 70-85 cm are recorded as the F3 generation individual plant population selected for field testing.
  • the national standard for the above-mentioned powdery mildew grading standard is NY/T613-2002, which is generally at the peak of powdery mildew when wheat is heading; it is recorded in five levels of investigation. "1" level leaves have no visible symptoms. "2" level institutional leaves are affected. "3” level disease spots spread to the middle leaves. "4" level disease spots spread to flag leaves. "5" level disease spots spread to ears and awns; Natural disease in the field), a total of 275 strains were selected.
  • the selected F3 generation single-plant grains obtained in the above 4 were seeded with a counting plate with 80 grains, seed lines, 4 meters long, and Austrian on-demand planter to obtain F4 generation plant lines. Every 10 rows add 1 row each for high yield control and high quality control.
  • the F4 generation plant rows with the winter freezing injury grade less than or equal to 3 in the F4 generation plant row group were selected to obtain the winter freezing injury identification F4 generation plant row group; a total of 227 rows were selected.
  • the F4 generation plant populations whose powdery mildew disease grade and stripe rust disease grade were both less than grade 4 among the F4 generation plants identified by winter freezing injury were selected to obtain the disease resistance identification F4 generation plant population, and a total of 169 plants were selected.
  • F4 generation plant population for field character identification.
  • the whole row is completely plucked and divided into individual plants for threshing; each individual plant leaves 80 seeds, the rest of the row seeds are all mixed, and the thousand-seed weight is determined.
  • a total of 116 rows of plants with a thousand-grain weight greater than or equal to 38g were selected and recorded as the F4 generation plant-row population identified by the thousand-seed weight.
  • the selected F4 generation single-plant grains obtained in the above 5 were simultaneously sown and plot sown to obtain F5 generation plant population and F5 generation community population.
  • the plot is 4 meters long and 1.5 meters wide.
  • the high-yield control (Jinan 22) and the high-quality control (Jinan 17) were planted at the same time.
  • Lodging grade (refer to the national wheat variety test record standard): "1" level is not down; “2" level is slightly lodging, and the plant inclination angle is less than 30; “3" level is moderately lodging, and the plant inclination angle is 30-45; “4" Grade 5 lodging is more serious, and the plant is inclined at an angle of 45-60; “5" grade is severe, and the plant is inclined at an angle of 60 or more.
  • the F5 generation community population derived from the combination of Ji 954072 and Jimai 17 in winter was grade 3 for freezing damage and 440,000 ears per mu (400,000 for high-quality control and 450,000 for high-yield control); lodging was grade 2.
  • the grains of the F5 generation plot population that meet the field selection criteria were harvested, and the dough characteristics and the quality of the bread were measured by the farinometer and the bread processing quality (refer to the ICC standard, No. 115 and "Determination of water absorption and rheological characteristics of wheat flour dough Quality Instrument Method” GB/T14614-2006.).
  • the stable time of the F5 generation community from the combination of Ji 954072 and Jimai 17 was 40 minutes, and the bread score was 89.5.
  • the F5 generation cell population from the combination of Ji 954072 and Jimai 17 is a quality identification cell group.
  • the plot yield of the F5 generation plot population from the combination of Ji954072 and Jimai 17 was 5.28 kg; the plot yield of the high-quality control was 4.88 kg; the plot yield of the high-yield control was 5.78 kg.
  • the F5 generation community from the combination of Ji 954072 and Jimai 19 meets the above criteria and is selected as the F5 generation community.
  • the F5 generation plant group was harvested in the whole row, and the output was counted.
  • the yields of F5 generations from the combination of Ji 954072 and Jimai 17 that meet the above criteria were 11.3 kg, 10.85 kg, 12.51 kg, and 11.32 kg, respectively.
  • the selected plants and rows are recorded as F5 generation as selected plants and rows.
  • the F5 generation single plant derived from the combination of Ji 954072 and Jimai 17, which satisfies both the identification of plot A and the identification of plant B at the same time, is selected as the target variety and named Jimai 44.
  • the new wheat line Jimai 44 was taken out of the nursery and participated in various levels of regional trials. Currently participating in the second year Shandongzhou high-fertilizer regional test and national varieties comparison test. This line has the characteristics of stable spring growth, raised flag leaves, compact plant type, good stalk elasticity, and strong lodging resistance. The plant height is about 80cm, early maturity, and the maturity period is 2-3 days earlier than that of Jimai 22. Large ears , Rectangular, long awn, white grain, horny, full grain.

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Abstract

本发明公开了一种小麦产量与品质协同提高的劣汰选优选育方法。本发明提供了一种选育产量与品质提高的目标小麦品种的方法，其结合了系谱法和混合法的优势，提供了利用分子标记结合表型鉴定的全世代决选、各世代按不同标准选取的方法，与衍生系统法不同，即保留了系谱法可考查株系历史和亲缘关系的可追溯性，又保留了混合法操作简单、遗传多样性丰富的特点。组配组合前亲本背景清楚，组合组配目的性强，选择过程简单、目标清晰、容易操作，尤其对没有育种经验者而言，更易掌握，为快速选育优质高产小麦新品种提供技术支撑。

Description

一种小麦产量与品质协同提高的劣汰选优选育方法 技术领域
本发明涉及生物技术领域，尤其涉及一种小麦产量与品质协同提高的劣汰选优选育方法。
背景技术
小麦一直是我国北方人民的主要口粮。随着耕地面积不断减少，提高单产成为保证总产增加的唯一途径，进而保障我国粮食安全。面对人民生活水平的提高和膳食结构的改变，我国小麦生产开始进行战略性调整，从单纯产量型向优质高产高效型转变。由于小麦产量与品质一直是大家公认的矛盾体，如何较好协调二者的关系，成为亟待解决的问题。虽然，我国育种家也曾提出过一些优质强筋小麦的选取指标，但基本都是局部改进，未见有同时提高质与量选取方法的系统报告。
小麦是常规自花授粉作物，目前国内外在杂种后代选取中通常采用系谱法、混合法、衍生系统法、单籽粒传及双单倍体等方法进行，国内以系谱法应用最为广泛。然而，随着人工成本增加和育种规模不断扩大，上述方法均较难适应育种现状。
发明内容
为了选育高产量与品质提高的目标小麦品种，本发明提供了如下技术方案：
本发明的目的是提供一种选育产量与品质提高的目标小麦品种的方法。
本发明提供的方法包括如下步骤：
1)从多个亲本组合中选取符合如下标准的亲本组合，作为多个候选亲本组合；所述标准为所述亲本组合中的2个亲本均含有至少1个高品质相关蛋白；
2)将多个所述候选亲本组合分别进行杂交，得到F 0代杂交粒；点播所述F 0杂交粒，得到F 1代单行群体；再从所述F 1代单行群体中选取单株产量大于优质对照品种或大于等于高产对照品种,或比高产对照品种产量减产在5％以内的单株作为入选F 1代单株；
3)收集所有所述入选F 1代单株的种子，过筛选取大于8目筛子的种子进行小区播种，得到F 2代小区群体；从所述F 2代小区群体中选取符合如下标准3)-1至3)-3的小区，作为入选F 2代小区；
3)-1，条锈病病级小于4级且冬季冻害小于4级的小区；3)-2，小区产量大于优质对照品种或大于等于高产对照品种,或比高产对照品种产量减产在5％以内的小区；3)-3，千粒重大于等于35克的小区；
4)将所述入选F 2代小区单株的籽粒进行点播，得到F3代群体；从所述F3代群体中选取符合如下4)-1至4)-3标准的单株，得到入选F3代单株；
4)-1，含有与高品质相关蛋白的单株；4)-2、单株分蘖成穗不少于5个，棍棒或方型穗、旗叶上举(旗叶夹角在10-30度之间为宜)、白粉病小于4级条锈病小于4级、株高70-85厘米的单株；4)-3、单株籽粒蛋白质含量大于等于13％；
5)将所述入选F 3代单株的籽粒进行点播，得到F4代株行群体；从所述F4代株行群体中选取符合如下5)-1至5)-5标准的单株，得到入选F4代株行；
5)-1，冬季冻害等级小于等于3级；5)-2，白粉病病级和条锈病病级均小于4级；5)-3，株高在75-80厘米；5)-4千粒重大于等于38g的株行；5)-5株行单株揉混特性鉴定中揉混仪峰值时间不低于2分钟，8分钟带宽不低于10％；
6)将所述入选F4代株行的单株种子同时点播和小区播种，得到F5代株行群体和F5代小区群体；选取同时符合株行群体筛选标准和小区筛选标准的单株，为目标小麦品种；
所述小区筛选标准为如下6)-1至6)-3：
6)-1，冬季冻害小于等于3级、亩穗数大于优质对照或大于等于高产对照，倒伏小于等于2级；
6)-2，小区品质鉴定中面团特性稳定时间不低于8分钟，面包评分不低于80；
6)-3，小区产量高于优质对照或大于等于高产对照品种,或比高产对照品种产量减产在5％以内；
所述株行群体筛选标准为如下6)-4至6)-5：
6)-4，产量大于优质对照品种或大于等于高产对照品种,或比高产对照品种产量减产在5％以内的株行；
6)-5，含有与高品质相关蛋白的单株。
上述方法中，产量与品质提高的目标小麦品种为产量与品质高于高产对照或优质对照。
上述高产对照为当地麦区的高产区试对照品种，在本发明的实施例中具体为济麦22；上述优质对照为当地麦区的优质区试对照品种，在本发明的实施例中具体为济南17。
上述方法中，所述小区播种均为条播；
或，所述点播均为株行行播。
上述方法中，步骤1)中，所述从多个亲本组合中选取符合如下标准的亲本组合的方法为：将多个亲本组合分别进行蛋白水平鉴定和分子水平鉴定，选取组合中任一亲本含有至少1个高品质相关蛋白，且该组合中任一亲本的cDNA中含有所述高品质相关蛋白的编码DNA分子；
或，步骤4)和步骤6)中选取含有与高品质相关蛋白的单株均通过分子水平鉴 定；
所述蛋白水平鉴定为通过SDS电泳检测所述高品质相关蛋白的分子量；具体方法如下：提取样本的总蛋白，电泳检测分子量，得到高品质相关蛋白大小的条带，则该样本中含有高品质相关蛋白；
所述分子水平鉴定为用高品质相关蛋白对应的分子标记扩增所述高品质相关蛋白。
具体方法如下：用所述高品质相关蛋白对应的分子标记扩增待测样本，得到分子标记对应的PCR产物大小，则待测样本含有高品质相关蛋白的编码DNA分子，从而确定待测样本含有高品质相关蛋白。
上述方法中，所述高产对照为当地麦区的高产区试对照品种；
或，所述优质对照为当地麦区的优质区试对照品种。
上述方法中，步骤2)中，所述点播F 0杂交粒为将40粒所述F 0杂交粒按照2-4米行长点播；
步骤3)中，所述小区播种采用的小区长4米、宽1.5米，播种6行；
步骤4)中，所述点播为单穗60行，每行80粒；
步骤5)中，所述点播为单穗60行，每行80粒。
上述方法中，所述高品质相关蛋白为麦谷蛋白亚基11(Genbank No:LT626208.1.2016.10.13)、7+8(Genbank No:JF736013.1.2011.9.19)、JF736014.1.2011.9.19)和5+10(Genbank No:AY804129.2.2007.3.8、AY863056.1.2005.1.19)中任一种。
上述方法中，所述7+8高分子量麦谷蛋白亚基对应的分子标记为By8分子标记；
所述5+10高分子量麦谷蛋白亚基对应的分子标记为Dx5分子标记；
所述By8分子标记由序列1所示的单链DNA分子或其衍生物和序列2所示的单链DNA分子或其衍生物组成；
所述Dx5分子标记由序列3所示的单链DNA分子或其衍生物和序列4所示的单链DNA分子或其衍生物组成。
上述方法中，所述单链DNA分子或其衍生物为将单链DNA分子经过一个或几个核苷酸的取代和/或缺失和/或添加且与所述单链DNA分子具有相同功能的单链DNA分子。
上述方法中，所述亲本组合为济954072和济麦17组合或济954072和济麦19组合。
上述方法中，所述小区产量按照如下方法计算：先从某所述小区中选取单株条锈病病级小于等于1、单穗长度不小于8cm、棍棒或方型穗、株高70-85厘米的单株，记作单穗入选单株，统计该小区中所有单穗入选单株的籽粒重量，再收获该小区中除去单穗入选单株其他单株籽粒，统计其他单株的籽粒重量；再将单穗入选单株的籽粒 重量和其他单株的籽粒重量相加，得到该小区的产量；
上述方法中，所述F 1代单行群体的单株产量为F 1代单行群体产量/F 1代单行播种粒数；
上述方法中，所述籽粒蛋白质含量采用DA7200多功能近红外分析仪进行籽粒品质测定。
本发明结合了系谱法和混合法的优势，提供了利用分子标记结合表型鉴定的全世代决选、各世代按不同标准选取的方法，与衍生系统法不同，即保留了系谱法可考查株系历史和亲缘关系的可追溯性，又保留了混合法操作简单、遗传多样性丰富的特点。组配组合前亲本背景清楚，组合组配目的性强，选择过程简单、目标清晰、容易操作，尤其对没有育种经验者而言，更易掌握，为快速选育优质高产小麦新品种提供技术支撑。
附图说明
图1为HMW-GS的SDS-PAGE电泳图谱，从左至右：川麦56(标准对照)、中国春(标准对照)、蜀麦482(标准对照)、济南17、济麦954072。
图2为Dx5亚基的分子标记检测；从左至右:1：Marker；2：济954072；3：济南17；4：济麦19；5-13：其他品种。
图3为By8亚基的分子标记检测；从左至右：1：Marker；2：济954072；3：济南17；4-7：其他品种；8：济麦19；9-15：其他品种。
图4为8目铁筛。
图5为3M胶带选穗示意图。
图6为数粒板数粒80粒。
图7为济麦44的HMW-GS SDS-PAGE电泳图谱。
具体实施方式
下述实施例中所使用的实验方法如无特殊说明，均为常规方法。
下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
试验材料：济954072、济南17、济麦19及其它亲本材料。
济954072(贾金川,徐士清,孙妍,潘梅昌,徐海洋.优质小麦新品种比较试验[J].中国种业,2003,(12):35)；
济南17由山东鲁研良种有限公司出售；
济麦19由山东鲁研良种有限公司出售；
实施例1、一种产量与品质提高的小麦的选育方法
以下用2种组合的方式为例进行筛选：
组合1：济954072和济南17配制的组合
组合2：济954072和济麦19配制的组合。
一、产量与品质提高的小麦的选育方法
1、鉴定亲本组合是否含有与产量或品质相关的蛋白
1)蛋白水平检测
分别提取济954072、济南17、济麦19组织的麦谷蛋白(纪军,刘冬成,王静,等.一种小麦高、低分子量麦谷蛋白亚基的提取方法[J].遗传,2008,30(1):123-126.)，SDS-PAGE检测。
以川麦56和中国春作为标准对照。
济954072、济南17、济麦19的HMW-GS的SDS-PAGE电泳结果如图1所示，从左至右：川麦56(标准对照)、中国春(标准对照)、蜀麦482(标准对照)、济南17、济麦954072，济954072含1、7+9、5+10高分子量麦谷蛋白亚基；济南17含1、7+8、4+12高分子量麦谷蛋白亚基；济麦19含1、7+8、2+12。
其中与高品质相关的麦谷蛋白亚基分别为1(Genbank No:LT626208.1.2016.10.13)、7+8(Genbank No:JF736013.1.2011.9.19)、JF736014.1.2011.9.19)和5+10(Genbank No:AY804129.2.2007.3.8、AY863056.1.2005.1.19)。
济954072、济南17、济麦19均含有2个与高品质相关的麦谷蛋白亚基，培育比其品质更好的小麦，需要含有3个与高品质相关的麦谷蛋白亚基：1、7+8、5+10高分子量麦谷蛋白亚基。
2)分子水平检测
分别提取济954072、济南17、济麦19叶片组织的基因组DNA，用表1中的5+10高分子量麦谷蛋白亚基分子标记Dx5和7+8高分子量麦谷蛋白亚基的By8分子标记(表1)进行PCR扩增。
表1 为Dx5、By8分子标记信息及扩增条件
Figure PCTCN2019112083-appb-000001
Dx5分子标记扩增结果如图2所示，从左至右:1：Marker；2：济954072；3：济 南17；4：济麦19；5-13：其他品种；可以看出，济954072扩增到450bp即为含有5+10高分子量麦谷蛋白亚基编码DNA分子；而济南17、济麦19经过Dx5扩增，没有目标片段的获得。
By8分子标记扩增结果如图3所示，从左至右：1：Marker；2：济954072；3：济南17；4-7：其他品种；8：济麦19；9-15：其他品种；可以看出，济南17扩增到527bp即为含有7+8高分子量麦谷蛋白亚基；济麦19扩增到527bp即为含有7+8高分子量麦谷蛋白亚基；而济954072经过By8扩增，没有目标片段的获得。
该结果与SDS-PAGE检测结果一致，这2个标记也用于下面叙述的后代分子标记辅助选取检测。
因此，济954072和济南17组合、济954072和济麦19组合预期都是要选育出单株具有1、7+8、5+10高分子量麦谷蛋白亚基组合，也就是将济954072中的5+10高分子量谷蛋白亚基分别转移到济南17及济麦19中替换4+12及2+12亚基。
2、杂交F1代的获得和选育入选组合
1)点播获得杂交F1代及选育如下F1代单株
以济954072为母本，以济南17为父本进行杂交，得到F 0杂交粒。
每个济954072和济南17组合做4个杂交穗。
统计收获的杂交粒数，如果杂交粒数少于40粒、或籽粒太瘪则于以淘汰。
将上述40粒F 0杂交粒按照2米行长全部点播(5公分1粒)，得到济954072和济南17组合的F 1代单行群体。
以济954072为母本，以济麦19为父本进行杂交，得到F 0杂交粒。
每个济954072和济麦19组合做4个杂交穗。
统计收获的杂交粒数，如果杂交粒数少于40粒、或籽粒太瘪则于以淘汰。
将上述40粒F 0杂交粒按照2米行长全部点播(5公分1粒)，得到济954072和济麦19组合的F 1代单行群体。
2)单株产量进行杂交F1代选取
分别计算上述获得2种组合的F 1代单行群体的单株产量，选取单株产量大于优质对照品种(济南17)或大于等于高产对照品种(济麦22),或比高产对照品种产量减产在5％以内的F 1代单株，记作入选F1代单株。
具体方法如下：
分别将上述获得2种组合的F 1代单行群体全部收获后称重，计算单株产量＝单行群体产量/单行播种粒数，结果济954072/济南17组合的F 1单株产量为11.3克，济954072/济麦19组合F 1单株产量为10.5克。以种植高产对照(当地麦区的高产区试对照品种济麦22)和优质对照(当地麦区的优质区试对照品种济南17)各40粒按照 2米行长全部点播(5公分1粒)，作为对照，统计对照的单株产量，结果对照品种济麦22的单株产量为12.8克，对照品种济南17的单株产量为10.2克。
比较后，得到入选F1代单株。源于济954072和济麦14的F1代以及源于济954072和济麦17的F1代均入选。
3、小区播种获得F 2代及选育入选F 2代小区
1)小区播种获得F 2代
将上述源于2种组合的入选F1代单株的种子，分别用8目筛子(如图4)进行过筛，得到大于8目的F1代单株种子(目的是去除千粒重轻的种子)；
分别称取120克源于2种组合的大于8目的F1代单株种子进行小区播种，小区长4米、宽1.5米，6行，奥地利小区播种机条播，接近或高于生产种植密度，得到源于2种组合的F 2代小区群体。
同时种植高产对照小麦(济麦22)及优质对照小麦(济南17)，得到对照济麦22小区和对照济南17小区。
2)F 2代选取
从F 2代小区群体中选取符合如下标准(1)至(3)的小区，作为入选F 2代小区:(1)，条锈病病级小于4级且冬季冻害小于4级的小区；(2)，小区产量大于优质对照品种或大于等于高产对照品种,或比高产对照品种产量减产在5％以内的小区；(3)，千粒重大于等于35克的小区；具体如下：
(1)田间选取
A、条锈病
分别在源于2种组合的F 2代小区群体的四周种辉县红(张宏,任志龙,胡银岗,等.陕麦139抗条锈病基因遗传分析[J].作物学报,2010,36(1):109-114.)诱发行，拔节期进行条锈病接种采用扫抹法接种。具体做法是用吐温20(Tween20)水溶液(0.05％，V/V)喷洒叶片，使雾状水滴均匀分布在叶面上，然后将初繁好的混合条锈菌生理小种CYR29和CYR32直接扫抹接种。
条锈病一般分为以下几级(NY/T 1443.1-2007):“0”近免疫：完全无症状,或偶有极小淡色斑点，有各种类型枯死班,无孢子堆；“1”高度抵抗:夏孢子堆很少,周围有明显枯斑；“2”中度抵抗:夏孢子堆少而分散，但外形正常，周围有退绿或枯死斑；“3”中度感染:夏孢子堆较多,外形正常,周围有退绿现象；“4”高度感染:夏孢子堆很多，外形正常，周围无退绿现象。
源于济954072和济麦17组合的F 2代小区群体的条锈病级别为3；
源于济954072和济麦19组合的F 2代小区群体的条锈病级别为3。
B、冬季冻害
观察源于2种组合的F 2代小区群体过冬的冬季冻害级别，冬季冻害分五级(国标号NY/T 2283.2-2012)，“1”级无冻害；“2”级叶尖受冻发黄；“3”级叶片冻死一半；“4”级叶片全枯；“5”级植株冻死。
源于济954072和济麦17组合的F 2代小区群体的冬季冻害级别为3；
源于济954072和济麦19组合的F 2代小区群体的冬季冻害级别为3。
选取冬季冻害小于4级，且条锈病病级小于4级的F 2代小区群体，为田间选取F 2代小区群体。
(2)小区产量选取
a、单穗选取
上述1)得到的源于2种组合的F 2代小区群体进行单穗选取，选取单株条锈病病级小于等于1、单穗长度不小于8cm、棍棒或方型穗、株高70-85厘米的单株，记作单穗入选单株(用3M蓝色标签选穗，图5)，统计该F2代小区中所有单穗入选单株的籽粒重量。
b、小区产量
分别机械收获该F2代小区中除去单穗入选单株的其他单株籽粒。
计算F 2代小区产量＝该F2代小区中所有单穗入选单株的籽粒重量+该F2代小区中除去单穗入选单株的其他单株籽粒。
源于济954072和济麦17组合的F 2代小区产量为5.21千克；
源于济954072和济麦19组合的F 2代小区产量为4.79千克；
统计对照济麦22小区产量和对照济南17小区产量，分别为5.46千克和5.02千克。
选取小区产量大于或等于高产对照小麦(济麦22)或优质对照小麦(济南17)的F 2代小区，得到小区产量选取F 2代小区群体。
据上述产量结果，源于济954072和济麦17组合的F2代小区结果符合，予以保留，源于济954072和济麦19组合的F2代小区结果不符合，予以淘汰，结果入选源于济954072和济麦17组合的F2代小区为小区产量选取F 2代小区群体。
(3)千粒重
统计源于2种组合的F 2代小区群体的千粒重，源于济954072和济麦17组合的F 2代小区的千粒重为43克，源于济954072和济麦19组合的F 2代小区的千粒重为42克。
选取千粒重大于等于35克的小区，记作千粒重选取F 2代小区群体。
将符合上述(1)、(2)和(3)的小区，即满足条锈病病级小于4级且冬季冻害小于4级的小区、小区产量大于或等于高产对照品种或优质对照品种的小区、千粒重大于等于35克的小区，为源于济954072和济麦17组合的F2代小区；记作入选F 2代小区；
4、F3代的获得和筛选
1)F3代的获得
将上述3选取的入选F 2代小区的所有单株的单穗下一年度种小区，单穗60行，每行80粒，用数粒板数80粒(如图6)，奥地利点播机点播；得到F3代群体。
整理好顺序后，各小区每10行加高产对照和优质对照各1行。
2)F3代的选取
从F3代群体中选取符合如下(1)至(3)标准的单株，得到入选F3代单株：(1)，含有与高品质相关蛋白的单株；(2)、单株分蘖成穗不少于5个，棍棒或方型穗、旗叶上举(旗叶夹角在10-30度之间)、白粉病小于4级条锈病小于4级、株高70-85厘米的单株；(3)、单株籽粒蛋白质含量大于等于13％；具体如下：
(1)分子鉴定选取含有与高品质相关蛋白的单株；
过冬前F3代群体单株挂牌取单株叶片，常规方法提取DNA作为模板，分别用Dx5分子标记(目的片段为450bp)、By8分子标记(目的片段为527bp)进行检测，含有Dx5分子标记目的片段和By8分子标记目的片段的单株，即为含有1、7+8、5+10亚基的单株，记作分子鉴定入选F3代群体单株，共计382株入选。
(2)田间检测
将上述分子鉴定入选F3代群体单株按照如下标准选取：选取单株分蘖成穗不少于5个，棍棒或方型穗、旗叶上举(旗叶夹角在10-30度之间)、白粉病小于4级、条锈病小于4级、株高70-85厘米的单株，记作田间检测入选F3代单株群体。
上述白粉病分级标准的国标为NY/T613-2002，一般在小麦抽穗时白粉病盛发期；分五级调查记载。“1”级叶片上无肉眼可见症状“2”级基层叶片发病“3”级病斑蔓延至中部叶片“4”级病斑蔓延至剑叶“5”级病斑蔓延至穗及芒；采用田间自然发病)，共计275株入选。
(3)籽粒品质鉴定
收获上述(2)得到的田间检测入选F3代群体单株的籽粒，用DA7200多功能近红外分析仪进行籽粒品质测定，原则为“淘劣”而不是“选优”，判定指标为：选取单株籽粒蛋白质含量大于等于13％的F3代群体单株，即为入选F3代单株籽粒，共计156株入选。
5、F4代的获得及入选F4代单株的选取
1)F4代的获得
将上述4得到的入选F3代单株籽粒用数粒板数80粒，种株行，4米行长，奥地利点播机点播，得到F4代株行群体。每10行加高产对照和优质对照各1行。
2)F4代的筛选
从所述F4代株行群体中选取符合如下(1)-(5)至标准的单株，得到入选F4代株行；
(1)冬季冻害等级小于等于3级；(2)白粉病病级和条锈病病级均小于4级；(3)株高在75-80厘米；(4)千粒重大于等于38g的株行；(5)株行单株揉混特性鉴定中揉混仪峰值时间不低于2分钟，8分钟带宽不低于10％；具体如下：
(1)冬前调查冬季冻害
选取F4代株行群体中冬季冻害等级小于等于3级的F4代株行，得到冬季冻害鉴定F4代株行群体；，共计227株行入选。
(2)抗病性
选取冬季冻害鉴定F4代株行中白粉病病级和条锈病病级均小于4级的F4代株行群体，得到抗病性鉴定F4代株行群体，共计169株行入选。
(3)田间性状
选取抗病性鉴定F4代株行群体中株高在75-80厘米的株行，得到田间性状鉴定F4代株行群体，共计138株行入选。
(4)千粒重鉴定
将田间性状鉴定F4代株行群体挂牌。整行全拔，分单株脱粒；每个单株留80粒，其余该行种子全部混合，测定千粒重。选取株行千粒重大于等于38g的株行，记作千粒重鉴定F4代株行群体，共计116株行入选。
(5)揉混特性鉴定
取千粒重鉴定F4代株行群体各单行的200克籽粒，磨粉、利用揉混仪进行揉混特性测定；选取揉混仪峰值时间不低于2分钟，8分钟带宽不低于10％的单株。
符合上述入选标准的F4代群体单株，为入选F4代单株，共计55株行入选。
6、F5代的获得及鉴定
1)F5代的获得
将上述5获得的入选F4代单株籽粒同时行播和小区播种，得到F5代株行群体和F5代小区群体。小区长4米，宽1.5米。同时种植高产对照(济麦22)及优质对照(济南17)。
2)F5代的鉴定
选取同时符合株行群体筛选标准和小区筛选标准的单株，为目的品种；具体如下：
A、小区筛选
(1)、田间选取
选取冬季冻害小于等于3级、亩穗数大于优质对照或等于高产对照，且倒伏小于等于2级的小区，作为田间选取小区；
倒伏性等级(参照国家小麦品种试验记载标准)：“1”级未倒；“2”级倒伏轻微，植株倾斜角度小于30；“3”级中等倒伏，植株倾斜角度30-45；“4”级倒伏较严重,植株倾斜角度45-60；“5”级倒伏严重,植株倾斜角度在60以上。
源于济954072和济麦17组合F5代小区群体冬季冻害为3级、亩穗数为44万，(优质对照为40万，高产对照为45万)；倒伏为2级。
(2)品质鉴定
收获上述符合田间选取标准的F5代小区群体的籽粒，进行面团特性粉质仪及面包加工品质测定(参照ICC标准，No.115和《小麦粉面团的物理特性吸水量和流变学特性的测定粉质仪法》GB/T14614-2006进行。)。
选取面团特性稳定时间不低于8分钟，面包评分不低于80的F5代组合。
源于济954072和济麦17组合F5代小区群体稳定时间为40分钟，且面包评分为89.5。
因此，源于济954072和济麦17组合F5代小区群体为品质鉴定小区群体。
(3)产量检测
选取小区产量(产量计算方法同前)大于优质对照或大于等于高产对照,或比高产对照品种产量减产在5％以内的组合。
源于济954072和济麦17组合F5代小区群体的小区产量为5.28千克；优质对照的小区产量为4.88千克；高产对照的小区产量为5.78千克。
因此，源于济954072和济麦19组合F5代小区群体符合上述标准，为入选F5代小区。
B、株行筛选
(1)株行产量检测
F5代株行群体整行收获，统计产量。
选取整行产量大于优质对照品种或大于等于高产对照品种,或比高产对照品种产量减产在5％以内的株行。
源于济954072和济麦17组合F5代株行群体符合上述标准的株行的产量分别为11.3千克、10.85千克、12.51千克、11.32千克。
按照上述标准选取株行，记作F5代入选株行。
(2)株行分子标记检测
上述F5代入选株行分别用Dx5和By8分子标记检测，选取含有7+8和5+10亚基(由于1亚基在最初亲本中就有，后代肯定有，一般不进行鉴定)的株行。
结果源于济954072和济麦17组合的F5代株行含有7+8和5+10亚基，为入选株行；
选取同时满足A小区鉴定和B株行鉴定的条件为源于济954072和济麦17组合的F5代单株，即为目的品种，命名为济麦44。
二、检测小麦新品系济麦44
将小麦新品系济麦44出圃，参加各级区域试验。目前正参加第二年山东省高肥区试及国家品种比较试验。该品系具有春季生长稳健，旗叶上举，株型较紧凑，茎秆弹性好，抗倒伏能力强的特点，株高80cm左右，早熟，熟期较济麦22提前2-3天，大穗、长方型，长芒，白粒，角质，籽粒饱满。2014-2015年参加山东省预备试验，统一品质检测结果：籽粒蛋白质含量15.4％，面团稳定时间41.8min，最大抗延阻力800BU，达到国家优质强筋小麦标准，2015-2016年参加山东省第一年区域试验，统一品质检测：籽粒蛋白质含量14.7％，面团稳定时间39.5min，达到国家优质强筋小麦标准。产量较对照品种济麦22增产2.3％，综合特性表现优异，有极高推广价值。分子标记及SDS-PAGE检测，表明其含有1、7+8、5+10亚基(图7)。

Claims (9)

  1. 一种选育产量与品质提高的目标小麦品种的方法，包括如下步骤：
    1)从多个亲本组合中选取符合如下标准的亲本组合，作为多个候选亲本组合；所述标准为所述亲本组合中的2个亲本均含有至少1个高品质相关蛋白；
    2)将多个所述候选亲本组合分别进行杂交，得到F 0代杂交粒；点播所述F 0杂交粒，得到F 1代单行群体；再从所述F 1代单行群体中选取单株产量大于优质对照品种或大于等于高产对照品种,或比高产对照品种产量减产在5％以内的单株作为入选F 1代单株；
    3)收集所有所述入选F 1代单株的种子，过筛选取大于8目筛子的种子进行小区播种，得到F 2代小区群体；从所述F 2代小区群体中选取符合如下标准3)-1至3)-3的小区，作为入选F 2代小区；
    3)-1，条锈病病级小于4级且冬季冻害小于4级的小区；3)-2，小区产量大于优质对照品种或大于等于高产对照品种,或比高产对照品种产量减产在5％以内的小区；3)-3，千粒重大于等于35克的小区；
    4)将所述入选F 2代小区单株的籽粒进行点播，得到F3代群体；从所述F3代群体中选取符合如下4)-1至4)-3标准的单株，得到入选F3代单株；
    4)-1，含有与高品质相关蛋白的单株；4)-2、单株分蘖成穗不少于5个，棍棒或方型穗、旗叶上举、白粉病小于4级条锈病小于4级、株高70-85厘米的单株；4)-3、单株籽粒蛋白质含量大于等于13％；
    5)将所述入选F 3代单株的籽粒进行点播，得到F4代株行群体；从所述F4代株行群体中选取符合如下5)-1至5)-5标准的单株，得到入选F4代株行；
    5)-1，冬季冻害等级小于等于3级；5)-2，白粉病病级和条锈病病级均小于4级；5)-3，株高在75-80厘米；5)-4千粒重大于等于38g的株行；5)-5株行单株揉混特性鉴定中揉混仪峰值时间不低于2分钟，8分钟带宽不低于10％；
    6)将所述入选F4代株行的单株种子同时点播和小区播种，得到F5代株行群体和F5代小区群体；选取同时符合株行群体筛选标准和小区筛选标准的单株，为目标小麦品种；
    所述小区筛选标准为如下6)-1至6)-3：
    6)-1，冬季冻害小于等于3级、亩穗数大于优质对照或等于高产对照，倒伏小于等于2级；
    6)-2，小区品质鉴定中面团特性稳定时间不低于8分钟，面包评分不低于80；
    6)-3，小区产量大于优质对照品种或大于等于高产对照品种,或比高产对照产量减产在5％以内；
    所述株行群体筛选标准为如下6)-4至6)-5：
    6)-4，产量大于优质对照品种或大于等于高产对照品种,或比高产对照品种产量减产在5％以内的株行；
    6)-5，含有与高品质相关蛋白的单株。
  2. 根据权利要求1所述的方法，其特征在于：
    所述小区播种均为条播；
    或，所述点播均为株行行播。
  3. 根据权利要求1或2所述的方法，其特征在于：
    步骤1)中，所述从多个亲本组合中选取符合如下标准的亲本组合的方法为：将多个亲本组合分别进行蛋白水平鉴定和分子水平鉴定，选取组合中任一亲本含有至少1个高品质相关蛋白，且该组合中任一亲本的cDNA中含有所述高品质相关蛋白的编码DNA分子；
    或，步骤4)和步骤6)中选取含有与高品质相关蛋白的单株均通过分子水平鉴定；
    所述蛋白水平鉴定为通过SDS电泳检测所述高品质相关蛋白的分子量；
    所述分子水平鉴定为用与所述高品质相关蛋白对应的分子标记扩增所述高品质相关蛋白。
  4. 根据权利要求1-3中任一所述的方法，其特征在于：
    所述高产对照为当地麦区的高产区试对照品种；
    或，所述优质对照为当地麦区的优质区试对照品种。
  5. 根据权利要求1-4中任一所述的方法，其特征在于：
    步骤2)中，所述点播F 0杂交粒为将40粒所述F 0杂交粒按照2-4米行长点播；
    步骤3)中，所述小区播种采用的小区长4米、宽1.5米，播种6行；
    步骤4)中，所述点播为单穗60行，每行80粒；
    步骤5)中，所述点播为单穗60行，每行80粒。
  6. 根据权利要求1-4中任一所述的方法，其特征在于：
    所述高品质相关蛋白为麦谷蛋白亚基1、麦谷蛋白亚基7+8和麦谷蛋白亚基5+10中任一种。
  7. 根据权利要求7所述的方法，其特征在于：
    所述7+8高分子量麦谷蛋白亚基对应的分子标记为By8分子标记；
    所述5+10高分子量麦谷蛋白亚基对应的分子标记为Dx5分子标记；
    所述By8分子标记由序列1所示的单链DNA分子或其衍生物和序列2所示的单链DNA分子或其衍生物组成；
    所述Dx5分子标记由序列3所示的单链DNA分子或其衍生物和序列4所示的单链 DNA分子或其衍生物组成。
  8. 根据权利要求7所述的方法，其特征在于：
    所述单链DNA分子或其衍生物为将单链DNA分子经过一个或几个核苷酸的取代和/或缺失和/或添加且与所述单链DNA分子具有相同功能的单链DNA分子。
  9. 根据权利要求1-8中任一所述的方法，其特征在于：
    所述亲本组合为济954072和济麦17组合或济954072和济麦19组合。
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