CN115786565A - SNP molecular marker linked with wheat spikelet number QTL and application thereof - Google Patents
SNP molecular marker linked with wheat spikelet number QTL and application thereof Download PDFInfo
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Abstract
The invention discloses an SNP molecular marker linked with wheat spikelet number QTL and application thereof, belonging to the fields of molecular biology and crop genetic breeding. The KASP-3D-1 molecular marker is an SNP molecular marker with polymorphism of G/A, and is co-located on the short arm of the wheat 3D chromosome with the wheat spikelet number QTL QSns.sau-MC-3D.1. The KASP-3D-1 molecular marker is represented by a nucleotide sequence shown in SEQ ID NO: 1-3. The molecular marker KASP-3D-1 provided by the invention is tightly linked with the spikelet number QTL QSns.sau-MC-3D.1 on the wheat 3D, and can be used for positioning the trait of the spikelet number of the wheat, so that plants with less spikelet number are eliminated in the breeding process, the breeding work efficiency is improved, and a certain basis is provided for the genetic analysis research of the spikelet number of the wheat.
Description
Technical Field
The invention relates to the field of molecular biology and crop genetic breeding, in particular to a SNP molecular marker linked with wheat spikelet number QTL QSns.sau-MC-3D.1 and application thereof.
Background
Common wheat (AABBDD, triticum aestivum L.) is one of the most important grain crops in the world, and according to the statistics of grain and agricultural organizations in the United nations, the annual yield of wheat must be increased by 1.6% by 2050 years so as to ensure the grain demand of 91 hundred million people in the world. With the increase of the population of the world and the reduction of the planting area, the increase of the food yield has important strategic significance for solving the problems of food supply and safety in the future.
The wheat yield is controlled by multiple genes and has complex quantitative characters, and the wheat yield has the characteristics of low heritability, large environmental influence, high selection difficulty and the like. The wheat yield = spike number × spike grain number × grain weight, and these constituent factors are more adaptive to the environment and more stable in heredity than the yield. The ear number depends to a large extent on the number of spikelets, which therefore has an important influence on yield. To date, researchers have identified spikelet number QTLs on all 21 chromosomes of wheat using parental populations such as recombinant inbreds, backcrosses and dihaploids. For example, zhai Huijie et al (2016) used a recombinant inbred population to detect a major QTL on the 1B chromosome that controls spikelet number that accounts for 30.75% of phenotypic variation. The SNP marker Kukri _ c11327_977 (Zhang Cheng et al 2022) which is closely linked to SNS was found on chromosome 1A. In addition, some genes related to spikelet number have been reported, such as the homology-based cloned genes trs1/WFZP-A (Du Dejie et al 2021), VRN-A3/FT-A1 (Yan Liuling et al 2006), Q (Justin Faris et al 2003), taTB1-4A (Laura Dixon et al 2018), PPD-A1 (James Belales et al 2007) and WAPO1 by map-based cloning (Saarah Kuzay rt al 2019). Although many spikelet number-related QTL/genes have been reported in wheat, the major and stable spikelet number QTL identified in multiple environments is still limited. Therefore, the identification of the stable major genetic locus for controlling the number of spikelets has very important significance for clarifying the genetic basis of wheat yield traits and improving the wheat yield.
The traditional breeding method of wheat has the problems of time consumption, high cost, low return and the like, and molecular markers assist in breeding, do not depend on phenotype selection, are not influenced by factors such as environment and gene interaction, and directly select genotypes, so that the breeding efficiency can be greatly improved. Single Nucleotide Polymorphism (SNP) refers to a DNA sequence Polymorphism caused by a change in DNA at a specific Nucleotide position in a genome, such as a transition, a transversion, an insertion, or a deletion. The technology is that known sequence information is utilized to compare and search SNP sites, specific primers are designed by utilizing the discovered variation sites to carry out PCR amplification on genome DNA or cDNA, specific polymorphic products based on the SNP sites are obtained, and finally, the electrophoresis technology is utilized to analyze the polymorphism of the products. The SNP markers have the advantages of large quantity and wide distribution; uneven distribution among individual genes and the entire genome; SNP allele frequencies are easily estimated.
Competitive Allele Specific PCR (KASP) is a novel genotyping technology with low cost and high throughput characteristics developed by LGC company (Laboratory of the Government Chemist) (http:// www.lggeneomics.com). The technology carries out accurate biallelic genotyping on SNP and InDel sites through Specific matching of terminal bases of primers, and is widely applied to molecular marker-assisted selection of grain crops such as rice, wheat and soybean.
In the previous research, although a great deal of research has been conducted on QTL positioning of wheat spikelet number, at present, few closely linked molecular markers which are related to the wheat spikelet number and can be used for actual molecular marker-assisted selective breeding are available. Therefore, QTL or gene related to the number of spikelets is obtained through research, the number of spikelets is increased by utilizing the modern molecular biology technology, and then the number of grains per spike is increased, and finally the purpose of breeding a new wheat variety with high yield and high quality is achieved, which is significant in the wheat breeding work.
Disclosure of Invention
The invention aims to provide an SNP molecular marker linked with wheat spikelet number QTL and application thereof, which are used for solving the problems in the prior art, the molecular marker is a co-dominant marker, the detection is accurate and efficient, the amplification is convenient and stable, the trait of the wheat spikelet number can be positioned, plants with small spikelet number are eliminated in the breeding process, the breeding work efficiency is improved, and a certain foundation is provided for the genetic analysis research of the wheat spikelet number.
Based on the above purposes, the applicant utilizes the natural mutant 'msf' with multiple spikelets and multiple florets as female parent and wheat variety (national examined variety) 'Chuannong 16' as male parent for hybridization to obtain hybrid F 1 ，F 1 Selfing the single plant to obtain F 2 At F 2 Using single-event propagation up to F 6 And generating a recombinant inbred line containing 198 single plants to form a genetic mapping population. The number of spikelets of the recombinant inbred line group is investigated and identified, plant DNA of parent msf, chuannong 16 and the recombinant inbred line group is extracted, and a wheat 16K SNP liquid chip is used for positioning the QTL of the spikelet number. The development of the wheat 16K SNP liquid-phase chip product depends on the scientific research team of Kangzhen academy of science and technology university of agriculture and forestry in northwest, takes 20 parts of resequencing data of a subject group as the basis, utilizes 1,520 parts of multi-platform genotyping data of germplasm resources collected in the world, published resequencing and exon capturing data to carry out SNP screening, and uses Borid GenoBaitThe s technology was developed and optimized to finally retain 14,868 segments of mSNP (37,669 SNP markers) to construct a GBW16K product (hereinafter referred to as a 16K SNP chip). The 37,669 SNPs are evenly distributed on 21 chromosomes, with an average of 1,794 markers per chromosome. The 16K SNP chip greatly reduces the use cost of molecular assisted breeding, and the price of the product is 20 percent lower than that of other products of the same type. The method is suitable for molecular marker assisted breeding, whole genome selective breeding genetic similarity analysis, genetic relationship analysis variety protection and authenticity identification for germplasm resource genotype identification.
According to the 16K SNP chip data, a genetic map is constructed by utilizing JoinMap4.0. Combining spikelet number table data of a population, using an integral complex Interval Mapping-ADD (ICIM-ADD) in QTL IcMapping 4.1, under the condition that a threshold LOD is not less than 2.5, using BLUP (best unbiased prediction) values of 5 ecological points and 5 ecological point spikelets in two years of 2020-2022 to detect the spikelet number QTL, positioning a stably expressed wheat spikelet number main effect QTL QSL QSns-MC-3 D.1 in an 8cM Interval on a short arm of a 3D chromosome, further carrying out compact map and obtaining a molecular marker tightly linked with the spikelet number QTL-MC-3 D.1, positioning side winged markers by using a 16K SNP chip data positioning result, screening SNP markers positioned in an excavation Interval, further developing a high-efficiency linear SNP molecular marker with the SNP between the SNP parent sites, and further developing a KASP molecular marker. Totally designing 15 pairs of 45 KASP primers, and finally obtaining the marker KASP-3D-1 which is tightly linked with the spikelet number QTL QSns.sau-MC-3D.1. The invention discovers that the small spike number QTL of wheat QSns-sau-MC-3D.1 comes from a female parent 'msf', the QTL is positioned on a 3D short arm of a wheat chromosome, and the physical position of a genome version of RefSeqv2.1 in Chinese spring is 44357639-6439055 bp.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides a KASP-3D-1 molecular marker linked with wheat spikelet number QTL QSns.sau-MC-3D.1, wherein the KASP-3D-1 molecular marker is an SNP molecular marker with G/A polymorphism, and is co-located on a wheat 3D chromosome short arm with the wheat spikelet number QTL QSns.sau-MC-3D.1.
Preferably, the KASP-3D-1 molecular marker is represented by the nucleotide sequence shown in SEQ ID NO: 1-3.
The invention also provides a primer group, which comprises two specific primers and a universal primer for amplifying the KASP-3D-1 molecular marker, wherein the nucleotide sequences of the two specific primers are shown as SEQ ID NO. 1-2, and the nucleotide sequence of the universal primer is shown as SEQ ID NO. 3.
Preferably, the 5 'ends of the two specific primers are modified with different fluorophores, or the 3' ends of the two specific primers are modified with different fluorophores. More preferably, the fluorescent group includes, but is not limited to, FIFC, FAM, TET, HEX, JOE, TAMRA, BHQ.
The invention also provides a kit containing the primer group.
The invention also provides an application of the KASP-3D-1 molecular marker, the primer group or the kit in the regulation of the wheat spikelet number quantity character.
The invention also provides an application of the KASP-3D-1 molecular marker, the primer group or the kit in wheat seed resource improvement, or cultivation of multiple-spikelet type wheat or high-yield wheat, or in wheat molecular assisted breeding.
The invention also provides application of the KASP-3D-1 molecular marker, the primer group or the kit in screening wheat varieties or strains with increased spikelet numbers.
The invention also provides a method for screening a multi-spikelet number type wheat strain by utilizing the KASP-3D-1 molecular marker, which comprises the following steps:
taking the genome DNA of a wheat plant to be detected as a template, and utilizing the nucleotide sequence shown in SEQ ID NO:1-3, performing fluorescent quantitative PCR amplification, judging whether the small spike number QTL QSns-MC-3D.1 is contained according to fluorescence, and screening to obtain a multi-small-spike type wheat strain.
Preferably, if the fluorescence is marked by the primer shown by SEQ ID NO.1, the wheat plant to be detected is judged to contain spikelet number QTL QSns.sau-MC-3D.1, and the corresponding wheat plant to be detected is a multiple-spikelet number type wheat plant line.
The invention discloses the following technical effects:
the invention discloses a molecular marker KASP-3D-1 located on a wheat 3D chromosome and linked with the number of spikelets of wheat, wherein the molecular marker is a flanking marker of the spikelet number QTL QSns.sau-MC-3D.1 on the short arm of the wheat 3D chromosome, and the linkage degree is high. The marker can be used for detecting the spikelet number QTL on the 3D chromosome of the wheat, rapidly screening plants with the locus, and further facilitating the molecular assisted breeding of high-yield wheat. The molecular marker KASP-3D-1 provided by the invention is tightly linked with the spikelet number QTL QSns.sau-MC-3D.1 on the wheat 3D, and can be used for positioning the trait of the spikelet number of the wheat, so that plants with less spikelet number are eliminated in the breeding process, the breeding work efficiency is improved, and a certain basis is provided for the genetic analysis research of the spikelet number of the wheat.
The invention discloses a spikelet number QTL QSns-sau-MC-3D.1 from wheat 'msf' for the first time, which is positioned on a short arm of a wheat 3D chromosome and can obviously increase the spikelet number of the wheat. The QTL has higher utilization value in wheat yield (small spike number regulation) breeding; the invention discloses a molecular marker KASP-3D-1 for accurately detecting the spikelet number QTL QSns-sau-MC-3D.1 of wheat 'msf' based on a fluorescent quantitative PCR platform for the first time, and the molecular marker is a codominant marker, and has the advantages of accurate and efficient detection and convenient and stable amplification. The molecular marker KASP-3D-1 is remarkably related to the spikelet number QTL QSns, sau-MC-3D.1, presents coseparation marker characteristics, has high accuracy for molecular marker-assisted selection, improves the selection and identification efficiency of wheat varieties with multiple spikelets and adapting to different environments, and has high success rate.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a map of the location of wheat spikelet QTL QSns. Sau-MC-3D.1 on the 3D chromosome;
FIG. 2 shows the fluorescence reading result of the molecular marker KASP-3D-1 detection of the recombinant inbred line strain plant of 'msf' x 'Chuannong 16' of the present invention; wherein FAM (orange, 'msf') fluorescence is a strain with a large number of spikelets, HEX (blue, 'Chuannong 16') fluorescence is a strain with a small number of spikelets, and black fluorescence is blank control;
FIG. 3 shows the fluorescence reading results of the molecular marker KASP-3D-1 detection of F2 strain plant of the wheat 'msf' x wheat strain 'Sichuan wheat 969' of the present invention; among them, FAM (orange, 'msf') fluorescence is a line with a large number of spikelets, HEX (blue, 'hollyhock 969') fluorescence is a line with a small number of spikelets, and black fluorescence is a blank control.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but rather as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the documents are cited. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including but not limited to.
Example 1 obtaining of wheat spikelet QTL QSns. Sau-MC-3D.1 and its molecular marker KASP-3D-1
(1) 'msf' is a natural multi-spikelet multi-floret mutant material, is collected in Chongzhou base field of Sichuan province in 5 months in 2017, is identified as a natural mutant due to the special multi-spikelet and multi-floret characteristics and is inconsistent with all surrounding materials, seeds are stored in Sichuan agriculture university wheat institute, and the applicant promises that the seed resource is open to the outside in 20 years from the application date. 'Chuannong 16' is a national wheat variety cultivated by Sichuan university of agriculture.
The wheat 'msf' is used as a female parent, a wheat variety 'Chuannong 16' is used as a male parent for hybridization to obtain a hybrid F1, F1 generation single plants are subjected to selfing to obtain F2, a single spike propagation method is used for F2 till F6 generation, a recombinant selfing line containing 198 lines is obtained, and a genetic mapping population is formed.
(2) And (3) identifying the number phenotype of the spikelets of the recombinant inbred line population: analyzing and identifying the number of spikelets of the recombinant inbred line in the wax ripening period of the wheat, eliminating marginal effect, respectively collecting five single plants with consistent growth vigor, calculating the number of the spikelets of the main spike, and obtaining an average value which represents the number of the spikelets of the plant line.
(3) 16K SNP liquid chip analysis
a) DNA extraction
And (3) extracting parent ' msf ', chuannong 16' and recombinant inbred line population plant DNA by using a CTAB method.
b) Genotyping analysis
The extracted DNA was subjected to quality testing using a ultramicro spectrophotometer (Thermo Fisher Scientific, manufactured in USA) and, when qualified, sent to the company for genotyping, in this study the genotyping of the parental and mapping populations was performed using the 16K SNP chip of Shijiazhuang Boriedi Biotechnology Ltd (http:// www. Mol. Lbere. Com).
c) Construction of linkage map
According to the 16K SNP chip data, a genetic map is constructed by utilizing JoinMap4.0. Combining spikelet number table data of a population, detecting the spikelet number QTL by using BLUP (best linear unbiased prediction) values of 5 ecological points and 5 ecological point spikelets in 2020-2022 two years under the condition that an integral complex Interval Mapping-ADD (ICIM-ADD) in QTL IciMapping 4.1 is used, setting a threshold LOD to be more than or equal to 2.5, positioning the major QTL of the spikelet number of wheat QSs, namely QSs-MC-3D.1, and calculating the position of the QSs-MC-3D.1 and the genetic distance between molecular markers.
d) Comparison of spikelet number sites and acquisition of closely linked molecular markers
There are also many QTLs associated with spikelets and located on the 3D chromosome in previous research reports. For example, QTsn.cau-3D.3 is located at 3.99Mb on the 3D chromosomal break, the close linkage marker is CAP11_ c3914_325 (ZHai Huijie et al.2016), and the QTL1935 physical location is within 110.04-129.55Mb on the short arm of the 3D chromosome, overlapping with QSns.cd-3D (Chen Dan et al.2017). Two spikelet-related SNPs, T/C (Zhai Huijie et al.2016) and C/T (Sun Congwei et al.2017), were located at 512.68Mb and 600.26Mb, respectively, on the long arm of the 3D chromosome. Although these and spikelet number QTLs/SNPs are located on the 3D chromosome, they are far from the spikelet number QTL QSns. Sau-MC-3D.1 identified in this study, and therefore QSns. Sau-MC-3D.1 is likely to be a new and stable major QTL identified in multi-environment.
Based on sequence information of flanking markers, DNAMAN is utilized to design 15 pairs of KASP primers (45 in total; see table 1), and genotyping is carried out on parent 'msf' and 'Chuannong 16' to obtain polymorphic sites, and finally, the markers KASP-3D-1 (G/A) are determined to be closely linked with spikelet number QTL QSns.sau-MC-3D.1.
TABLE 1 KASP primer sequences
1 pair of molecular markers KASP-3D-1 are finally obtained from the designed 15 pairs of flanking marker KASP primers, and are closely linked with the spikelet number QTL QSns.sau-MC-3D.1, and the result is shown in figure 1. And the typing was good among the 66 inbred lines randomly chosen (FIG. 2).
Example 2 application of KASP molecular marker KASP-3D-1 to selection of control spike number QTL QSns. Sau-MC-3D.1
(1) 'msf' is a natural multi-spikelet multi-floret mutant material, and 'Shumai 969' is a powdery mildew resistant strain with good plant type, both provided by wheat research institute of Sichuan university of agriculture. Constructing a segregation population F by using multiple spikelets and multiple florets mutant wheat 'msf' as a female parent and using a common wheat variety 'Sichuan wheat 969' with few spikelets as a male parent 2 66 lines were randomly selected among the progeny lines.
(2) The KASP-3D-1 labeling detection is carried out on the obtained 66 strains, and the specific method comprises the following steps:
extracting genome DNA of 66 strains; taking the DNA fragment as a template, taking a specific primer pair of a molecular marker KASP-3D-1 as a primer to carry out PCR amplification and carry out fluorescence reading, wherein the primer is as follows:
FAM tag upstream primer: (FAM tag sequence underlined)
5'-GAAGGTGACCAAGTTCATGCTAGAGTCCAAGGGAGAGTCCGG-3'(SEQ ID NO.1)；
HEX tag upstream primer: (wave line part is HEX tag sequence)
5'-
AGAGTCCAAGGGAGAGTCCGA-3'(SEQ ID NO.2)；
A universal downstream primer:
5'-ACGCAAAGATGAACCAAGCTC-3'(SEQ ID NO.3)。
the amplification system of the PCR amplification is as follows: 5. Mu.L Master Mix; mu.L of mixed primers (three primers SEQ ID No.1, SEQ ID No.2 and SEQ ID No.3 added 60. Mu.L, 60. Mu.L and 120. Mu.L, respectively, at a concentration of 10 ng/. Mu.L, and ddH added thereto) 2 O230. Mu.L was mixed and used as a mixed primer); 5ng of template DNA; adding double distilled water to a total amount of 10 mu L; at the same time, at least 3 independent blanks of DNA template replaced by double distilled water are added.
The procedure for the PCR amplification described above was: pre-denaturation at 94 ℃ for 15min; denaturation at 94 ℃ for 20s and renaturation/elongation at 63 ℃ for 60s for 10 cycles; denaturation at 94 ℃ for 20s, and renaturation/elongation at 55 ℃ for 1min, for 26 cycles; after completion, fluorescence readings were taken for 1min at 37 ℃.
The fluorescence reading results are shown in fig. 3, and the genotype of the plant in which FAM (orange) fluorescence consistent with 'msf' was detected was designated as a, the plant was a line with multiple panicles, the plant exhibited HEX (blue) fluorescence similar to 'hollyhock 969' was designated as B, and the plant was a line with fewer panicles. The genotype (the genotype identified by the molecular marker method of the invention) of each strain and the field phenotypic value of spikelet number are shown in table 2.
Table 2'msf' x 'Shumai 969' F 2 The corresponding result of the genotype and phenotype of the strain KASP-3D-1
The results in table 2 show that the average spikelet number of plants of the same type of 'msf' as containing spikelet number QTL qsns. Sau-MC-3d.1 is 25.14, which is significantly higher (P < 0.01) than the spikelet number of plants of type 'hollyhock 969' (average spikelet number 22.06). The actual result is consistent with the expected result, which shows that the small spike number QTL QSns.sau-MC-3D.1 of the invention really has the effect of obviously increasing the small spike number; meanwhile, the molecular marker KASP-3D-1 can be used for tracking and identifying the wheat spikelet number QTL QSns-sau-MC-3D.1.
The above-described embodiments are only intended to illustrate the preferred embodiments of the present invention, and not to limit the scope of the present invention, and various modifications and improvements made to the technical solution of the present invention by those skilled in the art without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.
Claims (10)
1. A KASP-3D-1 molecular marker linked with wheat spikelet number QTL QSns.sau-MC-3D.1 is characterized in that the KASP-3D-1 molecular marker is an SNP molecular marker with G/A polymorphism, and the molecular marker and the wheat spikelet number QTL QSns.sau-MC-3D.1 are co-located on the short arm of a wheat 3D chromosome.
2. The KASP-3D-1 molecular marker linked to wheat spikelet number QTL QSns. Sau-MC-3D.1 as claimed in claim 1, wherein said KASP-3D-1 molecular marker is represented by the nucleotide sequence shown in SEQ ID NO: 1-3.
3. A primer group, which is characterized by comprising two specific primers and a universal primer for amplifying the KASP-3D-1 molecular marker in claim 1, wherein the nucleotide sequences of the two specific primers are shown as SEQ ID NO. 1-2, and the nucleotide sequence of the universal primer is shown as SEQ ID NO. 3.
4. The primer set of claim 3, wherein the two specific primers are modified at their 5 'ends with different fluorophores or at their 3' ends with different fluorophores.
5. A kit comprising the primer set according to claim 3 or 4.
6. Use of a KASP-3D-1 molecular marker according to any one of claims 1 to 2, or a primer set according to any one of claims 3 to 4, or a kit according to claim 5 for modulating the wheat spikelet number trait.
7. A KASP-3D-1 molecular marker according to any one of claims 1 to 2, or a primer set according to any one of claims 3 to 4, or a kit according to claim 5, for use in wheat seed resource improvement, or breeding of multiple-panicle wheat or high-yield wheat, or in wheat molecular assisted breeding.
8. Use of a KASP-3D-1 molecular marker according to any one of claims 1 to 2, or a primer set according to any one of claims 3 to 4, or a kit according to claim 5, for screening a wheat variety or line with an increased number of spikelets.
9. A method of screening multiple panicle number wheat lines using the KASP-3D-1 molecular marker of claim 1, comprising the steps of:
taking the genome DNA of a wheat plant to be detected as a template, and utilizing the nucleotide sequence shown as SEQ ID NO:1-3, performing fluorescent quantitative PCR amplification, judging whether the small spike number QTL QSns-MC-3D.1 is contained according to fluorescence, and screening to obtain a multi-small-spike type wheat strain.
10. The method of claim 9, wherein the wheat plant to be tested is a multiple spike wheat line if the fluorescence of the primer marker of SEQ ID NO.1 is determined to contain spike number QTL QSns.sau-MC-3D.1.
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| CN116640878A (en) * | 2023-07-12 | 2023-08-25 | 四川农业大学 | Molecular marker of wheat floret number OTL per spike developed based on novel mode of multi-single-combined-standard-evidence and application thereof |
| CN116769961A (en) * | 2023-07-28 | 2023-09-19 | 四川农业大学 | Wheat spike number per spike QTL linkage molecular marker developed by multi-sieve-mixing-determining four-step method and application |
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| CN116640878A (en) * | 2023-07-12 | 2023-08-25 | 四川农业大学 | Molecular marker of wheat floret number OTL per spike developed based on novel mode of multi-single-combined-standard-evidence and application thereof |
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| CN116769961A (en) * | 2023-07-28 | 2023-09-19 | 四川农业大学 | Wheat spike number per spike QTL linkage molecular marker developed by multi-sieve-mixing-determining four-step method and application |
| CN116769961B (en) * | 2023-07-28 | 2024-01-26 | 四川农业大学 | Wheat spike number per spike QTL linkage molecular marker developed by multi-sieve-mixing-determining four-step method and application |
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