CN109825621B - Wheat spikelet number QTL (quantitative trait locus) linked SNP (single nucleotide polymorphism) molecular marker and application thereof - Google Patents
Wheat spikelet number QTL (quantitative trait locus) linked SNP (single nucleotide polymorphism) molecular marker and application thereof Download PDFInfo
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Abstract
The invention discloses a wheat spikelet number QTL linked SNP molecular marker and application thereof, wherein the SNP molecular marker KASP-1 is positioned on a 2D chromosome short arm of a RefSeqv1.0 genome version, the front and back 100bp sequences are shown as SEQ ID NO.46, the polymorphism is C/T, and the SNP molecular marker can be obtained by amplifying primers shown as SEQ ID NO. 1-3. The molecular marker can accurately track the wheat spikelet number QTL QSns-sau-2 D.1, and predict the spikelet number characteristic of wheat, thereby facilitating molecular breeding. The invention also discloses a method for identifying the wheat spikelet number QTL QSns-2 D.1 molecular marker, which can enhance the accuracy of spikelet number prediction by utilizing the provided method so as to rapidly screen out a wheat variety or a strain with increased spikelet number for breeding and greatly accelerate the breeding process of a wheat high-yield variety.
Description
Technical Field
The invention relates to the field of molecular biology and crop genetic breeding, in particular to a wheat spikelet number QTL QSns.sau-2D.1 linked SNP molecular marker and application thereof.
Background
Wheat is one of the most important grain crops in the world and is also an important grain crop in China, the perennial planting area of the wheat is more than 2000 ten thousand hectares, and the wheat occupies about 27 percent of the area of all the grain crops; the total yield exceeds 1 hundred million tons and accounts for about 22 percent of the yield of all food crops. Wheat yield is dependent on grain weight, grain per ear and ear per unit area. The grain number of the panicle is the most active factor in yield composition factors, the degree of adjustment is large, and the grain number of the panicle is determined by the number of small panicle, the number of differentiated florets and the floret setting rate. The small spike number is the premise of forming the small flower number, and the increased small spike number is the basis for improving the grain number of the spike.
The wheat yield traits are complex Quantitative traits, are controlled by a plurality of Quantitative Trait Loci (QTL), and have the characteristics of low heritability, large environmental influence and high selection difficulty, so that the traditional breeding method has the problems of long time, large consumption, high cost and small achievement in the breeding process. The molecular marker assists breeding, is not dependent on phenotype selection, namely is not influenced by factors such as environment, gene interaction, gene and environment interaction and the like, but directly selects genotypes, so that the breeding efficiency can be greatly improved.
Single Nucleotide Polymorphism (SNP) refers to a DNA sequence Polymorphism caused by a change such as a transition, a transversion, an insertion, or a deletion at a specific Nucleotide position in DNA in a genome. The technology is that known sequence information is utilized to compare and search SNP sites, and then specific primers are designed by utilizing the discovered variation sites to carry out PCR amplification on genome DNA or cDNA, so as to obtain specific polymorphic products based on the SNP sites, and finally, the polymorphism of the products is analyzed by utilizing the electrophoresis technology. 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.
KASP is a novel genotyping technology with low cost and high throughput characteristics by competitive Allele Specific PCR (KASP) developed by LGC (Laboratory of the Goverment Chemist) (http:// www.lgcgenomics.com), carries out accurate double-Allele genotyping on SNP and In Del sites by Specific matching of terminal bases of primers, and is widely applied to molecular marker-assisted selection of crops such as rice, wheat, soybean and the like.
Heretofore, researchers have carried out QTL positioning on spikelet number and found that QTL related to the spikelet number widely exists in wheat, but at present, the number of closely linked molecular markers which are related to the trait of the spikelet number of wheat and can be used for actual molecular breeding is not large. Therefore, QTL or gene related to the number of spikelets is obtained through research, the number of spikelets is increased by utilizing molecular biology technology, the number of grains per spike is further increased, the purpose of breeding new wheat varieties with increased yield is finally achieved, and the method has great significance in wheat breeding work.
Disclosure of Invention
The invention aims to provide a molecular marker closely linked with wheat spikelet number QTL QSns-sau-2 D.1.
The invention also aims to provide the molecular-labeled fluorescent quantitative PCR primer.
The third purpose of the invention is to provide the application of the molecular marker closely linked with the wheat spikelet number QTL QSns.sau-2 D.1.
The purpose of the invention is realized by the following technical scheme:
based on the purposes, the applicant utilizes a wheat strain '20828' with multiple spikelets as a female parent and a wheat strain 'CN 16' as a male parent for hybridization to obtain a hybrid F1，F1Selfing the single plant to obtain F2At F2Using single ear propagation up to F8And thirdly, obtaining a recombinant inbred line containing 199 lines to form a genetic mapping population. The phenotype of the spikelet number of the recombinant inbred line group is identified, the DNA of the parents '20828', 'CN 16' and the plant of the recombinant inbred line group is extracted, and the wheat 55K SNP chip is used for positioning the QTL of the spikelet number of the wheat in the research. The wheat 55K SNP chip is an economic medium-density SNP chip developed on the basis of a wheat 660KSNP chip. The chip contains about 55,000 wheat SNP markers which are uniformly distributed on 21 chromosomes, and each chromosome averagely contains 2600 markers, the average genetic distance between the markers is about 0.1cM, and the average physical distance is less than 300Kb, so that the chip is suitable for general germplasm resource diversity analysis, genetic mapping and new gene discovery, comparative genome analysis, and variety registration and identification (fingerprint analysis).
According to 55K SNP chip data, a genetic map is constructed by using JoinMap4.0. Combining with the spikelet number table data of the population, detecting the QTL by using a BlUP (best linear unbiased prediction) value of 8 ecological points and 8 ecological point spikelets in 2016 + 2018 three years under the condition that an integral Composite Interval Mapping-ADD (ICIM-ADD) in QTL IciMapping 4.0 is used and the LOD is set to be more than or equal to 2.5, stably expressed wheat spikelet number major QTL QSns-sau-2 D.1 is positioned in the 1cM interval on the short arm of the 2D chromosome, physically locating the flanking markers and screening the genes located in the interval every 1Mbp, obtaining 40 genes by co-screening, cloning the genes in parent '20828' and 'CN 16', 15 pairs of 45 KASP primers (table 1) are designed for obtaining polymorphic sites and developing molecular markers, and finally the obtained SNP molecular marker KASP-1 is closely linked with the spikelet number QTL QSns.sau-2 D.1.
The wheat spikelet number QTL QSns-2 D.1 comes from a female parent '20828', the QTL is positioned on a wheat chromosome 2D short arm, and the physical position of a RefSeqv1.0 genome version is 33735919. The invention provides application of the wheat spikelet number QTL QSns.sau-2D.1 in regulation of wheat spikelet number characters, wherein the wheat spikelet number QTL QSns.sau-2D.1 is tightly linked with a molecular marker KASP-1 and can be obtained by amplifying 3 primers with nucleotide sequences shown as SEQ ID No.1, 2 and 3 respectively. Specifically, the wheat spikelet number QTL QSns-2 D.1 can improve the wheat spikelet number. Spikelet number QTL qsns. sau-2d.1 significantly increased spikelet number, with an average LOD value of 19.86, accounting for approximately 30.78% of phenotypic variation.
Further, the invention provides an SNP molecular marker KASP-1 of wheat spikelet number QTL QSns.sau-2D.1, which is closely linked with wheat spikelet number QTL QSns.sau-2D.1, is positioned on the 2D chromosome short arm of RefSeqv1.0 genome version, has a genetic distance of less than 1cM with QSns.sau-2D.1, has a sequence of 100bp before and after the sequence as shown in SEQ ID NO.46, is positioned at the 101bp of the sequence as shown in SEQ ID NO.46, and has polymorphism of C/T.
The molecular marker KASP-1 of the wheat spikelet number QTL QSns.sau-2D.1 is obtained by PCR amplification of 3 primer pairs with nucleotide sequences shown in SEQ ID No. 1-3. The 3 primers comprise 2 upstream primers and 1 universal downstream primer, and the 5' ends of the 2 upstream primers are respectively modified with different fluorescent groups; the nucleotide sequences of the 3 primers are respectively shown as SEQ ID NO.1, 2 and 3.
The invention provides application of the molecular marker KASP-1 in molecular assisted breeding of crops.
The invention provides application of the molecular marker KASP-1 in cultivation of wheat with multiple spikelet number characters.
The invention also provides a specific primer pair of the SNP molecular marker KASP-1 for detecting the wheat spikelet number QTL QSns.sau-2D.1, wherein the sequences of the upstream primer and the downstream primer are respectively shown as SEQ ID NO. 1-3.
The invention provides application of the specific primer pair in improvement of wheat germplasm resources.
The invention provides application of the specific primer pair in the preparation of a material with multiple small ears of wheat.
Kits containing the above specific primer pairs also fall within the scope of the present invention.
The invention provides a molecular marking method for identifying wheat spikelet number QTL QSns.sau-2D.1, which comprises the steps of taking DNA of a material to be identified as a template, carrying out fluorescent quantitative PCR amplification by using a fluorescent quantitative PCR primer, carrying out genotype typing by using an amplification result, and identifying a plant capable of reading fluorescence modified by the primer SEQ ID NO.2 as a plant containing the wheat spikelet number QTL QSns.sau-2 D.1.
Specifically, the application includes the following steps:
1) extracting the genome DNA of a plant to be detected;
2) taking the genome DNA of a plant to be detected as a template, and performing PCR amplification reaction and reading a fluorescence value in a CFX96Real-Time System by using a primer of an amplification molecular marker KASP-1;
3) and detecting the fluorescence of the PCR amplification product, wherein if the fluorescence modified by the primer SEQ ID NO.2 can be read, the plant to be detected is the wheat resource with the characteristic of multiple spikelets.
The amplification system of the PCR amplification is as follows: 5 μ L Master Mix, three primers SEQ ID No: 1. 2 and 3 at a concentration of 10 ng/. mu.L, 120. mu.L, and 300. mu.L, respectively, and ddH2O 460. mu.L were added and mixed to be used as mixed primers, 1.4. mu.L of the mixed primers, 5ng of the template DNA, and double distilled water were added to a total amount of 10. mu.L, and at least 3 independent blanks were added in place of the DNA template in the double distilled water.
The procedure of the PCR amplification is as follows: pre-denaturation at 94 ℃ for 15 min; denaturation at 94 ℃ for 20s and renaturation/elongation at 61 ℃ for 60s for 10 cycles; denaturation at 94 ℃ for 20s and renaturation/elongation at 55 ℃ for 60s for 26 cycles; after completion, fluorescence readings were taken.
The invention discloses a molecular marker KASP-1 which is positioned on a wheat 2D chromosome and linked with the wheat spikelet number, wherein the molecular marker is a flanking marker of the spikelet number QTL QSns-sau-2 D.1 on the long arm of the wheat 2D chromosome, and the linkage degree is high. The marker can be used for detecting the spikelet number QTL on the 2D chromosome of wheat, rapidly screening plants with the locus, and further facilitating molecular assisted breeding of high-yield wheat. The molecular marker KASP-1 provided by the invention is tightly linked with the spikelet number QTL QSns.sau-2D.1 on the wheat 2D, 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 foundation is provided for the research of the spikelet number gene of the wheat.
The invention has the beneficial effects that: the invention discloses a spikelet number QTL QSns-sau-2 D.1 from wheat 20828' for the first time, which is positioned on a short arm of a wheat 2D chromosome and can obviously increase the spikelet number of the wheat. The QTL has higher utilization value in wheat yield (small ear number regulation) breeding. Spikelet number QTL qsns. sau-2d.1 significantly increased spikelet number, with an average LOD value of 19.86, accounting for approximately 30.78% of phenotypic variation.
The invention discloses a molecular marker KASP-1 for accurately detecting the spikelet number QTL QSns.sau-2D.1 of wheat 20828' based on a fluorescent quantitative PCR platform for the first time, and the molecular marker KASP-1 is a codominant marker, and has the advantages of accurate and efficient detection and convenient and stable amplification. The molecular marker KASP-1 disclosed by the invention is remarkably related to the spikelet number QTL QSns.sau-2D.1, presents the characteristic of a coseparation marker, has high accuracy when being used 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
FIG. 1 shows the location of the wheat ear number QTL QSns.sau-2D.1 on the 2D chromosome in example 1 of the present invention.
FIG. 2 shows the fluorescence readings of the molecular marker KASP-1 detection of the recombinant inbred line strain plant of '20828' × 'CN 16' in example 1 of the present invention; wherein, HEX (square, '20828') fluorescence is a strain with a large spikelet number, FAM (round, 'CN 16') fluorescence is a strain with a small spikelet number; the triangle fluorescence is a heterozygous strain; the diamond fluorescence is blank.
FIG. 3 shows the fluorescence reading results of the detection of molecular marker KASP-1 in the plant molecular marker of the recombinant inbred line strain of '20828' x common wheat strain 'SY 95-71' in example 2 of the present invention; wherein, HEX (square, '20828') fluorescence is a strain with a large spikelet number, FAM (round, 'SY 95-71') fluorescence is a strain with a small spikelet number; the triangle fluorescence is a heterozygous strain; the diamond fluorescence is blank.
Detailed Description
The following examples further illustrate the present invention but are not to be construed as limiting the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention.
The wheat germplasm resources used in the embodiment of the invention are all from a planting resource library of Lanxiujin researchers at the wheat research institute of Sichuan university of agriculture.
Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1 obtaining of wheat spikelet number QTL QSns. sau-2D.1 and its molecular marker KASP-1
(1) Hybridizing the wheat strain '20828' as female parent and the wheat strain 'CN 16' as male parent to obtain hybrid F1，F1Selfing the single plant to obtain F2At F2Using single ear propagation up to F8And thirdly, obtaining a recombinant inbred line containing 199 lines to form a genetic mapping population.
(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 mature period of the wheat, removing the single plants at two ends of each row, respectively collecting five single plants with consistent growth vigor, selecting a main spike, calculating the number of spikelets, and obtaining an average value which represents the number of spikelets of the plant line.
(3)55K SNP chip analysis
a) DNA extraction: the DNA of parent '20828', 'CN 16' and the plant of the recombinant inbred line population is extracted by a CTAB method.
b) The extracted DNA was subjected to quality testing using an ultramicro spectrophotometer and was sent to the company for genotyping after qualification, and the genotyping of the parental and mapping groups in this study was performed by a 55K SNP chip developed by the Biotechnology Ltd of Beijing Boao Crystal science, http:// www.capitalbiotech.com, which is commercially available.
c) Constructing a linkage map: according to 55K SNP chip data, a genetic map is constructed by using JoinMap4.0. Combining spikelet number table data of a population, detecting the QTL by using BLUP (best linear unbiased prediction) values of 8 ecological points and 8 ecological point spikelets in three years of 2016-2018 under the condition that threshold LOD is not less than 2.5 by using an inclusion complex Interval Mapping-ADD (ICIM-ADD) in QTL IcImMapping 4.0, locating the wheat spikelet number QTL QSns-2 D.1, and calculating the position of the QSns-2 D.1 and the genetic distance between molecular markers.
d) Densification of genetic map and acquisition of closely linked molecular markers: in order to compact a genetic map and obtain a molecular marker tightly linked with the spikelet number QTL QSns.sau-2D.1, a flanking marker is physically positioned and genes positioned in an interval are screened by utilizing a 55K SNP chip data positioning result, the genes are cloned at a parent of '20828' and 'CN 16', polymorphic sites are obtained and molecular markers are developed, a KASP primer is designed by utilizing DNMAN (45 primers are designed totally, 15 pairs of KASP primers are designed, and the sequence of the primers is shown in a table 2), and finally the SNP marker KASP-1 and the spikelet number QTL QSns.sau-2D.1 are tightly linked.
TABLE 115 pairs of KASP primer sequences
e) And (6) carrying out analysis. 2 molecular markers are finally obtained from 15 pairs of KASP primers, wherein KASP-1 is closely linked with spikelet number QTL QSns.sau-2 D.1. The results are shown in FIGS. 1 and 2.
Example 2 application of molecular marker KASP-1 in selection control of spikelet number QTL QSns
(1) A common wheat line '20828' with a large number of spikelets is used as a female parent, a common wheat line 'SY 95-71' with a small number of spikelets is used as a male parent to construct a recombinant inbred line, and 80 lines are randomly selected from the progeny lines.
(2) KASP-1 marker detection is carried out on the obtained 80 strains, and the specific method comprises the following steps: extracting the DNA of 80 strains; taking the DNA fragment as a template, taking a specific primer pair of a molecular marker KASP-1 as a primer to perform PCR amplification and perform fluorescence reading, wherein the primer is as follows:
primer on FAM tag: (FAM tag sequence underlined)
5’-GAAGGTGACCAAGTTCATGCTTATAAACCGGTCGAACTCGC-3’(SEQ ID No.1)
Primers on HEX tag: (wave line part HEX tag sequence)
5’-GAAGGTCGGAGTCAACGGATTTATAAACCGGTCGAACTCGT-3’(SEQ ID No.2)
A universal downstream primer:
5’-TGGTGCTTCTCCTTGGCGAG-3’(SEQ ID No.3)
the amplification system of the PCR amplification is as follows: 5 μ L Master Mix, three primers SEQ ID No: 1. 2 and 3 at a concentration of 10 ng/. mu.L, 120. mu.L, and 300. mu.L, respectively, and ddH2O 460. mu.L were added and mixed to be used as mixed primers, 1.4. mu.L of the mixed primers, 5ng of the template DNA, and double distilled water were added to a total amount of 10. mu.L, and at least 3 independent blanks were added in place of the DNA template in the double distilled water.
The procedure of the PCR amplification is as follows: pre-denaturation at 94 ℃ for 15 min; denaturation at 94 ℃ for 20s and renaturation/elongation at 61 ℃ for 60s for 10 cycles; denaturation at 94 ℃ for 20s and renaturation/elongation at 55 ℃ for 60s for 26 cycles; after completion, fluorescence readings were taken.
As a result of the fluorescence reading (see FIG. 3), the genotype of the plant in which HEX (blue) fluorescence consistent with that of '20828' was detected was designated as A, the genotype was designated as a multi-tassel type strain, the genotype was designated as B, the genotype was designated as a FAM (orange) fluorescence similar to that of 'SY 95-71', and the genotype was designated as a few-tassel type strain. The BLUP values for each line genotype and spikelet number of 6 ecopoints are shown in table 2. The average spikelet number of plants of the same type as '20828' containing spike number QTL QSns. sau-2D.1 was 25.51, which was significantly higher than the spike number of plants of the type 'SY 95-71' (average 22.45). The actual result is consistent with the expected result, which shows that the spikelet number QTL QSns-sau-2 D.1 of the invention has the effect of obviously increasing the spikelet number; meanwhile, the molecular marker KASP-1 can be used for identifying the spikelet number QTL QSns-sau-2 D.1 by tracking.
TABLE 2 '20828' × 'SY 95-71' results of genotype-1 and phenotype correspondence for recombinant inbred line KASP-1
While the invention has been described in detail in the foregoing by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that certain modifications and improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
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gtatttttcc ttctcccgtt gtaacgcacg ggcatgtttg ctagttatgt cataaaaagg 60
agaaatagta atttataaac cggtcgaact cgcgcagacg ctctggtgct tctccttggc 120
gagccaggcg tcttccatgg cgtagtagtt attcatcaat tcggacagtc cggccatggt 180
tctcggccgg tgatggatga g 201
Claims (8)
1. Wheat spikelet number QTLQSns.sau-2D.1A linked SNP molecular marker, wherein said SNP molecular marker is KASP-1, located on the short 2D chromosome arm of the genomic version of refseqv1.0, andQSns.sau-2D.1the genetic distance between the SNP molecular markers is less than 1cM, the sequences of the front and back 100bp are shown as SEQ ID NO.46, and the polymorphism of the SNP molecular markers is C/T; it can be obtained by amplifying the following 3 primers: the 3 primers comprise 2 upstream primers and 1 universal downstream primer, and the 5' ends of the 2 upstream primers are respectively modified with different fluorescent groups; the nucleotide sequences of the 3 primers are respectively shown as SEQ ID NO.1, 2 and 3.
2. QTL for detecting spikelet number of wheatQSns.sau-2D.1The specific primer combination of the linked SNP molecular markers is characterized by comprising 3 primers with nucleotide sequences shown as SEQ ID No. 1-3, wherein the 3 primers comprise 2 upstream primers and 1 universal downstream primer, the nucleotide sequences of the 2 upstream primers respectively correspond to SEQ ID No.1 and SEQ ID No.2, and the 5' ends of the upstream primers are respectively modified with different fluorescent groups.
3. A kit comprising the specific primer combination according to claim 2.
4. The SNP molecular marker according to claim 1 or the specific primer combination or primer set according to claim 2QTL for identifying wheat spikelet number by using kit according to claim 3QSns.sau-2D.1The use of (1).
5. Use of the SNP molecular marker according to claim 1, or the specific primer combination according to claim 2, or the kit according to claim 3 for identifying high-yielding wheat.
6. The use of the SNP molecular marker of claim 1, the specific primer combination of claim 2, or the kit of claim 3 for improving wheat germplasm resources, or for creating high-yield wheat materials with a large number of spikelets, or for molecular assisted breeding of wheat.
7. The application of any one of claims 4 to 6, wherein the genomic DNA of a plant sample to be tested is used as a template, fluorescent quantitative PCR amplification is carried out by using fluorescent quantitative PCR primers, genotyping is carried out by using an amplification result, and a plant which can read the fluorescence modified by the primers SEQ ID NO.2 is identified as containing wheat spikelet number QTLQSns.sau-2D.1The plant of (1).
8. The use of claim 7, wherein the amplification reaction system of the fluorescence quantitative PCR: adding 120 muL, 120 muL and 300 muL and adding ddH2O 460 muL to the 5 muL Master Mix and the 3 primers SEQ ID NO.1, 2 and 3 according to the concentration of 10 ng/muL respectively, mixing, and using the mixture as a mixed primer, adding 1.4 muL, 5ng template DNA and double distilled water to the total amount of 10 muL, and adding at least 3 independent blanks for replacing DNA templates with double distilled water;
fluorescent quantitative PCR procedure: pre-denaturation at 94 ℃ for 15 min; denaturation at 94 ℃ for 20s and renaturation/elongation at 61 ℃ for 60s for 10 cycles; denaturation at 94 ℃ for 20s and renaturation/elongation at 55 ℃ for 60s for 26 cycles;
genotyping the results obtained; and detecting the fluorescence of the PCR amplification product, wherein if the fluorescence modified by the primer SEQ ID NO.2 can be read, the plant to be detected is the wheat resource with the characteristic of multiple spikelets.
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| CN110184381B (en) * | 2019-06-25 | 2023-07-25 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | SNP locus related to wheat small ear number per ear and application thereof |
| CN111118191B (en) * | 2019-09-30 | 2022-04-22 | 山西农业大学 | KASP molecular marker of wheat head top spikelet fructicity main effect QTL and application thereof |
| CN111187852B (en) * | 2020-01-16 | 2021-09-21 | 中国科学院遗传与发育生物学研究所 | SNP (single nucleotide polymorphism) site related to wheat yield and application thereof |
| CN111560452B (en) * | 2020-03-18 | 2023-05-12 | 淮阴师范学院 | Identification of wheat TaDEP1 gene transcription enhancement cis-acting element, molecular marker and application thereof |
| CN111647677B (en) * | 2020-04-26 | 2022-07-08 | 四川农业大学 | Molecular marker closely linked with wheat grain filling rate QTL QGfr. sicau-6D and application |
| CN111763759B (en) * | 2020-07-23 | 2021-10-08 | 四川农业大学 | Wheat tillering angle QTL QTA-2B linked molecular marker and application thereof |
| CN112779348B (en) * | 2020-12-31 | 2022-05-20 | 四川农业大学 | Major QTL site of ear number per unit area of wheat, KASP primer closely linked with major QTL site and application of KASP primer |
| CN112760402B (en) * | 2021-01-29 | 2024-01-26 | 江苏里下河地区农业科学研究所 | KASP primer group for detecting wheat spike number and biomass density and application thereof |
| CN112779350A (en) * | 2021-02-07 | 2021-05-11 | 四川农业大学 | Molecular marker closely linked with wheat spikelet grain number QTLQGns |
| CN114410817B (en) * | 2022-01-10 | 2022-11-01 | 河北师范大学 | Wheat spikelet number per spike character related SNP site and application thereof |
| CN116064583B (en) * | 2022-10-08 | 2024-01-26 | 青海省农林科学院 | Highland barley spike length regulating gene and KASP molecular marker and application thereof |
| 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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